BRPI0806354A2 - plantas oleaginosas transgências, sementes, óleos, produtos alimentìcios ou análogos a alimento, produtos alimentìcios medicinais ou análogos alimentìcios medicinais, produtos farmacêuticos, bebidas fórmulas para bebês, suplementos nutricionais, rações para animais domésticos, alimentos para aquacultura, rações animais, produtos de sementes inteiras, produtos de óleos misturados, produtos, subprodutos e subprodutos parcialmente processados - Google Patents

plantas oleaginosas transgências, sementes, óleos, produtos alimentìcios ou análogos a alimento, produtos alimentìcios medicinais ou análogos alimentìcios medicinais, produtos farmacêuticos, bebidas fórmulas para bebês, suplementos nutricionais, rações para animais domésticos, alimentos para aquacultura, rações animais, produtos de sementes inteiras, produtos de óleos misturados, produtos, subprodutos e subprodutos parcialmente processados Download PDF

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BRPI0806354A2
BRPI0806354A2 BRPI0806354-0A BRPI0806354A BRPI0806354A2 BR PI0806354 A2 BRPI0806354 A2 BR PI0806354A2 BR PI0806354 A BRPI0806354 A BR PI0806354A BR PI0806354 A2 BRPI0806354 A2 BR PI0806354A2
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Howard G Damude
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Du Pont
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Abstract

PLANTAS OLEAGINOSAS TRANSGêNICAS, SEMENTES, óLEOS, PRODUTOS ALIMENTìCIOS OU ANáLOGOS A ALIMENTO, PRODUTOS ALIMENTìCIOS MEDICINAIS OU ANáLOGOS ALIMENTìCIOS MEDICINAIS, PRODUTOS FARMACêUTICOS, BEBIDAS, FóRMULAS PARA BEBêS, SUPLEMENTOS NUTRICIONAIS, RAçõES PARA ANIMAIS DOMéSTICOS, ALIMENTOS PARA AQUACULTURA, RAçõES ANIMAIS, PRODUTOS DE SEMENTES INTEIRAS, PRODUTOS DE óLEOS MISTURADOS, PRODUTOS, SUBPRODUTOS E SUBPRODUTOS PARCIALMENTE PROCESSADOS. São descritas plantas oleaginosas que foram transformadas para produzir ácido araquidónico, construções recombinantes utilizadas nessas transformações, métodos de produção de ácido araquidónico em plantas e usos de óleos e sementes obtidos dessas plantas transformadas em uma série de aplicações alimentícias e alimentos.

Description

"PLANTAS OLEAGINOSAS TRANSGÊNICAS, SEMENTES, ÓLEOS, PRODUTOS ALIMENTÍCIOS OU ANÁLOGOS A ALIMENTO, PRODUTOS ALIMENTÍCIOS MEDICINAIS OU ANÁLOGOS ALIMENTÍCIOS MEDICINAIS, PRODUTOS FARMACÊUTICOS, BEBIDAS, FÓRMULAS PARA BEBÊS, SUPLEMENTOS NUTRICIONAIS, RAÇÕES PARA ANIMAIS DOMÉSTICOS, ALIMENTOS PARA AQUACULTURA, RAÇÕES ANIMAIS, PRODUTOS DE SEMENTES INTEIRAS, PRODUTOS DE ÓLEOS MISTURADOS, PRODUTOS, SUBPRODUTOS E SUBPRODUTOS PARCIALMENTE PROCESSADOS"
O presente pedido reivindica o benefício do Pedido Provisório Norte-Americano n° 60/889373, depositado em doze de fevereiro de 2007, cujo teor integral é incorporado ao presente como referência.
Campo da Invenção
A presente invenção encontra-se no campo da biotecnologia. Mais especificamente, a presente invenção refere-se a plantas oleaginosas que foram transformadas para produzir altos níveis de ácido araquidônico (um ácido graxo ômega 6) e teor reduzido de ácidos graxos ômega 3.
Antecedentes da Invenção
Duas famílias principais de ácidos graxos póli-insaturados (PUFAs) são os ácidos graxos ômega 3 tais como ácido eicosapentaenóico (EPA) e os ácidos graxos ômega 6 tais como ácido araquidônico (ARA; cis- 5,8,11,14-eicosatetraenoico). ARA é um precursor importante na produção de eicosanoides (tais como prostaglandinas, tromboxanos, prostaciclina e leucotrienos). Além disso, ARA é reconhecido como: (1) um ácido graxo póli- insaturado de cadeia longa essencial (PUFA); (2) o principal ácido graxo ômega 6 encontrado no cérebro humano; e (3) um componente importante de leite de peito e muitas fórmulas para bebês, com base no seu papel no desenvolvimento visual e neurológico inicial. Os adultos obtêm ARA facilmente de alimentos tais como carne, ovos e leite. Os adultos podem também sintetizar ARA a partir de ácido gamalinolênico da alimentação, embora ineficientemente. Fontes comerciais de óleo de ARA são tipicamente produzidas a partir de óleo de peixe purificado e altamente refinado ou fermentação (utilizando, por exemplo, micro-organismos dos gêneros Mortierella (fungo filamentoso), Entomophthora, Pythium e Porphyridium (alga vermelha)). Mais notadamente, a MarteK Biosciences Corporation (Columbia MD) produz um óleo de fungos que contém ARA (ARASCO®; vide a Patente Norte-Americana n° 5.658.767) que é substancialmente livre de EPA e que é derivado de Mortierella alpina ou Pythium insidiuosum. Um dos principais mercados para este óleo são fórmulas para bebês.
Infelizmente, existem várias desvantagens associadas à produção comercial de PUFAs (tais como ARA) a partir de fontes naturais. As fontes naturais de PUFAs, tais como peixe, tendem a possuir composições de óleo altamente heterogêneas. Os óleos obtidos dessas fontes podem necessitar de extensa purificação para separar um ou mais PUFAs desejados ou produzir um óleo que seja enriquecido com um ou mais PUFAs. Fontes naturais também são objeto de flutuações incontroláveis de disponibilidade. Estoques de peixe podem sofrer variação natural ou podem ser esgotados devido à pesca excessiva. Óleos de peixe possuem sabores e odores desagradáveis que podem ser de separação economicamente difícil, se não impossível, do prouto desejado e podem tornar esses produtos inaceitáveis como suplementos alimentícios. Óleos animais e, particularmente, óleos de peixe podem acumular poluentes ambientais. O clima e doenças podem causar flutuações dos rendimentos das fontes vegetais e de peixes.
O fornecimento em expansão de ARA de peixe e de síntese química não é suficiente para as necessidades comerciais. É de interesse, portanto, encontrar meios alternativos que permitam a produção de quantidades comerciais de ARA. A biotecnologia, utilizando uma planta oleaginosa, oferece uma via atraente de produção de ARA de forma segura e eficiente para o seu custo.
Os pedidos copendentes do Cessionário do Depositante incluem os seguintes: (1) Patente PCT n0 WO 04/071467 (publicada em 26 de agosto de 2004; Pasta do Advogado n° BB-1538) refere-se à produção de PUFAs em plantas; (2) Patente PCT n0 WO 04/071178 (publicada em 26 de agosto de 2004; Pasta do Advogado n° BB-1531) refere-se a promotores de anexina e seu uso na expressão de transgenes em plantas; (3) Patente PCT n0 WO 05/047479 (publicada em 26 de maio de 2005; Pasta do Advogado n° CL-2432) descreve uma delta-15 dessaturase de Fusarium moniliforme\ (4) Pedido Norte- Americano n° 11/601.563 (depositado em dezesseis de novembro de 2006, publicado em 24 de maio de 2007; Pasta do Advogado n° BB-1562) descreve uma delta-9 elongase de Eulgena gracilis; (5) Pedido de Patente Norte- Americano n° 11/737772 (depositado em vinte de abril de 2007; Pasta do Advogado n° BB-1566) descreve uma delta-8 dessaturase de Pavlova Iutheri; (6) Pedido de Patente Norte-Americano n° 11/748629 (depositado em quinze de maio de 2007, publicado em vinte de dezembro de 2007; Pasta do Advogado n° CL-3486) descreve uma delta-5 dessaturase de Eulgena gracilis: e (7) Pedido de Patente Norte-Americano n° 11/876115 (depositado em 22 de outubro de 2007; Pasta do Advogado n° BB-1574) descreve uma delta-8 dessaturase de Tetruetreptia pomquetensis CCMP 1491.
Descrição Resumida da Invenção
Em uma primeira realização, a presente invenção refere-se a uma planta oleaginosa transgênica que produz sementes maduras nas quais o perfil total de ácido graxo de sementes compreende pelo menos 10,0% de ácido araquidônico e menos de 5% de ácidos graxos ômega 3 totais que contêm pelo menos dezoito átomos de carbono e pelo menos quatro uniões duplas.
Em uma segunda realização, a presente invenção refere-se a uma planta oleaginosa transgênica que produz sementes maduras nas quais o perfil total de ácido graxo de sementes compreende pelo menos 10,0% de ácido araquidônico e menos de 1% de ácidos graxos ômega 3 totais que contêm pelo menos dezoito átomos de carbono e pelo menos quatro uniões duplas.
Em uma terceira realização, a presente invenção refere-se a uma planta oleaginosa transgênica que produz sementes maduras nas quais o perfil total de ácido graxo de sementes compreende pelo menos 10,0% de ácido araquidônico e menos de 5% de ácido gamalinolênico.
Em uma quarta realização, a presente invenção refere-se a uma planta oleaginosa transgênica que produz sementes maduras nas quais o perfil total de ácido graxo de sementes compreende pelo menos 10,0% de ácido araquidônico e até 1%, inclusive, de ácido gamalinolênico.
Em uma quinta realização, a presente invenção refere-se a uma planta oleaginosa transgênica que produz sementes maduras nas quais o perfil total de ácido graxo de sementes compreende pelo menos 10,0% de ácido araquidônico, até 1%, inclusive, de ácido gamalinolênico e menos de 1% de ácidos graxos ômega 3 totais que contêm pelo menos dezoito átomos de carbono e pelo menos quatro uniões duplas.
A presente invenção também se refere a sementes obtidas a partir dessas plantas oleaginosas transgênicas, bem como óleo obtido dessas sementes. Plantas oleaginosas que podem ser transformadas podem ser selecionadas a partir do grupo que consiste de soja, espécies de Brassica, girassol, milho, algodão, Iinho e açafrão.
Os óleos de acordo com a presente invenção podem ser incorporados em um produto alimentício ou análogo a alimento, alimento medicinal ou análogo de alimento medicinal, produto farmacêutico, bebida, fórmula para bebês, suplemento nutricional, ração para animais domésticos, ração animal ou alimento para aquacultura.
Os produtos alimentícios que incorporam esses óleos podem ser selecionados a partir do grupo que consiste de uma partícula alimentícia seca por pulverização, partícula alimentícia seca por congelamento, produtos de carne, alimento de cereal, alimento de lanche, produto assado, alimento extrudado, alimento frito, alimento saudável, alimento lácteo, análogos de carne, análogos de queijo, análogos de leite, ração para animais domésticos, ração animal ou alimento para aquacultura.
As sementes obtidas dessa planta oleaginosa transgênica podem ser incorporadas em uma ração para animais domésticos, alimento para aquacultura e/ou ração animal.
Também são de interesse produtos de sementes inteiras elaborados com sementes de acordo com a presente invenção, produtos de óleos misturados que compreendem um óleo de acordo com a presente invenção, produtos elaborados por meio da hidrogenação, fracionamento, interesterificação ou hidrólise de um óleo de acordo com a presente invenção, subproduto(s) fabricado(s) durante a produção de um óleo de acordo com a presente invenção e subprodutos parcialmente processados fabricados durante a produção de um óleo de acordo com a presente invenção.
DEPOSITOS BIOLOGICOS
O plasmídeo a seguir foi depositado junto à Coleção Norte- Americana de Cultivos de Tipos (ATCC), 10801 University Boulevard, Manassas VA 20110-2209 e possui a designação, Número de Acesso e data de depósito a seguir (Tabela 1). Tabela 1
Depósito de ATCC
<table>table see original document page 7</column></row><table>
Breve Descrição das Figuras e Listagem de Seqüências
A presente invenção pode ser compreendida mais completamente a partir da descrição detalhada a seguir e das figuras e Listagem de Seqüências anexas, que fazem parte do presente pedido.
A Fig. 1 é um processo de ácido graxo de ômega 3 e ômega 6 representativo que fornece a conversão de ácido mirístico por meio de vários intermediários de ácido docosa-hexanoico.
A Fig. 2 é um mapa de plasmídeo pKR1084 (vide também SEQ ID N° 7).
A Fig. 3 é um mapa de plasmídeo BC-Tpom_ALS (vide também SEQ ID N° 10).
A Fig. 4 é um mapa de plasmídeo BC-Tpom_ALSrev (vide também SEQ ID N° 11).
A Fig. 5 é um mapa de plasmídeo BC-Tpom_Ann-EgD5_ALS (vide também SEQ ID N° 17).
A Fig. 6 é um mapa de plasmídeo BC-Tpom_Ann-EgD5_ALSrev (vide também SEQ ID N° 18).
A Fig. 7 é um mapa de plasmídeo KS263 (vide também SEQ ID N° 20).
A Fig. 8 é um mapa de plasmídeo BC-ELfad3_ALS (vide também SEQ ID N° 27). A Fig. 9 é um mapa de plasmídeo BC-Tpom_Ann-ELfad3_ALS (vide também SEQ ID N0 29).
A Fig. 10 é um mapa de plasmídeo HPfad3ABA'_ALS (vide também SEQ ID N0 39).
A Fig. 11 é um mapa de plasmídeo BC-Tpom_Ann- HPfad3ABA'_ALS (vide também SEQ ID N0 26).
A Fig. 12 exibe combinações de construção para gerar ácido araquidônico (ARA) em soja.
A Fig. 13 exibe um cromatograma do perfil de lipídios de um extrato celular de Euglena gracilis conforme descrito nos Exemplos.
A Fig. 14 é um mapa do plasmídeo pKKE2 (vide também SEQ ID N° 40).
A Fig. 15 é um mapa do plasmídeo pKR973 (vide também SEQ ID N° 41).
A Fig. 16 é um mapa do plasmídeo pKR1005 (vide também SEQ ID N° 44).
A Fig. 17 exibe os perfis de ácido graxo para embriões do evento 4838-4-18 que possui os níveis mais altos de ácido araquidônico (vide Exemplo 13).
A Fig. 18 exibe um cromatograma do perfil de lipídios de um extrato celular de Euglena anabaena conforme descrito nos Exemplos. A Fig. 19 é uma mapa do plasmídeo pKR1183. A Fig. 20 é uma mapa do plasmídeo pKR1253. A Fig. 21 é uma mapa do plasmídeo pKR1255. A Fig. 22 é uma mapa do plasmídeo pKR1189.
A Fig. 23 é uma mapa do plasmídeo pKR1229. A Fig. 24 é uma mapa do plasmídeo pKR1249. A Fig. 25 é uma mapa do plasmídeo pKR1193. A Fig. 26 exibe os perfis de ácidos graxos para cinco eventos que possuem o teor médio de ALA mais baixo (média dos cinco embriões somáticos de soja analisados) junto com um evento (2148-3-8-1) que possui um perfil de ácido graxo típico de embriões do tipo selvagem para este experimento. Os ácidos graxos são identificados como 16:0 (palmitato), 18:0 (ácido esteárico), 18:1 (ácido oleico), LA e ALA. Composições de ácidos graxos são expressas como um percentual em peso (% peso) do total de ácidos graxos.
A Fig. 27 exibe os perfis de ácidos graxos para cinco eventos que possuem o teor médio de DGLA mais alto (média de cinco embriões somáticos de soja analisados). Os ácidos graxos são identificados como 16:0 (palmitato), 18:0 (ácido esteárico), 18:1 (ácido oleico), LA, ALA, EDA, ERA, DGLA e ETA. As composições de ácidos graxos são expressas na forma de percentual em peso (% em peso) do total de ácidos graxos.
A Fig. 28 exibe os perfis médios de ácidos graxos (média de dez embriões somáticos de soja) para vinte eventos que possuem o ARA mais alto. Os ácidos graxos são identificados como 16:0 (palmitato), 18:0 (ácido esteárico), 18:1 (ácido oleico), LA, ALA, EDA, SCI, DGLA, ARA, ERA, JUN, ETA e EPA. As composições de ácido graxo são expressas na forma de percentual em peso (% em peso) de ácidos graxos totais. Os ácidos graxos relacionados como "outros" incluem: 18:2 (5,9), 18:3 (5,9,12), STA1 20:0, 20:1 (11), 20:2 (7,11) ou 20:2 (8,11) e DPA.
A Fig. 29 exibe os perfis reais de ácidos graxos para cada embrião somático de soja de um evento (AFS 5416-8-1-1) que contém um teor médio de ARA de 17,0% e teor médio de EPA de 1,5%. Os ácidos graxos são identificados como 16:0 (palmitato), 18:0 (ácido esteárico), 18:1 (ácido oleico), LA, ALA, EDA, SCI, DGLA, ARA, ERA, JUN, ETA e EPA; as composições de ácidos graxos são expressas na forma de percentual em peso (% em peso) do total de ácidos graxos. Os ácidos graxos relacionados como "outros" incluem: 18:2 (5,9), 18:3 (5,9,12), STA, 20:0, 20:1 (11), 20:2 (7,11) ou 20:2 (8,11) e DPA.
A Fig. 30 exibe os perfis médios de ácidos graxos (média de nove ou dez embriões somáticos de soja) para vinte eventos que possuem o ARA mais alto. Os ácidos graxos são identificados como 16:0 (palmitato), 18:0 (ácido esteárico), 18:1 (ácido oleico), LA, ALA, EDA, SCI, DGLA, ARA, ERA, JUN, ETA e EPA; as composições de ácidos graxos são expressas na forma de percentual em peso (% em peso) do total de ácidos graxos. Os ácidos graxos relacionados como "outros" incluem: 18:2 (5,9), 18:3 (5,9,12), STA, 20:0,20:1 (11), 20:2 (7,11) ou 20:2 (8,11) e DPA.
A Fig. 31 exibe os perfis de lipídios de semente de Arabidopsis a granel T2 para os dezenove eventos transformados. Os ácidos graxos são identificados como 16:0 (palmitato), 18:0 (ácido esteárico), 18:1 (ácido oleico), LA, ALA, 20:1 (ácido eicosenoico), EDA, DGLA, ERA, ETA e EPA. As composições de ácidos graxos são expressas na forma de percentual em peso (% peso) do total de ácidos graxos. Os ácidos graxos relacionados como "outros" incluem: 18:2 (5,9), 18:3 (5,9,12), STA, 20:0 (ácido araquídico), 20:2 (7,11) ou 20:2 (8,11), SCI e JUP.
A Fig. 32 exibe os perfis de lipídios de sementes de Arabidopsis T2 individuais transformadas com pKR1193 para expressão de uma delta-9 elongase de Euglena anabaena c om uma delta-8 dessaturase de Euglena anabaena e uma delta-5 dessaturase de Euglena anabaena em Arabidopsis. Os ácidos graxos são identificados como 16:0 (palmitato), 18:0 (ácido esteárico), 18:1 (ácido oleico), LA, GLA, ALA, STA, 20:1 (ácido eicosenoico), EDA, DGLA, ERA, ETA, EPA e DPA; e composições de ácidos graxos relacionadas na Fig. 32 são expressas como um percentual em peso (% em peso) do total de ácidos graxos. Para a Fig. 32, os ácidos graxos relacionados como "outros" incluem: 18:3 (5,9,12), 20:0 (ácido araquídico), 20:2 (7,11) ou 20:2 (8,11)e 22:0.
As descrições de seqüências resumem a Listagem de Seqüências anexa ao presente. A Listagem de Seqüências contém códigos de uma letra para caracteres de seqüências de nucleotídeos e os códigos de uma e três letras para aminoácidos conforme definido nos padrões IUPAC-IUB descritos em Nucleic Acids Research 13: 3021-3030 (1985) e no Biochemical Journal 219 (2): 345-373 (1984).
SEQ ID Nº 1 é a seqüência de nucleotídeos de KS129. SEQ ID Nº 2 é a seqüência de nucleotídeos de pKR457. SEQ ID Nº 3 é a seqüência de nucleotídeos de pKR606. SEQ ID Nº 4 é a seqüência de nucleotídeos de pKR277. SEQ ID Nº 5 é a seqüência de nucleotídeos de pKR804. SEQ ID Nº 6 é a seqüência de nucleotídeos de pKR953. SEQ ID Nº 7 é a seqüência de nucleotídeos de pKR1084. SEQ ID Nº 8 é a seqüência de nucleotídeos de pKR1002. SEQ ID Nº 9 é a seqüência de nucleotídeos de pKR226. SEQ ID Nº 10 é a seqüência de nucleotídeos de BC-TpomALS. SEQ ID Nº 11 é a seqüência de nucleotídeos de BC- Tpom_ALSrev.
SEQ ID Nº 12 é a seqüência de nucleotídeos de pKR132. SEQ ID Nº 13 é a seqüência de nucleotídeos do oligonucleotídeo
EgD5-5Not.
SEQ ID Nº 14 é a seqüência de nucleotídeos do oligonucleotídeo
EgD5-3Not.
SEQ ID Nº 15 é a seqüência de nucleotídeos de pKR268. SEQ ID Nº 16 é a seqüência de nucleotídeos de Ann-EgD5. SEQ ID Nº 17 é a seqüência de nucleotídeos de BC-Tpom_Ann-
EgD5_ALS. SEQ ID N0 18 é a seqüência de nucleotídeos de BC-Tpom_Ann- EgD5_ALSrev.
SEQ ID N0 19 é a seqüência de nucleotídeos de KS133. SEQ ID N0 20 é a seqüência de nucleotídeos de KS263. SEQ ID N0 21 é a seqüência de nucleotídeos de ELfad3-5Not. SEQ ID N0 22 é a seqüência de nucleotídeos de Elfad3-3Not. SEQ ID N0 23 é a seqüência de nucleotídeos do conjunto de ELVISLIVES/fad3 ELfad3Not.
SEQ ID N0 24 é a seqüência de nucleotídeos de pKR179. SEQ ID N0 25 é a seqüência de nucleotídeos de BC-ELfad3. SEQ ID N0 26 é a seqüência de nucleotídeos de BC-Tpom Ann- HPfad3ABA'_ALS.
SEQ ID N0 27 é a seqüência de nucleotídeos de BC-ELfad3_ALS. SEQ ID N0 28 é a seqüência de nucleotídeos de Ann-ELfad3. SEQ ID N0 29 é a seqüência de nucleotídeos de BC-Tpom_Ann-
ELfad3_ALS.
SEQ ID N0 30 é a seqüência de nucleotídeos de HPfad3-1. SEQ ID N0 31 é a seqüência de nucleotídeos de HPfad3-2. SEQ ID N0 32 é a seqüência de nucleotídeos de HPfad3AB. SEQ ID N0 33 é a seqüência de nucleotídeos de HPfad3-3. SEQ ID N0 34 é a seqüência de nucleotídeos de HPfad3-4. SEQ ID N0 35 é a seqüência de nucleotídeos de HPfad3A'. SEQ ID N0 36 é a seqüência de nucleotídeos de HPfad3ABA'. SEQ ID N0 37 é a seqüência de nucleotídeos de BC-HPfad3ABA'. SEQ ID N0 38 é a seqüência de nucleotídeos de Ann-
HPfad3ABA'.
SEQ ID N0 39 é a seqüência de nucleotídeos de HPfad3ABA' ALS. SEQ ID N° 40 é a seqüência de nucleotídeos de pKKE2. SEQ ID N° 41 é a seqüência de nucleotídeos de pKR973. SEQ ID N° 42 é a seqüência de nucleotídeos do primer de oligonucleotídeo oEugEL1-1. SEQ ID N° 43 é a seqüência de nucleotídeos do primer de oligonucleotídeo oEugEL1-2. SEQ ID N° 44 é a seqüência de nucleotídeos de pKR1005. SEQ ID N° 45 é a seqüência de nucleotídeos de pKR906.
Tabela 2
Resumo de Seqüências de Ácidos Nucleicos e Aminoácidos de Vários Genes de Processos Biossintéticos de PUFA
<table>table see original document page 13</column></row><table> SEQ ID N0 60 é a seqüência de nucleotídeos do oligonucleotídeo
SMART IV.
SEQ ID N0 61 é a seqüência de nucleotídeos do primer adaptador de kit 3'-RACE da Invitrogen.
SEQ ID N0 62 é a seqüência de nucleotídeos de TpomNot-5.
SEQ ID N0 63 é a seqüência de nucleotídeos de TpomNot-3.
SEQ ID N0 64 é a seqüência de nucleotídeos de pLKF114-10.
SEQ ID N0 65 é a seqüência de nucleotídeos de pKR271.
SEQ ID N0 66 é a seqüência de nucleotídeos de pKR886r.
SEQ ID N0 67 é a seqüência de nucleotídeos de pKR72.
SEQ ID N0 68 é a seqüência de nucleotídeos de oCon-1.
SEQ ID N0 69 é a seqüência de nucleotídeos de oCon-2. SEQ ID N0 70 é a seqüência de nucleotídeos do conjunto Kti/Notl/Kti3'Salb3' modificado.
SEQ ID N0 71 é a seqüência de nucleotídeos de pKR970.
SEQ ID N0 72 é a seqüência de nucleotídeos de PvDES5'Not-1. SEQ ID N0 73 é a seqüência de nucleotídeos de PvDES3'Not-1. SEQ ID N0 74 é a seqüência de nucleotídeos de pLF113. SEQ ID N0 75 é a seqüência de nucleotídeos de pKR287. SEQ ID N0 76 é a seqüência de nucleotídeos de pKR1155.
SEQ ID N0 77 é a seqüência de nucleotídeos de pKR952. SEQ ID N0 78 é a seqüência de nucleotídeos de primer M13F. SEQ ID N0 79 é a seqüência de aminoácidos de Pavlova sp. CCMP459 C20-PUFA Elongase (Acesso NCBI n° AAV33630 (Gl 54307108; CDS AY630573).
SEQ ID N0 80 é a seqüência de nucleotídeos da extremidade 5' do inserto de cDNA de eeg1c.pk016.e6.f.
SEQ ID N0 81 é a seqüência de nucleotídeos de eeg1c.pk016.e6.f.
SEQ ID N0 82 é a seqüência de nucleotídeos da região de códon de eeg1c.pk016.e6.f.
SEQ ID N0 83 é a seqüência de aminoácidos deduzida de SEQ ID N0 82, também denominada seqüência de aminoácidos da DHA sintase 1 de Euglena gracilis (EgDHAsynI).
SEQ ID N0 84 é a seqüência de aminoácidos da dessaturase de ácido graxo delta-4 de Euglena gracilis.
SEQ ID N0 85 é a seqüência de aminoácidos da C20PUFA elongase 2 de Ostreococcus tauri.
SEQ ID N0 86 é a seqüência de aminoácidos da C20PUFA elongase 2 de Thalassiosira pseudonana.
SEQ ID N0 87 é a seqüência de nucleotídeos da delta-4 dessaturase de Thraustochytrium aureum.
SEQ ID N0 88 é a seqüência de aminoácidos da delta-4 dessaturase de Schizoehytrium aggregatum.
SEQ ID N0 89 é a seqüência de aminoácidos da delta-4 dessaturase de Thalassiosira pseudonana.
SEQ ID N0 90 é a seqüência de aminoácidos da delta-4 dessaturase de Isoehrysis galbana.
SEQ ID N0 91 é a seqüência de nucleotídeos do domínio EgDHAsynI C20 elongase.
SEQ ID N0 92 é a seqüência de aminoácidos do domínio EgDHAsynI C20 elongase (EgDHAsyn1C20Elo1).
SEQ ID N0 93 é a seqüência de aminoácidos do motivo NG de KNGK.
SEQ ID N0 94 é a seqüência de aminoácidos do motivo NG de PENGA. SEQ ID N° 95 é a seqüência de aminoácidos do motivo NG de PENGA.
SEQ ID N° 96 é a seqüência de aminoácidos do motivo NG de PCENGTV.
SEQ ID N° 97 é a seqüência de nucleotídeos do Iigante rico em prolina EgDHAsynI.
SEQ ID N° 98 é a seqüência de aminoácidos do Iigante rico em prolina EgDHAsynI.
SEQ ID N° 99 é a seqüência de nucleotídeos de pLF121-1.
SEQ ID N° 100 é a seqüência de nucleotídeos de pLF121-2.
SEQ ID N° 101 é a seqüência de nucleotídeos da região de códon de EaD9Elo1.
SEQ ID N° 102 é a seqüência de nucleotídeos da região de códon de EaD9Elo2.
SEQ ID N° 103 é a seqüência de aminoácidos de EaD9Elo1.
SEQ ID N° 104 é a seqüência de aminoácidos de EaD9Elo2.
SEQ ID N° 105 é a seqüência de nucleotídeos de pLF119.
SEQ ID N° 106 é a seqüência de nucleotídeos da região de códon de EaD5Des1.
SEQ ID N° 107 é a seqüência de aminoácidos de EaD5Des1.
SEQ ID N° 108 é a seqüência de nucleotídeos de EaD9-5Bbs.
SEQ ID N° 109 é a seqüência de nucleotídeos de EaD9-3fusão.
SEQ ID N° 110 é a seqüência de nucleotídeos de EgDHAsynI Ligação-5fusão.
SEQ ID N° 111 é a seqüência de nucleotídeos de MWG511.
SEQ ID N° 112 é a seqüência de nucleotídeos de EgD9elo- EgDHAsynI Ligação.
SEQ ID N° 113 é a seqüência de nucleotídeos de pLF124. SEQ ID N° 114 é a seqüência de nucleotídeos de KS366.
SEQ ID N° 115 é a seqüência de nucleotídeos de pKR1777.
SEQ ID N° 116 é a seqüência de nucleotídeos de pKR1179.
SEQ ID N° 117 é a seqüência de nucleotídeos de pKR1183.
SEQ ID N° 118 é a seqüência de nucleotídeos de pKR1237.
SEQ ID N° 119 é a seqüência de nucleotídeos de pKR1252.
SEQ ID N° 120 é a seqüência de nucleotídeos de pKR1253.
SEQ ID N° 121 é a seqüência de nucleotídeos de oligonucleotídeo oEAd5-1-1.
SEQ ID N° 122 é a seqüência de nucleotídeos de oligonucleotídeo oEAd5-1-2.
SEQ ID N° 123 é a seqüência de nucleotídeos de pKR1136.
SEQ ID N° 124 é a seqüência de nucleotídeos de pKR1139.
SEQ ID N° 125 é a seqüência de nucleotídeos de pKR561.
SEQ ID N0 126 é a seqüência de nucleotídeos de oligonucleotídeo HPfad3A'-2.
SEQ ID N° 127 é a seqüência de nucleotídeos de oligonucleotídeo HPfad3ABA'-2.
SEQ ID N° 128 é a seqüência de nucleotídeos de pLF129.
SEQ ID N° 129 é a seqüência de nucleotídeos de pKR1189.
SEQ ID N° 130 é a seqüência de nucleotídeos de pKR1209.
SEQ ID N° 131 é fad3c CDS.
SEQ ID N° 132 é a seqüência de aminoácidos de fad3C.
SEQ ID N° 133 é a seqüência de nucleotídeos de oligonucleotídeo fad3c-5.
SEQ ID N° 134 é a seqüência de nucleotídeos de oligonucleotídeo fad3c-3.
SEQ ID N° 135 é a seqüência de nucleotídeos de pKR1213. SEQ ID N0 136 é a seqüência de nucleotídeos de pKR1218. SEQ ID N0 137 é a seqüência de nucleotídeos de pKR1210. SEQ ID N0 138 é a seqüência de nucleotídeos de pKR1219. SEQ ID N0 139 é a seqüência de nucleotídeos de pKR1225. SEQ ID N0 140 é a seqüência de nucleotídeos de pKR1229. SEQ ID N0 141 é a seqüência de nucleotídeos de pKR1249. SEQ ID N0 142 é a seqüência de nucleotídeos de delta-8 dessaturase CDS de Euglena gracilis (Eg5).
SEQ ID N0 143 é a seqüência de nucleotídeos do primer específico de vetor pDonor222Eg5-1.
SEQ ID N0 144 é a seqüência de nucleotídeos de D8DEG3-1. SEQ ID N0 145 é a seqüência de nucleotídeos de D8DEG3-2. SEQ ID N0 146 é a seqüência de nucleotídeos de D8DEG3-3. SEQ ID N0 147 é a seqüência de nucleotídeos de D8DEG3-4. SEQ ID N0 148 é a seqüência de nucleotídeos de pHD23-1.
SEQ ID N0 149 é a seqüência de aminoácidos de delta-8 dessaturase de Euglena gracilis (Acesso NCBI n° AAD45877).
SEQ ID N0 150 é a seqüência de nucleotídeos de pLF118-3. SEQ ID N0 151 é a seqüência de nucleotídeos da região de códon de EaD8Des3.
SEQ ID N0 152 é a seqüência de aminoácidos de EaD8Des3. SEQ ID N0 153 é a seqüência de nucleotídeos de oligonucleotídeo EaD8-5.
SEQ ID N0 154 é a seqüência de nucleotídeos de oligonucleotídeo EaD8-3.
SEQ ID N0 155 é a seqüência de nucleotídeos de pLF120-3. SEQ ID N0 156 é a seqüência de nucleotídeos de oEAd9el1-1. SEQ ID N0 157 é a seqüência de nucleotídeos de oEAd9el1-2. SEQ ID N0 158 é a seqüência de nucleotídeos de pKR1137. SEQ ID N0 159 é a seqüência de nucleotídeos de pKR1140. SEQ ID N0 160 é a seqüência de nucleotídeos de pKR1173. SEQ ID N0 161 é a seqüência de nucleotídeos de pKR393. SEQ ID N0 162 é a seqüência de nucleotídeos de pKR407. SEQ ID N0 163 é a seqüência de nucleotídeos de pKR1176. SEQ ID N0 164 é a seqüência de nucleotídeos de pKR1178. SEQ ID N0 165 é a seqüência de nucleotídeos de oligo oKti5 SEQ ID N0 166 é a seqüência de nucleotídeos de oligo oKti6 SEQ ID N0 167 é a seqüência de nucleotídeos de pKR193. SEQ ID N0 168 é a seqüência de nucleotídeos de pKR1174. SEQ ID N0 169 é a seqüência de nucleotídeos de pKR1186. SEQ ID N0 170 é a seqüência de nucleotídeos de pKR1193. SEQ ID N0 171 é a seqüência do primer de SeqE. SEQ ID N0 172 é a seqüência do primer de SeqW.
Descrição Detalhada da Invenção
O relatório descritivo de cada referência mencionada no presente é integralmente incorporado ao presente como referência.
Conforme utilizado no presente e nas reivindicações anexas, as formas no singular "um", "uma", "o" e "a" indicam referência ao plural, a menos que o contexto indique claramente em contrário. Desta forma, por exemplo, referência a "uma planta" inclui uma série dessas plantas e referência a "uma célula" inclui uma ou mais células e seus equivalentes conhecidos dos técnicos no assunto e assim por diante.
No contexto do presente relatório descritivo, utiliza-se uma série de termos e abreviações. São fornecidas as definições a seguir.
O termo "EgD9elo" designa uma delta-9 elongase isolada de Euglena gracilis (SEQ ID N0 46 e 47; vide o pedido copendente do Cessionário do Depositante que possui Pedido Norte-Americano n° 11/601.563 (depositado em dezesseis de novembro de 2006, que foi publicado em 24 de maio de 2007; Pasta do Advogado n° BB-1562).
O termo "PavD8" designa uma enzima delta-8 dessaturase isolada de Pavlova Iutheri (SEQ ID N0 48 e 49; vide também o pedido copendente do Cessionário do Depositante que possui Pedido de Patente Norte-Americano n° 11/737772 (depositado em vinte de abril de 2007; Pasta do Advogado n° BB-1566).
O termo "TpomD8" designa uma enzima delta-8 dessaturase isolada de Tetruetreptia pomquetensis CCMP1491 (SEQ ID N0 50 e 51; vide também o pedido copendente do Cessionário do Depositante que possui Pedido de Patente Norte-Americano n° 11/876115 (depositado em 22 de outubro de 2007; Pasta do Advogado n° BB-1574).
O termo "MaD5" designa uma enzima delta-5 dessaturase isolada de Mortierella alpina (SEQ ID N0 52 e 53; vide também Acesso GenBank n° AF067654 e Patente Norte-Americana n° 6.075.183).
O termo "EgD5" designa uma enzima delta-5 dessaturase isolada de Euglena gracilis (SEQ ID N0 54 e 55; vide também o Pedido Copendente de Depositante que possui Pedido de Patente Norte-Americano n° 11/748629 (depositado em quinze de maio de 2007, publicado em vinte de dezembro de 2007; Pasta do Advogado n° CL-3486).
O termo "SdD17" designa uma enzima delta-17 dessaturase isolada de Saprolegnia diclina (SEQ ID N0 56 e 57).
O termo "FmD15" designa uma enzima delta-15 dessaturase isolada de Fusarium moniliforme (SEQ ID N0 58 e 59; vide também o Pedido Copendente do Cessionário do Depositante que possui Patente PCT n0 WO 05/047479 (publicada em 26 de maio de 2005; Pasta do Advogado n0 CL- 2432). "Quadro de leitura aberta" é abreviado ORF.
"Reação em cadeia de polimerase" é abreviada PCR.
"Coleção Norte-Americana de Cultivos de Tipos" é abreviada ATCC.
"Ácido(s) graxo(s) póli-insaturado(s)" é(são) abreviado(s) PUFA(s).
"Triacilgliceróis" são abreviados TAGs.
A expressão "ácido graxos" designa ácidos alifáticos de cadeia longa (ácidos alcanoicos) com comprimentos de cadeia variáveis de cerca de C12 a C22 (embora sejam conhecidos ácidos com comprimento de cadeia mais longo e mais curto). Os comprimentos de cadeia predominantes são de Ci6 a C22· Detalhes adicionais referentes à diferenciação entre "ácidos graxos saturados" e "ácidos graxos insaturados", "ácidos graxos monoinsaturados" e "ácidos graxos póli-insaturados" (ou "PUFAs") e "ácidos graxos ômega 6" (n-6 ou n-6) e "ácidos graxos ômega 3" (n-3 ou n-3) são fornecidos na Patente PCT n0 WO 04/101757.
Ácidos graxos ômega 3 são uma família de ácidos graxos póli- insaturados que contêm em comum uma união dupla carbono-carbono na posição ômega 3. A expressão ômega 3 ("n-3", "ω-3") significa que a primeira união dupla existe como a terceira união carbono-carbono a partir da extremidade metila terminal (ômega) da cadeia de carbono. Ácidos graxos ômega 3 importantes em nutrição são os seguintes: ácido alfalinolênico (ALA), ácido eicosapentaenoico (EPA) e ácido docosa-hexanoico (DHA). O corpo humano não pode sintetizar ácidos graxos ômega 3 novos, mas pode sintetizar todos os demais ácidos graxos ômega 3 necessários a partir do ácido graxo ômega 3 mais simples ALA. ALA é, portanto, um nutriente essencial que deve ser obtido de alimentos e os outros ácidos graxos ômega 3 que podem ser sintetizados a partir dele no interior do corpo ou obtidos de alimentos às vezes são também denominados nutrientes essenciais. A Tabela 3 abaixo relaciona ácidos graxos ômega 3.
Tabela 3
Ácidos Graxos Ômega 3
<table>table see original document page 22</column></row><table>
Ácidos graxos ômega 6 são ácidos graxos em que a expressão "ômega 6" significa que a primeira união dupla na cadeia principal de carbono do ácido graxo ocorre na posição ômega menos 6; ou seja, o sexto carbono a partir da extremidade do ácido graxo. Ácido Iinoleico (18:2), o ácido graxo ômega 6 de cadeia mais curta, é um ácido graxo essencial. Ácido araquidônico (20:4) é um ácido graxo n-6 fisiologicamente significativo e é o precursor de prostaglandinas e outras moléculas fisiologicamente ativas. A Tabela 4 define os ácidos graxos ômega 6.
Tabela 4
Ácidos Graxos Ômega 6
<table>table see original document page 22</column></row><table> <table>table see original document page 23</column></row><table>
Os ácidos graxos são descritos no presente por meio de um sistema de notação simples de "X:Y", em que X é o número de átomos de carbono (C) no ácido graxo específico e Y é o número de uniões duplas. O número após a designação de ácido graxo indica a posição da união dupla a partir da extremidade carboxila do ácido graxo com o afixo "c" para a configuração eis da união dupla (tal como ácido palmítico (16:0), ácido esteárico (18:0), ácido oleico (18:1, 9c), ácido petroselínico (18:1, 6c), LA (18:2, 9c, 12c), GLA (18:3, 6c, 9c, 12c) e ALA (18:3, 9c, 12c, 15c)). A menos que especificado em contrário, 18:1, 18:2 e 18:3 designam ácidos graxos oleico, LA e ALA, respectivamente. Caso não seja descrito especificamente em contrário, considera-se que as uniões duplas possuem a configuração eis. As uniões duplas em 18:2 (9,12), por exemplo, seriam consideradas na configuração eis.
A nomenclatura utilizada para descrever PUFAs no presente relatório descritivo é exibida abaixo na Tabela 5. Na coluna intitulada "Notação Abreviada", o sistema de referência ômega é utilizado para indicar o número de carbonos, o número de uniões duplas e a posição da união dupla mais próxima ao carbono ômega, a partir do carbono ômega (que é numerado 1 com este propósito). O restante da tabela resume os nomes comuns de ácidos graxos ômega 3 e ômega 6 e seus precursores, as abreviações que serão utilizadas ao longo de todo o restante do presente relatório descritivo e o nome químico de cada composto. Tabela 5
Nomenclatura de Ácidos Graxos Póli-Insaturados ε Precursores
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A expressão "ácido araquidônico" ("ARA") designa um ácido graxo ômega 6 que possui a fórmula química C20H32O2. Também recebe o nome 20:4 (n-6). O seu nome químico sistemático é descrito acima na Tabela 5. É um componente alimentício essencial para mamíferos. O ácido livre é o precursor da biossíntese de prostaglandinas, tromboxanos, derivados de ácido hidroxieicosatetraenoico que incluem leucotrienos. Dentro das células, o ácido é encontrado na forma esterificada como um componente acila principal de fosfolipídios de membranas. Pouco ou nenhum ARA é encontrado nas plantas. O termo ARA, da forma utilizada no presente, engloba o ácido livre e seus derivados, tais como sua forma esterificada.
A expressão "produção de ARA em alto nível" designa uma planta oleaginosa transgênica que produz sementes maduras nas quais o perfil total de ácido graxo da semente compreende pelo menos 10% de ARA, pelo menos 15% de ARA, pelo menos 20% de ARA ou pelo menos 25% de ARA. A forma estrutural do ARA não é limitadora; desta forma, o ARA pode existir, por exemplo, no perfil de ácido graxo de semente na forma de ácidos graxos livres ou em formas esterificadas tais como acilgliceróis, fosfolipídios, sulfolipídios ou glicolipídios.
A expressão "teor reduzido de ácidos graxos ômega 3" designa uma planta oleaginosa transgênica que produz sementes maduras nas quais o perfil de ácido graxo de semente total compreende menos de 5% do total de ácidos graxos ômega 3 ou, de maior preferência, menos de 1% do total de ácidos graxos ômega 3 ou qualquer percentual inteiro de 5% a 1%, tal como 4%, 3% e 2%.
A expressão "teor reduzido de GLA" designa uma planta oleaginosa transgênica que produz sementes maduras em que o perfil de ácido graxo de semente total compreende menos de 5% de GLA total ou, de maior preferência, até 1% de GLA total, inclusive, ou qualquer percentual inteiro de 5% a 1%, tal como 4%, 3% e 2%.
Um processo metabólico ou processo biossintético, em um sentido bioquímico, pode ser considerado como uma série de reações químicas que ocorrem no interior de uma célula, catalisado por enzimas, para atingir a formação de um produto metabólico a ser utilizado ou armazenado pela célula, ou o início de um outro processo metabólico (denominado em seguida etapa de geração de fluxo). Muitos desses processos são elaborados e envolvem uma modificação etapa por etapa da substância inicial para moldá-la em um produto que possui a estrutura química exata desejada.
A expressão "processo biossintético de PUFA" designa um processo metabólico que converte ácido oleico em LA, EDA, GLA, DGLA, ARA, ALA, STA, ETrA1 ETA, EPA1 DPA e DHA. Este processo é bem descrito na literatura (vide, por exemplo, a Patente PCT n0 WO 06/052870). De forma simplista, este processo envolve o alongamento da cadeia de carbono por meio da adição de átomos de carbono e dessaturação da molécula por meio da adição de uniões duplas, por uma série de enzimas de dessaturação e alongamento especiais (ou seja, "enzimas do processo biossintético de PUFA") presentes na membrana de retículo endoplasmático. Mais especificamente, "enzima do processo biossintético de PUFA" designa qualquer das enzimas a seguir (e genes que codificam as mencionadas enzimas) associadas à biossíntese de PUFA, que incluem: uma delta-4 dessaturase, delta-5 dessaturase, delta-6 dessaturase, delta-12 dessaturase, delta-15 dessaturasse, delta-17 dessaturase, delta-9 dessaturase, delta-8 dessaturase, delta-9 elongase, C14/16 elongase, Cie/18 elongase, C18/20 elongase e C20/22 elongase.
A expressão "processo biossintético de ácido graxo ômega 3/ômega 6" designa um conjunto de genes que, quando expressos sob as condições apropriadas, codificam enzimas que catalisam a produção de ácidos graxos ômega 3 e ômega 6. Tipicamente, os genes envolvidos no processo biossintético de ácido graxo ômega 3/ômega 6 codificam enzimas de processo biossintético de PUFA. Um processo representativo é ilustrado na Fig. 1, fornecendo a conversão de ácido mirístico por meio de diversos intermediários de DHA, o que demonstra como ácidos graxos ômega 3 e ômega 6 podem ser produzidos a partir de uma fonte comum. O processo é naturalmente dividido em duas porções, em que uma porção gerará ácidos graxos ômega 3 e a outra porção, ácidos graxos ômega 6.
O termo "funcional", da forma utilizada no presente em contexto com o processo biossintético de ácidos graxos ômega 3 e ômega 6, indica que alguns dos genes (ou todos) no processo expressam enzimas ativas, o que resulta em catálise in vivo ou conversão de substrato. Dever-se-á compreender que "processo biossintético de ácido graxo ômega 3/ômega 6" ou "processo biossintético de ácido graxo ômega 3/ômega 6 funcional" não indica que todos os genes de enzimas do processo biossintético de PUFA sejam necessários, pois uma série de produtos de ácido graxo necessitará apenas da expressão de um subconjunto dos genes deste processo.
A expressão "processo de delta-9 elongase/delta-8 dessaturase" designa um processo biossintético de produção de PUFAs de cadeia longa. Este processo compreende, no mínimo, uma delta-9 elongase e uma delta-8 dessaturase, de forma a permitir a biossíntese de DGLA e/ou ETA a partir de LA e ALA, respectivamente. Com a expressão de outras dessaturases e elongases, ARA, EPA1 DPA e DHA podem também ser sintetizados. Este processo pode ser vantajoso em algumas realizações, pois a biossíntese de GLA e/ou STA é excluída.
A expressão "ácido graxo intermediário" designa qualquer ácido graxo produzido em um processo metabólico de ácido graxo que possa ser adicionalmente convertido em um produto ácido graxo desejado neste processo por meio da ação de outras enzimas de processos metabólicos. Ao produzir-se, por exemplo, EPA utilizando o processo de delta-9 elongase/delta- 8 dessaturase, EDA, ETrA, DGLA, ETA e ARA podem ser produzidos e são considerados "ácidos graxos intermediários", pois esses ácidos graxos podem ser adicionalmente convertidos em EPA por meio da ação de outras enzimas de processos metabólicos.
A expressão "subproduto ácido graxo" designa qualquer ácido graxo produzido em um processo metabólico de ácido graxo que não seja o produto de ácido graxo pretendido do processo nem um "ácido graxo intermediário" do processo. Ao produzir-se EPA utilizando, por exemplo, o processo de delta-9 elongase/delta-8 dessaturase, podem também ser produzidos ácido sciadônico (SCI) e ácido juniperônico (JUP) por meio da ação de uma delta-5 dessaturase sobre EDA ou ETrA1 respectivamente. Eles são considerados "subprodutos ácidos graxos", pois nenhum pode ser adicionalmente convertido em EPA por meio da ação de outras enzimas de processos metabólicos.
O termo "fad3" designa um gene que codifica (ou seja, causa a produção de) uma delta-15 ou ômega-3 dessaturase. Especificamente, a enzima delta-15 dessaturase converte ácido Iinoleico em ácido alfalinolênico. FAD3 e/ou fad3 designa qualquer dos três genes fad3 de soja.
O primeiro gene ômega-3 dessaturase vegetal identificado foi fad3 de Arabidopsis (Yadav et al, Plant Physiol. 103: 467-476 (1993)) e pelo menos três versões deste gene foram identificadas no genoma de soja, GmFAD3A (Acesso GenBank n° AY204710), GmFAD3B (Acesso GenBank n° AY204711 e L22964) e GmFAD3C (Acesso GenBank n° AY204712) (Bilyeu et al, Crop Sei. 43: 1833-1838 (2003); Anai et al, Plant Sei. 168: 1615-1623 (2005)). Grãos de soja que contêm níveis reduzidos de ALA (alfalinolênico baixo ou "baixo lin") causados por mutações em um ou mais dos genes fad3 foram identificados (analisados em Bilyeu et al, Crop Sei. 46: 1913-1918 (2006)). Mutações em GmFAD3A e GmFAD3C na linhagem de soja CX1512-44, com níveis de ALA de cerca de 3% e em todos os três genes FAD3, gerando grãos de soja com níveis de ALA de cerca de 1% dos ácidos graxos totais (tais como linhagem A29 (Ross et al, Crop Sei. 40: 383-386 (2000)) foram caracterizadas no nível molecular (vide Bilyeu et al, 2006)).
As expressões "regular para baixo" ou "regulagem para baixo", da forma utilizada no presente, designam redução ou decréscimo do nível de expressão de um gene ou polinucleotídeo.
A regulagem para baixo de qualquer combinação dos genes fad3 em uma planta de soja elaborada geneticamente utilizando métodos descritos no presente faz com que a planta de soja produza sementes maduras que possuem um perfil de óleo composto de baixos níveis de ácido linolênico (soja de baixo lin). Por outro lado, o genoma de uma planta de soja poderá ser elaborado para aumentar a expressão de genes fad3, de forma a resultar em um perfil de óleo em que o nível de ácido linolênico constitui cerca de 50% do total de ácidos graxos no óleo obtido a partir dessas plantas de soja transgênica.
Os termos "triacilglicerol", "óleo" e "TAGs" designam lipídios neutros compostos de três resíduos acila graxos esterificados em uma molécula de glicerol (e esses termos serão utilizados de forma intercambíável ao longo de todo o presente relatório descritivo). Esses óleos podem conter PUFAs de cadeia longa, bem como ácidos graxos saturados e insaturados mais curtos e ácidos graxos saturados com cadeia mais longa. Desta forma, "biossíntese de óleo" designa, genericamente, a síntese de TAGs na célula.
"Percentual (%) de PUFAs no total de frações de óleo e lipídio totais" designa o percentual de PUFAs com relação ao total de ácidos graxos nessas frações. A expressão "fração de lipídio total" ou "fração de lipídio" refere-se à soma de todos os lipídios (ou seja, neutros e polares) em um organismo oleaginoso, de forma a incluir os lipídios que estão localizados na fração de fosfatidilcolina (PC), fração de fosfatidiletanolamina (PE) e fração de triacilglicerol (TAG ou óleo). Os termos "lipídio" e "óleo" serão utilizados, entretanto, de forma intercambiável ao longo de todo o relatório descritivo.
As expressões "eficiência de conversão" e "percentual de conversão de substrato" designam a eficiência com que uma enzima específica (tal como uma dessaturase) pode converter substrato em produto. A eficiência de conversão é medida de acordo com a fórmula a seguir: ([produto]/[substrato + produto]) * 100, em que "produto" inclui o produto imediato e todos os produtos no processo dele derivados.
"Dessaturase" é um polipeptídeo que pode dessaturar, ou seja, introduzir uma união dupla em um ou mais ácidos graxos para produzir um ácido graxo ou precursor de interesse. Apesar do uso do sistema de referência ômega ao longo de todo o relatório descritivo para designar ácidos graxos específicos, é mais conveniente indicar a atividade de uma dessaturase por meio de contagem a partir da extremidade carboxila do substrato utilizando o sistema delta. São de interesse específico no presente delta-8 dessaturases que dessaturarão um ácido graxo entre o oitavo e o nono átomo de carbono numerados a partir da extremidade carboxil-terminal da molécula e que podem, por exemplo, catalisar a conversão de EDA em DGLA e/ou ETrA em ETA. Outras dessaturases de ácidos graxos úteis incluem, por exemplo: (1) delta-5 dessaturases que catalisam a conversão de DGLA em ARA e/ou ETA em EPA; (2) delta-6 dessaturases que catalisam a conversão de LA em GLA e/ou ALA em STA; (3) delta-4 dessaturases que catalisam a conversão de DPA em DHA; (4) delta-12 dessaturases que catalisam a conversão de ácido oleico em LA; (5) delta-15 dessaturases que catalisam a conversão de LA em ALA e/ou GLA em STA; (6) delta-17 dessaturases que catalisam a conversão de ARA em EPA e/ou DGLA em ETA; e (7) delta-19 dessaturases que catalisam a conversão de ácido palmítico em ácido palmitoleico (16:1) e/ou ácido esteárico em ácido oleico (18:1). Na técnica, delta-15 e delta-17 dessaturases também são ocasionalmente denominadas "ômega-3 dessaturases", "w-3 dessaturases" e/ou "n-3 dessaturases", com base na sua capacidade de conversão de ácidos graxos ômega 6 nos seus correspondentes ômega 3 (tal como conversão de LA em ALA e ARA em EPA, respectivamente). Em algumas realizações, é mais desejável determinar empiricamente a especificidade de uma dessaturase de ácido graxo específica por meio de transformação de um hospedeiro apropriado com o gene para a dessaturase de ácido graxo específica e determinação do seu efeito sobre o perfil de ácido graxo do hospedeiro.
A expressão "sistema de elongase" designa uma suíte de quatro enzimas que são responsáveis pelo alongamento de uma cadeia de carbono de ácidos graxos para produzir um ácido graxo que é dois carbonos mais longo que o substrato de ácido graxo sobre o qual age o sistema de elongase. Mais especificamente, o processo de alongamento ocorre em associação com sintase de ácido graxo, por meio do quê CoA é o veículo acila (Lassner et al, Plant Cell 8: 281-292 (1996)). Na primeira etapa, que se descobriu ser específica de substrato e também limitadora da velocidade, malonil-CoA é condensado com um acil-CoA de cadeia longa para gerar dióxido de carbono (CO2) e β-cetoacil-CoA (em que a porção acila tenha sido alongada em dois átomos de carbono). Reações subsequentes incluem redução para β- hidroxiacil-CoA, desidratação em um enoil-CoA e uma segunda redução para gerar o acil-CoA alongado. Exemplos de reações catalisadas por sistemas de elongase são a conversão de GLA em DGLA1 STA em ETA1 LA em EDA, ALA > em ETRA e EPA em DPA.
Para os propósitos do presente, uma enzima que catalisa a primeira reação de condensação (ou seja, conversão de malonil-CoA e acil- CvoA de cadeia longa em β-cetoacil-CoA) será denominada genericamente uma "elongase". Geralmente, a seletividade de substrato de elongases é um tanto ampla mas segregada pelo comprimento de cadeia e pelo grau de insaturação. Consequentemente, elongases podem possuir especificidades diferentes. Uma C14/16 elongase, por exemplo, utilizará um substrato Ci4 (tal como ácido mirístico), uma Ci6/ie elongase utilizará um substrato Ci6 (tal como palmitato), uma C18/20 elongase utilizará um substrato Cie (tal como GLA, STA) e uma C20/22 elongase utilizará um substrato C20 (tal como EPA). De forma similar, uma "delta-9 elongase" pode ser capaz de catalisar a conversão de LA em EDA e/ou ALA em ETrA. É importante observar que algumas elongases possuem ampla especificidade e, portanto, uma única enzima pode ser capaz de catalisar várias reações de elongase. Desta forma, uma delta-9 elongase pode também agir, por exemplo como uma C16/18 elongase, C18/20 elongase e/ou C20/22 elongase e pode possuir especificidades alternativas, mas não preferidas, para ácidos graxos delta-5 e delta-6, tais como EPA e/ou GLA, respectivamente.
Da forma utilizada no presente, "ácido nucleico" indica um polinucleotídeo e inclui polímero de fita simples ou dupla de bases desoxirribonucleotídeo ou ribonucleotídeo. Os ácidos nucleicos podem também incluir fragmentos e nucleotídeos modificados. Desta forma, as expressões "polinucleotídeo", "seqüência de ácido nucleico", "seqüência de nucleotídeos" ou "fragmento de ácido nucleico" são utilizadas de forma intercambiável e são um polímero de RNA ou DNA que possui fita simples ou dupla, que contenha opcionalmente bases de nucleotídeos sintéticas, não naturais ou alteradas. Nucleotídeos (normalmente encontrados na sua forma de 5'-monofosfato) são designados por uma denominação de uma letra conforme segue: "A" por adenilato ou desoxiadenilato (para RNA ou DNA1 respectivamente), "C" para citidilato ou desoxicitidilato, "G" para guanilato ou desoxiguanilato, "U" para uridilato, "T" para desoxitimidilato, "R" para purinas (A ou G), "Y" para pirimidinas (C ou Τ), "K" para G ou Τ, "H" para A, C ou Τ, "I" para inosina e "N" para qualquer nucleotídeo.
As expressões "subfragmento que é funcionalmente equivalente" e "subfragmento funcionalmente equivalente" são utilizadas de forma intercambiável no presente. Estas expressões designam uma parte ou subsequência de um fragmento de ácido nucleico isolado no qual a capacidade de alteração da expressão genética ou produção de um certo fenótipo é retida, seja o fragmento ou subfragmento codificador ou não o codificador de uma enzima ativa. O fragmento ou subfragmento pode ser utilizado, por exemplo, no projeto de genes quiméricos para produzir o fenótipo desejado em uma planta transformada. Genes quiméricos podem ser projetados para uso em supressão ligando-se um fragmento de ácido nucleico ou seu subfragmento, codifique ou não uma enzima ativa, na orientação com ou sem sentido com relação a uma seqüência promotora de plantas.
A expressão "domínio conservado" ou "motivo" indica um conjunto de aminoácidos conservados em posições específicas ao longo de uma seqüência alinhada de proteínas relacionadas evolucionariamente. Embora aminoácidos em outras posições possam variar entre proteínas homólogas, aminoácidos que são altamente conservados em posições específicas indicam aminoácidos que são essenciais na estrutura, estabilidade ou atividade de uma proteína. Como são identificados pelo seu alto grau de conservação em seqüências alinhadas de uma família de homólogos de proteína, eles podem ser utilizados como identificadores ou "assinaturas" para determinar se uma proteína com uma seqüência recém determinada pertence a uma família de proteínas identificada anteriormente.
As expressões "homologia", "homólogo", "substancialmente similar" e "substancialmente correspondente" são utilizadas no presente de forma intercambiável. Elas designam fragmentos de ácidos nucleicos em que modificações em uma ou mais bases de nucleotídeos não afetam a capacidade do fragmento de ácido nucleico de mediar a expressão genética ou produzir um certo fenótipo. Estas expressões também designam modificações dos fragmentos de ácidos nucleicos de acordo com a presente invenção, tais como exclusão ou inserção de um ou mais nucleotídeos que não alterem substancialmente as propriedades funcionais do fragmento de ácido nucleico resultante com relação ao fragmento não modificado inicial. Compreende-se, portanto, como apreciarão os técnicos no assunto, que a presente invenção engloba mais que as seqüências de exemplos específicas.
Além disso, os técnicos no assunto reconhecem que seqüências de ácido nucleico substancialmente similares englobadas pela presente invenção também são definidas pela sua capacidade de hibridização (sob condições moderadamente estringentes, tais como 0,5 X SSC, 0,1% SDS, 60 °C) com as seqüências exemplificadas no presente, ou qualquer parte das seqüências de nucleotídeos descritas no presente e que são funcionalmente equivalentes a qualquer das seqüências de ácido nucleico descritas no presente. As condições de estringência podem ser ajustadas para selecionar fragmentos moderadamente similares, tais como seqüências homólogas de organismos com relacionamento distante, até fragmentos altamente similares, tais como genes que duplicam enzimas funcionais de organismos intimamente relacionados. Lavagens após a hibridização determinam as condições de estringência.
A expressão "hibridiza-se seletivamente" inclui referência a hibridização, sob condições de hibridização estringente, de uma seqüência de ácido nucleico em uma seqüência alvo de ácido nucleico especificada até um grau detectavelmente maior (tal como pelo menos duas vezes sobre o fundo) que a sua hibridização em seqüências de ácido nucleico não alvo e na exclusão substancial de ácidos nucleicos não alvo. Seqüências que se hibridizam seletivamente possuem tipicamente pelo menos cerca de 80% de identidade de seqüências, 90% de identidade de seqüências ou até 100% de identidade de seqüências, inclusive (ou seja, totalmente complementares), entre si.
A expressão "condições estringentes" ou "condições de hibridização estringentes" inclui referência a condições sob as quais uma sonda será hibridizada seletivamente na sua seqüência de sinal. Condições estringentes são dependentes de seqüência e serão diferentes em diferentes circunstâncias. Ao controlar-se a estringência das condições de hibridização e/ou lavagem, podem ser identificadas seqüências alvo que são 100% complementares à sonda (sondagem homóloga). Alternativamente, as condições de estringência podem ser ajustadas para permitir alguma falta de coincidência em seqüências, de tal forma que sejam detectados graus mais baixos de similaridade (sondagem heteróloga). Geralmente, uma sonda possui menos de cerca de mil nucleotídeos de comprimento, opcionalmente menos de quinhentos nucleotídeos de comprimento.
Tipicamente, as condições estringentes serão aquelas em que a concentração de sal é de menos de cerca de 1,5 M de íon de Na, tipicamente cerca de 0,01 a 1,0 M de concentração de íons de Na (ou outros sais) sob pH 7,0 a 8,3 e a temperatura é de pelo menos cerca de 30 °C para sondas curtas (tais como dez a cinqüenta nucleotídeos) e pelo menos cerca de 60 °C para sondas longas (tais como mais de cinqüenta nucleotídeos). As condições estringentes podem também ser atingidas com a adição de agentes desestabilizantes tais como formamida. Exemplos de condições de baixa estringência incluem hibridização com uma solução de tampão de 30 a 35% de formamida, 1 M NaCI1 1% SDS (dodecil sulfato de sódio) a 37 °C e lavagem em 1X a 2X SSC (2OX SSC = 3,0 M NaCI/0,3 M citrato trissódico) a 50 até 55 0C. Exemplos de condições de estringência moderadas incluem hibridização em 40 a 45% de formamida, 1 M de NaCI1 1% SDS a 37 °C e lavagem em 0,5X a 1X SSC a 55 até 60 °C. Exemplos de condições de alta estringência incluem hibridização em 50% formamida, 1 M de NaCI1 1% SDS a 37 0C e lavagem em 0,1XSSCa 60 até 65 °C.
A especificidade é tipicamente a função de lavagens pós- hibridização, em que os fatores críticos são a resistência iônica e a temperatura da solução final de lavagem. Para híbridos de DNA e DNA1 a Tm pode ser aproximada a partir da equação de Meinkoth et al, Anal. Biochem. 138: 267- 284 (1984): Tm = 81,5 °C + 16,6 (log M) + 0,41 (%GC) - 0,61 (% forma) - 500/I; em que M é a molaridade de cátions monovalentes, %GC é o percentual de nucleotídeos de citosina e guanosina no DNA, % forma é o percentual de formamida na solução de hibridização e L é o comprimento do híbrido em pares de bases. A Tm é a temperatura (sob resistência iônica e pH definidos) em que 50% de uma seqüência alvo complementar hibridizam-se em uma sonda perfeitamente coincidente. A Tm é reduzida em cerca de 1 °C para cada 1% de falta de coincidência; desta forma, Tm e as condições de hibridização e/ou lavagem podem ser ajustadas para hibridização em seqüências com a identidade desejada. Caso se deseje seqüências com > 90% de identidade, por exemplo, a Tm pode ser reduzida em 10 °C. Geralmente, são selecionadas condições estringentes como sendo cerca de 5 °C mais baixas que o ponto de fusão (Tm) térmico para a seqüência específica e o seu complemento em uma resistência iônica e pH definidos. Condições severamente estringentes podem utilizar, entretanto, uma hibridização e/ou lavagem 1, 2, 3 ou 4°C mais baixas que o ponto de fusão (Tm) térmico; condições moderadamente estringentes podem utilizar uma hibridização e/ou lavagem 6, 7, 8, 9 ou 10°C mais baixas que o ponto de fusão (Tm) térmico; condições de baixa estringência podem utilizar uma hibridização e/ou lavagem 11, 12, 13, 14, 15 ou 20°C mais baixa que o ponto de fusão (Tm) térmico. Utilizando a equação, composições de hibridização e lavagem e a Tm desejada, os técnicos comuns no assunto compreenderão que são inerentemente descritas variações da estringência de soluções de hibridização e/ou lavagem. Caso o grau desejado de falta de coincidência resulte em Tm de menos de 45 °C (solução aquosa) ou 32°C (solução de formamida), prefere-se aumentar a concentração de SSC de forma que se possa utilizar uma temperatura mais alta. Um extenso guia de hibridização de ácidos nucleicos é encontrado em Tijssen, Laboratory Techniques in Biochemistry and Molecular Biology - Hybridization with Nucleic Acid Probes, Parte I, Capítulo 2, Overview of Principies of Hybridization and the Strategy of Nucleic Acid Probe Assays, Elsevier, Nova lorque (1993); e Current Protocols in Molecular Biology, Capítulo 2, Ausubel et al, Eds., Greene Publishing and Wiley-Interscience, Nova Iorque (1995). As condições de hibridização e/ou lavagem podem ser aplicadas por pelo menos dez, trinta, sessenta, noventa, 120 ou 240 minutos.
"Identidade de seqüências" ou "identidade", no contexto de seqüências de ácidos nucleicos ou polipeptídeos, designa as bases de ácidos nucleicos ou resíduos de aminoácidos nas duas seqüências que são idênticas quando alinhadas para correspondência máxima ao longo de uma janela de comparação especificada.
Desta forma, "percentual de identidade de seqüências" designa o valor determinado por meio de comparação de duas seqüências alinhadas idealmente ao longo de uma janela de comparação, em que a parte da seqüência de polipeptídeos ou polinucleotídeos na janela de comparação pode compreender adições ou exclusões (ou seja, lacunas) em comparação com a seqüência de referência (que não compreende adições nem exclusões) para alinhamento ideal das duas seqüências. O percentual é calculado determinando-se a quantidade de posições em que ocorre o resíduo de aminoácidos ou base de ácido nucleico idêntico nas duas seqüências para gerar o número de posições coincidentes, dividindo-se o número de posições coincidentes pelo número total de posições na janela de comparação e multiplicando-se os resultados por 100 para gerar o percentual de identidade de seqüências. Exemplos úteis de percentuais de identidades de seqüências incluem, mas sem limitar-se a 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% ou 95%, ou qualquer percentual inteiro de 50 a 100%. Estas identidades podem ser determinadas utilizando qualquer dos programas descritos no presente.
Alinhamentos de seqüências e cálculos de similaridade ou identidade percentual podem ser determinados utilizando uma série de métodos de comparação projetados para detectar seqüências homólogas que incluem, mas sem limitações, o programa Megalign® da suíte de computação bioinformática LASERGENE (DNASTAR Inc., Madison Wl). Dentro do contexto do presente pedido, compreender-se-á que, ao utilizar-se software de análise de seqüências para análise, os resultados da análise serão baseados nos "valores padrão" do programa indicado, a menos que especificado em contrário. Da forma utilizada no presente, "valores padrão" indicarão qualquer conjunto de valores ou parâmetros carregados originalmente com o software durante a sua primeira inicialização.
O "método Clustal V de alinhamento" corresponde ao método de alinhamento denominado Clustal V (descrito por Higgins e Sharp, CABIOS. 5: 151-153 (1989); Higgins1 D. G. et al (1992), Comput. AppL Biosci. 8: 189-191) e encontrado no programa MegAlign® da suíte de computação bioinformática LASERGENE (DNASTAR Inc., Madison Wl). Para múltiplos alinhamentos, os valores padrão correspondem a PENALIDADE DO INTERVALO = 10 e PENALIDADE DO COMPRIMENTO DO INTERVALO = 10. Os parâmetros padrão para alinhamentos em pares e cálculo do percentual de identidade de seqüências de proteínas utilizando o método Clustal são COMPRIMENTO DE PALAVRA = 1, PENALIDADE DO INTERVALO = 3, VISUALIZAÇÃO DO MELHOR RESULTADO = 5 e ESPAÇAMENTO EM DIAGONAIS = 5. Para ácidos nucleicos, esses parâmetros são COMPRIMENTO DE PALAVRA = 1, PENALIDADE DO INTERVALO = 5, VISUALIZAÇÃO DO MELHOR RESULTADO = 4 e ESPAÇAMENTO EM DIAGONAIS = 4. Após o alinhamento das seqüências utilizando o programa Clustal V, é possível obter um "percentual de identidade" observando-se a tabela "distâncias de seqüências" no mesmo programa.
"Método BLASTN de alinhamento" é um algoritmo fornecido pelo Centro Nacional de Informações Biotecnológicas (NCBI) para comparar seqüências de nucleotídeos utilizando parâmetros padrão.
Os técnicos no assunto compreendem bem que muitos níveis de identidade de seqüências são úteis na identificação de polipeptídeos de outras espécies, em que esses polipeptídeos possuem função ou atividade similar ou idêntica. Exemplos úteis de percentuais de identidades incluem, mas sem limitar-se a 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% ou 95%, ou qualquer percentual inteiro de 50% a 100%. De fato, qualquer identidade de aminoácido inteira de 50% a 100% pode ser útil na descrição da presente invenção, tal como 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% ou 99%. Além disso, é de interesse qualquer complemento parcial ou de comprimento total desse fragmento de nucleotídeo isolado.
"Gene" designa um fragmento de ácido nucleico que expressa uma proteína específica, que inclui seqüências reguladoras anteriores (seqüências não codificadoras 5') e posteriores (seqüências não codificadoras 3') à seqüência codificadora. "Gene nativo" indica um gene encontrado na natureza com suas próprias seqüências reguladoras. "Gene quimérico" designa qualquer gene que não é um gene nativo, que compreende seqüências reguladoras e de codificação que não são encontradas juntas na natureza. Consequentemente, um gene quimérico pode compreender seqüências reguladoras e seqüências de codificação que são derivadas de fontes diferentes, ou seqüências reguladoras e seqüências de codificação derivadas da mesma fonte, mas dispostas de maneira diferente da encontrada na natureza. Gene "exógeno" indica um gene normalmente não encontrado no organismo hospedeiro, mas que é introduzido no organismo hospedeiro por meio de transferência genética. Os genes exógenos podem compreender genes nativos inseridos em um organismo não nativo ou genes quiméricos. Um "transgene" é um gene que foi introduzido no genoma por meio de um procedimento de transformação.
O termo "genoma", conforme aplicado a uma célula vegetal, engloba não apenas DNA cromossômico encontrado no interior do núcleo, mas DNA de organelas encontrado em componentes subcelulares (tais como mitocôndrias e plastídeos) da célula.
"Gene otimizado por códons" é um gene que possui a sua freqüência de uso de códons projetada para imitar a freqüência de uso de códons preferido da célula hospedeira. "Alelo" é uma das diversas formas alternativas de um gene que ocupa um dado local sobre um cromossomo. Quando todos os alelos presentes em um dado local sobre um cromossomo forem idênticos, aquela planta é homozigótica naquele local. Caso os alelos presentes em um dado local sobre um cromossomo difiram, aquela planta é heterozigótica naquele local.
"Seqüência codificadora" indica uma seqüência de DNA que codifica uma seqüência de aminoácidos específica. "Seqüências reguladoras" indicam seqüências de nucleotídeos localizadas acima (seqüências não codificadoras 5'), na própria ou abaixo (seqüências não codificadoras 3') de uma seqüência codificadora e que influenciam a transcrição, estabilidade ou processamento de RNA ou tradução da seqüência codificadora associada. As seqüências reguladoras podem incluir, mas sem limitar-se a promotores, seqüências líderes de tradução, introns, seqüências de reconhecimento de poliadenilação, locais de processamento de RNA, locais de união de efetores e estruturas de circuito de haste.
"Promotor" designa uma seqüência de DNA capaz de controlar a expressão de uma seqüência codificadora ou RNA funcional. A seqüência promotora consiste de elementos próximos e mais distantes no fluxo, com estes últimos freqüentemente denominados amplificadores. Consequentemente, um "amplificador" é uma seqüência de DNA que pode estimular a atividade promotora e pode ser um elemento inato do promotor ou um elemento heterólogo inserido para amplificar o nível ou a especificidade de tecido de um promotor. Os promotores podem ser derivados em sua totalidade de um gene nativo ou ser compostos de diferentes elementos derivados de diferentes promotores encontrados na natureza, ou podem mesmo compreender segmentos de DNA sintéticos. Os técnicos no assunto compreendem que diferentes promotores podem dirigir a expressão de um gene em diferentes tecidos ou tipos de células, em diferentes estágios de desenvolvimento, ou em resposta a diferentes condições ambientais. Reconhece-se ainda que, como na maior parte dos casos as fronteiras exatas de seqüências reguladoras não foram totalmente definidas, fragmentos de DNA com alguma variação podem possuir atividade promotora idêntica. Os promotores que fazem com que um gene seja expresso na maior parte dos tipos de células na maior parte das vezes são comumente denominados "promotores constitutivos". Novos promotores de diversos tipos úteis em células vegetais estão constantemente sendo descobertos; numerosos exemplos podem ser encontrados na compilação de Okamuro, J. K. e Goldberg, R. B., Biochemistry of Plants 15: 1-82 (1989).
"Seqüência líder de tradução" designa uma seqüência de polinucleotídeos localizada entre a seqüência promotora de um gene e a seqüência codificadora. A seqüência líder de tradução está presente no mRNA totalmente processado após a seqüência inicial de tradução. A seqüência líder de tradução pode afetar o processamento do transcrito primário em mRNA, a estabilidade do mRNA ou a eficiência da tradução. Foram descritos exemplos de seqüências líderes de tradução (Turner, R. e Foster, G. D. (1995), Mol. BiotechnoL 3: 225-236 (1995)).
"Seqüências não-codificadoras 3"', "término de transcrição" ou "seqüências de término" designam seqüências de DNA em local posterior a uma seqüência codificadora e incluem seqüências de reconhecimento de poliadenilação e outras seqüências codificadoras de sinais reguladores capazes de afetar o processamento de mRNA ou a expressão genética. O sinal de poliadenilação é normalmente caracterizado por afetar a adição de traços de ácido poliadenílico à extremidade 3' do precursor de mRNA. A utilização de diferentes seqüências não-codificadoras 3' é exemplificada por Ingelbrecht, I. L. et al, Plant Ce//1: 671-680 (1989).
"Transcrito de RNA" designa o produto resultante da transcrição catalisada por RNA polimerase de uma seqüência de DNA. Quando o transcrito de RNA for uma cópia complementar perfeita da seqüência de DNA1 ele é denominado transcrito primário. Um transcrito de RNA é denominado RNA maduro quando for uma seqüência de RNA derivada do processamento pós- transcricional do transcrito primário. "RNA mensageiro" ou "mRNA" designa o RNA que não contém introns e que pode ser traduzido em proteína pela célula. "cDNA" designa DNA que é complementar a um modelo de mRNA e sintetizado a partir dele utilizando a enzima transcriptase reversa. O cDNA pode ser de fita única ou convertido em forma de fita dupla utilizando o fragmento Klenow de DNA polimerase I. RNA "com sentido" designa transcrito de RNA que inclui o mRNA e pode ser traduzido em proteína em uma célula ou in vitro.
"RNA sem sentido" designa um transcrito de RNA que é complementar a um mRNA ou transcrito primário alvo, no todo ou em parte, e que bloqueia ou reduz a expressão de um gene alvo (Patente Norte-Americana n° 5.107.065). A complementaridade de um RNA sem sentido pode dar-se com qualquer parte do transcrito genético específico, ou seja, na seqüência não- codificadora 5', seqüência não-codificadora 3', introns ou na seqüência codificadora. "RNA funcional" designa RNA sem sentido, RNA de ribozima ou outro RNA que pode não ser traduzido, mas que ainda tenha efeito sobre processos celulares. As expressões "complemento" e "complemento reverso" são utilizadas de forma intercambiável no presente com relação a transcritos de mRNA e destinam-se a definir o RNA sem sentido da mensagem.
A expressão "ligado operativamente" designa a associação de seqüências de ácido nucleico sobre um fragmento de ácido nucleico isolado, de tal forma que a função de um seja regulada pelo outro. Um promotor é ligado operativamente a uma seqüência codificadora, por exemplo, quando for capaz de regular a expressão daquela seqüência codificadora (ou seja, a seqüência codificadora encontra-se sob o controle transcricional do promotor). As seqüências codificadoras podem ser ligadas operativamente a seqüências reguladoras em orientação com sentido ou sem sentido. Em outro exemplo, as regiões de RNA complementar de acordo com a presente invenção podem ser ligadas operativamente, seja direta ou indiretamente, 5' ao mRNA alvo, ou 3' ao mRNA alvo, ou no mRNA alvo, ou uma primeira região complementar é 5' e seu complemento é 3' para o mRNA alvo.
As técnicas padrão de DNA recombinante e clonagem molecular utilizadas no presente são bem conhecidas na técnica e são descritas de forma mais completa em Sambrook, J., Fritsch, E. F. e Maniatis, T., Molecular Cloning: A Laboratory Manual·, Cold Spring Harbor Laboratory: Cold Spring Harbor NY (1989). Os métodos de transformação são bem conhecidos dos técnicos no assunto e são descritos abaixo.
"PCR" ou "reação em cadeia de polimerase" é um método de síntese de grandes quantidades de segmentos de DNA específicos e consiste de uma série de ciclos repetitivos (Perkin Elmer Cetus Instruments, Norwalk CT). Tipicamente, o DNA de fita dupla é desnaturado por calor, os dois primers complementares às fronteiras 3' do segmento alvo são combinados sob baixa temperatura e estendidos em seguida em uma temperatura intermediária. Um conjunto destas três etapas consecutivas é denominado "ciclo".
O termo "recombinante" designa uma combinação artificial de dois segmentos de seqüência de outra forma separados, por meio, por exemplo, de síntese química ou da manipulação de segmentos isolados de ácidos nucleicos por meio de técnicas de engenharia genética.
Os termos "plasmídeo", "vetor" e "conjunto" designam um elemento cromossômico adicional que freqüentemente conduz genes que não fazem parte do metabolismo central da célula e, normalmente, encontram-se na forma de fragmentos de DNA com fita dupla. Esses elementos podem ser seqüências de reprodução autônoma, seqüências integrantes de genomas, seqüências de fagos ou nucleotídeos, lineares ou circulares, de um DNA ou RNA de fita simples ou dupla, derivado de qualquer fonte, no qual uma série de seqüências de nucleotídeos foi unida ou recombinada em uma construção exclusiva que é capaz de introduzir um fragmento promotor e seqüência de DNA para um produto genético selecionado junto com uma seqüência não traduzida 3' apropriada em uma célula. "Conjunto de transformação" designa um vetor específico que contém um gene exógeno e contém elementos adicionais ao gene exógeno que facilitam a transformação de uma célula hospedeira específica. "Conjunto de expressão" designa um vetor específico que contém um gene exógeno e que contém elementos adicionais ao gene exógeno que permitem expressão aprimorada daquele gene em um hospedeiro exógeno (ou seja, para um fragmento de ácido nucleico discreto para o qual pode-se mover uma seqüência de ácido nucleico ou fragmento).
As expressões "construção recombinante", "construção de expressão", "construção quimérica", "construção" e "construção de DNA recombinante" são utilizadas de forma intercambiável no presente. Uma construção recombinante compreende uma combinação artificial de fragmentos de ácido nucleico, tais como seqüências reguladoras e de codificação que não são encontradas juntas na natureza. Uma construção recombinante pode compreender, por exemplo, seqüências reguladoras e seqüências de codificação que são derivadas de diferentes fontes ou seqüências reguladoras e seqüências de codificação derivadas da mesma fonte, mas dispostas de forma diferente da encontrada na natureza. Esta construção pode ser utilizada por si própria ou pode ser utilizada em conjunto com um vetor. Caso seja empregado um vetor, a seleção do vetor depende do método a ser utilizado para transformar as células hospedeiras, como é bem conhecido dos técnicos no assunto. Pode ser utilizado, por exemplo, um vetor de plasmídeo. Os técnicos no assunto conhecem bem os elementos genéticos que necessitam estar presentes sobre o vetor a fim de transformar, selecionar e propagar com sucesso células hospedeiras que compreendem qualquer dos fragmentos de ácidos nucleicos isolados de acordo com a presente invenção. Os técnicos no assunto também reconhecerão que eventos diferentes e independentes de transformação resultarão em níveis e padrões diferentes de expressão (Jones et al., EMBO J. 4: 2411-2418 (1985); De Almeida et al, MoL Geri. Genetics 218: 78-86 (1989)) e, portanto, que diversos eventos devem ser selecionados a fim de obter linhagens que exibam o nível e padrão desejado de expressão. Esta seleção pode ser atingida por meio de análise Southern de DNA1 análise Northern de expressão de mRNA, análise de imunomanchas de expressão de proteínas ou análise fenotípica, entre outras.
O termo "expressão", da forma utilizada no presente, designa a elaboração de um produto final funcional (tal como um mRNA ou uma proteína (seja precursora ou madura)).
O termo "introduzido" indica o fornecimento de um ácido nucleico (tal como construção de expressão) ou proteína em uma célula. Introduzido indica referência à incorporação de um ácido nucleico em uma célula eucariótica ou procariótica em que o ácido nucleico pode ser incorporado ao genoma da célula e inclui referência ao fornecimento transitório de um ácido nucleico ou proteína para a célula. Introduzido inclui referência a métodos de transformação estável ou transitória, bem como cruzamento sexual. Desta forma, "introduzido" no contexto de inserção de um fragmento de ácido nucleico (tal como uma construção de expressão/construção de DNA recombinante) em uma célula indica "transfecção", "transformação" ou "transdução" e inclui referência à incorporação de um fragmento de ácido nucleico em uma célula eucariótica ou procariótica em que o fragmento de ácido nucleico pode ser incorporado ao genoma da célula (tal como cromossomo, plasmídeo, plastídeo ou DNA de mitocôndria), convertido em uma réplica autônoma ou expresso transitoriamente (tal como mRNA transfectado).
Proteína "madura" indica um polipeptídeo processado após a tradução (ou seja, do qual qual(is)quer pré ou pró-peptídeo(s) presente(s) no produto de tradução primária tenha(m) sido removido(s)). Proteína "precursora" indica o produto primário de tradução de mRNA (ou seja, com pré e pró- peptídeos ainda presentes). Pré e pró-peptídeos podem ser mas não se limitam a sinais de localização intracelular.
O termo "multizima" indica um polipeptídeo isolado que possui pelo menos duas atividades enzimáticas independentes e separáveis.
Preferencialmente, a multizima compreende uma primeira atividade enzimática ligada a uma segunda atividade enzimática.
A expressão "proteína de fusão" é utilizada de forma intercambiável com o termo "multizima". Desta forma, "proteína de fusão" designa um polipeptídeo isolado que possui pelo menos duas atividades enzimáticas independentes e separáveis.
A expressão "gene de fusão" indica um polinucleotídeo ou gene que codifica uma multizima. Um gene de fusão pode ser construído por meio de ligação de pelo menos dois fragmentos de DNA entre si, em que cada fragmento de DNA codifica uma atividade enzimática independente e separada. Um exemplo de gene de fusão é descrito abaixo no Exemplo 21 (gene de fusão Híbrido 1 - HGLA Sintase) que foi construído por meio de ligação da delta-9 elongase de Euglena anabaena (EaD9Elo1; SEQ ID N0 101) e a delta-8 dessaturase de Tetruetreptia pomquetensis CCMP1491 (TpomD8; SEQ ID N0 50), utilizando o Iigante rico em prolina DHAsintase 1 de Euglena gracilis (EgdHAsynIligação; SEQ ID N0 97).
"DHA sintase" é um outro exemplo de multizima. Especificamente, uma DHA sintase compreende uma C20 elongase ligada a uma delta-4 dessaturase. O termo "ligação" designa a união ou ligação de pelo menos dois polipeptídeos que possuem atividades enzimáticas independentes e separáveis.
O termo "ligante" designa a união ou ligação entre dois ou mais polipeptídeos, em que cada qual possui atividades enzimáticas independentes e separáveis.
A ligação utilizada para formar uma multizima é compreendida, no mínimo, por uma união de polipeptídeo simples. Em um outro aspecto, a ligação pode ser compreendida por um resíduo de aminoácido, tal como prolina, ou um polipeptídeo. Pode ser desejável que, caso a ligação seja um polipeptídeo, ela contenha pelo menos um resíduo de aminoácido prolina.
Um exemplo de ligante é exibido em SEQ ID N0 97 (o ligante rico em prolina EgDHAsynI).
A origem do promotor selecionado para dirigir a expressão da seqüência de codificação de multizima não é importante desde que possua atividade de transcrição suficiente para expressar mRNA traduzível para os fragmentos de ácidos nucleicos desejados no tecido hospedeiro desejado no momento certo. Podem ser utilizados promotores heterólogos ou não heterólogos (ou seja, endógenos) para a prática da presente invenção. Os promotores apropriados incluem, por exemplo, mas sem limitações: a subunidade prime alfa de promotor beta conglicinina, o promotor inibidor de tripsina Kunitz 3, o promotor de anexina, o promotor de glicinina Gy1, a subunidade beta de promotor beta conglicinina, o promotor de P34/Gly Bd m 30K, o promotor de albumina, o promotor Leg A1 e o promotor Leg A2.
"Transformação estável" indica a transferência de um fragmento de ácido nucleico para o genoma de um organismo hospedeiro, incluindo genomas nucleares e de organelas, o que resulta em herança geneticamente estável. Por outro lado, "transformação transitória" indica a transferência de um fragmento de ácido nucleico para o núcleo ou uma organela que contém DNA de um organismo hospedeiro, resultando em expressão genética sem integração nem herança estável. Os organismos hospedeiros que contêm os fragmentos de ácidos nucleicos transformados são denominados organismos "transgênicos".
Da forma utilizada no presente, "transgênico" designa uma planta ou célula que compreende no seu genoma um polinucleotídeo heterólogo. Preferencialmente, o polinucleotídeo heterólogo é integrado de forma estável no interior do genoma, de forma que o polinucleotídeo seja passado para gerações sucessivas. O polinucleotídeo heterólogo pode ser integrado ao genoma isoladamente ou como parte de uma construção de expressão. Transgênico é utilizado no presente para incluir qualquer célula, linhagem celular, calo, tecido, parte de planta ou planta, cujo genótipo foi alterado pela presença de ácido nucleico heterólogo que inclui os transgênicos inicialmente alterados desta forma, bem como os criados por meio de cruzamentos sexuais ou propagação assexuada a partir do transgênico inicial. O termo "transgênico" conforme utilizado no presente não engloba a alteração do genoma (cromossômico ou extracromossômico) por meio de métodos de cultivo de plantas convencionais ou de eventos de ocorrência natural, tais como fertilização cruzada aleatória, infecção viral não recombinante, transformação bacteriana não recombinante, transposição não recombinante ou mutação espontânea.
"Inibição sem sentido" designa a produção de transcritos de RNA sem sentido capazes de suprimir a expressão da proteína alvo. "Cossupressão" designa a produção de transcritos de RNA com sentido capazes de suprimir a expressão de genes exógenos ou endógenos idênticos ou substancialmente similares (Patente Norte-Americana n° 5.231.020). Construções de cossupressão em plantas foram anteriormente projetadas concentrando-se na sobre-expressão de uma seqüência de ácido nucleico que possui homologia para um mRNA endógeno, na orientação com sentido, o que resulta na redução de todo o RNA que possui homologia para a seqüência sobre- expressa (Vaucheret et al, Plant J. 16: 651-659 (1998); e Gura1 Nature 404: 804-808 (2000)). A eficiência geral deste fenômeno é baixa e a extensão da redução de RNA é amplamente variável. Trabalhos mais recentes descreveram o uso de estruturas de "grampo de cabelo" que incorporam, no todo ou em parte, uma seqüência de codificação de mRNA em orientação complementar que resulta em uma potencial estrutura de "circuito de haste" para o RNA expresso (Patente PCT n0 WO 99/53050, publicada em 21 de outubro de 1999; Patente PCT n0 WO 02/00904, publicada em três de janeiro de 2002). Isso aumenta a freqüência de cossupressão nas plantas transgênicas recuperadas. Uma outra variação descreve o uso de seqüências virais de plantas para dirigir a supressão ou "silenciamento" de seqüências de codificação de mRNA próximas (Patente PCT n0 WO 98/36083, publicada em 20 de agosto de 1998). Nenhum destes dois fenômenos de cossupressão foi elucidado mecanicamente, embora evidências genéticas tenham começado a desemaranhar esta complexa situação (Elmayan et al, Plant Cell 10: 1747-1757 (1998)). "Regulagem para baixo" pode ser realizada por meio de qualquer das tecnologias de cossupressão, RNAi, RNA sem sentido ou miRNA (micro RNA). miRNAs são agentes reguladores que consistem de cerca de 22 ribonucleotídeos. miRNAs são altamente eficientes na inibição da expressão de genes endógenos. Vide, por exemplo, Javier et al, Nature 425: 257-263 (2003), incorporado ao presente como referência.
O termo "euglenofíceo" indica um grupo de flagelos fotossintéticos ou incolores unicelulares ("euglenoides") encontrados vivos em água doce, água salgada, solo e ambientes parasíticos. A classe é caracterizada por unicélulas solitárias, em que a maior parte nada livremente e possui dois flagelos (um dos quais pode ser não emergente) decorrentes de uma invaginação anterior conhecida como reservatório. Euglenoides fotossintéticos contêm um a vários cloroplastas verdes-grama, que variam de pequenos discos até placas expandidas ou fitas. Os euglenoides incolores dependem da osmotrofia ou fagotrofia para assimilação de nutrientes. Cerca de mil espécies foram descritas e classificadas em cerca de quarenta gêneros e seis ordens. Exemplos de euglenofíceos incluem, mas sem limitar-se de nenhuma forma aos gêneros a seguir: Eutreptiella e Tetruetreptia.
O termo "planta" indica plantas inteiras, órgãos de plantas, tecidos vegetais, sementes, células de plantas, sementes e sua prole. As células vegetais incluem, sem limitações, células de sementes, cultivos em suspensão, embriões, regiões de meristema, tecido de calo, folhas, raízes, brotos, gametófitos, esporófitos, pólen e microesporos.
"Prole" compreende qualquer geração subsequente de uma planta.
A presente invenção refere-se à produção de ácido araquidônico em plantas oleaginosas. Mais especificamente, a presente invenção refere-se a sementes de Arabidopsis e soja transgênicas que contenham pelo menos 10% de ácido araquidônico e um teor reduzido de ácidos graxos ômega 3. A presente invenção também se refere a sementes de Arabidopsis e soja transgênicas que contenham pelo menos 10% de ácido araquidônico, teor reduzido de ácidos graxos ômega 3 e teor reduzido de ácido gamalinolênico.
A presente invenção refere-se a plantas oleaginosas transgênicas que sejam capazes de produzir sementes maduras nas quais o perfil total de ácido graxo da semente compreende pelo menos 10% de ácido araquidônico (ARA, 20:4, ômega 6) e menos de 5% do total de ácidos graxos ômega 3 que contêm pelo menos dezoito átomos de carbono e pelo menos quatro uniões duplas. Também é de interesse uma planta oleaginosa transgênica que produz sementes maduras nas quais o perfil total de ácido graxo da semente compreende pelo menos 10,0% de ácido araquidônico e menos de 1% do total de ácidos graxos ômega 3 que contêm pelo menos dezoito átomos de carbono e pelo menos quatro uniões duplas.
Em um outro aspecto, a presente invenção refere-se a uma planta oleaginosa transgênica que produz sementes maduras em que o perfil total de ácido graxo das sementes compreende pelo menos 10,0% de ácido araquidônico e menos de 5% de ácido gamalinolênico.
Em ainda outro aspecto, a presente invenção refere-se a uma planta oleaginosa transgênica que produz sementes maduras em que o perfil total de ácido graxo das sementes compreende pelo menos 10,0% de ácido araquidônico e até 1% de ácido gamalinolênico, inclusive.
Em mais um aspecto, a presente invenção refere-se a uma planta oleaginosa transgênica que produz sementes maduras em que o perfil total de ácido graxo das sementes compreende pelo menos 10,0% de ácido araquidônico, até 1% de ácido gamalinolênico, inclusive, e menos de 1% de ácidos graxos ômega 3 totais que possuem pelo menos dezoito átomos de carbono e pelo menos quatro uniões duplas.
O acúmulo desse ácido graxo póli-insaturado (PUFA) específico pode ser realizado por meio da introdução de qualquer um dentre dois processos biossintéticos de ácido graxo ômega 3/ômega 6 funcionais diferentes. O primeiro processo utiliza, entre outras, proteínas com atividades de delta-6 dessaturase, C18/20 elongase e delta-5 dessaturase no hospedeiro para expressão recombinante em alto nível, em que o óleo de ARA também compreende GLA
O outro processo utiliza, entre outros, proteínas com atividades de delta-9 elongase, delta-8 dessaturase e delta-5 dessaturase, de forma a permitir a produção de um óleo de ARA que é isento de qualquer GLA A produção de altos níveis de ARA com um teor reduzido de ácidos graxos ômega 3 (ou seja, EPA) pode ser realizada, mas sem limitações, por meio de qualquer dos processos acima em combinação com a regulagem para baixo do gene fad3 que codifica uma enzima que possui atividade de delta-15 dessaturase. Além disso, o cruzamento de grãos de soja que expressam processo de produção de ARA com grãos de soja "baixo lin" transgênicos ou não transgênicos pode ser utilizado para reduzir o teor de ácidos graxos ômega 3 tais como EPA. Adicionalmente, a introdução de qualquer processo em uma planta hospedeira que naturalmente não realiza altos níveis de ALA tal como milho, girassol ou açafrão pode também ser utilizada para produzir ARA com baixo teor de ácidos graxos ômega 3.
"Delta-15 dessaturase" designa uma dessaturase de ácido graxo que catalisa a formação de uma união dupla entre as posições de carbono 3 e 4 (numeradas a partir da extremidade metila) (ou seja, as que correspondem às posições de carbono 15 e 16 (numeradas a partir do carbono carbonila) de uma cadeia acila graxa com dezoito carbonos de comprimento e posições de carbono 13 e 14 (numeradas a partir do carbono carbonila) de uma cadeia acila graxa com dezesseis carbonos de comprimento. A regulagem para baixo do gene fad3 é descrita na Patente Norte-Americana n° 5.952.544 emitida para Browse et al em quatorze de setembro de 1999.
PUFAs tais como ARA ou seus derivados, elaborados por meio da metodologia descrita no presente, podem ser utilizados como substitutos ou suplementos alimentares, particularmente fórmulas para bebês, para pacientes que sofrem alimentação intravenosa ou para evitar ou tratar a desnutrição. Alternativamente, os PUFAs purificados (ou seus derivados) podem ser incorporados em óleos de cozinha, gorduras ou margarinas formuladas, de tal forma que, durante o uso normal, o consumidor receberia a quantidade desejada para suplementação alimentar. Os PUFAs podem também ser incorporados em fórmulas para bebês, suplementos nutricionais ou outros produtos alimentícios e podem encontrar uso como agentes anti-inflamatórios ou redutores do colesterol. Opcionalmente, as composições podem ser empregadas para uso farmacêutico (humano ou veterinário). Neste caso, os PUFAs são geralmente administrados oralmente, mas podem ser administrados por qualquer via por meio da qual possam ser absorvidos com sucesso, tal como parenteral (por exemplo, subcutânea, intramuscular ou intravenosa), retal, vaginal ou tópica (tal como na forma de loção ou unguento para a pele).
A suplementação de dietas humanas ou animais com PUFAs produzidos por meios recombinantes pode resultar em níveis mais altos dos PUFAs adicionados, bem como seus metabólitos. Tratamento com EPA pode resultar, por exemplo, não apenas em níveis mais altos de EPA, mas também produtos subsequentes de EPA tais como eicosanoides (ou seja, prostaglandinas, leucotrienos, tromboxanos). Mecanismos reguladores complexos podem tornar desejável a combinação de diversos PUFAs ou a adição de diferentes conjugados de PUFAs, a fim de evitar, controlar ou superar esses mecanismos para atingir os níveis desejados de PUFA(s) específico(s) em um indivíduo.
Biossíntese DE ARA (ácido GRAXO ómega 6)
O processo metabólico em que ácido oleico é convertido em ARA envolve o alongamento da cadeia de carbono por meio da adição de átomos de carbono e dessaturação da molécula por meio da adição de uniões duplas. Isso requer uma série de enzimas de alongamento e dessaturação especiais. Conforme exibido na Fig. 1 e descrito abaixo, entretanto, existem dois processos alternativos.
Especificamente, os dois processos exigem a conversão iniciai de ácido oleico em LA (18:2), o primeiro dos ácidos graxos ômega 6, por uma delta-12 dessaturase. Em seguida, utilizando o processo delta-6 dessaturase/delta-6 elongase para biossíntese de ARA, os PUFAs são formados conforme segue: (1) LA é convertido em GLA pela atividade de uma delta-6 dessaturase; (2) GLA é convertido em DGLA pela ação de uma C18/20 elongase; e (3) DGLA é convertido em ARA pela ação de uma delta-5 dessaturase.
Alternativamente, o "processo de delta-9 elongase/delta-8 dessaturase" pode ser utilizado para formação de ARA conforme segue: (1) LA é convertido em EDA por uma delta-9 elongase; (2) EDA é convertido em DGLA por uma delta-8 dessaturase; e (3) DGLA é convertido em ARA por uma delta-5 dessaturase.
Os técnicos no assunto serão capazes de identificar os vários possíveis genes que codificam cada uma das enzimas desejadas para biossíntese de ARA. Seqüências de dessaturase e elongase úteis podem ser derivaas de qualquer fonte, tal como isoladas de uma fonte natural (de bactérias, algas, fungos, plantas, animais etc.), produzidas por meio de um processo semissintético ou sintetizadas como novas. Embora a fonte específica dos genes dessaturase e elongase introduzidos no hospedeiro não seja crítica, as considerações para a seleção de um polipeptídeo específico que possui atividade de dessaturase ou elongase incluem: (1) a especificidade de substrato do polipeptídeo; (2) se o polipeptídeo ou um de seus componentes é uma enzima limitadora da velocidade; (3) se a dessaturase ou elongase é essencial para a síntese de um PUFA desejado; e/ou (4) cofatores exigidos pelo polipeptídeo. O polipeptídeo expresso possui preferencialmente parâmetros compatíveis com o ambiente bioquímico da sua localização na célula hospedeira (vide a Patente PCT n0 WO 04/101757 para detalhes adicionais). Em realizações adicionais, também será útil considerar a eficiência de conversão de cada dessaturase e/ou elongase específica. Mais especificamente, como cada enzima raramente funciona com 100% de eficiência para converter substrato em produto, o perfil final de lipídios de óleos não purificados produzidos em uma célula hospedeira será tipicamente uma mistura de vários PUFAs que consistem do ácido graxo ômega 3/ômega 6 desejado, bem como vários PUFAs intermediários acima no fluxo. Desta forma, a consideração da eficiência de conversão de cada enzima também é uma variável ao otimizar-se a biossíntese de um ácido graxo desejado que deve ser considerado à luz do perfil de lipídios final desejado do produto.
Com cada uma das considerações acima em mente, possíveis genes que possuem as atividades de dessaturase e elongase apropriadas podem ser identificados de acordo com a literatura disponível ao público (tal como GenBank), literatura de patentes e análise experimental de micro- organismos que possuem a capacidade de produzir PUFAs. Os Números de Acesso GenBank a seguir, por exemplo, designam exemplos de genes disponíveis ao público úteis na biossíntese de ARA: AY131238, Y055118, AY055117, AF296076, AF007561, L11421, NM_031344, AF465283, AF465281, AF110510, AF465282, AF419296, AB052086, AJ250735, AF126799, AF126798 (δ6 dessaturases); AF390174 (δ9 elongase); AF139720 (58 dessaturase); AF199596, AF226273, AF320509, AB072976, AF489588, AJ510244, AF419297, AF07879, AF067654, AB022097 (δ5 dessaturases); AAG36933, AF110509, AB020033, AAL13300, AF417244, AF161219, AY332747, X86736, AF240777, AB007640, AB075526, AP002063 (512 dessaturases); AF338466, AF438199, E11368, E11367, D83185, U90417, AF085500, AY504633, NM_069854, AF230693 (59 dessaturases); e NP_012339, NP_009963, NP_013476, NP_599209, BAB69888, AF244356, AAF70417, AAF71789, AF390174, AF428243, NP_955826, AF206662, AF268031, ΑΥ591335, ΑΥ591336, ΑΥ591337, ΑΥ591338, ΑΥ605098, ΑΥ605100, ΑΥ630573 (elongases Cum, Ci6/ie e C18/20)· De forma similar, a literatura de patentes fornece muitas seqüências de DNA adicionais de genes (e/ou detalhes referentes a vários dos genes acima e seus métodos de isolamento) envolvidos na produção de PUFA (tais como WO 02/077213 (09 elongases); WO 00/34439 e WO 04/057001 (δ8 dessaturases); US 5.968.809 (δ6 dessaturases); US 5.972.664 e US 6.075.183 (δ5 dessaturases); WO 94/11516, US 5.443.974, WO 03/099216 e WO 05/047485 (512 dessaturases); WO 91/13972 e US 5.057.419 (δ9 dessaturases); e WO 00/12720, US 6.403.349, US 6.677.145, US 2002/0139974 A1, US 2004/0111763 (CU/16, C16/18 e C18/20 elongases)). Cada uma dessas patentes e pedidos é integralmente incorporada ao presente como referência.
Contempla-se que a discussão acima destina-se a ser um exemplo e não limitadora. Consequentemente, numerosos outros genes que codificam (1) delta-6 dessaturases, C18/20 elongases e delta-5 dessaturases (e, opcionalmente, outros genes que codificam delta-9 dessaturases, delta-12 dessaturases, C14/16 elongases e/ou C16/18 elongases); ou (2) delta-9 elongases, delta-8 dessaturases e delta-5 dessaturases (e, opcionalmente, outros genes que codificam delta-9 dessaturases, delta-12 dessaturases, Cun6 elongases e/ou Cie/18 elongases) derivadas de fontes diferentes seriam apropriados para modificar o genoma de uma planta oleaginosa.
Às vezes, pode ser desejável minimizar os subprodutos ácidos graxos. A relativa abundância de subprodutos ácidos graxos poderá ser reduzida aumentando-se a atividade total de delta-8 dessaturase. Uma abordagem para minimizar os subprodutos ácidos graxos seria a expressão de mais de uma delta-8 dessaturase (ou seja, a mesma delta-8 dessaturase ou diferente). A presença, por exemplo, de ácido sciadônico (SCI) e/ou ácido juniperônico (JUP) (comumente empregados nos lipídios de sementes de gimnospermas (Wolff et al, Lipids 35 (1): 1-22 (2000)), tais como os da família Pinaceae (pinheiro)) poderá ser considerada como subproduto ácido graxo de um processo de delta-6 dessaturase/delta-6 elongase ou um processo de delta- 9 elongase/delta-8 dessaturase. Embora se considere que esses ácidos graxos possuam diversas propriedades aprimoradoras da saúde (Nakane et al, Biol. Pharm. Buli. 23: 758-761 (2000)), a sua presença como subproduto ácido graxo em um processo de PUFA elaborado, tal como em uma safra de oleaginosa, pode não ser desejável, dependendo da aplicação.
A expressão "processo de delta-6 desaturase/delta-6 elongase" também designa um processo biossintético de produção de PUFAs de cadeia longa. Este processo compreende, no mínimo, uma delta-6 dessaturase e uma delta-6 elongase, de forma a permitir a biossíntese de DGLA e/ou ETA a partir de LA e ALA, respectivamente. Com a expressão de outras dessaturases e elongases, podem também ser sintetizados ARA, EPA, DPA e DHA. Ocasionalmente, uma delta-6 elongase pode alongar ácidos graxos diferentes do ácido graxo pretendido. Delta-6 elongases geralmente convertem GLA em DGLA, por exemplo, mas algumas delta-6 elongases podem também converter substratos indesejados, tais como LA ou ALA em EDA ou ETrA, respectivamente. Em um processo de delta-6 dessaturase/delta-6 elongase, EDA e ETrA seriam considerados "subprodutos ácidos graxos", conforme definido abaixo. A adição de uma delta-8 dessaturase a um processo de delta-6 dessaturase/delta-6 elongase forneceria um meio de conversão dos "subprodutos ácidos graxos" EDA e ETrA de volta para os "ácidos graxos intermediários" (conforme definido abaixo) DGLA e ETA, respectivamente. Sistemas de expressão em plantas, conjuntos, vetores ε transformação Um promotor é uma seqüência de DNA que dirige a maquinaria celular de uma planta para a produção de RNA a partir da seqüência de codificação contígua abaixo no fluxo (3') do promotor. A região promotora influencia a velocidade, estágio de desenvolvimento e tipo de célula em que é elaborado o transcrito de RNA do gene. O transcrito de RNA é processado para produzir mRNA que serve de modelo para a tradução da seqüência de RNA na seqüência de aminoácidos do polipeptídeo codificado. A seqüência líder não traduzida 5' é uma região do mRNA acima no fluxo da região de codificação de proteína que pode desempenhar um papel no início e tradução do mRNA. O sinal de término de transcrição e poliadenilação 3' é uma região não traduzida abaixo no fluxo da região de codificação de proteína que funciona na célula vegetal para causar o término da transcrição de RNA e a adição de nucleotídeos de poliadenilato à extremidade 3' do RNA.
A origem do promotor selecionado para dirigir a expressão da seqüência de codificação não é importante, desde que possua suficiente atividade de transcrição suficiente para realizar a presente invenção por meio da expressão de mRNA traduzível para os fragmentos de ácido nucleico desejados no tecido hospedeiro desejado no momento correto. Podem ser utilizados promotores heterólogos ou não heterólogos (ou seja, endógenos) para a prática da presente invenção. Promotores apropriados incluem, por exemplo, mas sem limitações: a subunidade prime alfa de promotor de beta conglicinina, o promotor inibidor de tripsina Kunitz 3, o promotor de anexina, o promotor de glicinina Gy1, a subunidade beta de promotor beta conglicinina, o promotor de P34/Gly Bd m 30K, o promotor de albumina, o promotor Leg A1 e o promotor Leg A2.
O promotor de anexina, ou P34, é descrito na Patente PCT n0 WO 04/071178 (publicada em 26 de agosto de 2004). O nível de atividade do promotor de anexina é comparável com o de muitos promotores fortes conhecidos, tais como: (1) o promotor de CaMV 35S (Atanassova et al, Plant Mol. Biol. 37: 275-285 (1998); Battraw e Hall1 Plant Mol. Biol. 15: 527-538 <1990); Holtorf et al, Plant Mol. Biol. 29: 637-646 (1995); Jefferson et al, EMBO J. 6: 3901-3907 (1987); Wllmink et al, Plant Mol. Biol. 28: 949-955 (1995)); (2) promotores de oleosina de Arabidopsis (Plant et al, Plant Mol. Bioi 25: 193-205 (1994); Li1 dissertação de Ph.D na Universidade A&M do Texas, págs. 107-128
(1997)); (3) os promotores de proteína de extensão de ubiquitina de Arabidopsis (Callis et al, J. Bioi Chem. 265 (21): 12486-93 (1990)); (4) um promotor do gene ubiquitina de tomate (Rollfinke et al, Gene 211 (2): 267-76
(1998)); (5) um promotor de proteína de choque de calor de soja (Schoffl et al, Moi Gen. Genet. 217 (2-3): 246-53 (1989)); e (6) um promotor de gene de H3 histona de milho (Atanassova et al, Plant Moi Bioi 37 (2): 275-85 (1989)).
Uma outra característica útil do promotor de anexina é o seu perfil de expressão em sementes em desenvolvimento. O promotor de anexina é mais ativo no desenvolvimento de sementes em estágios iniciais (antes de dez dias após a polinização) e é em grande parte tranqüilo em estágios posteriores. O perfil de expressão do promotor de anexina é diferente de vários promotores específicos de sementes, tais como promotores de proteína de armazenagem de sementes, que freqüentemente fornecem atividade mais alta nos últimos estágios de desenvolvimento (Chen et al, Dev. Genet. 10: 112-122 (1989); Ellerstrom et al, Plant Moi Biol. 32: 1019-1027 (1996); Keddie et al, Plant Mol. Bioi 24: 327-340 (1994); Plant et al (acima); Li (acima)). O promotor de anexina possui um perfil de expressão mais convencional, mas permanece diferente de outros promotores específicos de sementes conhecidos. Desta forma, o promotor de anexina será um candidato muito atraente quando se desejar a sobre-expressão ou supressão de um gene em embriões em um estágio de desenvolvimento inicial. Pode ser desejável, por exemplo, sobre-expressar um gene que regule o desenvolvimento inicial de embriões ou um gene envolvido no metabolismo antes da maturação da semente.
Após a identificação de um promotor apropriado para a expressão de uma seqüência de codificação, o promotor é ligado operativamente em seguida em uma orientação com sentido utilizando meios convencionais bem conhecidos dos técnicos no assunto.
Os métodos de clonagem molecular e DNA recombinante padrão utilizados no presente são bem conhecidos na técnica e descritos mais completamente em Sambrook1 J. et al em Molecular Cloning: A Laboratory Manual, segunda edição; Cold Spring Harbor Laboratory Press: Cold Spring Harbor, Nova Iorque 1989 (a seguir, "Sambrook et al, 1989") ou Ausubel, F. M. Brent, R., Kíngston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A. e Struhl1 K., Eds., em Current Protocols in Molecular Biology, John Wiley & Sons: Nova lorque, 1990 (a seguir, "Ausubel et al, 1990"). Um gene de fusão pode ser construído, por exemplo, por meio de ligação de pelo menos dois fragmentos de DNA em quadro, de forma a não introduzir um códon de parda (fusão em quadro). O gene de fusão resultante será tal que cada fragmento de DNA codifica uma atividade enzimática separada e independente.
Após a elaboração da construção recombinante, ela pode ser introduzida em uma célula vegetal selecionada por meio de métodos bem conhecidos dos técnicos comuns no assunto (tais como transfecção, transformação e eletroporação). As células de plantas oleaginosas são as células vegetais preferidas. A célula vegetal transformada é cultivada em seguida e regenerada sob condições apropriadas que permitam a expressão do PUFA de cadeia longa que é recuperado opcionalmente em seguida e purificado.
As construções recombinantes de acordo com a presente invenção podem ser introduzidas em uma célula vegetal; ou, alternativamente, cada construção pode ser introduzida em células vegetais separadas.
A expressão em uma célula vegetal pode ser realizada de forma transitória ou estável, conforme descrito acima.
Os PUFAs de cadeia longa desejados podem ser expressos em sementes. Também de acordo com a presente invenção, encontram-se sementes ou partes de plantas obtidas a partir dessas plantas transformadas.
As partes de plantas incluem tecidos diferenciados e não diferenciados que incluem, mas sem limitar-se aos seguintes: raízes, hastes, brotos, folhas, pólen, sementes, tecido de tumores e várias formas de células e cultivo (tais como células isoladas, protoplastas, embriões e tecido de calo). O tecido vegetal pode estar em uma planta ou em cultivo de órgão, tecido ou célula vegetal.
A expressão "órgão de planta" designa um tecido vegetal ou um grupo de tecidos que constituem uma parte morfológica e funcionalmente distinta de uma planta. O termo "genoma" designa o seguinte: (1) o complemento completo de material genético (genes e seqüências não de codificação) que está presente em cada célula de um organismo, vírus ou organela; e/ou (2) um conjunto completo de cromossomos herdados na forma de uma unidade (haploide) de um pai.
Desta forma, a presente invenção também se refere a um método de transformação celular, que compreende a transformação de células com a construção recombinante de acordo com a presente invenção e seleção das células transformadas com uma das construções recombinantes de acordo com a presente invenção.
Também é de interesse um método de produção de plantas transformadas que compreende a transformação de uma célula vegetal com os polinucleotídeos de acordo com a presente invenção e regeneração de plantas a partir da célula vegetal transformada.
Os métodos de transformação de dicotiledôneas (principalmente utilizando Agrobacterium tumefaciens) e obtenção de plantas transgênicas foram publicados, dentre outros, para: algodão (Patente Norte-Americana n° 5.004.863; Patente Norte-Americana n° 5.159.135); soja (Patente Norte- Americana n° 5.569.834; Patente Norte-Americana n° 5.416.011); Brassica (Patente Norte-Americana n° 5.463.174); amendoim (Cheng et al, Plant Cell Rep. 15: 653-657 (1996); McKentIy et al, Plant Cell Rep. 14: 699-703 (1995)); mamão (Ling, K. et al, Bio/Technology 9: 752-758 (1991)); e ervilha (Grant et al, Plant Cell Rep. 15: 254-258 (1995)). Para uma análise de outros métodos comumente utilizados de transformação vegetal, vide Newell, C. A. (Mol. Biotechnol. 16: 53-65 (2000)). Um desses métodos de transformação utiliza Agrobacterium rhizogenes (Tepfler, M. e Casse-Delbart, F., Microbioi Sei. 4: 24-28 (1987)). A transformação de grãos de soja utilizando fornecimento direto de DNA foi publicada utilizando fusão de PEG (Patente PCT n0 WO 92/17598), eletroporação (Chowrira, G. M. et al, Mol. Biotechnol. 3: 17-23 (1995); Christou, P. et al, Proc. Natl. Acad. Sei. U. S. A. 84: 3962-3966 (1987)); microinjeção e bombardeamento de partículas (McCabe, D. E. et al, Bio/Technology 6: 923 (1988); Christou et al, PlantPhysiol. 87: 671-674 (1988)).
Existe uma série de métodos de regeneração de plantas a partir de tecido vegetal. O método de regeneração específico dependerá do tecido vegetal inicial e da espécie vegetal específica a ser regenerada. A regeneração, desenvolvimento e cultivo de plantas de transformadores de protoplastas vegetais isolados ou de vários explantes transformados é bem conhecida na técnica (Weissbach e Weissbach, em Methods for Plant Molecular Biology (Eds.), Acadêmico: San Diego CA (1988)). Este processo de regeneração e crescimento inclui tipicamente as etapas de seleção de células transformadas e cultivo das células individualizadas por meio dos estágios habituais de desenvolvimento embriônico através do estágio de plantículas enraizadas. Embriões transgênicos e sementes são regenerados de forma similar. Os brotos enraizados transgênicos resultantes são plantados em seguida em um meio de crescimento vegetal apropriado tal como solo. Preferencialmente, as plantas regeneradas são autopolinizadas para fornecer plantas transgênicas homozigóticas. Caso contrário, o pólen obtido das plantas regeneradas é cruzado em plantas cultivadas de sementes de linhagens agronomicamente importantes. Por outro lado, o pólen das plantas dessas linhagens importantes é utilizado para polinizar plantas regeneradas. Uma planta transgênica de acordo com a presente invenção que contém um polipeptídeo desejado é cultivada utilizando métodos bem conhecidos dos técnicos no assunto.
Além dos procedimentos discutidos acima, os praticantes são familiares com os materiais de recurso padrão que descrevem procedimentos e condições específicas para: construção, manipulação e isolamento de macromoléculas (tais como moléculas de DNA1 plasmídeos etc.); geração de fragmentos de DNA recombinantes e construções de expressão recombinantes; e seleção e isolamento de clones. Vide, por exemplo, Sambrook et al, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor: Nova Iorque (1989); Maliga et al, Methods in Plant Molecular Biology, Cold Spring Harbor: Nova Iorque (1995); Birren et al, Genome Analysis: Detecting Genes, vol. 1, Cold Spring Harbor: Nova Iorque (1998); Birren et al, Genome Analysis: Analyzing DNA, Vol. 2, Cold Spring Harbor: Nova Iorque (1998); Plant Molecular Biology: A Laboratory Manual, eds. Clark, Springer: Nova Iorque (1997).
Exemplos de plantas oleaginosas incluem, mas sem limitar-se a: soja, espécies de Brassica, girassol, milho, algodão, linho e açafrão. Sementes obtidas dessas plantas também se encontram dentro do escopo da presente invenção, bem como óleo obtido a partir dessas sementes.
A seleção de combinação de conjuntos utilizados para a produção de ARA depende, em parte, do perfil de PUFA e/ou perfil de dessaturase/elongase das células de plantas oleaginosas a serem transformadas. Purificação ε processamento de óleos de PUFA
PUFAs podem ser encontrados nas plantas hospedeiras na forma de ácidos graxos livres ou em formas esterificadas tais como acilgliceróis, fosfolipídios, sulfolipídios ou glicolipídios e podem ser extraídos das células hospedeiras por meio de uma série de meios bem conhecidos na técnica. Uma análise de métodos de extração, análise de qualidade e padrões de aceitabilidade para lipídios de levedura é o de Z. Jacobs (Criticai Reviews in Biotechnology, 12 (5/6): 463-491 (1992)). Uma rápida análise do processamento abaixo no fluxo também é disponível por meio de A. Singh e O. Ward (Adv. Appl. Microbiol., 45: 271-312 (1997)).
De forma geral, os meios de purificação de PUFAs podem incluir extração com solventes orgânicos, sonicação, extração de fluido supercrítica (utilizando, por exemplo, dióxido de carbono), saponificação e meios físicos tais como prensas ou suas combinações. Um refere-se aos ensinamentos da Patente PCT n° WO 04/101757 para detalhes adicionais. Métodos de isolamento de óleos de semente são bem conhecidos na técnica (Young et al, Processing of Fats and Oils, em The Lipid Handbook, Gunstone et al, Eds., Capítulo 5, págs. 253-257; Chapman & Hall: Londres (1994)). Óleo de soja é produzido, por exemplo, utilizando uma série de etapas que envolvem a extração e purificação de um produto óleo comestível da semente que contém óleo. Óleos de soja e subprodutos de soja são produzidos utilizando as etapas generalizadas exibidas na Tabela 6.
Tabela 6
Etapas Generalizadas de Produção de Óleo de Soja ε Subprodutos
<table>table see original document page 65</column></row><table> <table>table see original document page 66</column></row><table>
Mais especificamente, as sementes de soja são limpas, temperadas, descascadas e transformadas em flocos, de forma a aumentar a eficiência da extração de óleo. A extração de óleo normalmente é realizada por meio de extração em solvente (tal como hexano), mas pode também ser atingida por meio de uma combinação de extração em solvente e/ou sob pressão física. O óleo resultante é denominado óleo bruto. O óleo bruto pode ter sua resina retirada por meio da hidratação de fosfolipídios e outros complexos de lipídios neutros e polares que facilitam a sua separação da fração de triglicéride sem hidratação (óleo de soja). As gomas de Iecitina resultantes podem ser adicionalmente processadas para fabricar produtos de lecitina comercialmente importantes utilizados em uma série de produtos alimentícios e industriais como agentes de emulsificação e liberação (ou seja, antiadesivos). Óleo com resina retirada pode ser adicionalmente refinado para a remoção de impurezas (principalmente ácidos graxos livres, pigmentos e gomas residuais). O refinamento é realizado por meio da adição de um agente cáustico que reage com ácido graxo livre para formar sabão e hidrata fosfatídeos e proteínas no óleo bruto. Utiliza-se água para lavar traços de sabão formados durante o refinamento. O subproduto de sabão pode ser utilizado diretamente em rações animais ou acidulado para recuperar os ácidos graxos livres. A coloração é removida por meio de adsorção com uma terra branqueadora que remove a maior parte dos compostos de clorofila e carotenoides. O óleo refinado pode ser hidrogenado, de forma a resultar em gorduras com várias texturas e propriedades de fusão. Pode-se utilizar winterização (fracionamento) para remover estearina do óleo hidrogenado por meio de cristalização sob condições de resfriamento cuidadosamente controladas. A desodorização (principalmente por meio de destilação de vapor a vácuo) é a última etapa e é projetada para remover compostos que forneçam odor ou sabor ao óleo. Outros subprodutos valiosos tais como tocoferóis e esteróis podem ser removidos durante o processo de desodorização. Destilado desodorizado que contém esses subprodutos pode ser vendido para a produção de vitamina E natural e outros produtos farmacêuticos com alto valor. Gorduras e óleos refinados, embranquecidos (hidrogenados, fracionados) e desodorizados podem ser embalados e vendidos diretamente ou adicionalmente processados em produtos mais especializados. Uma referência mais detalhada ao processamento de sementes de soja, produção de óleo de soja e utilização de subprodutos pode ser encontrada em Erickson1 Practical Handbook of Soybean Processing and Utilization, The American Oil Chemists' Society e United Soybean Board (1995). O óleo de soja é líquido à temperatura ambiente porque contém teor relativamente baixo de ácidos graxos saturados em comparação com óleos tais como manteiga de cacau, semente de palma, palma e coco.
Os óleos vegetais que contêm PUFAs que foram refinados e/ou purificados podem ser hidrogenados, de forma a resultarem em gorduras com diversas texturas e propriedades de fusão. Muitas gorduras processadas (incluindo espalhantes, gorduras de confeitaria, manteigas duras, margarinas, coberturas de cozimento etc.) necessitam de graus variáveis de solidez à temperatura ambiente e somente podem ser produzidas por meio de alteração das propriedades físicas do óleo fonte. Isso é mais comumente atingido por meio de hidrogenação catalítica.
A hidrogenação é uma reação química na qual adiciona-se hidrogênio às uniões duplas de ácido graxo insaturado com o auxílio de um catalisador tal como níquel. Óleo de soja com alto teor de ácido oleico, por exemplo, contém ácidos graxos oleico, LA e linolênico insaturados e cada um deles pode ser hidrogenado. A hidrogenação possui dois efeitos principais. Em primeiro lugar, a estabilidade oxidativa do óleo aumenta como resultado da redução do teor de ácido graxo insaturado. Em segundo lugar, as propriedades físicas do óleo são alteradas porque as modificações de ácido graxo aumentam o ponto de fusão, resultando em uma gordura sólida ou semissólida à temperatura ambiente.
Existem muitas variáveis que afetam a reação de hidrogenação que, por sua vez, altera a composição do produto final. As condições de operação, que incluem pressão, temperatura, tipo e concentração de catalisador, agitação e projeto de reator, encontram-se dentre os parâmetros mais importantes que podem ser controlados. Podem ser utilizadas condições de hidrogenação seletivas para hidrogenar os ácidos graxos mais insaturados em detrimento dos menos insaturados. Emprega-se freqüentemente hidrogenação muito leve ou de passagem para aumentar a estabilidade de óleos líquidos. Hidrogenação adicional converte um óleo líquido em uma gordura fisicamente sólida. O grau de hidrogenação depende das características de fusão e desempenho desejadas e projetadas para o produto final específico. Gorduras líquidas (utilizadas na fabricação de produtos de cozimento, gorduras sólidas e gorduras utilizadas para operações de fritura e torração comerciais) e matéria prima para a fabricação de margarina encontram-se dentre os inúmeros produtos de gordura e óleo possíveis atingidos por meio de hidrogenação. Uma descrição mais detalhada de hidrogenação e produtos hidrogenados pode ser encontrada em Patterson1 H. B. W., Hydrogenation of Fats and Oils: Theory and Practice, The American Oil Chemists' Society (1994).
Óleos hidrogenados tornaram-se um tanto controversos devido à presença de isômeros de ácidos graxos trans resultantes do processo de hidrogenação. A ingestão de grandes quantidades de isômeros trans foi relacionada com efeitos prejudiciais à saúde, que incluem razões mais altas entre lipoproteínas de baixa densidade e de alta densidade no plasma sangüíneo e aumento do risco de doenças cardíacas coronarianas.
ÓLEOS QUE CONTÉM PUFA PARA USO EM ALIMENTOS
As composições nutricionais incluem alimentos ou preparações para consumo humano que incluem para consumo enteral ou parenteral que, quando consumidos, (a) nutrem ou constroem tecidos ou fornecem energia; e/ou (b) mantêm, restauram ou sustentam situação nutricional ou função metabólica adequada. Essas composições compreendem pelo menos um óleo ou ácido produzido de acordo com a presente invenção e podem apresentar-se em uma forma sólida ou líquida. Além disso, a composição pode incluir macronutrientes comestíveis, vitaminas e sais minerais em quantidades apropriadas para atingir o propósito pretendido. As quantidades variarão, por exemplo, caso o uso se destine a bebês, crianças ou adultos normais e saudáveis que possuam necessidades especiais, tais como os que acompanham certas condições metabólicas.
Exemplos de macronutrientes que podem ser adicionados à composição incluem, mas sem limitar-se a gorduras comestíveis, carbo- hidratos e proteínas. Exemplos de gorduras comestíveis apropriadas incluem, mas sem limitar-se a óleo de coco, óleo de soja e mono e diglicérides. Glicose, lactose comestível e amido hidrolisado podem ser mencionados como exemplos de carbo-hidratos. Exemplos de proteínas apropriadas incluem, mas sem limitações, proteínas de soja, soro de eletrodiálise, leite desnatado de eletrodiálise, soro de leite ou os hidrolisados dessas proteínas. Exemplos de vitaminas e sais minerais apropriados incluem, mas sem limitações, cálcio, fósforo, potássio, sódio, cloreto, magnésio, manganês, ferro, cobre, zinco, selênio, iodo e complexo de Vitaminas A, E, D, C e B.
Exemplos de composições nutricionais incluem, mas sem limitações, fórmulas para bebês, suplementos alimentares e composições de re-hidratação.
O mercado sustenta atualmente uma série de produtos alimentícios e alimentos que incorporam ácidos graxos de ômega 3 e/ou ômega 6 (particularmente ARA, EPA e DHA). Contempla-se que os óleos de sementes e plantas e óleos de sementes alterados de acordo com a presente invenção que compreendem PUFAs funcionarão em alimentos e produtos alimentícios para fornecer os benefícios à saúde das formulações atuais. Em comparação com outros óleos vegetais, acredita-se que os óleos de acordo com a presente invenção funcionem de forma similar a outros óleos em aplicações alimentícias do ponto de vista físico (óleos parcialmente hidrogenados tais como óleo de soja são amplamente utilizados, por exemplo, como ingredientes de espalhantes moles, margarina e gorduras para fritura e cozimento).
Óleos de sementes e plantas e óleos de sementes alteradas que contêm ácidos graxos de ômega 3 e/ou ômega 6, conforme descrito no presente, serão apropriados para uso em uma série de alimentos e produtos alimentícios que incluem, mas sem limitar-se a: análogos de alimentos, produtos de carne, produtos de cereais, alimentos assados, alimentos de lanche e produtos lácteos. Além disso, os óleos de sementes e plantas e óleos de sementes alteradas do presente podem ser utilizados em formulações para fornecer benefício à saúde em alimentos medicinais que incluem nutricionais medicinais, suplementos alimentícios, fórmulas para bebês e produtos farmacêuticos. Os técnicos no assunto de processamento de alimentos e formulação de alimentos compreenderão como a quantidade e a composição dos óleos vegetais podem ser adicionadas ao alimento ou produto alimentício. Essa quantidade será indicada no presente como quantidade "eficaz" e dependerá do alimento ou produto alimentício, do alimento que o produto se destina a suplementar ou da condição média que o alimento medicinal ou nutricional medicinal destina-se a corrigir ou tratar.
Análogos alimentícios podem ser elaborados utilizando processos bem conhecidos dos técnicos no assunto. Podem ser mencionados análogos de carne, análogos de queijo, análogos de leite e similares. Análogos de carne fabricados com grãos de soja contêm proteína de soja ou tofu e outros ingredientes misturados entre si para simular vários tipos de carnes. Essas alternativas de carne são vendidas como alimentos congelados, enlatados ou secos. Normalmente, eles podem ser utilizados da mesma forma que os alimentos que substituem. Alternativas de carne fabricadas com grãos de soja são excelentes fontes de proteína, ferro e vitaminas B. Exemplos de análogos de carne incluem, mas sem limitar-se a análogos de hambúrguer, análogos de salsicha, análogos de toucinho e similares.
Análogos de alimentos podem ser classificados como imitação ou substitutos, dependendo das suas características funcionais e de composição. Uma imitação de queijo, por exemplo, necessita apenas relembrar o queijo que se destina a substituir. Um produto pode geralmente ser denominado substituto de queijo, entretanto, somente se for nutricionalmente equivalente ao queijo que está substituindo e atender às exigências mínimas de composição para aquele queijo. Desta forma, substituto de queijo freqüentemente conterá níveis de proteína mais altos que imitações de queijo e será fortificado com vitaminas e sais minerais.
Análogos de leite ou produtos alimentícios não lácteos incluem, mas sem limitar-se a imitações de leite e sobremesas congeladas não lácteas (tais como as fabricadas com grãos de soja e/ou produtos de proteína de soja).
Os produtos de carne englobam uma ampla variedade de produtos. Nos Estados Unidos, "carne" inclui "carnes vermelhas" produzidas a partir de bois, porcos e carneiros. Além das carnes vermelhas, existem produtos de aves que incluem galinhas, perus, gansos, galinhas d'Angola, patos e os peixes e frutos do mar. Existe uma ampla variedade de produtos de carne processada e temperada: frescos, curados e fritos, curados e cozidos. Lingüiças e salsichas são exemplos de produtos de carne processada. Desta forma, a expressão "produtos de carne", da forma utilizada no presente, inclui, mas sem limitações, produtos de carne processada.
Um produto alimentício de cereal é um produto alimentício derivado do processamento de grãos de cereal. Grãos de cereal incluem qualquer planta da família das gramíneas que gere grãos (sementes) comestíveis. Os cereais mais populares são cevada, milho, milho branco, aveia, quinoa, arroz, centeio, sorgo, triticale, trigo e arroz selvagem. Exemplos de produtos alimentícios de cereais incluem, mas sem limitar-se a: grãos inteiros, grãos triturados, flocos, farinha, farelo, gérmen, cereais matinais, alimentos extrudados, macarrão e similares.
Produtos assados compreendem quaisquer dos produtos alimentícios de cereais mencionados acima que foram assados ou processados de forma comparável a assados (ou seja, para secar ou endurecer submetendo-se ao calor). Exemplos de produtos assados incluem, mas sem limitações: pães, bolos, roscas, barras, macarrão, migalhas de pão, lanches assados, minibiscoitos, minibolachas, minicookies e minipretzels. Conforme mencionado acima, os óleos de acordo com a presente invenção podem ser utilizados como ingredientes.
Produtos alimentícios de lanche compreendem quaisquer dos produtos alimentícios descritos acima ou abaixo.
Produtos alimentícios fritos compreendem quaisquer dos produtos alimentícios descritos acima ou abaixo que tenham sido fritos.
Produtos alimentícios saudáveis são quaisquer produtos alimentícios que forneçam benefícios à saúde. Muitos produtos alimentícios derivados de oleaginosas podem ser considerados alimentos saudáveis.
As bebidas podem apresentar-se em forma líquida ou em pó seco.
Pode-se mencionar, por exemplo, bebidas não carbonatadas tais como sucos de frutas, frescas, congeladas, enlatadas ou concentradas; bebidas de leite integral ou aromatizado etc. Fórmulas nutricionais para adultos e bebês são bem conhecidas na técnica e disponíveis comercialmente (tais como Similac®, Ensure®, Jevity® e Alimentum® da Divisão de Produtos Ross, Abbott Laboratories).
Fórmulas para bebês são líquidos ou pós reconstituídos administrados a bebês e crianças jovens. "Fórmula para bebês" é definida no presente como um produto nutricional enteral que pode ser substituído por leite de peito humano na alimentação de bebês e é tipicamente composto de um percentual desejado de gordura misturada com percentuais desejados de carbo-hidratos e proteínas em uma solução aquosa (vide, por exemplo, a Patente Norte-Americana n° 4.670.285). Com base nos estudos mundiais de composição, bem como níveis especificados por grupos de especialistas, leite de peito humano médio contém tipicamente cerca de 0,20% a 0,40% do total de ácidos graxos (considerando cerca de 50% de calorias de gordura); e, geralmente, a razão entre DHA e ARA variaria de cerca de 1:1 a 1:2 (vide, por exemplo, formulações de Enfamil LIPIL® (Mead Johnson & Company) e Similac Advance® (Divisão de Produtos Ross, Abbott Laboratories)). Fórmulas para bebês detêm um papel especial a desempenhar na alimentação de bebês, pois freqüentemente são a única fonte de nutrientes para bebês; e, embora a amamentação ainda seja a melhor nutrição para os bebês, as fórmulas para bebês são uma alternativa suficientemente próxima para que os bebês não apenas sobrevivam, mas desenvolvam-se.
Produtos lácteos são produtos derivados do leite. Um análogo de leite ou produto não lácteo é derivado de uma fonte diferente de leite, tal como leite de soja, conforme discutido acima. Estes produtos incluem, mas sem limitações: leite integral, leite desnatado, produtos de leite fermentado tais como iogurte ou leite azedo, creme, manteiga, leite condensado, leite desidratado, clareador de café, creme de café, sorvete, queijo etc.
Produtos alimentícios adicionais que poderão incluir os óleos que contêm PUFA de acordo com a presente invenção são, por exemplo, gomas de mascar, confeitos e confeitos congelados, gelatinas e pudins, doces duros e moles, doces de frutas e geleias, açúcar branco granulado, substitutos de açúcar, molhos doces, coberturas e xaropes, bem como misturas em pó misturadas secas.
ÓLEOS QUE CONTÊM PUFA PARA USO EM PRODUTOS ALIMENTÍCIOS SAUDÁVEIS.
ALIMENTOS MEDICINAIS E PRODUTOS FARMACÊUTICOS Produtos alimentícios saudáveis são quaisquer produtos alimentícios que forneçam benefícios à saúde e incluem alimentos funcionais, alimentos medicinais, nutricionais medicinais e suplementos alimentares.
"Alimentos medicinais" são alimentos administrados sob a supervisão de um médico e destinados à administração alimentar específica de doenças para as quais são estabelecidas exigências nutricionais distintas.
Além disso, os óleos de plantas e sementes e óleos de sementes alteradas de acordo com a presente invenção podem ser utilizados em composições farmacêuticas padrão (os óleos que contêm PUFA de cadeia longa poderão ser facilmente incorporados, por exemplo, em qualquer dos produtos alimentícios mencionados acima, de forma a produzir um alimento funcional ou medicinal). Formulações mais concentradas que compreendem PUFAs incluem cápsulas, pós, pastilhas, géis, cápsulas gelatinosas, concentrados líquidos e emulsões que podem ser utilizadas como um suplemento alimentício em seres humanos ou animais diferentes de seres humanos.
Desta forma, uma composição farmacêutica poderá compreender um ou mais dos ácidos graxos e/ou óleos resultantes, bem como um veículo, adjuvante ou carreador farmaceuticamente aceitável padrão, bem conhecido e não tóxico, tal como solução salina tamponada com fosfato, água, etanol, polióis, óleos vegetais, agente umectante ou uma emulsão, tal como uma emulsão de água e óleo. A composição pode apresentar-se em uma forma liquida ou sólida.
Possíveis vias de administração incluem oral, retal, parenteral, tópica etc. A via de administração dependerá do efeito desejado.
A dosagem a ser administrada ao paciente pode ser determinada pelos técnicos comuns no assunto. Os fatores a serem considerados incluem, mas sem limitações, peso do paciente, idade do paciente, situação de imunidade do paciente etc.
A composição pode apresentar-se em uma série de formas tais como solução, dispersão, suspensão, emulsão ou pó estéril que é reconstituído em seguida. Desta forma, as suspensões, além dos compostos ativos, podem conter agentes formadores de suspensão tais como álcoois isoestearílicos etoxilados, polioxietileno sorbitol e sorbitan ésteres, celulose microcristalina, meta-hidróxido de alumínio, bentonita, agar-agar e tragacanto ou misturas de substâncias e similares.
Formas de dosagem sólidas tais como pastilhas e cápsulas podem ser preparadas utilizando métodos bem conhecidos na técnica. Óleos e ácidos graxos de acordo com a presente invenção podem ser empastilhados, por exemplo, com bases de pastilhas convencionais, tais como lactose, sacarose e amido de milho, em combinação com aglutinantes tais como acácia, amido de milho ou gelatina, desintegrantes ou estearato de magnésio. As cápsulas podem ser preparadas por meio de incorporação desses excipientes em uma cápsula de gelatina junto com antioxidantes e óleo/ácido graxo desejado. Os termos "dose" e "porção" são utilizados de forma intercambiável no presente e designam a quantidade de uma composição farmacêutica ou nutricional ingerida por um paciente em um ambiente isolado e projetada para fornecer quantidades eficazes dos componentes desejados.
É possível que essa composição possa ser utilizada para fins cosméticos. Ela pode ser adicionada a composições cosméticas previamente existentes, de maneira a formar uma mistura, ou pode ser utilizada como uma composição isolada.
ÓLEOS QUE CONTÊM PUFA PARA USO EM RAÇÕES ANIMAIS E APLICAÇÕES
VETERINÁRIAS
Rações animais são definidas genericamente no presente como produtos destinados a uso como alimentos ou para mistura em ração para animais diferentes de seres humanos. Os óleos de sementes e plantas e óleos de sementes alteradas de acordo com a presente invenção podem ser utilizados como ingredientes em várias rações animais.
Mais especificamente, embora sem limitações no presente, espera-se que os óleos de acordo com a presente invenção possam ser utilizados em produtos de rações para animais domésticos, produtos alimentícios para ruminantes e aves e produtos alimentícios para aquacultura. Produtos alimentícios para animais domésticos são os produtos destinados a ser fornecidos a animais domésticos (tais como cães, gatos, pássaros, répteis i e roedores). Estes produtos podem incluir o cereal e produtos alimentícios saudáveis acima, bem como carne e subprodutos de carne, produtos de proteína de soja, produtos de grama e feno (tais como alfafa, rabo-de-gato, aveia ou capim-cevadinha, legumes). Produtos alimentícios para aves e ruminantes são aqueles em que o produto destina-se a alimentação animal (tal como por perus, galinhas, bois e porcos). Como ocorre com as rações para animais domésticos acima, esses produtos podem incluir cereais e produtos alimentícios saudáveis, produtos de proteína de soja, carne e subprodutos de carne e produtos de grama e feno, conforme relacionado acima. Produtos alimentícios para aquacultura (ou "alimentos aquáticos") são os produtos destinados a uso em aquacultura, ou seja, referentes à propagação, cultivo ou criação de organismos e/ou animais aquáticos em água doce ou salgada.
Dever-se-á apreciar que as composições farmacêuticas e nutricionais descritas acima podem ser utilizadas com relação a animais, pois os animais podem experimentar as mesmas necessidades e condições dos seres humanos.
Exemplos
A presente invenção é adicionalmente definida nos Exemplos a seguir, nos quais as partes e percentuais são em peso e os graus são Celsius, a menos que indicado em contrário. Dever-se-á compreender que estes Exemplos, embora indiquem realizações preferidas da presente invenção, são fornecidos unicamente como forma de ilustração. A partir da discussão acima e destes Exemplos, os técnicos no assunto podem determinar as características essenciais da presente invenção e, sem abandonar seu espírito e escopo, podem elaborar várias mudanças e modificações da presente invenção para adaptá-la a vários usos e condições. Desta forma, diversas modificações da presente invenção além das exibidas e descritas no presente serão evidentes para os técnicos no assunto a partir do relatório descritivo acima. Essas modificações também se destinam a enquadrar-se dentro do escopo das reivindicações anexas.
Métodos gerais
Os métodos de clonagem molecular e DNA recombinante padrão utilizados nos Exemplos são bem conhecidos na técnica e descritos por: (1) Sambrook, J., Fritsch, E. F. e Maniatis1 T., Molecular Cloning: A Laboratory Manual; Cold Spring Harbor Laboratory: Cold Spring Harbor, Nova Iorque (1989) (Maniatis); (2) T. J. Silhavy, M. L. Bennan e L. W. Enquist, Experiments with Gene Fusions; Cold Spring Harbor Laboratory: Cold Spring Harbor, Nova lorque (1984); e (3) Ausubel, F. M. et al, Current Protocols in Molecular Biology, publicado pela Greene Publishing Assoc. e Wiley-Interscience (1987).
Os materiais e métodos apropriados para a manutenção e o crescimento de cultivos microbianos são bem conhecidos na técnica. Os métodos apropriados para uso nos exemplos a seguir podem ser encontrados conforme descrito em Manual of Methods for General Bacteriology (Phillipp Gerhardt, R. G. E. Murray, Ralph N. Costilow1 Eugene W. Nester, Willis A. Wood1 Noel R. Krieg e G. Briggs Phillips, Eds.), American Society for Microbiology: Washington DC (1994)); ou por Thomas D. Brock em Biotechnology: A Textbook of Industrial Microbiology, segunda edição, Sinauer Associates: Sunderland MA (1989). Todos os reagentes, enzimas de restrição e materiais utilizados para o crescimento e manutenção de células microbianas foram obtidos por meio da Aldrich Chemicals (Milwaukee Wl), DIFCO Laboratories (Detroit Ml), GIBCO/BRL (Gaithersburg MD) ou Sigma Chemical Company (St. Louis MO), a menos que especificado em contrário. Realizou-se clonagem molecular geral de acordo com métodos padrão (Sambrook et al, acima). A seqüência de DNA foi gerada em um sequenciador automático ABI utilizando tecnologia de término de tingimento (US 5.366.860, EP 272.007) empregando uma combinação de vetor e primers específicos de inserto. A edição de seqüências foi realizada em Sequencher (Gene Codes Corporation, Ann Arbor Ml). Todas as seqüências representam a cobertura de pelo menos duas vezes nas duas direções. Comparações de seqüências genéticas foram realizadas utilizando software DNASTAR (DNA Star1 Inc.).
O significado das abreviações é o seguinte: "seg" indica segundo(s), "min" indica minuto(s), "h" indica hora(s), "d" indica dia(s), "μΙ" indica microlitro(s), "ml" indica mililitro(s), "I" indica litro(s), "μΜ" indica micromolar, "mM" indica milimolar, "M" indica molar, "mmol" indica milimol(es), 'Vmol" indica micromol(es), "g" indica grama(s), >g" indica micrograma(s), "ng" indica nanograma(s), "U" indica unidade(s), "bp" indica par(es) de bases e "kB" indica quilobase(s).
Exemplo 1
Transformação de Cultivos de Embriões Somáticos de Soja com Vetores
de Expressão de Soja Condições de cultivo
Cultivos de suspensão embriogênica de soja (cv. Jack) são mantidos em 35 ml de meio líquido SB196 (abaixo) em um agitador giratório, 150 rpm, 26 0C com luzes fluorescentes brancas frias em um fotoperíodo de 16:8 horas de dia/noite com intensidade de luz de 60 a 85 pE/m2/s. Os cultivos são subcultivados a cada sete dias a duas semanas por meio de inoculação de cerca de 35 mg de tecido em 35 ml de SB196 líquido novo (o intervalo de subcultivo preferido é a cada sete dias).
Cultivos de suspensão embriogênica de soja são transformados com os plasmídeos de expressão de soja por meio do método de bombardeamento de disparador de partículas (Klein et al, Naturel 327: 70 (1987)) utilizando um instrumento DuPont Biolistic PDS1000/HE (retroajuste de hélio) para todas as trnsformações.
INICIO DE CULTIVO DE SUSPENSAO EMBRIOGENICA DE SOJA
Os cultivos de soja são iniciados duas vezes por mês com cinco a sete dias entre cada início. Vagens com sementes imaturas de plantas de soja disponíveis são picadas em 45 a 55 dias após o plantio. As sementes são removidas das vagens e colocadas em uma caixa magenta esterilizada. As sementes de soja são esterilizadas por meio da sua agitação por quinze minutos em uma solução de Clorox a 5% com uma gota de sabão marfim (ou seja, 95 ml de água destilada em autoclave mais 5 ml de Clorox e uma gota de sabão, bem misturados). As sementes são enxaguadas utilizando duas garrafas de um litro de água destilada estéril e as com menos de 4 mm são colocadas sobre lâminas de microscópio individuais. A extremidade pequena da semente é cortada e os cotilédones prensados para fora do revestimento de semente. Os cotilédones são transferidos para placas contendo meio SB1 (25 a 30 cotilédones por placa). As placas são embaladas com fita de fibra e mantidas a 26 0C com luzes fluorescentes brancas frias em um fotoperíodo de 16:8 horas de dia/noite em intensidade de luz de 60 a 80 pE/m2/s por oito semanas, com uma alteração de meios após quatro semanas. Após incubação sobre meio SB1, embriões secundários são cortados e colocados em meios líquidos SB196 por sete dias.
PREPARACAO DE DNA PARA BOMBARDEAMENTO
Um plasmídeo intacto ou um fragmento de plasmídeo de DNA que contém os genes de interesse e o gene marcador selecionável são utilizados para bombardeamento. Fragmentos de plasmídeos de expressão de soja, cuja construção é descrita no presente, são obtidos por meio de isolamento de gel de plasmídeos digeridos. Em cada caso, 100 μ9 de DNA de plasmídeo são utilizados em 0,5 ml da mistura de enzimas específica descrita abaixo. Os plasmídeos são digeridos com Ascl (100 unidades) em Tampão NE 4 (20 mM de Tris-acetato, 10 mM de acetato de magnésio, 50 mM de acetato de potássio, 1 mM de ditiotreitol, pH 7,9), 100 μg/ml de BSA e 5 mM de betamercaptoetanol a 37°C por uma hora e meia. Os fragmentos de DNA resultantes são separados por meio de eletroforese de gel sobre 1% SeaPIaque GTG agarose (BioWhitaker Molecular Applications) e os fragmentos de DNA que contêm os conjuntos genéticos são cortados do gel de agarose. DNA é purificado a partir da agarose utilizando a enzima de digestão de GELase seguindo o protocolo do fabricante.
Uma parcela de 50 μΙ de água destilada estéril contendo 3 mg de partículas de ouro (3 mg de ouro) foi adicionada a 30 μl de uma solução de 10 μg/μl de solução de DNA (fragmento de DNA preparado conforme descrito no presente), 25 μΙ de 5 M CaCb e 20 μl de 0,1 M espermidina. A mistura é agitada por três minutos no nível 3 de um agitador por turbilhonamento e centrifugada por dez segundos em um microcentrifugador de bancada. O sobrenadante é removido, seguido por uma lavagem com 400 μl de etanol a 100% e uma outra centrifugação rápida. Os 400 μl de etanol são removidos e a pelota é novamente suspensa em 40 μΙ de etanol a 100%. Cinco microlitros de suspensão de DNA foram liberados para cada disco suspenso do disco de instrumento Biolistic PDS1000/HE. Cada parcela de 5 μl continha cerca de 0,375 mg de ouro por bombardeamento (ou seja, por disco).
Preparação de tecidos ε bombardeamento com DNA
Cerca de 150 a 200 mg de cultivos de suspensão embriônica com sete dias de idade são colocados em uma placa petri de 60 χ 15 mm estéril vazia e a placa é coberta com tela plástica. A câmara é evacuada a vácuo de 68 a 71 cm de mercúrio e o tecido é bombardeado uma ou duas vezes por Λ placa com pressão de ruptura da membrana definida em 1100 psi. O tecido é colocado a cerca de 8,9 cm da tela de retenção/parada.
Seleção de embriões transformados
Embriões transformados são selecionados utilizando clorossulfurona (quando o gene acetolactato sintase (ALS) for utilizado como o marcador selecionável).
Após o bombardeamento, o tecido foi colocado em meios SB196 novos e cultivado conforme descrito acima. Seis a oito dias após o bombardeamento, o SB196 é trocado com SB196 novo contendo 100 ng/ml de clorossulfurona. Os meios de seleção são renovados uma vez por semana. Quatro a seis semanas após a seleção, tecido verde transformado é observado crescendo a partir de conjuntos embriogênicos necróticos não transformados. Tecido verde isolado é removido e inoculado em placas com múltiplas cavidades para gerar novos cultivos de suspensão embriogênica transformados e propagados de forma clonal.
Amadurecimento dos embriões
Conjuntos embriogênicos transformados da transformação de produção são cultivados por quatro a seis semanas em placas com múltiplas cavidades conforme descrito acima a 26 0C em SB196 sob lâmpadas fluorescentes brancas frias (Phillips Econowatt branca fria F40/CW/RS/EW) e Agro (Philips F40 Agro) (40 watts) em um fotoperíodo de 16:8 horas com intensidade da luz de 90 a 120 μΕ/πι28. Após esse período, conjuntos de embriões são removidos para um meio agar sólido, SB166, por uma a duas semanas e subcultivados em seguida para meio SB103 por três a quatro semanas para amadurecer os embriões. Após o amadurecimento sobre placas em SB103, os embriões individuais são removidos dos conjuntos, secos e selecionados conforme as alterações nas suas composições de ácidos graxos conforme descrito no Exemplo 2. Quando desejado, são obtidas plantas a partir de alguns eventos conforme descrito abaixo.
Dissecação ε germinação de embriões
Embriões individuais amadurecidos são dissecados por meio de sua colocação em uma placa petri pequena vazia (60 χ 15 mm) por cerca de quatro a sete dias. As placas são vedadas com fita de fibra (criando uma pequena câmara de umidade). Os embriões dissecados são plantados em meio SB71-4 onde são mantidos em germinação sob as mesmas condições de cultivo descritas acima. As plantículas germinadas são removidas do meio de germinação, completamente enxaguadas com água, plantadas em seguida em Redi-Earth em uma bandeja de embalagem de 24 células e cobertas com uma cúpula plástica transparente. Após uma a duas semanas, a cúpula é removida e as plantas são endurecidas por uma semana adicional. Caso as plantículas pareçam duras, elas são transplantadas para um vaso de 25,4 cm de Redi- Earth com até três plantículas por vaso. Após dez a dezesseis semanas, as sementes maduras são colhidas, picadas e analisadas para determinar o teor de ácidos graxos conforme descrito no presente.
Receitas de meios:
SB196 - Meio de proliferação líquido FN Lite (por litro)
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Soluções padrão FN Lite
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MEIO SOLIDO SB1 (POR LITRO)
1 pacote de sais MS (Gibco/BRL - Cat. N° 11117-066) 1 ml de vitaminas B5 1000X padrão 31,5 g de glicose
2 ml de 2,4-D (20 mg/l de concentração final)
pH 5,7
8 g de TC agar
Meio sólido SB199 (por litro)
1 pacote de sais MS (Gibco/BRL - Cat. n° 11117-066)
1 ml de vitaminas B5 1000X padrão
30 g de sacarose
4 ml de 2,4-D (concentração final de 40 mg/l)
pH 7,0
g de gelrita
Meio sólido SB166 (por litro)
1 pacote de sais MS (Gibco/BRL - Cat. N0 11117-066)
1 ml de vitaminas B5 1000X padrão
60 g de maltose
750 mg de MgC^ hexa-hidrato
5 g de carvão ativado
pH 5,7
2 g de gelrita
Meio sólido SB103 (por litro)
1 pacote de sais MS (Gibco/BRL - Cat. N0 11117-066)
1 ml de vitaminas B5 1000X padrão
60 g de maltose
750 mg de MgCb hexa-hidrato
pH 5,7
2 g de gelrita
Meio sólido SB 71-4 (por litro)
1 garrafa de sais B5 de Gamborg com sacarose (Gibco/BRL - cat. N° 21153-036)
pH 5,7
5 g de TC Agar
Padrão 2.4-D
Obter pré-fabricado da Phytotech Cat. No. D 295 - concentração de 1 mg/ml.
Vitaminas B5 padrão (por 100 ml)
Armazenar parcelas a -20 0C: -10 mg demioinositol -100 mg de ácido nicotínico -100 mg de piridoxina HCI -1 g de tiamina
Caso a solução não se dissolva com rapidez suficiente, aplicar nível baixo de calor por meio da placa sob agitação quente.
SB 228 - Histodiferenciação ε maturação de soja (SHaM) (por litro)
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Ajustar volume para 900 ml. pH 5,8 Autoclave
Adicionar a meios resfriados (< 30 0C):
* Glutamina (concentração final de 30 mM) 4%: 110 ml.
* Obs.: O volume final será de 1010 ml após a adição de glutamina.
Como glutamina degrada-se com relativa rapidez, pode ser preferível a adição imediata antes do uso dos meios. A validade é de duas semanas após a adição de glutamina; os meios base podem ser mantidos por mais tempo sem glutamina.
Macro FN-lite para SHAM 10X - Estoque n° 1 (por litro)
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Trazar para o volume Trazer para o volume Autoclave
FeEDTA 10OX- Estoque n° 3 (por litro)
<table>table see original document page 88</column></row><table>
EDTA deve ser completamente dissolvido antes da adição de ferro.
Trazer para o volume
A solução é fotossensível. A(s) garrafa(s) deverá(ão) ser embalada(s) em folha metálica para omissão de luz. Autoclave
Ca 10OX - Estoque n° 4 (por litro)
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Trazer para volume
Autoclave
Vitamina B51000X - Estoque n° 5 (por litro)
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Trazer para volume Armazenar congelado 4% Glutamina - estoque n° 6 (por litro)
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Adicionar gradualmente mediante agitação e aplicação de baixo calor. Não exceder 35 0C. Trazer para volume Filtrar e esterilizar Armazenar congelado* * Obs.: aquecer estoque congelado em banho a 31 0C para dissolver completamente os cristais.
Estoque de clorossulfona 1 mg/ml em 0,01 N Hidróxido de Amônio
Exemplo 2
Análise de ácidos graxos de embriões somáticos de soja ε sementes de soja Embriões de soja somáticos maduros são um bom modelo para embriões zigóticos. Embora no estado de embrião globular em cultivo líquido, embriões de soja somáticos contêm quantidades muito baixas de triacilglicerol ou proteínas de armazenagem típicas de embriões de soja zigóticos em amadurecimento. Neste estágio de desenvolvimento, a razão entre triacilglicéride total e lipídio polar total (fosfolipídios e glicolipídio) é de cerca de 1:4, como é típico de embriões de soja zigóticos no estágio de desenvolvimento a partir dos quais foi iniciado o cultivo de embriões somáticos. Também no estágio globular, os mRNAs para as proteínas de sementes proeminentes, subunidade α de β-conglicinina, inibidor de tripsina Kunitz 3 e Iectina de sementes são essencialmente ausentes. Mediante transferência para meios livres de hormônios para permitir a diferenciação para o estado de embriões somáticos em amadurecimento, triacilglicerol torna-se a classe de lipídio mais abundante. Da mesma forma, mRNAs para subunidade α de β-conglicinina, inibidor de tripsina Kunitz 3 e Iectina de semente tornam-se mensagens muito abundantes na população de mRNA total. Com base nisso, o sistema de embriões de soja somáticos comporta-se de forma muito similar a embriões de soja zigóticos em amadurecimento in vivo e, portanto, é um sistema modelo bom e rápido para análise dos efeitos fenotípicos de modificação da expressão de genes no processo de biossíntese de ácidos graxos (vide a Patente PCT n° WO 02/00904, Exemplo 3). De forma mais importante, o sistema de modelo também prevê a composição de ácido graxo de sementes de plantas derivadas de embriões transgênicos.
Cultivo de suspensão embriogênica de soja (cv. Jack) é transformado conforme descrito no Exemplo 1. Um subconjunto de embriões de soja gerados a partir de cada evento (dez embriões por evento) é colhido, tomado em ampolas GC de vidro e metil ésteres de ácido graxo são preparados por meio de transesterificação. Para a transesterificação, 50 μΙ de hidróxido de trimetilsulfônio (TMSH) e 0,5 ml de hexano são adicionados aos embriões em ampolas de vidro e incubados por trinta minutos à temperatura ambiente mediante agitação. Metil ésteres de ácidos graxos (5 μΙ injetados de camada de hexano) são separados e quantificados utilizando um Cromatógrafo de Gás Hewlett-Packard 6890 equipado com uma coluna capilar de sílica fundida Omegawax 320 (Catálogo n° 24152, Supelco Inc.). A temperatura do forno é programada para manter-se em 220 0C por 2,6 minutos, aumentar para 240 0C a 20 °C/min e mantida em seguida por 2,4 minutos adicionais. Gás veículo é fornecido por um gerador de hidrogênio Whatman. Os tempos de retenção são comparados com os de metil ésteres de padrões disponíveis comercialmente (Nu-Check Prep., Inc.).
Os ácidos graxos são identificados como 16:0 (palmitato), 18:0 (ácido esteárico), 18:1 (ácido oleico), LA, 18:2 (5,9), GLA, ALA, STA, 20:0 (ácido eicosanoico), 20:1 (11), 20:2 (7,11) ou 20:2 (8,11), EDA, SCI, DGLA, ARA, ERA, JUN, ETA, EPA e DPA; e as composições de ácidos graxos são expressas na forma de percentual em peso (% peso) do total de ácidos graxos.
Da mesma forma, as sementes são colhidas e um pequeno floco é retirado de parte de cada semente (do lado diretamente oposto ao eixo embriônico) utilizando uma lâmina de barbear. Os flocos de sementes são analisados para determinar ácidos graxos, conforme descrito acima. Os nomes de sementes são designados por uma série de cinco números separados por hífens, em que os três primeiros números indicam um evento específico, o quarto número indica a planta e o quinto número indica a semente analisada.
Exemplo 3
Crescimento de Euglena gracius. Síntese de cDNA ε Construção de Biblioteca
Euglena gracilis foi obtido por meio do laboratório do Dr. Richard Triemer na Universidade do Estado de Michigan (East Lansing Ml). De 10 ml de cultivo em crescimento ativo, uma parcela de 1 ml foi transferida para 250 ml de meio Euglena gracilis (Eg) em uma garrafa de vidro de 500 ml. Meio Eg foi elaborado combinando-se 1 g de acetato de sódio, 1 g de extrato de carne de boi (U126-01, Difco Laboratories, Detroit Ml), 2 g de triptona Bacto® (0123- 17-3, Difco Laboratories), 2 g de extrato de levedura Bacto® (0127-17-9, Difco Laboratories) em 970 ml de água. Após esterilização com filtro, 30 ml de sobrenadante de solo e água (15-3790, Carolina Biological Supply Company, Burlington NC) foram adicionados assepticamente para gerar o meio Eg final. Cultivos de Euglena gracilis foram cultivados a 23 °C com um ciclo de 16 h de luz e 8 h de escuro por duas semanas sem agitação.
Após duas semanas, 10 ml de cultivo foram removidos para análise de lipídios e centrifugados a 1800 χ g por cinco minutos. A pelota foi lavada uma vez com água e novamente centrifugada. A pelota resultante foi seca por cinco minutos a vácuo, novamente suspensa em 100 μL de hidróxido de trimetilsulfônio (TMSH) e incubada à temperatura ambiente por quinze minutos com agitação. Em seguida, adicionou-se 0,5 ml de hexano e as ampolas foram incubadas por quinze minutos à temperatura ambiente com agitação. Metil ésteres de ácido graxo (5 μΙ injetados da camada de hexano) foram separados e quantificados utilizando um Cromatógrafo de Gás Hewlett- Packard 6890 equipado com uma coluna capilar de sílica fundida Omegawax 320 (Supelco Inc., Cat. n° 24152). A temperatura do forno foi programada para manutenção a 220 0C por 2,7 min, aumento para 240 0C a 20 °C/min e manutenção em seguida por 2,3 minutos adicionais. Gás veículo foi fornecido por um gerador de hidrogênio Whatman. Os tempos de retenção foram comparados com os de metil ésteres de padrões disponíveis comercialmente (Nu-Chek Prep, Inc., Cat. n° U-99-A) e o cromatograma resultante é exibido na Fig. 13.
O cultivo por duas semanas restante (240 ml) foi peletizado por meio de centrifugação a 1800 χ g por dez minutos, lavado uma vez com água e novamente centrifugado. O RNA total foi extraído da pelota resultante utilizando o reagente RNA STAT-60® (TEL-TEST, Inc., Friendswood TX) e seguindo o protocolo fornecido pelo fabricante (utilizar 5 ml de reagente, RNA dissolvido em 0,5 ml de água). Desta forma, foi obtido 1 mg de RNA total (2 mg/ml) a partir da pelota. O mRNA foi isolado de 1 mg de RNA total utilizando o Kit de Purificação de mRNA (Amersham Biosciences, Piscataway NJ) seguindo o protocolo fornecido pelo fabricante. Desta forma, foram obtidos 85 pg de mRNA.
Uma biblioteca de cDNA foi gerada utilizando o Kit de Construção de Bibliotecas de cDNA Cloneminer® (Cat. n° 18249-029, Invitrogen Corporation, Carlsbad CA) e seguindo o protocolo fornecido pelo fabricante (Versão B, 25-0608). Utilizando o método sem radiomarcação, cDNA foi sintetizado a partir de 3,2 pg de mRNA (descrito acima) utilizando o primer Biotin-atfB2-Oligo(dT). Após a síntese da primeira e da segunda fita, o adaptador attBI foi adicionado, ligado e o cDNA foi fracionado de tamanho utilizando cromatografia de coluna. DNA das frações 7 e 8 (tamanho que varia de cerca de 800 a 1500 bp) foi concentrado, recombinado em pDONR® 222 e transformado em células resistentes a fagos E. coli ElectroMAX® DH10B® T1 (Invitrogen Corporation). A biblioteca de Euglena gracilis foi denominada eeglc.
Exemplo 4
CONSTRUÇÃO DE VETOR DE EXPRESSÃO DE SOJA PKR1084 PARA COEXPRESSÃO DA DELTA-9 ELONGASE DE EUGLENA GRACILIS E DA DELTA-5 DESSATURASE DE MORTIERELLA ALPINA
O fragmento Notl de pKS129 (SEQ ID N° 1, que é descrito na Patente PCT n° WO 04/071467), que contém o MaD5 (SEQ ID N° 52) foi clonado no local Notl de pKR457 (SEQ ID N° 2, que é descrito na Patente PCT n° WO 05/047479), para gerar pKR606 (SEQ ID N° 3).
O vetor pKR606 (SEQ ID N° 3) foi digerido com BsMII e, após enchimento para suavizar as extremidades, o fragmento que contém o conjunto Gy1/MaD5/legA2 foi clonado no local /V°°NgoMI preenchido de pKR277 (SEQ ID N° 4, que é descrito na Patente PCT n° WO 04/071467) para produzir pKR804 (SEQ ID N° 5).
Um clone da biblioteca de cDNA de Euglena (eeglc; vide Exemplo 3), denominado eeg1c.pk001.n5.f, que contém a delta-9 elongase de Euglena gracilis (EgD9elo; SEQ ID N° 46; vide também o Pedido Norte- Americano n° 11/601.563 (depositado em dezesseis de novembro de 2006 e publicado em 24 de maio de 2007; Pasta do Advogado n° BB-1562)) foi utilizado como modelo para amplificar EgD9elo com primers de oligonucleotídeos oEugEL1-1 (SEQ ID N° 42) e oEugEL1-2 (SEQ ID N° 43), utilizando Polimerase de DNA VentR® (Cat. n° M0254S, New England Biolabs Inc., Beverly MA), seguindo o protocolo do fabricante. O fragmento de DNA resultante foi clonado no vetor de clonagem pCR-Blunt® utilizando o Kit de Clonagem de PCR Zero Blunt® (Invitrogen Corporation), seguindo o protocolo do fabricante, para produzir pKR906 (SEQ ID N° 45).
O plasmídeo pKR906 (SEQ ID N0 45) foi digerido com Notl e o fragmento que contém o EgD9elo foi clonado no plasmídeo pKR132 (SEQ ID N0 12, que é descrito na Patente PCT n0 WO 2004/071467) para gerar pKR953 (SEQ ID N0 6).
O plasmídeo pKR953 (SEQ ID N0 6) foi digerido com Psfl e o fragmento que contém o EgD9elo foi clonado no local Sbfl de pKR804 (SEQ ID N° 5) para gerar pKR1084 (SEQ ID N0 7, Fig. 2).
Desta forma, a delta-5 dessaturase de Mortierella alpina (MaD5) foi expressa com a delta-9 elongase de Euglena gracilis (EgD9elo) atrás de promotores fortes específicos de sementes.
Exemplo 5
Crescimento de Tetruetreptia pomquetensis CCMP1491. síntese de cDNA ε clonagem de delta-8 dessaturase Células CCMP1491 de Tetruetreptia pomquetensis (de um litro de cultivo) foram adquiridas do Centro Nacional Provasoli-Guillard de Cultivo de Fitoplâncton Marinho (CCMP) (Laboratório Bigelow de Ciências Oceânicas, West Boothbay Harbor, Maine). RNA total foi isolado utilizando o reagente trizol (Invitrogen, Carlsbad CA), de acordo com o protocolo do fabricante. A pelota celular foi novamente suspensa em 0,75 ml de reagente trizol, misturada com 0,5 ml de esferas de vidro de 0,5 mm e homogeneizada em um minibatedor de esferas Biospec (Bartlesville OK) no ambiente mais alto por três minutos. A mistura foi centrifugada em uma centrífuga Eppendorf por trinta segundos a 14000 rpm para remover fragmentos e esferas de vidro. O sobrenadante foi extraído com 150 μΙ de 24:1 clorofórmio:álcool isoamílico. A fase aquosa superior foi utilizada para isolamento de RNA.
Para isolamento de RNA1 a fase aquosa foi misturada com 0,375 ml de álcool isopropílico e mantida em incubação à temperatura ambiente por cinco minutos. RNA precipitado foi recolhido por meio de centrifugação a 8000 rpm e 4 ° por cinco minutos. A pelota foi lavada uma vez com 0,7 ml de etanol a 80% e seca em ar. Desta forma, foram obtidos 95 pg de RNA total a partir de Tetruetreptia pomquetensis CCMP1491.
RNA total (0,95 pg de RNA total em 1 μΙ) foi utilizado como modelo para sintetizar cDNA de fita dupla. Utilizou-se o Kit de Construção de Bibliotecas de cDNA Creator® Smart® da BD Bioscience Clontech (Paio Alto CA). RNA total (1 μΙ) foi misturado com 1 μΙ de oligonucleotídeo SMART IV (SEQ ID N0 60), 1 μΙ do PrimerAdaptador do kit 3-RACE da Invitrogen (SEQ ID N0 61) e 2 μΙ de água. A mistura foi aquecida a 75 0C por cinco minutos e resfriada em seguida sobre gelo por cinco minutos. À mistura, adicionou-se 2 μΙ de 5X tampão de primeira fita, 1 μΙ de 20 mM DTT1 1 μΙ de mistura de dNTP (10 mM cada de dATP, dCTP, dGTP e dTTP) e 1 μΙ de transcriptase reversa PowerScript. A amostra foi incubada a 42 0C por uma hora. Os cDNAs de primeira fita resultantes foram utilizados em seguida como modelo de amplificação.
CCMP1491 de Tetruetreptia pomquetensis (TpomD8; SEQ ID N0 50; vide também o pedido copendente do Cessionário do Depositante que possui Pedido de Patente Norte-Americano n° 11/876115 (depositado em 22 de outubro de 2007; Pasta do Advogado n° BB-1574)) foi amplificado a partir do cDNA com os primers de oligonucleotídeos TpomNot-5 (SEQ ID N0 62) e TpomNot-3 (SEQ ID N0 63) utilizando Taq polimerase (Invitrogen Corporation) seguindo o protocolo do fabricante.
cDNA de Tetruetreptia pomquetensis CCMP1491 (1 μΙ) foi combinado com 50 pmol de TpomNot-5 (SEQ ID N0 62), 50 pmol de TpomNot-3 (SEQ ID N0 63), 1 μΙ de mistura de nucleotídeos PCR (10 mM, Promega, Madison Wl), 5 μΙ de 10X tampão PCR (Invitrogen Corporation), 1,5 μΙ de MgCI2 (50 mM, Invitrogen Corporation), 0,5 μΙ de Taq polimerase (Invitrogen Corporation) e água até 50 μΙ. As condições de reação foram de 94 0C por três minutos, seguidas por 35 ciclos de 94 0C por 45 segundos, 55 0C por 45 segundos e 72 0C por um minuto. O PCR foi finalizado a 72 0C por sete minutos e mantido em seguida a 4 0C. A reação de PCR foi analisada por meio de eletroforese de gel de agarose sobre 5 μΙ e observou-se uma faixa de DNA com peso molecular de cerca de 1,3 kb.
Os 45 μl de produto restantes foram separados por meio de eletroforese de gel de agarose e o DNA1 purificado utilizando o Kit de Recuperação de DNA de Gel Zymoclean® (Zymo Research, Orange CA) seguindo o protocolo do fabricante. O DNA resultante foi clonado no Vetor Easy pGEM®-T (Promega) seguindo o protocolo do fabricante para gerar pLF114-10 (SEQIDN0 64).
Exemplo 6
Construção de vetores de expressão de soja para expressão da delta-8 dessaturase de TETRUETREPTIA POMQUETENSIS CCMP1491
O fragmento de Psíl, que contém o conjunto Ann/SdD17/BD30 de pKR271 (SEQ ID N0 65; que é descrito na Patente PCT n0 WO 04/071467) é clonado no local SbfI de pKR226 (SEQ ID N0 9, que também é descrito na Patente PCT n0 WO 04/071467) para produzir o vetor pKR886r (SEQ ID N0 66).
Um plasmídeo inicial pKR72 (Acesso ATCC n° PTA-6019; SEQ ID N0 67, seqüência de 7085 bp), um derivado de pKS123 que foi descrito anteriormente na Patente PCT n0 WO 02/008269, contém o gene fosfotransferase de higromicina B (HPT) (Gritz, L. e Davies, J., Gene 25: 179- 188 (1983)), ladeado pelo promotor T7 e término de transcrição (conjunto T7prom/HPT/T7term) e uma origem de reprodução bacteriana (ori) para seleção e reprodução em bactérias (por exemplo, E. coli). Além disso, pKR72 também contém HPT, ladeado pelo promotor 35S (Odell et al, Nature 313: 810- 812 (1985)) e término de transcrição NOS 3' (Depicker et al, J. Mol. Appi Genet. 1: 561-570 (1982)) (conjunto 35S/HPT/NOS 3') para seleção em plantas tais como soja. pKR72 também contém um local de restrição de Noti, ladeado pelo promotor da subunidade α' de β-conglicinina (Beachy et al, EMBO J. 4: 3047-3053 (1985)) e a região de término de transcrição 3' do gene faseolina (Doyle et al, J. BioL Chem. 261: 9228-9238 (1986)), de forma a permitir forte expressão específica de tecido nas sementes de soja de genes clonados no local Nott.
O conjunto pcon/A/ofl/Phas3' no plasmídeo pKR72 (SEQ ID N0 67) é amplificado utilizando primers de oligonucleotídeos oCon-1 (SEQ ID N0 68) e oCon-2 (SEQ ID N0 69) utilizando DNA Polimerase VentR® (Catálogo n° M0254S, New England Biola bs Inc., Beverly MA) seguindo o protocolo do fabricante. O fragmento de DNA resultante é digerido com Xba\ e clonado no local Xibal de pUC19, para produzir pKR179 (SEQ ID N0 24).
TpomD8 é liberado do plasmídeo pLF114-10 (SEQ ID N0 64; vide Exemplo 5) por meio de digestão com NotI e é clonado no local Nott do plasmídeo pKR179 (SEQ ID N0 24) para produzir pKR1002 (SEQ ID N0 8).
O vetor pKR1002 (SEQ ID N0 8) é digerido com Psfl e o fragmento que contém o TpomD8 (SEQ ID N0 50) é clonado no local Sbft de pKR226 (SEQ ID N0 9; que é descrito na Patente PCT n0 WO 04/071467)) para gerar BC-Tpom_ALS (SEQ ID N0 10; Fig. 3) ou BC-Tpom_ALSrev (SEQ ID N0 11; Fig. 4), dependendo da orientação.
Desta forma, a delta-8 dessaturase de Tetruetmptia pomquetensis CCMP1491 (TpomD8) pode ser expressa atrás de um promotor específico de semente forte em um vetor de expressão de soja que possui o marcador de resistência a herbicida ALS e nenhuma delta-17 dessaturase de Saprolegnia diclina (SdD17) e/ou nenhuma delta-15 dessaturase de Fusarium moniliforme (FmD15).
Exemplo 7
Construção de vetores de expressão de soja para expressão da delta-8 dessaturasse de TETRUETREPTIA POMQUETENSIS CCMP1491 com a delta-5 dessaturase de EUGLENA GRACILIS
Um clone da biblioteca de cDNA de Euglena (eeglc; cuja construção é descrita no Exemplo 3), denominado eeg1c.pk016.n9.f, que contém a delta-5 dessaturase de Euglena gracilis (EgD5; SEQ ID N° 54; pedido copendente do Cessionário do Depositante que possui Pedido de Patente Norte-Americano n° 11/748629 (depositado em quinze de maio de 2007 e publicado em vinte de dezembro de 2007; Pasta do Advogado n° CL-3486)) é utilizado como modelo para amplificar EgD5 com primers de oligonucleotídeos EgD5-5Not (SEQ ID N° 13) e EgD5-3Not (SEQ ID N° 14) utilizando DNA Polimerase VentR® (Cat. n° M0254S, New England Biolabs Inc., Beverly MA) seguindo o protocolo do fabricante. O fragmento de DNA resultante é digerido com Nott e clonado no local Nott de pKR268 (SEQ ID N° 15; que é descrito na Patente PCT n° WO 04/071467) para gerar Ann-EgD5 (SEQ ID N° 16).
Ann-EgD5 (SEQ ID N° 16) é digerido com fís/WI e clonado no local fís/WI de BC-Tpom_ALS (SEQ ID N° 10) ou BC-Tpom_ALSrev (SEQ ID N° 11) para gerar BC-Tpom_Ann-EgD5_ALS (SEQ ID N° 17; Fig. 5) ou BC- Tpom_Ann-EgD5_ALSrev (SEQ ID N° 18, Fig. 6), respectivamente.
Exemplo 8
Construção de vetor de expressão de soja KS263 para regulagem para
BAIXO DE FAD3 DE SOJA UTILIZANDO REGIÕES DE COMPLEMENTARIDADE ELVISLIVES
Vetor de expressão de soja KS133 (SEQ ID N° 19), conforme descrito em EP 1297163 A2 (Patente PCT n0 WO 02/000904, publicada em três de janeiro de 2002) (cujos teores são incorporados ao presente como referência), contém 2x cópias das regiões complementares ELVISLIVES que ladeiam um local Nott. Os locais ELVISLIVES são ladeados pelo promotor KTi e seqüências de término KTi 3'.
O plasmídeo XF1, descrito na Patente PCT n0 WO 93/11245 (que foi publicada em dez de junho de 1993; e também Patente Norte-Americana n° 5.952.544) (cujos teores são incorporados ao presente como referência) contém o gene delta-15 dessaturase de soja (fad3) (Acesso GenBank n° L22964; também denominado GmFAD3B). O plasmídeo XF1 foi digerido com Hha\ e as extremidades foram suavizadas utilizando 3'->5' exonuclease. O DNA resultante foi digerido em seguida com EcoRI e preenchido em seguida. O fragmento de DNA que contém a região interna da delta-15 dessaturase de soja foi ligado em seguida ao local Not\ preenchido de KS133 para gerar KS263 (SEQ ID N0 20; Fig. 7). Desta forma, uma região da delta-15 dessaturase de soja, ladeada por regiões complementares ELVISLIVES, pode ser expressa atrás do promotor KTi específico de semente forte (também denominado KTi3).
Exemplo 9
Construção de vetores de expressão de soja para regulagem para baixo de Fad3 de soja utilizando regiões complementares ELVISLIVES
O conjunto ELVISLIVES/fad3 de KS263 (SEQ ID N0 20) é amplificado utilizando ELfad3-5Not (SEQ ID N0 21) e ELfad3-3Not (SEQ ID N0 22) para gerar um fragmento de DNA denominado ELfad3Not (SEQ ID N0 23). PCR é conduzido utilizando diversas temperaturas de combinação que variam de 45 0C a 72 0C com concentrações de DMSO que variam de 0 a 10%. Produtos de PCR são sequenciados para garantir a obtenção do fragmento desejado.
Fragmento de DNA ELfad3Not (SEQ ID N0 23) é digerido com Nofí e clonado no local Not\ do plasmídeo pKR179 (SEQ ID N0 24) para produzir BC-ELfad3 (SEQ ID N0 25).
O vetor BC-ELfad3 (SEQ ID N0 25) é digerido com Sbfi e o fragmento que contém o fragmento de DNA ELfad3Not é clonado no local SbfI de pKR226 (SEQ ID N0 9) para produzir BC-ELfad3_ALS (SEQ ID N0 27). Uma ilustração esquemática de BC-ELfad3_ALS (SEQ ID N0 27) é exibida na Fig. 8. Desta forma, uma região da delta-15 dessaturase de soja, ladeada por regiões complementares ELVISLIVES, pode ser expressa atrás do forte promotor de betaconglicinina específico de sementes.
Exemplo 10
Construção de vetores de expressão de soja para regulagem para baixo de FAP3 de soja utilizando regiões complementares ELVISLIVES e coexpressão com a delta-8 dessaturase de TETRUETREPTIA POMQUETENSIS CCMP1491:
Em uma construção similar à descrita acima, fragmento de DNA ELfad3Not (SEQ ID N0 23) é digerido com Nofí e clonado no local Notl de pKR268 (SEQ ID N0 15; Exemplo 7) para produzir Ann-ELfad3 (SEQ ID N0 28).
Ann-ELfad3 é digerido com BsiWI e clonado no local BsiWI de BC-Tpom_ALSrev (SEQ ID N0 11; Fig. 4) para gerar BC-Tpom_Ann- ELfad3_ALS (SEQ ID N0 29; Fig. 9). Desta forma, uma região da delta-15 dessaturase de soja, ladeada por regiões complementares de ELVISLIVES, pode ser expressa atrás do promotor de anexina específico de semente forte e coexpresso com o TpomD8.
Exemplo 11
Construção de vetores de expressão de soja para regulagem para baixo de fad3 de soja utilizando repetições invertidas de fad3
Uma parte da extremidade 5' do gene fad3 é amplificada a partir de XF1 (descrito no Exemplo 8) utilizando HPfad3-1 (SEQ ID N0 30) e HPfad3- 2 (SEQ ID N0 31) para gerar um fragmento de DNA denominado HPfad3AB (SEQ ID N0 32).
Uma parte da extremidade 3' do gene fad3 é amplificada a partir de XF1 (descrito acima) utilizando HPfad3-3 (SEQ ID N0 33) e HPfad3-4 (SEQ ID N0 34) para gerar um fragmento de DNA denominado HPfad3A' (SEQ ID N0 35).
HPfad3AB e HPfad3A' são combinados e amplificados com HPfad3-1 (SEQ ID N0 30) e HPfad3-4 (SEQ ID N° 34) para gerar HPfad3ABA' (SEQ ID N° 36).
HPfad3ABA' é digerido com Notl/Asp718 e é clonado no fragmento de local A/ofl/Asp718 do plasmídeo pKR179 (SEQ ID N° 24), que contém o promotor de betaconglicinina, para produzir BC-HPfad3ABA' (SEQ ID N° 37). O conjunto de expressão gerado não contém um término de transcrição.
O vetor BC-HPfad3ABA' (SEQ ID N0 37) é digerido com Sbfl e o fragmento que contém o fragmento de DNA de HPfad3ABA' é clonado no local Sbf1 de pKR226 (SEQ ID N° 9) para produzir BC-HPfad3ABA'_ALS (SEQ ID N° 39). Uma ilustração esquemática de BC-HPfad3ABA'_ALS (SEQ ID N° 39) é exibida na Fig. 10. Desta forma, uma região da delta-15 dessaturase de soja que forma uma estrutura de grampo de cabelo possa ser expressa atrás do promotor de betaconglicinina específico de semente forte.
Exemplo 12
Construção de vetores de expressão de soja para regulagem para baixo de Fad3 de soja utilizando grampos de cabelo de Fad3 ε coexpressão com a delta-8 dessaturase de Tetruetreptia pomquetensis CCMP1491
Em uma construção similar à descrita no Exemplo 11, HPfad3ABA' (SEQ ID N° 36) é digerido com Notl/Asp718 e clonado no fragmento de local A/ofl/Asp718 do plasmídeo pKR268 (SEQ ID N° 15), que contém promotor anexina de soja, para produzir Ann-HPfad3ABA' (SEQ ID N0 38). O conjunto de expressão gerado não contém um término de transcrição.
Ann-HPfad3ABA' é digerido com BsiWI e clonado no local BsiWI de BC-Tpom ALSrev (SEQ ID N0 11; Fig. 4) para gerar BC-Tpom_Ann- HPfad3ABA'_ALS (SEQ ID N0 26; Fig. 11). Desta forma, uma região da delta- 15 dessaturase de soja que forma uma estrutura de grampo de cabelo pode ser expressa atrás do promotor de anexina específico de semente forte e coexpresso com o TpomD8.
Exemplo 13
Crescimento de Pavlova lutheriíCCMP459). síntese de cDNA ε clonagem
de delta-8 dessaturase
Pavlova Iutheri (CCMP459) foi obtido a partir de CCMP e cultivado em frascos de 250 ml que contêm 50 ml de meio F/2-Si (fabricado utilizando Kit de Meio de Família F/2-KIT20F2 e Seqwater Filtrado-SEA2 de CCMP) a 26 0C com agitação a 150 rpm. Os cultivos foram transferidos para meio novo semanalmente utilizando diluição 1:4 (cultivo velho: meio novo).
Cultivos de 28 frascos (1400 ml) foram combinados, as células foram peletizadas por meio de centrifugação a 1800 χ g por dez minutos, lavados uma vez com água e novamente centrifugados. RNA total foi extraído da pelota resultante utilizando o reagente RNA STAT-60® (TEL-TEST, Inc., Friendswood TX) e seguindo o protocolo fornecido pelo fabricante. Desta forma, 2,6 mg de RNA total (2,6 mg/ml) foram obtidos a partir da pelota. O mRNA foi isolado a partir de 1,25 mg de RNA total utilizando o Kit de Purificação de mRNA (Amersham Biosciences, Piscataway NJ) seguindo o protocolo fornecido pelo fabricante. Desta forma, foram obtidos 112 pg de mRNA.
cDNA foi sintetizado a partir de 224 ng de mRNA utilizando o Sistema de Síntese de Primeira Fita SuperScript® para Kit RT-PCR (Invitrogen® Life Technologies, Carlsbad CA) com o primer oligo(dT) fornecido de acordo com o protocolo do fabricante. Após tratamento com RNase H de acordo com o protocolo, a delta-8 dessaturase de Pavlova Iutheri (PavD8; SEQ ID N° 48; descrita no pedido copendente do Cessionário do Depositante que possui o Pedido de Patente Norte-Americano n° 11/737772 (depositado em vinte de abril de 2007; Pasta de Advogado n° BB-1566) foi amplificada a partir do cDNA resultante com primers de oligonucleotídeos PvDES5'Not-1 (SEQ ID N0 72) e PvDES3'Not-1 (SEQ ID N0 73) utilizando as condições descritas abaixo.
cDNA (2 μΙ) da reação descrita acima foi combinado com 50 pmol de PvDES5'Not-1 (SEQ ID N0 72), 50 pmol de PvDES3"Not-1 (SEQ ID N0 73), 1 μl de mistura de nucleotídeos de PCR (10 mM, Promega, Madison Wl), 5 μΙ de 10X tampão PCR (Invitrogen Corporation), 1,5 μΙ de MgC^ (50 mM, Invitrogen Corporation), 0,5 μΙ de Taq polimerase (Invitrogen Corporation) e água até 50 μl. As condições de reação foram de 94 0C por três minutos, seguidas por 35 ciclos de 94 0C por 45 segundos, 55 0C por 45 segundos e 72 0C por um minuto. O PCR foi finalizado a 72 0C por sete minutos e, em seguida, mantido a 4°C. A reação de PCR foi analisada por meio de eletroforese de gel de agarose sobre 5 μΙ e foi observada uma faixa de DNA com peso molecular de cerca de 1,3 kb. Os 45 μΙ restantes de produto foram separados por meio de eletroforese de gel de agarose e o DNA, purificado utilizando o Kit de Recuperação de DNA de Gel Zymoclean® (Zymo Research, Orange CA), seguindo o protocolo do fabricante. O DNA resultante foi clonado no Vetor pGEM®-T Easy (Promega) para gerar pLF113 (SEQ ID N0 74).
Exemplo 14
Construção do vetor de expressão de soja pKR973 para coexpressão da delta-8 dessaturase de Pavlova luthericom a delta-9 elongase de Euglena gracilis ε a delta-5 dessaturase de Morteriella alpina
Vetor pKR287 (SEQ ID N0 75, que é descrito na Patente PCT n° WO 04/071467, publicada em 26 de agosto de 2004) contém a delta-5 dessaturase de Mortierella alpina (MaD5) (SEQ ID N0 52), ladeada pelo promotor de glicinina de soja Gy1 e pela região de término 3' de Iegumina A2 de ervilha (conjunto Gy1/MaD5/legA2). O vetor pKR287 foi digerido com Sbfl/BsiWI e o fragmento que contém o conjunto Gy1/MaD5/legA2 foi clonado no fragmento Sbfl/BsiWI de pKR277 (SEQ ID N0 4; que é descrito no Exemplo 4 e na Patente PCT n0 WO 04/071467) para produzir pK952 (SEQ ID N0 77).
Por meio de uma série de etapas de subclonagem, as seqüências que contêm locais de restrição Asp718 foram adicionadas às extremidades 5' e 3' do conjunto Kti/Notl/Kti3'Salb3' de pKR457 (SEQ ID N0 2; descrito no Exemplo 4) para gerar SEQ ID N0 70.
O plasmídeo pLF113 (SEQ ID N0 74; descrito no Exemplo 13) foi digerido com Not\ e o fragmento que contém o PavD8 foi clonado no local /Vofl do conjunto Kti/A/ofl/Kti3'Salb3' (SEQ ID N0 70) e, em seguida, o fragmento de DNA resultante foi digerido com Asp718 e clonado no local Sbfi de pKR952 (SEQ ID N0 77) para gerar pKR970 (SEQ ID N0 71).
O plasmídeo pKR953 (SEQ ID N0 6) foi digerido com Pstt e o fragmento que contém o EgD9elo foi clonado no local Sbfi de pKR970 (SEQ ID N0 71) para gerar pKR973 (SEQ ID N0 41, Fig. 15).
Desta forma, a delta-8 dessaturase de Pavlova Iutheri (PavD8) poderá ser coexpressa com a delta-5 dessaturase de Mortierella alpina (MaD5) e a delta-9 elongase de Euglena gracilis (EgD9elo) atrás de promotores fortes específicos de sementes.
Exemplo 15
Embriões de soja transformados com vetores de expressão de soja PKR1005 ε PKR973 que contém delta-8 dessaturase de Pavlova lutheri, delta-5 dessaturase de Mortierella alpina. delta-9 elongase de Euglena gracilis, delta-8 dessaturase de tetruetreptia pomquetensis e delta-17 dessaturase de saprolegnia dicuna pKR973 (SEQ ID N0 41; Fig. 15; descrito no Exemplo 14) contém Pav D8, MaD5 e EgD9elo. pKR1005 (SEQ ID N0 44; Fig. 16; descrito abaixo) contém TpomD8 e Sd 17.
O fragmento de Pstt, que contém o conjunto Ann/SdD17/BD30 de pKR271 (SEQ ID N0 65; que é descrito na Patente PCT n0 WO 04/071467), foi clonado no local SbfI de pKR226 (SEQ ID N0 9; que também é descrito na Patente PCT n0 WO 04/071467) para produzir o vetor pKR886r (SEQ ID N0 66; descrito no pedido copendente do Cessionário do Depositante que possui Pedido Norte-Americano n° 11/601.563 (depositado em dezesseis de novembro de 2006, publicado em 24 de maio de 2007; Pasta do Advogado n° BB-1562 e cujo conteúdo é incorporado ao presente como referência).
TpomD8 foi liberado do plasmídeo pLF114-10 (SEQ ID N0 64; vide Exemplo 5) por meio de digestão com Noti e foi clonado no local Noti do plasmídeo pKR179 (SEQ ID N0 24) para produzir pKR1002 (SEQ ID N0 8). O vetor pKR1002 (SEQ ID N0 8) foi digerido com Psfl e o fragmento que contém o TpomD8 (SEQ ID N0 50) foi clonado no local Sbfl de pKR886r (SEQ ID N0 66) para produzir pKR1005 (SEQ ID N0 44; Fig. 16; vide o pedido copendente do Cessionário do Depositante que possui Pedido de Patente Norte-Americano n° 11/876115 (depositado em 22 de outubro de 2007; Pasta do Advogado n° BB- 1574 e cujo conteúdo é incorporado ao presente como referência).
Cultivo de suspensão embriogênica de soja (cv. Jack) foi transformado com os fragmentos de /\scl de pKR1005 (SEQ ID N0 44; Fig. 16) e pKR973 (SEQ ID N0 41; Fig. 15; vide o pedido copendente do Cessionário do Depositante que possui Pedido de Patente Norte-Americano n° 11/737772 (depositado em vinte de abril de 2007; Pasta do Advogado n° BB-1566) (fragmentos que contêm os conjuntos de expressão), conforme descrito no presente. Um subconjunto de embriões de soja gerados a partir de cada evento (dez embriões por evento) foram colhidos, tomados em ampolas GC de vidro, metil ésteres de ácidos graxos (FAMEs) foram preparados por meio de transesterificação e analisados por meio de GC conforme descrito no Exemplo 10. Os tempos de retenção foram comparados com os de metil ésteres de padrões disponíveis comercialmente (Nu-Chek Prep., Inc.).
Desta forma, 373 eventos transformados com pKR1005 (SEQ ID N0 44; Fig. 16) e pKR973 (SEQ ID N0 41) (experimento denominado Cura 17) foram analisados. Dos 373 eventos analisados, foram identificados 56 que produziram ARA em pelo menos um embrião dentre dez analisados em uma abundância relativa de mais de 1,0% do total de ácidos graxos. Destes, foram analiados dois que produziram ARA em pelo menos um embrião dentre dez analisados em uma abundância relativa de mais de 10,0% do total de ácidos graxos. Altos níveis de ARA são prováveis devido à expressão relativa mais baixa do gene SdD17 nesses eventos.
Os perfis de ácidos graxos para embriões de um desses eventos (4828-4-18) que possuem o nível de ARA mais alto são exibidos na Fig. 17. O perfil de ácido graxo para um embrião (4828-4-18-4) foi descartado, pois nenhum sinal foi obtido. Neste caso, os níveis de ARA em embriões individuais são de até 20,3% do total de ácidos graxos, os níveis de EPA em embriões individuais são de menos de 5,5% do total de ácidos graxos e os níveis de GLA em embriões individuais são de menos de 1,2% do total de ácidos graxos. Os ácidos graxos são identificados como 16:0 (palmitato), 18:0 (ácido esteárico), 18:1 (ácido oleico), LA, GLA, ALA, EDA, DGLA, ARA, ERA, JUN1 ETA, EPA e DPA; e as composições de ácidos graxos relacionadas na Fig. 17 são expressas na forma de percentual em peso (% em peso) do total de ácidos graxos. Para a Fig. 17, os ácidos graxos relacionados como "outros" incluem: 18:2 (5,9), STA, 20:0, 20:1 (11), 20:2 (7,11) ou 20:2 (8,11) e SCI. Cada um desses ácidos graxos encontra-se presente em uma abundância relativa de menos de 2,2% do total de ácidos graxos.
Exemplo 16
Combinações de construções para a geração de ARA
Combinações de construções (fragmentos de Ascl que contêm conjuntos de genes) descritas na Fig. 12 são transformadas em cultivo de suspensão embriogênica de soja (cv. Jack), os embriões são amadurecidos e as plantas, geradas novamente conforme descrito para para produção no Exemplo 1. Os embriões e as sementes são analisados para determinar perfis de ácidos graxos conforme descrito no Exemplo 2. pKKE2 (SEQ ID N0 40; Fig. 14; Acesso ATCC n° PTA-4987), pKR1084 (SEQ ID N0 7) e pKR973 (SEQ ID N° 41, Fig. 15) são relacionados como Construção 1. pKR973 contém, além de EgD9elo e MaD5 descritos no presente, uma delta-8 dessaturase de Pavlova Iutheri (PavD8) (pedido copendente do Cessionário do Depositante que possui Pedido de Patente Norte-Americano n° 11/737772 (depositado em vinte de abril de 2007; Pasta do Advogado n° BB-1566, cujo teor é incorporado ao presente como referência).
Exemplo 17
Construção de vetores de expressão de soja alternativos para expressão de processos biossintéticos de ARA Além dos genes, construções ELVISLIVES, grampos de cabelo, promotores, terminais e conjuntos genéticos descritos no presente, os técnicos no presente podem apreciar que outras combinações de conjunto promotor/gene/terminal, promotor/grampo de cabelo e promotor/ELVISLIVES/terminal podem ser sintetizadas de forma similar, mas sem limitar-se à descrita no presente. Além disso, poderão ser geradas outras construções de grampos de cabelos que regulariam eficientemente para baixo fad3 quando expresso. De forma similar, pode ser desejável expressar outros genes de PUFA (tais como os descritos abaixo na Tabela 9) para coexpressão com qualquer dos genes, construções de ELVISLIVES, grampos de cabelo, promotores, terminais e conjuntos genéticos descritos no presente.
As Patentes PCT n0 WO 04/07167 e WO 04/071178 descrevem o isolamento de uma série de seqüências promotoras e términos de transcrição para uso em expressão específica em embrião em soja. Além disso, as Patentes PCT n0 WO 04/071467, WO 05/047479 e WO 06/012325 descrevem a síntese de diversas combinações de conjuntos de promotor/gene/terminal por meio da ligação de promotores individuais, genes e términos de transcrição entre si em combinações exclusivas. Geralmente, um local Not\ ladeado pelo promotor apropriado (tal como os relacionados, mas sem limitações, na Tabela 7) e um terminal de transcrição (tal como os relacionados, mas sem limitações, na Tabela 8) é utilizado para clonar o gene desejado ou construção de ELVISLIVES. Locais de Not\ podem ser adicionados a um gene de interesse, tais como os relacionados, mas sem limitações, na Tabela 9, utilizando amplificação por PCR com oligonucleotídeos projetados para introduzir locais NotI nas extremidades 5' e 3' do gene. O produto de PCR resultante é digerido em seguida com Notl e clonado em um conjunto promotor//Voíl/término apropriado. De forma similar, qualquer grampo de cabelo fad3 apropriado poderá ser clonado atrás de promotores tais como os relacionados, mas sem limitações, na Tabela 7. Geralmente, um local Notl na extremidade 5' do grampo de cabelo e um local Asp718 e Sbfl na extremidade 3' do grampo de cabelo é preferido embora outros locais de restrição possam ser utilizados. O grampo de cabelo é clonado em seguida nos locais Notl/Asp718 atrás de um promotor apropriado.
Além disso, as Patentes PCT n0 WO 04/071467, WO 05/047479 e WO 06/012325 descrevem, além disso, a ligação adicional de conjuntos genéticos individuais entre si em combinações exclusivas, junto com conjuntos de marcadores selecionáveis apropriados, a fim de obter a expressão fenotípica desejada. Embora isso seja realizado principalmente utilizando diferentes locais de enzimas de restrição, os técnicos no assunto podem desejada de promotor/gene/término de transcrição. Ao fazê-lo, qualquer combinação de promotor específico de embrião/gene/término de transcrição, promotor/ELVISLIVES/terminal de transcrição ou promotor/grampo de cabelo pode ser atingida. Os técnicos no assunto podem também apreciar que esses conjuntos podem ser localizados sobre fragmentos de DNA individuais ou sobre diversos fragmentos em que a coexpressão de genes, construções ELVISLIVES e/ou grampos de cabelo é o resultado da cotransformação de diversos fragmentos de DNA.
Tabela 7
Promotores Específicos de Sementes
<table>table see original document page 109</column></row><table> <table>table see original document page 110</column></row><table>
Tabela 9 Genes de Processo Biossintético de PUFA <table>table see original document page 110</column></row><table> <table>table see original document page 111</column></row><table>
Exemplo 18
Identificação de DHA sintase 1 (EgDHAsynD de biblioteca de cDNA de
Euglena gracius eeg1c
O Exemplo do presente descreve a clonagem de uma DHA sintase de Euglena gracilis. Este trabalho também é descrito no Pedido Provisório Norte-Americano n° 60/909790 (depositado em três de abril de 2007; Pasta do Advogado n° BB-1585), cujo conteúdo é incorporado ao presente como referência.
Clones da biblioteca de cDNA de Euglena (eeglc; vide Exemplo 3 e WO 2007/061845, publicado em 31 de maio de 2007 (cujo teor é incorporado ao presente como referência)) foram clocados em placas e o DNA foi isolado e sequenciado. Para o sequenciamento, os clones foram recuperados em primeiro lugar de cultivos de glicerol arquivados cultivados/congelados em placas de meios congelantes com 384 cavidades e inoculados com um tomador de colônia QPix automático (Genetix) em placas com 96 cavidades profundas que contêm LB + 50 pg/ml de canamicina. Após cultivo por vinte horas a 37 0C1 as células foram peletizadas por meio de centrifugação e armazenadas a -20 °C. Os plasmídeos foram isolados em seguida sobre um robô Eppendorf 5Prime, utilizando um método de minipreparação de Iise alcalina com formato de 96 cavidades modificado (Eppendorf PerfectPrep). Resumidamente, foi utilizado um filtro e condutor a vácuo para facilitar a remoção de fragmentos celulares após a precipitação de acetato. DNA de plasmídeo foi unido em seguida sobre uma segunda placa de filtro diretamente a partir do filtrado, lavado, seco e eluído.
Os plasmídeos foram sequenciados nas extremidades em placas com 384 cavidades, utilizando primer Universal M13F com primer de vetor (SEQ ID N0 78) e o kit de sequenciamento Prism ABI BigDye versão 3. Para a reação de sequenciamento, foram utilizados 100 a 200 ng de modelo e 6,4 pmol de primer e as condições de reação a seguir foram repetidas por 25 vezes: 96 °C por dez segundos, 50 °C por cinco segundos e 60 °C por quatro minutos. Após a limpeza com base em etanol, produtos de reação de sequenciamento de ciclos foram resolvidos e detectados sobre sequenciadores automáticos Perkin-EImerABI 3700.
Clones de cDNA que codificam homólogos de enzima de alongamento de ácidos graxos póli-insaturados C20 (C20-PUFA Elo) foram identificados por meio da condução de pesquisas de BLAST (Ferramenta de Busca e Alinhamento Local Básico; Altschul et al, J. Moi Biol. 215: 403-410 (1993)) em busca de similaridade para seqüências contidas no banco de dados "nr" de BLAST (que compreende todas as traduções de CDS GenBank não redundantes, seqüências derivadas do Banco de Dados de Proteínas Brookhaven com estrutura tridimensional, a última publicação importante do banco de dados de seqüências de proteínas SWISS-PROT, bancos de dados EMBL e DDBJ). As seqüências de cDNA de eeglc foram analisadas para determinar a similaridade com todas as seqüências de DNA disponíveis ao público contidas no banco de dados "nr" utilizando o algoritmo de BLASTN fornecido pelo Centro Nacional de Informações Biotecnológicas (NCBI). As seqüências de DNA foram traduzidas em todos os quadros de leitura e comparadas para determinar a similaridade para todas as seqüências de proteínas disponíveis ao público contidas no banco de dados "nr" utilizando o algoritmo BLASTX (Gish e States, Nat. Genet. 3: 266-272 (1993)) fornecido pelo NCBI. Para conveniência, o valor P (probabilidade) de observação de uma coincidência de uma seqüência de cDNA com uma seqüência contida nos bancos de dados pesquisados meramente ao acaso conforme calculado por meio de BLAST é relatado no presente como valor "pLog", que representa o negativo do Iogaritmo do valor P relatado. Consequentemente, quanto maior o valor pLog, maior a probabilidade de que a seqüência de cDNA e a encontrada por meio de BLAST representem proteínas homólogas.
A pesqgisa de BLASTX utilizando as seqüências de nucleotídeos do clone eeg1c.pk016.e6.f (também denominado pKR1049) revelou similaridade entre a proteína codificada pelo cDNA e o C20-PUFA Elo de Pavlova sp. CCMP459 (SEQ ID N0 79) (Acesso NCBI n° AAV33630 (Gl 54307108), local AAV33630, CDS AY630573; Pereira et al, Biochem. J. 384: 357-366 (2004)). A seqüência de uma parte do inserto de cDNA do clone eeg1c.pk016.e6.f é exibida em SEQ ID N0 80 (extremidade 5' do inserto de cDNA). Em seguida, a seqüência de inserto total de eeg1c.pk016.e6.f:fis foi obtida e é exibida em SEQ ID N0 81. A seqüência de codificação é definida em SEQ ID N0 82. A seqüência de aminoácidos deduzida correspondente é descrita em SEQ ID N0 83.
Conduziu-se sequenciamento de insertos completos utilizando um protocolo de transposição modificado. Clones identificados para FIS foram recuperados de estoques de glicerol arquivados na forma de colônias isoladas e DNA de plasmídeo foi isolado por meio de Iise alcalina. Modelos de plasmídeos foram transpostos por meio do kit de transposição do Sistema de Geração de Modelos (TGS II) (Finnzymes Oy1 Espoo, Finlândia), seguindo o protocolo do fabricante. O DNA transposto foi transformado em células eletrocompetentes EH10B (Edge BioSystems1 Gaithersburg MD) por meio de eletroporação. Diversos transformadores foram selecionados aleatoriamente a partir de cada reação de transposição, foi preparado o DNA de plasmídeo e modelos foram sequenciados conforme acima (ABI BigDye v3.1) no lado de fora do local de evento de transposição, utilizando os primers exclusivos SeqE (SEQ ID N° 171) e SeqW (SEQ ID N° 172).
A seqüência de aminoácidos descrita em SEQ ID N° 83 foi avaliada por meio de BLASTP. De forma interessante, concluiu-se que SEQ ID N° 83 é similar a ambos, C20-PUFA Elo e delta-4 dessaturase de ácido graxo. O terminal N de SEQ ID N° 83 (de cerca dos aminoácidos 16 a 268) gera um valor pLog de 60,30 (valor E de 5e-61; 124/258 aminoácidos idênticos; identidade de 48%) contra o CCMP459 C20-PUFA Elo de Pavlova sp. (SEQ ID N° 79). O terminal C de SEQ ID N° 83 (de cerca dos aminoácidos 253 a 793) gera um valor E de 0,0 (535/541 aminoácidos idênticos; 98% de identidade) contra a delta-4 dessaturase de ácido graxo de Euglena gracilis (SEQ ID N° 84) (Acesso NCBI n° AAQ19605 (Gl 33466346), local AAQ19605, CDS AY278558; Meyer et al, Biochemistry 42 (32): 9779-9788 (2003)). Avaliações e probabilidades de BLAST indicam que o fragmento de ácido nucleico do presente (SEQ ID N° 82) codifica uma multizima C20-PUFA Elo/delta-4 dessaturase de ácido graxo de Euglena gracilis, denominada no presente DHA sintase 1 de Euglena gracilis (EgDHAsyn 1).
A seqüência de aminoácidos de EgDHAsynI (SEQ ID N° 83) é 47,8% idêntica ao C20-PUFA Elo de CCMP459 de Pavlova sp. (SEQ ID N° 79) e 98,9% idêntica à delta-4 dessaturase de ácido graxo de Euglena gracilis (SEQ ID N° 84), utilizando o método de Jotun Hein. Cálculos de percentual de identidade de seqüências realizados por meio do método de Jotun Hein (Hein, J. J., Meth. Enz. 183: 626-645 (1990)) foram realizados utilizando o programa MegAlign® v6.1 da suíte de computação bioinformática LASARGENE (DNASTAR Inc., Madison Wl) com os parâmetros padrão para alinhamento em pares (COMPRIMENTO DE PALAVRA = 2).
A seqüência de aminoácidos de EgDHAsynI (SEQ ID N0 83) é 41,2% idêntica ao C20-PUFA Elo de CCMP459 de Pavlova sp. (SEQ ID N0 79) e 98,9% idêntica à delta-4 dessaturase de ácido graxo de Euglena gracilis (SEQ ID N0 84) utilizando o método Clustal V. Cálculos de percentual de identidade de seqüências realizados por meio do método Clustal V (Higgins, D. G. e Sharp, P. M., Comput. Appi Biosci. 5: 151-153 (1989); Higgins et al, Comput. AppL Biosci. 8: 189-191 (1992)) foram realizados utilizando o programa MegAlign® v6.1 da suíte de computação bioinformática LASARGENE (acima) com os parâmetros padrão para alinhamento em pares (COMPRIMENTO DE PALAVRA = 1, PENALIDADE DO INTERVALO = 3, VISUALIZAÇÃO DO MELHOR RESULTADO = 5, ESPAÇAMENTO EM DIAGONAIS = 5 e PENALIDADE DO COMPRIMENTO DO INTERVALO = 10).
O domínio C20 elongase de EgDHAsynI foi definido com base em comparação entre a seqüência de aminoácidos de EgDHAsynI (SEQ ID N°
83) e C20-PUFA Elo CCMP459 de Pavlova sp. (SEQ ID N° 79), PUFA elongase 2 de Ostreococcus tauri (SEQ ID N° 25 85) (Acesso NCBI n° AAV67798 (Gl 55852396), local AAV67798, CDS AY591336; Meyer et al, J. Lipid Res. 45 (10): 1899-1909 (2004)), PUFA elongase 2 de Thalassiosira pseudonana (SEQ ID N° 86) (Acesso NCBI n° AAV67800 (Gl 55852441), local AAV67800, CDS AY591338; Meyer et al, J. Lipid Res. 45 (10): 1899-1909 (2004)), delta-4 dessaturase de ácido graxo de Euglena gracilis (SEQ ID N°
84), delta-4 dessaturase de Thraustochytrium aureum (SEQ ID N° 87) (Acesso NCBI n° AAN75707 (Gl 25956288), local AAN75707, CDS AF391543), delta-4 dessaturase de Schizochytrium aggregatum (SEQ ID N° 88) (Patente PCT n° WO 2002/090493), delta-4 dessaturase de Thalassiosira pseudonana (SEQ ID N0 89) (Acesso NCBI n° AAX14506 (Gl 60173017), local AAX14506, CDS AY817156; Tonon et al, FEBS J. 272 (13): 3401-3412 (2005)) e delta-4 dessaturase de Isochrysis galbana (SEQ ID N0 90) (Acesso NCBI n° AAV33631 (Gl 54307110), local AAV33631, CDS AY630574; Pereira et al, Biochem. J. 384 (2): 357-366 (2004) e Patente PCT n0 WO 2002/090493). As seqüências de nucleotídeos e de aminoácidos correspondentes para o domínio C20 elongase de EgDHAsynI são descritas em SEQ ID N0 91 e SEQ ID N0 92, respectivamente.
No terminal C do domínio C20 elongase de EgDHAsynl, existe uma seqüência repetida que contém um motivo NG (ou seja, KNGK (SEQ ID N0 93), PENGA (SEQ ID N0 94), PENGA (SEQ ID N0 95), PCENGTV (SEQ ID N0 96)), denominada repetição NG. Embora o padrão ocorra com uma alta probabilidade de ocorrência,um varrimento da região NG repetida utilizando Prosite exibe o último motivo NG nessa região como um local de N-glicosilação potencial (NGTV). Após a região de repetição de NG, EgDHAsynl contém uma região rica em prolina (denominada ligante rico em prolina), que pode agir como um Iigante entre os domínios C20 elongase e delta-4 dessaturase. O ligante pode desempenhar um papel na manutenção dos domínios C20 elongase e delta-4 dessaturase na orientação estrutural adequada para permitir conversão eficiente de ácido eicosapentaenoico (EPA) em ácido docosa- hexaenoico (DHA). Embora o ligante rico em prolina seja definido como estendendo-se de P304 a V321 (com base na numeração para EgDHAsynl), a região de repetição de NG também é um pouco rica em prolina e pode também desempenhar um papel nessa função de ligante. As seqüências de nucleotídeos e de aminoácidos correspondentes para o ligante rico em prolina de EgDHAsynl são descritas em SEQ ID N0 97 e SEQ ID N0 98, respectivamente. EXEMPLO 19 IDENTIFICACAO DE UMA DELTA-9 ELONGASE DE UTEX 373 DE EUGLENA ANABAENA
O presente Exemplo descreve a síntese de uma biblioteca de cDNA de UTEX 373 de Euglena anabaena que inclui a geração de RNA1 síntese de cDNA, geração de uma biblioteca de cDNA e a identificação de uma delta-9 elongase. Este trabalho também é descrito no Pedido de Patente Provisório Norte-Americano n° 60/911925 (depositado em dezesseis de abril de 2007; Pasta do Advogado n° BB-1613), cujo conteúdo é incorporado ao presente como referência. Crescimento de UTEX 373 de Euglena anabaena ε preparação de RNA
UTEX 373 de Euglena anabaena foi obtido por meio do laboratório do Dr. Richard Triemer na Universidade do Estado de Michigan (East Lansing Ml). Cerca de 2 ml de cultivo foram removidos para análise de lipídios e centrifugados a 1800 x g por cinco minutos. A pelota foi lavada uma vez com água e novamente centrifugada. A pelota resultante foi seca por cinco minutos a vácuo, novamente suspensa em 100 μl de hidróxido de trimetilsulfônio (TMSH) e incubada à temperatura ambiente por quinze minutos mediante agitação. Após a incubação, foram adicionados 0,5 ml de hexano e as ampolas foram adicionalmente incubadas por quinze minutos à temperatura ambiente mediante agitação. Metil ésteres de ácidos graxos (5 μl injetados a partir da camada de hexano) foram separados e quantificados utilizando um Cromatógrafo de Gás Hewlett-Packard 6890 equipado com uma coluna capilar de sílica fundida Omegawax 320 (Supelco Inc., Cat. n° 24152). A temperatura do forno foi programada para manutenção a 170 °C por um minuto, aumento para 240 °C a 5 °C por minuto e manutenção em seguida por 1,0 minuto adicional. Gás veículo foi fornecido por um gerador de hidrogênio Whatman. Os tempos de retenção foram comparados com os de metil ésteres de padrões disponíveis comercialmente (Nu-Chek Prep, Inc., Cat. n° U-99-A) e o cromatograma resultante é exibido na Fig. 18. A presença de EDA, ERA, EPA e DHA no perfil de ácidos graxos, com a ausência de GLA e STA, sugeriu que Euglena anabaena utiliza o processo de delta-9 elongase e delta-8 dessaturase para biossíntese de LC-PUFA e seria uma boa fonte de genes biossintéticos de LC-PUFA tais como, mas sem limitações, delta-9 elongases.
Os 5 ml restantes de um cultivo em crescimento ativo foram transferidos para 25 ml de Meio AF-6 (Watanabe & Hiroki, NlES-CoIIection List ofStrains, quinta edição, National Institute for Environmental Studies1 Tsukuba, 127 págs. (2004)) em um frasco de vidro de 125 ml. Os cultivos de Euglena anabaena cresceram a 22 °C com um ciclo de dezesseis horas de luz e oito horas de escuro por duas semanas com agitação muito suave.
Após duas semanas, o cultivo (25 ml) foi transferido para 100 ml de meio AF-6 em uma garrafa de vidro de 500 ml e o cultivo cresceu por um mês conforme descrito acima. Após esse período, duas parcelas de 50 ml foram transferidas para duas garrafas de vidro de 500 ml separadas contendo 250 ml de meio AF-6 e os cultivos cresceram por dois meses conforme descrito acima (gerando um total de cerca de 600 ml de cultivo). Em seguida, os cultivos foram peletizados por meio de centrifugação a 1800 χ g por dez minutos, lavados uma vez com água e novamente centrifugados. O total de RNA foi extraído de uma das pelotas resultantes utilizando o reagente RNA STAT-60® (TEL-TEST Inc., Friendswood TX) e seguindo o protocolo fornecido pelo fabricante (utilizar 5 ml de reagente, RNA dissolvido em 0,5 ml de água). Desta forma, foram obtidos 340 pg de RNA total (680 pg/ml) da pelota. A pelota remanescente foi congelada em nitrogênio líquido e armazenada a -80 °C. O mRNA foi isolado de todos os 340 pg de RNA total utilizando o Kit de Purificação de mRNA (Amersham Biosciences, Piscataway NJ) seguindo o protocolo fornecido pelo fabricante. Desta forma, foram obtidos 9,0 pg de mRNA. Preparação de cDNA de Euglena anabaena ε geração de biblioteca de cDNA eug1c
Foi gerada uma biblioteca de cDNA utilizando o Kit de Construção de Bibliotecas de cDNA Cloneminer® (Cat. n° 18249-029, Invitrogen Corporation, Carlsbad CA) e seguindo o protocolo fornecido pelo fabricante (Versão B, 25-0608). Utilizando o método sem radiomarcação, cDNA foi sintetizado a partir de 5,12 pg de mRNA (descrito acima) utilizando o primer Biotin-aftB2-Oligo (dT). Após a síntese da primeira e do segunda fita, o adaptador attB1 foi adicionado, ligado e o cDNA teve seu tamanho fracionado utilizando cromatografia de coluna. DNA de frações foi concentrado, recombinado em pDONR® 222 e transformado em células resistentes a fagos T1 ElectroMAX® DH10B® de E. coli (Invitrogen Corporation). A biblioteca de Euglena anabaena foi denominada euglc.
A biblioteca de cDNA euglc foi colocada sobre placas LBKan (cerca de cem mil colônias), as colônias foram raspadas e DNA foi isolado utilizando Kit Spin Miniprep QIAprep® (Qiagen Inc., Valencia CA) seguindo o protocolo do fabricante. Desta forma, foi obtida uma sub-biblioteca de DNA de plasmídeos de euglc.
Levantamento de colônias
Cerca de 17.000 clones de biblioteca de cDNA euglc foram colocados em placas sobre três discos petri quadrados grandes (24 cm χ 24 cm) (Corning, Corning NY), cada qual contendo LB + 50 pg/ml de meios Agar de canamicina. As células foram cultivadas por uma noite a 37 0C e as placas foram resfriadas em seguida à temperatura ambiente.
Membrana de 0,45 pm Biodyne B (Cat. n° 60207, Pall Corporation, Pensacola FL) foi cortada até cerca de 22 cm χ 22 cm e a membrana foi cuidadosamente depositada sobre o Agar para evitar bolhas de ar. Após incubação por dois minutos à temperatura ambiente, a membrana foi marcada para orientação, erguida com uma pinça e colocada com o lado da colônia para cima sobre papel filtro embebido com 0,5 M hidróxido de sódio e 1,5 M cloreto de sódio. Após desnaturação por quatro minutos, o hidróxido de sódio foi neutalizado colocando-se a membrana sobre papel filtro embebido com 0,5 M Tris-HCI (pH 7,5) e 1,5 M cloreto de sódio por quatro minutos. Esta etapa foi repetida e a membrana foi rapidamente enxaguada em 2X tampão SSC (20X SSC em 3 M cloreto de sódio, 0,3 M citrato de sódio; pH 7,0) e seca em ar sobre papel filtro.
Hibridização
As membranas foram previamente hibridizadas a 65 °C em 200 ml de solução de hibridização por duas horas. A solução de hibridização continha 6X SSPE (20X SSPE é 3 M cloreto de sódio, 0,2 M fosfato de sódio, 20 mM de EDTA; pH 7,4); 5X reagente de Denhardt (100X reagente de Denhardt é 2% (p/v) Ficoll1 2% (p/v) polivinilpirrolidona, 2% (p/v) albumina de soro bovino acetilado), 0,5% dodecil sulfato de sódio (SDS), 100 pg/ml de DNA de esperma de salmão cortado e 5% sulfato de dextran.
Foi elaborada uma sonda de DNA utilizando um fragmento de DNA de Ncol/Notl purificado com gel de agarose que contém o gene delta-9 elongase de Euglena gracilis de pKR906 (SEQ ID N0 46; Exemplo 4) marcado com P32 dCTP utilizando o Sistema de Marcação de DNA RadPrime (Cat. n° 18428-011, Invitrogen, Carlsbad CA) seguindo as instruções do fabricante. P32 dCTP não incorporado foi separado utilizando uma coluna NICK (Cat. n° 17- 0855-02, Amersham Biosciences, Piscataway NJ) seguindo as instruções do fabricante. A sonda foi desnaturada por cinco minutos a 100 °C, colocada sobre gelo por três minutos e a metade foi adicionada à solução de hibridização.
A membrana foi hibridizada com a sonda por uma noite a 65 °C com suave agitação e lavada no dia seguinte por duas vezes com 2X SSC contendo 0,5% SDS (cinco minutos cada) e duas vezes com 0,2X SSC contendo 0,1% SDS (quinze minutos cada). Após a lavagem, hiperfilme (Cat. n° RPN30K, Amersham Biosciences, Piscataway NJ) foi exposto à membrana por uma noite a -80 °C.
Com base no alinhamento de placas com o hiperfilme exposto, colônias positivas foram tomadas utilizando a extremidade obtusa de uma pipeta de Pasteur em 1 ml de água e turbilhonadas. Diversas diluições foram realizadas e colocadas em placas sobre placas de Petri redondas pequenas (82 mm) contendo meios LB mais 50 pg/ml de canamicina para obter cerca de cem colônias isoladas em cavidades sobre uma única placa. Foram realizados levantamentos conforme descrito acima, exceto pelo uso de círculos de membrana NytranN (Cat. n° 10416116, Schleicher & Schuell, Keene NH) e conduziu-se hibridização em 100 ml utilizando a sonda rádio marcada restante. Desta forma, foram confirmados clones positivos.
Clones positivos individuais foram cultivados a 37 °C em LB + 50 pg/ml de meios líquidos de canamicina e plasmídeo foi purificado utilizando o Kit Spin Miniprep QIAprep® (Qiagen Inc.) seguindo o protocolo do fabricante.
Insertos de DNA foram sequenciados em terminais em placas com 384 cavidades conforme descrito no Exemplo 18 e as seqüências foram alinhadas e comparadas utilizando Sequencher® (Versão 4.2, Gene Codes Corporation, Ann Arbor Ml). Desta forma, os clones poderão ser categorizados em um dentre dois grupos distintos com base na seqüência de inserto (denominada EaD9Elo1 e EaD9Elo2). Os clones representativos que contêm o cDNA para cada classe de seqüências foram selecionados para estudo adicional e as seqüências para cada plasmídeo representativo (pLF121-1 e pLF121-2) são exibidas em SEQ ID N0 99 e SEQ ID N0 100, respectivamente. A seqüência exibida por um conjunto de NNNN's representa uma região da extremidade póli A que não foi sequenciada. As seqüências de codificação para EaD9Elo1 e EaD9Elo2 são exibidas em SEQ ID N0 101 e SEQ ID N0 102, respectivamente. As seqüências de aminoácidos correspondentes para EaD9Elo1 e EaD9Elo2 são exibidas em SEQ ID N0 103 e SEQ ID N0 104, respectivamente.
Exemplo 20
Identificação de uma delta-5 dessaturase de Euglena anabaena UTEX 373
O presente Exemplo descreve a identificação de uma delta-5 dessaturase de UTEX 373 de Euglena anabaena. Este trabalho também é descrito no Pedido Provisório Norte-Americano n° 60/915733 (depositado em três de maio de 2007; Pasta do Advogado n° BB-1614), cujo conteúdo é incorporado ao presente como referência.
Biblioteca de cDNA amplificada euglc foi colocada em placas e as colônias levantadas conforme descrito no Exemplo 19. Foi realizada uma sonda de DNA utilizando um fragmento de DNA Ncol/Notl purificado com gel de agarose que contém o gene delta-5 dessaturase de Euglena gracilis (EgD5; SEQ ID N° 54) de pDMW367, descrito anteriormente na Patente PCT n0 WO 2007/136877, publicada em 29 de novembro de 2007 (cujo conteúdo é incorporado ao presente como referência), marcado com P32. Os levantamentos de colônias foram sondados, os positivos foram identificados, confirmados e o DNA foi isolado e sequenciado exatamente conforme descrito no Exemplo 19.
Um clone representativo que contém um cDNA (pLF119) é exibido em SEQ ID N° 105 e o gene contido no cDNA foi denominado EaD5Des1. A seqüência de codificação para EaD5Des1 é exibida em SEQ ID N° 106. A seqüência de aminoácidos correspondente para EaD5Des1 é exibida em SEQ ID N° 107.
Exemplo 21
Construção de vetor de expressão de soja pKR1 183 para expressão de um gene de fusão de delta-9 elongase de Euglena anabaena ε delta-8 dessaturase CCMP1491 de Tetruetreptia pomquetensis (Híbrido 1 - HGLA Sintase)
Uma fusão em quadro entre a delta-9 elongase de Euglena anabaena (EaD9Elo1; SEQ ID N0 101), o Iigante rico em prolina DHA sintase 1 de Euglena gracilis (EgDHAsynI Ligação; SEQ ID N0 97) e a delta-8 dessaturase CCMP1491 de Tetmetreptia pomquetensis (TpomD8; SEQ ID N0 50; Exemplo 5; vide também o pedido copendente do Cessionário do Depositante que possui Pedido de Patente Norte-Americano n° 11/876115 (depositado em 22 de outubro de 2007; Pasta do Advogado n° BB-1574)) foi construída utilizando as condições descritas abaixo.
Uma fusão em quadro inicial entre EaD9Elo1 e EgDHAsynI Ligação (EaD9Elo-EgDHAsyn1 Ligação) foi realizada, ladeada por um local Nco\ na extremidade 5' e um local Not\ na extremidade 3', por meio de amplificação por PCR. EaD9Elo1 (SEQ ID N0 101) foi amplificado a partir de pLF121-1 (SEQ ID N0 99) com os oligonucleotídeos EaD9-5Bbs (SEQ ID N0 108) e EaD9-3fusão (SEQ ID N0 109), utilizando DNA Polimerase de Alta Fidelidade Phusion® (Cat. n° F553S, Finnzymes Oy1 Finlândia), seguindo o protocolo do fabricante. EgDHAsynI Ligação (SEQ ID N0 97) foi amplificado de forma similar a partir de pKR1049 (Exemplo 18) com os oligonucleotídeos EgDHAsynILigação-5fusão (SEQ ID N0 110) e MWG511 (SEQ ID N0 111). Os dois produtos de PCR resultantes foram combinados e novamente amplificados utilizando EaD9-5Bbs (SEQ ID N0 108) e MWG511 (SEQ ID N0 111) para formar EaD9Elo1 -EgDHAsynI Ligação. A seqüência de EaD9Elo1- EgDHAsynI Ligação é exibida em SEQ ID N0 112. EaD9Elo1-EgDHAsynI Ligação não contém um códon de parada em quadro acima no fluxo do local NotI na extremidade 3' e, portanto, um fragmento de DNA clonado no local Noti pode gerar uma fusão em quadro com EgD9elo-EgDHAsyn1 Ligação caso seja selecionado o quadro correto. EaD9Elo1-EgDHAsynILigação foi clonado no vetor de clonagem pCR-Blunt® utilizando o Kit de Clonagem de PCR Zero Blunt® (Invitrogen Corporation), seguindo o protocolo do fabricante, para produzir pLF124 (SEQ ID N0 113).
O plasmídeo KS366 (SEQ ID N0 114) contém locais de restrição de Nco\ e NotI exclusivos, ladeados pelo promotor para a subunidade a' de β- conglicinina (Beachy et al, EMBO J. 4: 3047-3053 (1985)) e a região de término de transcrição 3' do gene de faseolina (Doyle et al, J. Biol. Chem. 261: 9228-9238 (1986)). Diferentemente da substituição do local Noti exclusivo em pKR72 (SEQ ID N0 67) com um local de clonagem múltipla de NcoUNott exclusivo, o conjunto Bcon/Ncol/Notl/Phas3' em KS366 é idêntico ao encontrado em pKR72 (SEQ ID N0 67), exceto pelo fato de que os locais Hind\\\ Iadeadores foram substituídos por locais BamH\. O conjunto Bcon/Ncol/Notl/Phas3' de KS366 é clonado no local BamH\ de vetor pBluescript Il SK(+) (Stratagene).
O fragmento de DNA de BbsUNoti de pLF124 (SEQ ID N0 113), que contém EaD9Elo1-EgDHAsynI Ligação, foi clonado no fragmento de DNA de NcoUNoti de KS366 (SEQ ID N0 114) que contém o promotor para a subunidade a' de β-conglicinina, para produzir pKR1177 (SEQ ID N0 115).
O fragmento de DNA BamHI de pKR1177 (SEQ ID N0 115), que contém EaD9Elo1-EgDHAsynILigação, foi clonado no fragmento de DNA SamHI de pKR325, descrito anteriormente na Patente PCT n0 WO 2006/012325 (cujo conteúdo é incorporado ao presente como referência) para produzir pKR1179 (SEQ ID N0 116).
O fragmento Nott de pLF114-10 (Exemplo 5; SEQ ID N0 64) que contém TpomD8 foi clonado no fragmento Noti de pKR1179 (SEQ ID N0 116) para produzir pKR1183 (SEQ ID N0 117; Fig. 19). NaFig. 19, o gene de fusão (Híbrido 1 - HGLA sintase) é denominado EAd9ELONG-TPOMd8DS.
Exemplo 22
Construção de vetor de expressão de soja PKR1253 para expressão de um GENE DE FUSÃO DE DELTA-9 ELONGASE DE euglena anabaena E DELTA-8 DESSATURASE DE tetruetreptia pomquetensis CCMP1491 (HÍBRIDO 1 - HGLA SINTASE) COM UMA DELTA-5 DESSATURASE DE euglena gracilis
Por meio de uma série de etapas de subclonagem, um local Notl foi adicionado à extremidade 5' da delta-5 dessaturase de Euglena gracilis (EgD5; SEQ ID N0 54) de pDMW367 (Exemplo 20) e este fragmento de Notl que contém EgD5 foi clonado no local Notl de pKR457 (SEQ ID N0 2) para produzir pKR1237 (SEQ ID N0 118).
O fragmento Ascl de pKR1183 (SEQ ID N0 117; Exemplo 21), que contém o Híbrido 1 - HGLA sintase, foi clonado no fragmento Ascl de pKR277 (SEQ ID N0 4, que foi descrito anteriormente na Patente PCT n0 WO 2004/071467 (cujo conteúdo é incorporado ao presente como referência)) para produzir pKR1252 (SEQ ID N0 119).
O fragmento BsiWI de pKR1237 (SEQ ID N0 118), que contém o gene EgD5, foi clonado no local BsiWI de pKR1252 (SEQ ID N0 119) para produzir pKR1253 (SEQ ID N0 120; Fig. 20).
Exemplo 23
Construção de vetor de soja pKR1139 para expressão de uma delta-5 dessaturase de EUGLENA ANABAENA
O presente Exemplo descreve a clonagem de uma delta-5 dessaturase de UTEX 373 de Euglena anabaena em um vetor de expressão de soja. Este trabalho também é descrito no Pedido Provisório Norte-Americano n° 60/915733 depositado em três de maio de 2007; Pasta do Advogado n0 BB- 1614), cujo conteúdo é incorporado ao presente como referência.
EaD5Des1 (SEQ ID N0 101) foi amplificado a partir de pLF119 (SEQ ID N0 105, Exemplo 20) com oEAd5-1-1 (SEQ ID N0 121) e oEAd5-1-2 (SEQ ID N0 122), utilizando DNA Polimerase de Alta Fidelidade Phusion® (Cat. n° F553S, Finnzymes Oy1 Finlândia) seguindo o protocolo do fabricante. O produto de PCR resultante foi clonado no vetor de clonagem pCR-Blunt® utilizando o Kit de Clonagem PCR Zero Blunt® (Invitrogen Corporation), seguindo o protocolo do fabricante, para produzir pKR1136 (SEQ ID N0 123).
O fragmento de Notl para pKR1136 (SEQ ID N0 123) que contém o EaD5Des1 foi clonado no fragmento Notl de pKR974, descrito anteriormente na Patente PCT n° WO 2007/136877, publicada em 29 de novembro de 2007 (cujo conteúdo é incorporado ao presente como referência), para produzir pKR1139 (SEQ ID N° 124).
Exemplo 24
Construção do vetor de expressão de soja PKR1255 para expressão de um GENE DE FUSÃO DE DELTA-9 ELONGASE DE EUGLENA ANABAENA E DELTA-8 DESSATURASE CCMP1491 DE TETRUETREPTIA POMQUETENSIS (HÍBRIDO 1 - HGLA SlNTASE) COM UMA DELTA-5 DESSATURASE DE EUGLENA GRACIUS E UMA DELTA-5
DESSATURASE DE EUGLENA ANABAENA
O plasmídeo pKR1139 (SEQ ID N° 124; Exemplo 23) foi digerido com Sbfl e o fragmento que contém o EaD5Des1 foi clonado no local Sbfl de pKR1253 (SEQ ID N° 120; Exemplo 22) para produzir pKR1255 (SEQ ID N° 76; Fig- 21).
Exemplo 25
CONSTRUCAO DO VETOR DE EXPRESSAO DE SOJA PKR11589 PARQ REGULAGEM PARA BAIXO DA EXPRESSAO DE FAD3 DE SOJA
De forma similar ao descrito no Exemplo 11, foi projetado um vetor de expressão de soja para reduzir a expressão de fad3 em soja.
Um vetor inicial pKR561 (SEQ ID N° 125) foi montado por meio da inserção do fragmento BsiWI de pKR268 (SEQ ID N0 15; Exemplo 7) que contém o promotor de anexina no local BsiWI de pKR145, que é descrito na Patente PCT n° WO 04/071467.
Uma parte da extremidade 5' do gene fad3 foi amplificada a partir de XF1 (descrito no Exemplo 8) com DNA Polimerase de Alta Fidelidade Phusion® (Cat. n° F553S, Finnzymes Oy1 Finlândia) seguindo o protocolo do fabricante, utilizando HPfad3-1 (SEQ ID N° 30) e HPfad3-2 (SEQ ID N° 31) para produzir um fragmento de DNA denominado HPfad3AB (SEQ ID N0 32).
Uma parte da extremidade 3' do gene fad3 foi amplificada a partir de XF1 (descrito no Exemplo 8) com a DNA Polimerase de Alta Fidelidade Phusion®, utilizando HPfad3-3 (SEQ ID N0 33) e HPfad3-1 (SEQ ID N0 30) para produzir um fragmento de DNA denominado HPfad3A'-2 (SEQ ID N0 126).
HPfad3AB e HPfad3A'-2 foram combinados e amplificados utilizando DNA Polimerase de Alta Fidelidade Phusion® com HPfad3-1 (SEQ ID N0 30) para produzir HPfad3ABA'-2 (SEQ ID N0 127). HPfad3ABA'-2 (SEQ ID N0 127) é quase idêntico a HPfad3ABA' (SEQ ID N0 36), mas possui um local Notl na extremidade 5' e 3' do fragmento de DNA. O produto de PCR resultante foi clonado no vetor de clonagem pCR-Blunt® utilizando o Kit de Clonagem de PCR Zero Blunt® (Invitrogen Corporation), seguindo o protocolo do fabricante, para produzir pLF129 (SEQ ID N0 128).
O fragmento de Notl para pLF129 (SEQ ID N0 128) que contém o grampo de cabelo fad3 foi clonado no fragmento Notl de pK561 (SEQ ID N0 125) para produzir pKR1189 (SEQ ID N0 129; Fig. 22). Na Fig. 22, os domínios A e A' para fad3 são indicados pela designação TR1, enquanto o domínio B é
indicado por TR2.
Exemplo 26
Construção do vetor de expressão de soja PKR1249 para regulagem para baixo de FAD3 de soja e FAD3C de soja
O fragmento Notl/Hindlll de pLF129 (SEQ ID N0 128) que contém
os domínios TR1 e TR2 de fad3, conforme indicado na Fig. 22, foi clonado no fragmento de cadeia principal de Notl/Hindlll de pLF129 (SEQ ID N0 128) para produzir pKR1209 (SEQ ID N0 130).
A seqüência de codificação de GmFad3C (Acesso GenBank n° AY204712) (Bilyeu et al, Crop Sei. 43: 1833-1838 (2003); Anai et al, Plant Sei. 168: 1615-1623 (2005)) é exibida em SEQ ID N0 131 e a seqüência de aminoácidos correspondente é exibida em SEQ ID N0 132. Uma parte do gene fad3c foi amplificada a partir da biblioteca de cDNA de soja descrita na Patente PCT n° WO 93/11245 (que foi publicada em dez de junho de 1993; e também Patente Norte-Americana n° 5.952.544) (cujos conteúdos são incorporados ao presente como referência) com a DNA polimerase de Alta Fidelidade Phusion® (Cat. n° F553S, Finnzymes Oy1 Finlândia) seguindo o protocolo do fabricante, utilizando fad3c-5 (SEQ ID N° 133) e fad3c-3 (SEQ ID N° 134). O fragmento de DNA resultante foi clonado no vetor de clonagem PCR-Blunt® utilizando o Kit de Clonagem de PCR Zero Blunt® (Invitrogen Corporation), seguindo o protocolo do fabricante, para produzir pKR1213 (SEQ ID N° 135).
O fragmento EcoRV/Xhol de pKR1213 (SEQ ID N° 135) que contém o fragmento de fad3c foi clonado no local Notl (preenchido)/Xhol de pKR1209 (SEQ ID N° 130) para produzir pKR1218 (SEQ ID N° 136).
O fragmento Notl/Hindlll de pLF129 (SEQ ID N° 128) que contém o domínio TR1 somente a partir de fad3, conforme indicado na Fig. 22, foi clonado no fragmento de cadeia principal de Notl/Hindlll de pLE129 (SEQ ID N° 128) para produzir pKR1210 (SEQ ID N° 137).
O fragmento EcoRV/Xhol de pKR1213 (SEQ ID N° 135) que contém o fragmento de fad3c foi clonado no local Notl (preenchido)/Xhol de pKR1210 (SEQ ID N° 137) para produzir pKR1219 (SEQ ID N° 138).
O fragmento de Xhol (preenchido)/Hindlll de pKR1218 (SEQ ID N° 136) que contém o fragmento de fad3c bem como os domínios TR1 e TR2 de fad3 foi clonado no local Mlul (preenchido)/Hindlll de pKR1219 (SEQ ID N° 138) que contém o fragmento de fad3c bem como o domínio somente TR1 de fad3, para produzir pKR1225 (SEQ ID N° 139). Desta maneira, formou-se um novo grampo de cabelo que inclui fad3 e fad3c ladeado por locais Notl.
O fragmento Notl para pKR1225 (SEQ ID N° 139) que contém o novo grampo de cabelo que inclui fad3 e fad3c foi clonado no fragmento Notl de pK561 (SEQ ID N° 125; Exemplo 24) para produzir pKR1229 (SEQ ID N° 140; Fig. 23). Desta forma, o grampo de cabelo fad3/fad3c pode ser expresso a partir de um promotor forte específico de semente com seleção de higromicina em plantas.
O fragmento BsiWI para pKR1225 (SEQ ID N° 139) que contém o novo grampo de cabelo e inclui fad3 e fad3c foi clonado no fragmento BsiWI de pKR226 (SEQ ID N° 9; Exemplo 6) para produzir pKR1249 (SEQ ID N° 141; Fig. 24). Na Fig. 24, pKR1249 é marcado pKR1249_PHP33240. Desta forma, o grampo de cabelo fad3/fad3c pode ser expresso a partir de um promotor forte específico de semente com seleção de clorsulfurona (ALS) em plantas.
Exemplo 27
Identificação de delta-8 dessaturase de Euglena anabaena UTEX 373
O presente Exemplo descreve a identificação e clonagem de uma delta-8 dessaturase de UTEX 373 de Euglena anabaena. Este trabalho também é descrito no Pedido Provisório Norte-Americano n° 60/910831 (depositado em dez de abril de 2007; Pasta do Advogado n° BB-1615), cujo conteúdo é incorporado ao presente como referência.
IDENTIFICAÇÃO DE FRAGMENTOS DE CDNA QUE CODiFiCAM SUPOSTA DELTA-S dessaturase parcial
A sub-biblioteca de DNA de plasmídeo de euglc descrita no Exemplo 19 foi utilizada como modelo de degeneração de PCR utilizando primers degenerados com base na seqüência de nucleotídeos da delta-8 dessaturase de ácido graxo de Euglena gracilis (SEQ ID N° 142) e do primer específico de vetor pDonor222Eg5-1 (SEQ ID N° 143). Os quatro primers degenerados utilizados são exibidos na Tabela 10.
Tabela 10
oligonucleotídeos degenerados utilizados para amplificar uma parte dos Genes Delta-8 Dessaturase de UTEX 373 de Euglena anabaena
Primer Seqüência de nucleotídeos SEQ ID N° D8DEG3-1 rttrtgnckatctttccacca seq id n° 144 <table>table see original document page 130</column></row><table>
Um total de cinco reações foi configurada para a amostra de cDNA. A mistura de reação continha 1 μl de cDNA, 1 μl de cada um dentre o primer degenerado e específico de vetor (20 μΜ) e o PCR foi conduzido utilizando a DNA Polimerase de Alta Fidelidade Phusion® (Cat. n° F553S, Finnzymes Oy1 Finlândia) seguindo o protocolo do fabricante. Os fragmentos de DNA resultantes foram clonados no vetor de clonagem PCR-Blunt® utilizando o Kit de Clonagem de PCR Zero Blunt® (Invitrogen Corporation), seguindo o protocolo do fabricante.
DNA de plasmídeo dos clones resultantes foi purificado utilizando o Kit Spin Miniprep QIAprep® (Qiagen Inc.) seguindo o protocolo do fabricante, insertos de DNA foram sequenciados nas extremidades e as seqüências foram analisadas conforme descrito nos Exemplos 18 e 19.
Foi montada uma seqüência de consenso a partir das seqüências individuais obtidas e um clone representativo, denominado pHD23-1 (SEQ ID N° 148) que possui uma seqüência idêntica ao consenso foi selecionado para estudos adicionais.
A pesquisa BLASTX utilizando o inserto de seqüência de nucleotídeos de pHD23-1 revelou similaridade entre a proteína codificada pelo cDNA parcial e a seqüência de aminoácidos delta-8 dessaturase de Euglena gracilis (SEQ ID N0 149) (Acesso NCBI n° AAD45877 (Gl 5639724), local AAD45877, CDS AF 139720; Wallis e Browse, Arch. Biochem. Biophys. 365: 307-316 (1999)) e gerou um valor pLog de 63,4 (valor E de 4e-63). Isolamento de delta-8 dessaturases de comprimento total de UTEX 373 de Euglena anabaena
A biblioteca de cDNA amplificada euglc foi colocada em placas e as colônias foram levantadas conforme descrito no Exemplo 19.
Foi elaborada uma sonda de DNA utilizando um fragmento de DNA EcoRI purificado com gel de agarose que contém o fragmento de DNA parcial de delta-8 dessaturase de Euglena anabaena de pHD23-1 marcado com P32 dCTP. Os levantamentos de colônias foram sondados, os positivos foram identificados e confirmados e DNA foi isolado e sequenciado exatamente conforme descrito no Exemplo 19.
As seqüências foram alinhadas e comparadas utilizando Sequencher® (Versão 4.2, Gene Codes Corporation, Ann Arbor Ml) e, desta forma, os clones puderam ser classificados em um dentre quatro grupos distintos com base na seqüência de inserto (denominada EaD8Des1 a EaD8Des4). A seqüência para um clone representativo que contém EaD8Des3 (pLF118-3) é exibida em SEQ ID N0 150. A seqüência exibida por um conjunto de NNNN's representa uma região da extremidade póli A que não foi sequenciada. As seqüências de codificação e de aminoácidos para EaD8Des3 são exibidas em SEQ ID N0 151 e SEQ ID N0 152, respectivamente.
A fim de introduzir locais de restrição Notl e Ncol na extremidade 5' das seqüências de codificação e um local Notl na extremidade 3' das seqüências de codificação, EaD8Des3 (SEQ ID N0 151) foi amplificado a partir de pLF118-3 (SEQ ID N0 15) com os primers de oligonucleotídeos EaD8-5 (SEQ ID N0 153) e EaD8-3 (SEQ ID N0 154) utilizando a DNA Polimerase de Alta Fidelidade Phusion® (Cat. n° F553S, Finnzymes Oy, Finlândia) seguindo o protocolo do fabricante. O fragmento de DNA resultante foi clonado no vetor de clonagem PCR-Blunt® utilizando o Kit de Clonagem de PCR Zero Blunt® (Invitrogen Corporation), seguindo o protocolo do fabricante, para produzir pLF120-3 (SEQ ID N° 155).
EXEMPLO 28
CONSTRUCAO DE VETOR EXPRESSAO DE SOJA PKR1140 PARA EXPRESSAO DE DELTA-9 ELONGASE DE EUGLENA ANABAENA UTEX 373 (EADA9ELO1)
O presente Exemplo descreve a construção de um vetor de soja para a expressão de EaD9Elo1. Este trabalho também é descrito no Pedido Provisório Norte-Americano n° 60/911925 (depositado em dezesseis de abril de 2007; Pasta do Advogado n° BB-1613, cujo conteúdo é incorporado ao presente como referência).
A fim de introduzir locais de restrição de NotI e Nco\ na extremidade 5' das seqüências de codificação e um local Noti na extremidade 3' das seqüências de codificação, EaD9Elo1 foi amplificado por PCR. A seqüência de codificação para EaD9Elo1 (SEQ ID N° 101) foi amplificada a partir de pLF121-1 (SEQ ID N° 99) com os primers de oligonucleotídeos oEAd9el1-1 (SEQ ID N° 156) e oEAd9el1-2 (SEQ ID N° 157) utilizando DNA Polimerase de Alta Fidelidade Phusion® (Cat. n° F553S, Finnzymes Oy1 Finlândia) seguindo o protocolo do fabricante. Os fragmentos de DNA resultantes foram clonados no vetor de clonagem pCR-Blunt® utilizando o Kit de Clonagem de PCR Zero Blunt® (Invitrogen Corporation), seguindo o protocolo do fabricante, para produzir pKR1137 (SEQ ID N° 158).
EaD9Elo1 foi liberado de pKR1137 (SEQ ID N° 158) por meio de digestão com NotI e clonado no local Noti de pKR72 (SEQ ID N° 67) para produzir pKR1140 (SEQ ID N° 159).
EXEMPLO 29
CONSTRUCAO DE UM VETOR DE EXPRESSA DE ARABIDOPSIS PKR1193 PARA EXPRESSA DE UMA DELTA-9 ELONGASE DE EUGLENA ANABAENA COM UMA DELTA-8 DESSATURASE DE EUGLENA ANABAENA E UMA DELTA-5 DESSATURASE DE EUGLENA ANABAENA
O fragmento de Ascl de pKR1140 (SEQ ID N° 159) foi clonado no fragmento Ascl de pKR277 (SEQ ID N0 4) para produzir pKR1173 (SEQ ID N0 160).
O conjunto Gy1/Pavelo/legA2 foi liberado do plasmídeo pKR336 (descrito na Patente PCT n0 WO 04/071467; cujo conteúdo é incorporado ao presente como referência) por meio de digestão com Pstl/BamHI e clonado no local Pstl/BamHI de pKR268 (descrito na Patente PCT n0 WO 04/071467) para produzir pKR393 (SEQ ID N0 161). O gene Pavelo foi liberado de pKR393 (SEQ ID N0 161) por meio de digestão com Notl e o vetor foi novamente ligado para formar pKR407 (SEQ ID N0 162).
O fragmento Not\ de pLF120-3 (SEQ ID N0 155), que contém EaD8Des3, foi clonado no fragmento Nott de pKR407 (SEQ ID N0 162) para produzir pKR1176 (SEQ ID N0 163).
O fragmento Pstl de pKR1176 (SEQ ID N0 163), que contém EaD8Des3, foi clonado no fragmento Sbfl de pKR1173 (SEQ ID N0 160) para produzir pKR1178 (SEQ ID N0 164).
O conjunto pcon/A/ofI/Phas foi amplificado por meio de PCR a partir de pKS123, que é descrito nas Patentes PCT n0 WO 2004/071467 e WO 02/008269 (cujos conteúdos são incorporados ao presente como referência) utilizando os primers oKti5 (SEQ ID N0 165) e oKti6 (SEQ ID N0 166). O fragmento de PCR resultante foi digerido com BsiWl e clonado no local fís/WI de pKR124 (que é descrito na Patente PCT n0 WO 2004/071467, cujo conteúdo é incorporado ao presente como referência), que contém a origem bacteriana de reprodução e seleção, para produzir o plasmídeo pKR193 (SEQ ID N0 167).
O fragmento Nott de pKR1136 (SEQ ID N0 122; Exemplo 23) que contém EaD5Des1 foi clonado no fragmento Nott de pKR193 (SEQ ID N0 167) para produzir pKR1174 (SEQ ID N0 168).
O fragmento de BsiWI para pKR1174 (SEQ ID N0 168) que contém EaD5Des1 foi clonado no fragmento BsiWI de pKR1178 (SEQ ID N0 164) para produzir pKR1186 (SEQ ID N0 169).
O fragmento Ascl de pKR1186 (SEQ ID N° 169) que contém EaD9elo1 etc. foi clonado no local /\scl de pKR92 (que foi descrito anteriormente em WO 2007/061845 publicada em 31 de maio de 2007 para produzir pKR1193 (SEQ ID N° 170). Uma ilustração esquemática de pKR1193 é exibida na Fig. 25. Desta forma, EaD9Elo1 e EaD5Des1 foram expressos em Arabidopsis sob o controle do promotor de betaconglicinina de soja e EaD8Des3 foi expresso sob o controle do promotor Gy1 de glicinina de soja. O promotor de betaconglicinina de soja e o promotor Gy1 funcionam como promotores fortes específicos de sementes em Arabidopsis.
Exemplo 30
Regulagem para baixo dos genes fad3 ε fad3c de soja em embriões somáticos de soja por meio de transformação com PKR1189 ou PKR1229
O presente Exemplo descreve a transformação e a expressão em embriões somáticos de soja de pKR1189 (SEQ ID N0 129, Exemplo 25), que contém uma construção de grampo de cabelo íad3 ou pKR1229 (SEQ ID N0 140, Exemplo 26), que contém uma construção de grampo de cabelo fad3c e fad3. As duas construções também contêm o gene fosfotransferase de higromicina para seleção sobre higromicina.
Cultivo de suspensão embriogênica de soja (cv. Jack) foi transformado com pKR1189 (SEQ ID N0 129) ou pKR1229 (SEQ ID N0 140) e os embriões foram amadurecidos em meio líquido de maturação e histodiferenciação de soja (meios líquidos SHaM; Schmidt et al, Cell Biology and Morphogenesis, 24: 393 (2005)), conforme descrito anteriormente na Patente PCT n0 WO 2007/136877, publicada em 29 de novembro de 2007 (cujo conteúdo é incorporado ao presente como referência).
Após o amadurecimento em meios líquidos SHaM1 os embriões individuais foram removidos dos conjuntos, secos e selecionados para determinar alterações nas suas composições de ácido graxo conforme descrito no Exemplo 2. Em cada caso, um subconjunto de embriões de soja (ou seja, cinco embriões por evento) transformados com pKR1189 (SEQ ID N0 129) ou pKR1229 (SEQ ID N0 140) foi colhido e analisado.
Desta forma, foram analisados 41 eventos transformados com pKR1189 (SEQ ID N0 129; Experimento 2148) ou pKR1229 (SEQ ID N0 140; Experimento 2165). Os perfis de ácido graxo para os cinco eventos que possuem o teor médio mais baixo de ALA (média dos cinco embriões analisados) junto com um evento (2148-3-8-1) que possui um perfil de ácido graxo típico de embriões do tipo selvagem para este experimento são exibidos na Fig. 26. Na Fig. 26, os ácidos graxos são identificados como 16:0 (palmitato), 18:0 (ácido esteárico), 18:1 (ácido oleico), LA e ALA e composições de ácidos graxos são expressas na forma de percentual em peso (% peso) do total de ácidos graxos.
O teor de ALA em embriões somáticos que expressam uma construção de grampo de cabelo fad3 (evento número 2148, Fig. 26) ou uma construção de grampo de cabelo fad3c e fad3 (evento número 2165, Fig. 26) exibiu redução de pelo menos 50% em comparação com embriões do tipo selvagem típicos (Fig. 26). Isso é uma forte indicação de que a construção de grampo de cabelo é funcional para reduzir o teor de ALA em embriões de soja.
Exemplo 31
Embriões somáticos de soja transformados com pKR1 183 para expressão de um gene de fusão delta-9 elongase de euglena anabaena e delta-8 dessaturase de Tetruetreptia pomquetensis CCMP1491 (Híbrido 1 - HGLA sintase)
O presente Exemplo descreve a transformação e expressão em embriões somáticos de soja de pKR1183 (SEQ ID N0 117) que contêm o gene de fusão de delta-9 elongase de Euglena anabaena e delta-8 dessaturase de Tetruetreptia pomquetensis CCMP1491 (Híbrido 1 - HGLA sintase) e o gene higromicina fosfotransferase para seleção sobre higromicina.
Cultivo de suspensão embriogênica de soja (cv. Jack) foi transformado com pKR1183 (SEQ ID N0 117) e embriões foram amadurecidos em meio líquido de maturação e histodiferenciação de soja (meios líquidos SHaM; Schmidt et al, Cell Biology and Morphogenesis, 24: 393 (2005)), conforme descrito anteriormente na Patente PCT n0 WO 2007/136877 publicada em 29 de novembro de 2007 (cujo conteúdo é incorporado ao presente como referência).
Após amadurecimento em meios líquidos SHaM1 um subconjunto de embriões de soja (ou seja, quatro embriões por evento) transformados com pKR1183 (SEQ ID Nº 117) foi colhido e analisado conforme descrito no presente.
Desta forma, foram analisados vinte eventos transformados com pKR1183 (SEQ ID Nº 117; Experimento 2145). Os perfis de ácido graxo para os cinco eventos que contêm o teor médio de DGLA mais alto (média dos cinco embriões analisados) são exibidos na Fig. 27. Na Fig. 27, os ácidos graxos são identificados como 16:0 (palmitato), 18:0 (ácido esteárico), 18:1 (ácido oleico), LA, ALA, EDA, ERA, DGLA e ETA e composições de ácidos graxos são expressas na forma de percentual em peso (% em peso) do total de ácidos graxos.
Exemplo 32
Embriões de soja transformados com vetores de expressão de soja PKR1253 para expressão de um gene de fusão delta-9 elongase de euglena anabaena e delta-8 dessaturase de tetruetreptia pomquetensis CCMP1491 (Híbrido 1 - HGLA sintase) com uma delta-5 dessaturase de Euglena GRACILIS E PKR1249 PARA REGULAGEM PARA BAIXO DE FAD3 DE SOJA E FAd3C DE SOJA
Cultivo de suspensão embriogênica de soja (cv. Jack) foi transformado com os fragmentos Ascl de pKR1249 (SEQ ID N0 141, Exemplo 26) e pKR1253 (SEQ ID N0 120), conforme descrito no Exemplo 1. Um subconjunto de embriões de soja gerados a partir de cada evento (dez embriões por evento) foi colhido, tomado em ampolas GC de vidro e metil ésteres de ácidos graxos (FAMEs) foram preparados por meio de transesterificação e analisados por meio de GC conforme descrito no Exemplo 2. Os tempos de retenção foram comparados com os de metil ésteres de padrões disponíveis comercialmente (Nu-Chek Prep, Inc.).
Desta forma, foram analisados 142 eventos transformados com pKR1249 (SEQ ID N0 141; Exemplo 26) e pKR1253 (SEQ ID N0 120) (experimento denominado Cura 25). Dos 142 eventos analisados, foram identificados noventa que produziram ARA em pelo menos um embrião de dez analisados em uma abundância relativa de mais de 1,0% do total de ácidos graxos. Destes, foram identificados 64 que produziram ARA em pelo menos um embrião de dez analisados em abundância relativa de mais de 10,0% do total de ácidos graxos. Destes, foram identificados 44 eventos que produziram ARA em pelo menos um embrião de dez analisados em uma abundância relativa de mais de 20,0% do total de ácidos graxos.
Os perfis médios de ácido graxo (média de dez embriões) para vinte eventos que possuem o ARA mais alto são exibidos na Fig. 28. Os ácidos graxos são identificados como 16:0 (palmitato), 18:0 (ácido esteárico), 18:1 (ácido oleico), LA1 ALA, EDA, SCI, DGLA, ARA, ERA, JUN, ETA e EPA; e as composições de ácidos graxos relacionadas na Fig. 28 são expressas na forma de percentual em peso (% em peso) do total de ácidos graxos. Para a Fig. 28, os ácidos graxos relacionados como "outros" incluem: 18:2 (5,9), 18:3 (5,9,12), STA, 20:0, 20:1 (11), 20:2 (7,11) ou 20:2 (8,11) e DPA. Cada um desses ácidos graxos está presente em abundância relativa de menos de 2,0% do total de ácidos graxos. O total médio de ácido graxo ômega 3 (Total n-3) é a soma das médias de todos os ácidos graxos ômega 3.
Os perfis reais de ácido graxo para cada embrião de um evento (AFS 5416-8-1-1) que contêm teor médio de ARA de 17,0% e teor médio de EPA de 1,5% são exibidos na Fig. 29. Os ácidos graxos são identificados como 16:0 (palmitato), 18:0 (ácido esteárico), 18:1 (ácido oleico), LA, ALA, EDA, SCI, DGLA1 ARA, ERA, JUN, ETA e EPA; e composições de ácidos graxos relacionadas na Fig. 29 são expressas na forma de percentual em peso (% em peso) do total de ácidos graxos). Para a Fig. 29, os ácidos graxos relacionados como "outros" incluem: 18:2 (5,9), 18:3 (5,9,12), STA, 20:0, 20:1 (11), 20:2 (7,11) ou 20:2 (8,11) e DPA. Cada um desses ácidos graxos encontra-se presente em abundância relativa de menos de 2,0% do total de ácidos graxos. O total de ácidos graxos ômega 3 (total n-3) é a soma de todos os ácidos graxos ômega 3.
Como o teor de ALA é geralmente 1,5 a três vezes mais alto em embriões somáticos de soja que na semente (ou seja, 15% a 30% em embriões (vide, por exemplo, o embrião VVT típico na Fig. 26), dependendo das condições de maturação e do tempo, contra 7 a 10% em sementes (Bilyeu et al, 2005, Crop Sei. 45: 1830-1836)), espera-se que o teor de ômega 3 em geral e os teores de EPA especificamente sejam significativamente mais baixos em sementes que embriões somáticos.
Exemplo 33
Embriões de soja transformados com vetores de expressão de soja PKR1255 para expressão de um gene de fusão delta-9 elongase de euglena anabaena e delta-8 dessaturase de tetruetreptia pomquetensis CCMP1491 (Híbrido 1 - HGLA Sintase) com uma delta-5 dessaturase de Euglena gracius e uma delta-5 dessaturase de euglena anabaena e PKR1249 para regulagem para baixo de fad3 de soja e fad3c de soja
Cultivo de suspensão embriogênica de soja (cv. Jack) foi transformado com os fragmentos AscI de pKR1249 (SEQ ID N0 141; Exemplo 26) e pKR1255 (SEQ ID N0 124) conforme descrito no Exemplo 1. Um subconjunto de embriões de soja gerados a partir de cada evento (dez embriões por evento) foi colhido, tomado em ampolas GC de vidro e metil ésteres de ácidos graxos (FAMEs) foram preparados por meio de transesterificação e analisados por meio de GC conforme descrito no Exemplo 2. Os tempos de retenção foram comparados com os de metil ésteres de padrões disponíveis comercialmente (Nu-Chek Prep1 Inc.).
Desta forma, foram analisados 197 eventos transformados com pKR1249 (SEQ ID N0 141; Exemplo 26) e pKR1255 (SEQ ID N0 124) (experimento denominado Cura 26). Dos 197 eventos analisados, foram identificados 128 que produziram ARA em pelo menos um embrião de cada dez analisados em abundância relativa de mais de 1,0% do total de ácidos graxos. Destes, foram identificados 105 que produziram ARA em pelo menos um embrião de cada dez analisados em abundância relativa de mais de 10,0% do total de ácidos graxos. Destes, foram identificados 83 eventos que produziram ARA em pelo menos um embrião de cada dez analisados em abundância relativa de mais de 20,0% do total de ácidos graxos.
Os perfis médios de ácidos graxos (média de nove ou dez embriões) para vinte eventos que possuem o ARA mais alto são exibidos na Fig. 30. Os ácidos graxos são identificados como 16:0 (palmitato), 18:0 (ácido esteárico), 18:1 (ácido oleico), LA, ALA, EDA, SCI, DGLA, ARA, ERA, JUN1 ETA e EPA; e composições de ácidos graxos relacionadas na Fig. 30 são expressas na forma de percentual em peso (% peso) do total de ácidos graxos. Para a Fig. 30, os ácidos graxos relacionados como "outros" incluem: 18:2 (5,9), 18:3 (5,9,12), STA, 20:0, 20:1 (11), 20:2 (7,11) ou 20:2 (8,11) e DPA. Cada um desses ácidos graxos está presente em abundância relativa de menos de 2,0% do total de ácidos graxos. A média total de ácido graxo ômega 3 (total n-3) é a soma das médias de todos os ácidos graxos ômega 3.
Exemplo 34
Análise funcional de semente de Arabidopsis transformada com pKR1 193
PARA EXPRESSÃO DE DELTA-9 ELONGASE DE euglena anabaena COM UMA DELTA-8 DESSATURASE DE euglena anabaena E UMA DELTA-5 DESSATURASE DE euglena
anabaena em arabidopsis Um mutante duplo fed3/fee1 (Smith et al, Planta 217: 507-516 (2003) de Arabidopsis produz sementes em que o teor de ácido graxo 20:1 e ALA é de menos de 2,0%. As plantas de Arabidopsis mutantes duplas fadS/fael foram transformadas com pKR1193 (SEQ ID N0 170) e as plantas foram cultivadas, mantidas e a semente foi colhida conforme descrito anteriormente em WO 2007/061845, publicada em 31 de maio de 2007 (cujo conteúdo é incorporado ao presente como referência).
Semente T2 em segregação foi obtida de dezoito eventos individuais para cada uma e os perfis de lipídios de sementes T2 a granel para cada evento foram obtidos por meio de transesterificação com TMSH conforme descrito no Exemplo 2 com as modificações a seguir. Para cada evento, uma concha pequena cheia de sementes (cerca de 25 a 50 sementes cada concha) foi fragmentada em 50 μΙ de TMSH em um tubo Eppendorf de 1,5 ml. Após agitação em TMSH por quinze minutos, foram adicionados 400 μΙ de heptano e os tubos foram bem turbilhonados, agitados por quinze minutos adicionais e centrifugados a 13.000 χ g por um minuto. Após a agitação, a camada de heptano foi removida para ampolas GC de vidro e os metil ésteres de ácidos graxos foram analisados conforme descrito no Exemplo 2.
Os perfis de lipídios de semente a granel T2 para os dezenove eventos transformados são exibidos na Fig. 31. Os ácidos graxos são identificados como 16:0 (palmitato), 18:0 (ácido esteárico), 18:1 (ácido oleico), LA, ALA, 20:1 (ácido eicosenoico), EDA, DGLA, ERA, ETA e EPA; e as composições de ácidos graxos relacionadas na Fig. 31 são expressas na forma de percentual em peso (% em peso) do total de ácidos graxos. Para a Fig. 31, os ácidos graxos relacionados como "outros" incluem: 18:2 (5,9), 18:3 (5,9,12), STA1 20:0 (ácido araquídico), 20:2 (7,11) ou 20:2 (8,11), SCI e JUP. Cada um desses ácidos está presente em abundância relativa de menos de 2,0% do total de ácidos graxos.
Os perfis de lipídios de sementes T2 individuais (dez sementes por evento) para dois eventos transformados por fad3 e /ae1 (ou seja, ff 1194- 16 e 'ff1194-18), que possuem um teor de ARA a granel de 10,4% e 9,4%, respectivamente (Fig. 31) foram obtidos por meio de transesterificação com TMSH conforme descrito no presente, mas com as modificações a seguir. Uma semente foi fragmentada em 10 μΙ de TMSH em um tubo Eppendorf de 1,5 ml. Após agitação em TMSH por quinze minutos, foram adicionados 75 μΙ de heptano e os tubos foram bem turbilhonados, agitados por quinze minutos adicionais e centrifugados a 13.000 χ g por um minuto. Após centrifugação, a camada de heptano foi removida para arnpolas GC de vidro que contêm 200 μ! de insertos e os metil ésteres de ácidos graxos foram analisados conforme descrito no Exemplo 31.
Os perfis de lipídios de sementes individuais são exibidos na Fig. 32. Os ácidos graxos são identificados como 16:0 (palmitato), 18:0 (ácido esteárico), 18:1 (ácido oleico), LA, GLA, ALA, STA, 20:1 (ácido eicosenoico), EDA, DGLA, ERA, ETA, EPA e DPA; e as composições de ácidos graxos relacionadas na Fig. 32 são expressas na forma de percentual em peso (% em peso) do total de ácidos graxos. Para a Fig. 32, os ácidos graxos relacionados como "outros" incluem: 18:3 (5,9,12), 20:0 (ácido araquídico), 20:2 (7,11) ou 20:2 (8,11) e 22:0. Cada um desses ácidos está presente em abundância relativa de menos de 3,0% do total de ácidos graxos. Listagem de Seqüências
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caaaggatgt tatgatatat taacactcta tctatgcacc ttattgttct atgataaatt 180
tcctcttatt attataaatc atctgaatcg tgacggctta tggaatgctt caaatagtac 240
aaaaacaaat gtgtactata agactttcta aacaattcta accttagcat tgtgaacgag 300
acataagtgt taagaagaca taacaattat aatggaagaa gtttgtctcc atttatatat 360
tatatattac ccacttatgt attatattag gatgttaagg agacataaca attataaaga 420
gagaagtttg tatccattta tatattatat actacccatt tatatattat acttatccac 480
ttatttaatg tctttataag gtttgatcca tgatatttct aatattttag ttgatatgta 540
tatgaaaggg tactatttga actctcttac tctgtataaa ggttggatca tccttaaagt 600
gggtctattt aattttattg cttcttacag ataaaaaaaa aattatgagt tggtttgata 660
aaatattgaa ggatttaaaa taataataaa taacatataa tatatgtata taaatttatt 720
ataatataac atttatctat aaaaaagtaa atattgtcat aaatctatac aatcgtttag 780
ccttgctgga cgaatctcaa ttatttaaac gagagtaaac atatttgact ttttggttat 840
ttaacaaatt attatttaac actatatgaa attttttttt ttatcagcaa agaataaaat 900
taaattaaga aggacaatgg tgtcccaatc cttatacaac caacttccac aagaaagtca 960
agtcagagac aacaaaaaaa caagcaaagg aaatttttta atttgagttg tcttgtttgc 1020
tgcataattt atgcagtaaa acactacaca taaccctttt agcagtagag caatggttga 1080
ccgtgtgctt agcttctttt attttatttt tttatcagca aagaataaat aaaataaaat 1140
gagacacttc agggatgttt caacaagctt ggatccgtcg acggcgcgcc cgatcatccg 1200
gatatagttc ctcctttcag caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt 1260 tatgctagtt attgctcagc ggtggcagca gccaactcag cttcctttcg ggctttgtta 1320 gcagccggat cgatccaagc tgtacctcac tattcctttg ccctcggacg agtgctgggg 1380 cgtcggtttc cactatcggc gagtacttct acacagccat cggtccagac ggccgcgctt 1440 ctgcgggcga tttgtgtacg cccgacagtc ccggctccgg atcggacgat tgcgtcgcat 1500 cgaccctgcg cccaagctgc atcatcgaaa ttgccgtcaa ccaagctctg atagagttgg 1560 tcaagaccaa tgcggagcat atacgcccgg agccgcggcg atcctgcaag ctccggatgc 1620 ctccgctcga agtagcgcgt ctgctgctcc atacaagcca accacggcct ccagaagaag 1680 atgttggcga cctcgtattg ggaatccccg aacatcgcct cgctccagtc aatgaccgct 1740 gttatgcggc cattgtccgt caggacattg ttggagccga aatccgcgtg cacgaggtgc 1800 cggacttcgg ggcagtcctc ggcccaaagc atcagctcat cgagagcctg cgcgacggac 1860 gcactgacgg tgtcgtccat cacagtttgc cagtgataca catggggatc agcaatcgcg 1920 catatgaaat cacgccatgt agtgtattga ccgattcctt gcggtccgaa tgggccgaac 1980 ccgctcgtct ggctaagatc ggccgcagcg atcgcatcca tagcctccgc gaccggctgc 2040 agaacagcgg gcagttcggt ttcaggcagg tcttgcaacg tgacaccctg tgcacggcgg 2100 gagatgcaat aggtcaggct ctcgctgaat tccccaatgt caagcacttc cggaatcggg 2160 agcgcggccg atgcaaagtg ccgataaaca taacgatctt tgtagaaacc atcggcgcag 2220 ctatttaccc gcaggacata tccacgccct cctacatcga agctgaaagc acgagattct 2280 tcgccctccg agagctgcat caggtcggag acgctgtcga acttttcgat cagaaacttc 2340 tcgacagacg tcgcggtgag ttcaggcttt tccatgggta tatctccttc ttaaagttaa 2400 acaaaattat ttctagaggg aaaccgttgt ggtctcccta tagtgagtcg tattaatttc 2460 gcgggatcga gatctgatca acctgcatta atgaatcggc caacgcgcgg ggagaggcgg 2520 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 2580 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 2640 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 2700 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 2760 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 2820 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 2880 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc 2940 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 3000 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 3060 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 3120 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc 3180
tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 3240
caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 3300
atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 3360
acgttaaggg attttggtca tgacattaac ctataaaaat aggcgtatca cgaggccctt 3420
tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac 3480
ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc 3540
gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag 3600
agtgcaccat atggacatat tgtcgttaga acgcggctac aattaataca taaccttatg 3660
tatcatacac atacgattta ggtgacacta tagaacggcg cgccaagctt ttgatccatg 3720
cccttcattt gccgcttatt aattaatttg gtaacagtcc gtactaatca gttacttatc 3780
cttcccccat cataattaat cttggtagtc tcgaatgcca caacactgac tagtctcttg 3840
gatcataaga aaaagccaag gaacaaaaga agacaaaaca caatgagagt atcctttgca 3900
tagcaatgtc taagttcata aaattcaaac aaaaacgcaa tcacacacag tggacatcac 3960
ttatccacta gctgatcagg atcgccgcgt caagaaaaaa aaactggacc ccaaaagcca 4020
tgcacaacaa cacgtactca caaaggtgtc aatcgagcag cccaaaacat tcaccaactc 4080
aacccatcat gagccctcac atttgttgtt tctaacccaa cctcaaactc gtattctctt 4140
ccgccacctc atttttgttt atttcaacac ccgtcaaact gcatgccacc ccgtggccaa 4200
atgtccatgc atgttaacaa gacctatgac tataaatagc tgcaatctcg gcccaggttt 4260
tcatcatcaa gaaccagttc aatatcctag tacaccgtat taaagaattt aagatatact 4320
gcggccgcat gggaacggac caaggaaaaa ccttcacctg ggaagagctg gcggcccata 4380
acaccaagga cgacctactc ttggccatcc gcggcagggt gtacgatgtc acaaagttct 4440
tgagccgcca tcctggtgga gtggacactc tcctgctcgg agctggccga gatgttactc 4500
cggtctttga gatgtatcac gcgtttgggg ctgcagatgc cattatgaag aagtactatg 4560
tcggtacact ggtctcgaat gagctgccca tcttcccgga gccaacggtg ttccacaaaa 4620
ccatcaagac gagagtcgag ggctacttta cggatcggaa cattgatccc aagaatagac 4680
cagagatctg gggacgatac gctcttatct ttggatcctt gatcgcttcc tactacgcgc 4740
agctctttgt gcctttcgtt gtcgaacgca catggcttca ggtggtgttt gcaatcatca 4800
tgggatttgc gtgcgcacaa gtcggactca accctcttca tgatgcgtct cacttttcag 4860
tgacccacaa ccccactgtc tggaagattc tgggagccac gcacgacttt ttcaacggag 4 920
catcgtacct ggtgtggatg taccaacata tgctcggcca tcacccctac accaacattg 4 980 ctggagcaga tcccgacgtg tcgacgtctg agcccgatgt tcgtcgtatc aagcccaacc 5040 aaaagtggtt tgtcaaccac atcaaccagc acatgtttgt tcctttcctg tacggactgc 5100 tggcgttcaa ggtgcgcatt caggacatca acattttgta ctttgtcaag accaatgacg 5160 ctattcgtgt caatcccatc tcgacatggc acactgtgat gttctggggc ggcaaggctt 5220 tctttgtctg gtatcgcctg attgttcccc tgcagtatct gcccctgggc aaggtgctgc 5280 tcttgttcac ggtcgcggac atggtgtcgt cttactggct ggcgctgacc ttccaggcga 5340 accacgttgt tgaggaagtt cagtggccgt tgcctgacga gaacgggatc atccaaaagg 5400 actgggcagc tatgcaggtc gagactacgc aggattacgc acacgattcg cacctctgga 5460 ccagcatcac tggcagcttg aactaccagg ctgtgcacca tctgttcccc aacgtgtcgc 5520 agcaccatta tcccgatatt ctggccatca tcaagaacac ctgcagcgag tacaaggttc 5580 cataccttgt caaggatacg ttttggcaag catttgcttc acatttggag cacttgcgtg 5640 ttcttggact ccgtcccaag gaagagtagg C 5671 <210> 2 <211> 5252 <212> DNA <213> Seqüência Artificial <220> <223> plasmídeo pKR457 <220> <221> misc feature <222> (3872)..(3872) <223> η é a, c, g, ou t <400> 2 gtacgtgggc ggatcccccg ggctgcagga attcactggc cgtcgtttta caacgtcgtg 60 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca 120 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga 180 atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc 240 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac 300 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca 360 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga 420 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa 480 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt 540 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 600 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta 660 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag 720
taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca 780
gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta 840
aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc 900
gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc 960
ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca 1020
ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc 1080
acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca 1140
taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac 1200
tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg 1260
cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg 1320
ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg 1380
gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac 1440
gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc 1500
aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct 1560
aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc 1620
actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 1680
gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 1740
atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 1800
atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 1860
ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt 1920
gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa 1980
cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 2040
tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc 2100
cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct 2160
ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat 2220
gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc 2280
tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg 2340
ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc 2400
gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg 2460
cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca 2520 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact 2580
ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa 2640
acagctatga ccatgattac gccaagcttg catgcctgca ggtcgactcg acgtacgtcc 2700
tcgaagagaa gggttaataa cacatttttt aacattttta acacaaattt tagttattta 2760
aaaatttatt aaaaaattta aaataagaag aggaactctt taaataaatc taacttacaa 2820
aatttatgat ttttaataag ttttcaccaa taaaaaatgt cataaaaata tgttaaaaag 2880
tatattatca atattctctt tatgataaat aaaaagaaaa aaaaaataaa agttaagtga 2940
aaatgagatt gaagtgactt taggtgtgta taaatatatc aaccccgcca acaatttatt 3000
taatccaaat atattgaagt atattattcc atagccttta tttatttata tatttattat 3060
ataaaagctt tatttgttct aggttgttca tgaaatattt ttttggtttt atctccgttg 3120
taagaaaatc atgtgctttg tgtcgccact cactattgca gctttttcat gcattggtca 3180
gattgacggt tgattgtatt tttgtttttt atggttttgt gttatgactt aagtcttcat 3240
ctctttatct cttcatcagg tttgatggtt acctaatatg gtccatgggt acatgcatgg 3300
ttaaattagg tggccaactt tgttgtgaac gatagaattt tttttatatt aagtaaacta 3360
tttttatatt atgaaataat aataaaaaaa atattttatc attattaaca aaatcatatt 3420
agttaatttg ttaactctat aataaaagaa atactgtaac attcacatta catggtaaca 3480
tctttccacc ctttcatttg ttttttgttt gatgactttt tttcttgttt aaatttattt 3540
cccttctttt aaatttggaa tacattatca tcatatataa actaaaatac taaaaacagg 3600
attacacaaa tgataaataa taacacaaat atttataaat ctagctgcaa tatatttaaa 3660
ctagctatat cgatattgta aaataaaact agctgcattg atactgataa aaaaatatca 3720
tgtgctttct ggactgatga tgcagtatac ttttgacatt gcctttattt tatttttcag 3780
aaaagctttc ttagttctgg gttcttcatt atttgtttcc catctccatt gtgaattgaa 3840
tcatttgctt cgtgtcacaa atacaattta gntaggtaca tgcattggtc agattcacgg 3900
tttattatgt catgacttaa gttcatggta gtacattacc tgccacgcat gcattatatt 3960
ggttagattt gataggcaaa tttggttgtc aacaatataa atataaataa tgtttttata 4020
ttacgaaata acagtgatca aaacaaacag ttttatcttt attaacaaga ttttgttttt 4080
gtttgatgac gttttttaat gtttacgctt tcccccttct tttgaattta gaacacttta 4140
tcatcataaa atcaaatact aaaaaaatta catatttcat aaataataac acaaatattt 4200
ttaaaaaatc tgaaataata atgaacaata ttacatatta tcacgaaaat tcattaataa 4260
aaatattata taaataaaat gtaatagtag ttatatgtag gaaaaaagta ctgcacgcat 4320
aatatataca aaaagattaa aatgaactat tataaataat aacactaaat taatggtgaa 4380 tcatatcaaa ataatgaaaa agtaaataaa atttgtaatt aacttctata tgtattacac 4440
acacaaataa taaataatag taaaaaaaat tatgataaat atttaccatc tcataagata 4 500
tttaaaataa tgataaaaat atagattatt ttttatgcaa ctagctagcc aaaaagagaa 4560
cacgggtata tataaaaaga gtacctttaa attctactgt acttccttta ttcctgacgt 4620
ttttatatca agtggacata cgtgaagatt ttaattatca gtctaaatat ttcattagca 4680
cttaatactt ttctgtttta ttcctatcct ataagtagtc ccgattctcc caacattgct 4740
tattcacaca actaactaag aaagtcttcc atagcccccc aagcggccgc gacacaagtg 4800
tgagagtact aaataaatgc tttggttgta cgaaatcatt acactaaata aaataatcaa 4860
agcttatata tgccttccgc taaggccgaa tgcaaagaaa ttggttcttt ctcgttatct 4920
tttgccactt ttactagtac gtattaatta ctacttaatc atctttgttt acggctcatt 4980
atatccggtc tagaggatcc aaggccgcga agttaaaagc aatgttgtca cttgtcgtac 5040
taacacatga tgtgatagtt tatgctagct agctataaca taagctgtct ctgagtgtgt 5100
tgtatattaa taaagatcat cactggtgaa tggtgatcgt gtacgtaccc tacttagtag 5160
gcaatggaag cacttagagt gtgctttgtg catggccttg cctctgtttt gagacttttg 5220
taatgttttc gagtttaaat ctttgccttt gc 5252
<210> 3 <211> 6601 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo ρΚΗ606 <220>
<221> misc_feature <222> (5689) .. (5689) <223> η é a, c, g, ou t
<400> 3
ggccgcatgg gaacggacca aggaaaaacc ttcacctggg aagagctggc ggcccataac 60
accaaggacg acctactctt ggccatccgc ggcagggtgt acgatgtcac aaagttcttg 120
agccgccatc ctggtggagt ggacactctc ctgctcggag ctggccgaga tgttactccg 180
gtctttgaga tgtatcacgc gtttggggct gcagatgcca ttatgaagaa gtactatgtc 240
ggtacactgg tctcgaatga gctgcccatc ttcccggagc caacggtgtt ccacaaaacc 300
atcaagacga gagtcgaggg ctactttacg gatcggaaca ttgatcccaa gaatagacca 360
gagatctggg gacgatacgc tcttatcttt ggatccttga tcgcttccta ctacgcgcag 420
ctctttgtgc ctttcgttgt cgaacgcaca tggcttcagg tggtgtttgc aatcatcatg 480 ggatttgcgt gcgcacaagt cggactcaac cctcttcatg atgcgtctca cttttcagtg 540
acccacaacc ccactgtctg gaagattctg ggagccacgc acgacttttt caacggagca 600
tcgtacctgg tgtggatgta ccaacatatg ctcggccatc acccctacac caacattgct 660
ggagcagatc ccgacgtgtc gacgtctgag cccgatgttc gtcgtatcaa gcccaaccaa 720
aagtggtttg tcaaccacat caaccagcac atgtttgttc ctttcctgta cggactgctg 780
gcgttcaagg tgcgcattca ggacatcaac attttgtact ttgtcaagac caatgacgct 840
attcgtgtca atcccatctc gacatggcac actgtgatgt tctggggcgg caaggctttc 900
tttgtctggt atcgcctgat tgttcccctg cagtatctgc ccctgggcaa ggtgctgctc 960
ttgttcacgg tcgcggacat ggtgtcgtct tactggctgg cgctgacctt ccaggcgaac 1020
cacgttgttg aggaagttca gtggccgttg cctgacgaga acgggatcat ccaaaaggac 1080
tgggcagcta tgcaggtcga gactacgcag gattacgcac acgattcgca cctctggacc 1140
agcatcactg gcagcttgaa ctaccaggct gtgcaccatc tgttccccaa cgtgtcgcag 1200
caccattatc ccgatattct ggccatcatc aagaacacct gcagcgagta caaggttcca 1260
taccttgtca aggatacgtt ttggcaagca tttgcttcac atttggagca cttgcgtgtt 1320
cttggactcc gtcccaagga agagtaggcg gccgcgacac aagtgtgaga gtactaaata 1380
aatgctttgg ttgtacgaaa tcattacact aaataaaata atcaaagctt atatatgcct 1440
tccgctaagg ccgaatgcaa agaaattggt tctttctcgt tatcttttgc cacttttact 1500
agtacgtatt aattactact taatcatctt tgtttacggc tcattatatc cggtctagag 1560
gatccaaggc cgcgaagtta aaagcaatgt tgtcacttgt cgtactaaca catgatgtga 1620
tagtttatgc tagctagcta taacataagc tgtctctgag tgtgttgtat attaataaag 1680
atcatcactg gtgaatggtg atcgtgtacg taccctactt agtaggcaat ggaagcactt 1740
agagtgtgct ttgtgcatgg ccttgcctct gttttgagac ttttgtaatg ttttcgagtt 1800
taaatctttg cctttgcgta cgtgggcgga tcccccgggc tgcaggaatt cactggccgt 1860
cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc 1920
acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca 1980
acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct 2040
gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata 2100
gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct 2160
cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt 2220
ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata 2280
ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt 2340 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag 2400
acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca 24 60
tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc 2520
agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat 2580
cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc 2640
aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg 2700
gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc 2760
agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat 2820
aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga 2880
gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc 2940
ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc 3000
aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt 3060
aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc 3120
tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc 3180
agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca 3240
ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca 3300
ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt 3360
ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta 3420
acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg 3480
agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc 3540
ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag 3600
cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa 3660
gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc 3720
cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc 3780
gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta 3840
caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag 3900
aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct 3960
tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga 4020
gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc 4080
ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt 4140
atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg 4200 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg 4260
caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc 4320
cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc 4380
accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata 4440
acaatttcac acaggaaaca gctatgacca tgattacgcc aagcttgcat gcctgcaggt 4500
cgactcgacg tacgtcctcg aagagaaggg ttaataacac attttttaac atttttaaca 4560
caaattttag ttatttaaaa atttattaaa aaatttaaaa taagaagagg aactctttaa 4620
ataaatctaa cttacaaaat ttatgatttt taataagttt tcaccaataa aaaatgtcat 4680
aaaaatatgt taaaaagtat attatcaata ttctctttat gataaataaa aagaaaaaaa 4740
aaataaaagt taagtgaaaa tgagattgaa gtgactttag gtgtgtataa atatatcaac 4800
cccgccaaca atttatttaa tccaaatata ttgaagtata ttattccata gcctttattt 4860
atttatatat ttattatata aaagctttat ttgttctagg ttgttcatga aatatttttt 4920
tggttttatc tccgttgtaa gaaaatcatg tgctttgtgt cgccactcac tattgcagct 4980
ttttcatgca ttggtcagat tgacggttga ttgtattttt gttttttatg gttttgtgtt 5040
atgacttaag tcttcatctc tttatctctt catcaggttt gatggttacc taatatggtc 5100
catgggtaca tgcatggtta aattaggtgg ccaactttgt tgtgaacgat agaatttttt 5160
ttatattaag taaactattt ttatattatg aaataataat aaaaaaaata ttttatcatt 5220
attaacaaaa tcatattagt taatttgtta actctataat aaaagaaata ctgtaacatt 5280
cacattacat ggtaacatct ttccaccctt tcatttgttt tttgtttgat gacttttttt 5340
cttgtttaaa tttatttccc ttcttttaaa tttggaatac attatcatca tatataaact 5400
aaaatactaa aaacaggatt acacaaatga taaataataa cacaaatatt tataaatcta 5460
gctgcaatat atttaaacta gctatatcga tattgtaaaa taaaactagc tgcattgata 5520
ctgataaaaa aatatcatgt gctttctgga ctgatgatgc agtatacttt tgacattgcc 5580
tttattttat ttttcagaaa agctttctta gttctgggtt cttcattatt tgtttcccat 5640
ctccattgtg aattgaatca tttgcttcgt gtcacaaata caatttagnt aggtacatgc 5700
attggtcaga ttcacggttt attatgtcat gacttaagtt catggtagta cattacctgc 5760
cacgcatgca ttatattggt tagatttgat aggcaaattt ggttgtcaac aatataaata 5820
taaataatgt ttttatatta cgaaataaca gtgatcaaaa caaacagttt tatctttatt 5880
aacaagattt tgtttttgtt tgatgacgtt ttttaatgtt tacgctttcc cccttctttt 5940
gaatttagaa cactttatca tcataaaatc aaatactaaa aaaattacat atttcataaa 6000
taataacaca aatattttta aaaaatctga aataataatg aacaatatta catattatca 6060 cgaaaattca ttaataaaaa tattatataa ataaaatgta atagtagtta tatgtaggaa 6120
aaaagtactg cacgcataat atatacaaaa agattaaaat gaactattat aaataataac 6180
actaaattaa tggtgaatca tatcaaaata atgaaaaagt aaataaaatt tgtaattaac 6240
ttctatatgt attacacaca caaataataa ataatagtaa aaaaaattat gataaatatt 6300
taccatctca taagatattt aaaataatga taaaaatata gattattttt tatgcaacta 6360
gctagccaaa aagagaacac gggtatatat aaaaagagta cctttaaatt ctactgtact 6420
tcctttattc ctgacgtttt tatatcaagt ggacatacgt gaagatttta attatcagtc 6480
taaatatttc attagcactt aatacttttc tgttttattc ctatcctata agtagtcccg 6540
attctcccaa cattgcttat tcacacaact aactaagaaa gtcttccata gccccccaag 6600
c 6601
<210> 4 <211> 2577 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmídeo pKR277 <400> 4
agcttggatc tcctgcagga tctggccggc cggatctcgt acggatccgt cgacggcgcg 60
cccgatcatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag 120
gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt 180
cgggctttgt tagcagccgg atcgatccaa gctgtacctc actattcctt tgccctcgga 240
cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag 300
acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg 360
attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc 420
tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagccgcgg cgatcctgca 480
agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc 540
ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag 600
tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg 660
tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc 720
tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga 780
tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg 840
aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catagcctcc 900
gcgaccggct gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc 960 tgtgcacggc gggagatgca ataggtcagg ctctcgctga attccccaat gtcaagcact 1020 tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa 1080 ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa 1140 gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg 1200 atcagaaact tctcgacaga cgtcgcggtg agttcaggct tttccatggg tatatctcct 1260 tcttaaagtt aaacaaaatt atttctagag ggaaaccgtt gtggtctccc tatagtgagt 1320 cgtattaatt tcgcgggatc gagatctgat caacctgcat taatgaatcg gccaacgcgc 1380 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 1440 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 1500 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag 1560 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 1620 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 1680 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 1740 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag 1800 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 1860 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 1920 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 1980 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt 2040 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 2100 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 2160 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 2220 gaacgaaaac tcacgttaag ggattttggt catgacatta acctataaaa ataggcgtat 2280 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca 2340 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca 2400 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca 2460 gattgtactg agagtgcacc atatggacat attgtcgtta gaacgcggct acaattaata 2520 cataacctta tgtatcatac acatacgatt taggtgacac tatagaacgg cgcgcca 2577
<210> 5
<211> 6494
<212> DNA
<213> Seqüência Artificial <220> <223> plasmídeo pKR804 <220>
<221> misc_feature
<222> (3761)..(3761)
<223> η é a, c, g, ou t
<400>
ccggccggat ctcgtacgga tccgtcgacg gcgcgcccga tcatccggat atagttcctc 60 ctttcagcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat gctagttatt 120 gctcagcggt ggcagcagcc aactcagctt cctttcgggc tttgttagca gccggatcga 180 tccaagctgt acctcactat tcctttgccc tcggacgagt gctggggcgt cggtttccac 240 tatcggcgag tacttctaca cagccatcgg tccagacggc cgcgcttctg cgggcgattt 300 gtgtacgccc gacagtcccg gctccggatc ggacgattgc gtcgcatcga ccctgcgccc 360 aagctgcatc atcgaaattg ccgtcaacca agctctgata gagttggtca agaccaatgc 420 ggagcatata cgcccggagc cgcggcgatc ctgcaagctc cggatgcctc cgctcgaagt 480 agcgcgtctg ctgctccata caagccaacc acggcctcca gaagaagatg ttggcgacct 540 cgtattggga atccccgaac atcgcctcgc tccagtcaat gaccgctgtt atgcggccat 600 tgtccgtcag gacattgttg gagccgaaat ccgcgtgcac gaggtgccgg acttcggggc 660 agtccteggc ccaaagcatc agctcatcga gagcctgcgc gacggacgca ctgacggtgt 720 cgtccatcac agtttgccag tgatacacat ggggatcagc aatcgcgcat atgaaatcac 780 gccatgtagt gtattgaccg attccttgcg gtccgaatgg gccgaacccg ctcgtctggc 840 taagatcggc cgcagcgatc gcatccatag cctccgcgac cggctgcaga acagcgggca 900 gttcggtttc aggcaggtct tgcaacgtga caccctgtgc acggcgggag atgcaatagg 960 tcaggctctc gctgaattcc ccaatgtcaa gcacttccgg aatcgggagc gcggccgatg 1020 caaagtgccg ataaacataa cgatctttgt agaaaccatc ggcgcagcta tttacccgca 1080 ggacatatcc acgccctcct acatcgaagc tgaaagcacg agattcttcg ccctccgaga 1140 gctgcatcag gtcggagacg ctgtcgaact tttcgatcag aaacttctcg acagacgtcg 1200 cggtgagttc aggcttttcc atgggtatat ctccttctta aagttaaaca aaattatttc 1260 tagagggaaa ccgttgtggt ctccctatag tgagtcgtat taatttcgcg ggatcgagat 1320 ctgatcaacc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 1380 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 1440 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 1500 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 1560 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 1620 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 1680
gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 1740
ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 1800
cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 1860
ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 1920
actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 1980
tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca 2040
gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 2100
ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 2160
cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 2220
ttggtcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt 2280
ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt 2340
ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg 2400
tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg 2460
gacatattgt cgttagaacg cggctacaat taatacataa ccttatgtat catacacata 2520
cgatttaggt gacactatag aacggcgcgc caagcttgga tctcctgcag gatctggccg 2580
ggtacgtcct cgaagagaag ggttaataac acatttttta acatttttaa cacaaatttt 2640
agttatttaa aaatttatta aaaaatttaa aataagaaga ggaactcttt aaataaatct 2700
aacttacaaa atttatgatt tttaataagt tttcaccaat aaaaaatgtc ataaaaatat 2760
gttaaaaagt atattatcaa tattctcttt atgataaata aaaagaaaaa aaaaataaaa 2820
gttaagtgaa aatgagattg aagtgacttt aggtgtgtat aaatatatca accccgccaa 2880
caatttattt aatccaaata tattgaagta tattattcca tagcctttat ttatttatat 2940
atttattata taaaagcttt atttgttcta ggttgttcat gaaatatttt tttggtttta 3000
tctccgttgt aagaaaatca tgtgctttgt gtcgccactc actattgcag ctttttcatg 3060
cattggtcag attgacggtt gattgtattt ttgtttttta tggttttgtg ttatgactta 3120
agtcttcatc tctttatctc ttcatcaggt ttgatggtta cctaatatgg tccatgggta 3180
catgcatggt taaattaggt ggccaacttt gttgtgaacg atagaatttt ttttatatta 3240
agtaaactat ttttatatta tgaaataata ataaaaaaaa tattttatca ttattaacaa 3300
aatcatatta gttaatttgt taactctata ataaaagaaa tactgtaaca ttcacattac 3360
atggtaacat ctttccaccc tttcatttgt tttttgtttg atgacttttt ttcttgttta 3420
aatttatttc ccttctttta aatttggaat acattatcat catatataaa ctaaaatact 3480 aaaaacagga ttacacaaat gataaataat aacacaaata tttataaatc tagctgcaat 3540
atatttaaac tagctatatc gatattgtaa aataaaacta gctgcattga tactgataaa 3600
aaaatatcat gtgctttctg gactgatgat gcagtatact tttgacattg cctttatttt 3660
atttttcaga aaagctttct tagttctggg ttcttcatta tttgtttccc atctccattg 3720
tgaattgaat catttgcttc gtgtcacaaa tacaatttag ntaggtacat gcattggtca 3780
gattcacggt ttattatgtc atgacttaag ttcatggtag tacattacct gccacgcatg 3840
cattatattg gttagatttg ataggcaaat ttggttgtca acaatataaa tataaataat 3900
gtttttatat tacgaaataa cagtgatcaa aacaaacagt tttatcttta ttaacaagat 3960
tttgtttttg tttgatgacg ttttttaatg tttacgcttt cccccttctt ttgaatttag 4020
aacactttat catcataaaa tcaaatacta aaaaaattac atatttcata aataataaca 4080
caaatatttt taaaaaatct gaaataataa tgaacaatat tacatattat cacgaaaatt 4140
cattaataaa aatattatat aaataaaatg taatagtagt tatatgtagg aaaaaagtac 4200
tgcacgcata atatatacaa aaagattaaa atgaactatt ataaataata acactaaatt 4260
aatggtgaat catatcaaaa taatgaaaaa gtaaataaaa tttgtaatta acttctatat 4320
gtattacaca cacaaataat aaataatagt aaaaaaaatt atgataaata tttaccatct 4380
cataagatat ttaaaataat gataaaaata tagattattt tttatgcaac tagctagcca 4440
aaaagagaac acgggtatat ataaaaagag tacctttaaa ttctactgta cttcctttat 4500
tcctgacgtt tttatatcaa gtggacatac gtgaagattt taattatcag tctaaatatt 4560
tcattagcac ttaatacttt tctgttttat tcctatccta taagtagtcc cgattctccc 4620
aacattgctt attcacacaa ctaactaaga aagtcttcca tagcccccca agcggccgca 4680
tgggaacgga ccaaggaaaa accttcacct gggaagagct ggcggcccat aacaccaagg 4740
acgacctact cttggccatc cgcggcaggg tgtacgatgt cacaaagttc ttgagccgcc 4800
atcctggtgg agtggacact ctcctgctcg gagctggccg agatgttact ccggtctttg 4860
agatgtatca cgcgtttggg gctgcagatg ccattatgaa gaagtactat gtcggtacac 4920
tggtctcgaa tgagctgccc atcttcccgg agccaacggt gttccacaaa accatcaaga 4980
cgagagtcga gggctacttt acggatcgga acattgatcc caagaataga ccagagatct 5040
ggggacgata cgctcttatc tttggatcct tgatcgcttc ctactacgcg cagctctttg 5100
tgcctttcgt tgtcgaacgc acatggcttc aggtggtgtt tgcaatcatc atgggatttg 5160
cgtgcgcaca agtcggactc aaccctcttc atgatgcgtc tcacttttca gtgacccaca 5220
accccactgt ctggaagatt ctgggagcca cgcacgactt tttcaacgga gcatcgtacc 5280
tggtgtggat gtaccaacat atgctcggcc atcaccccta caccaacatt gctggagcag 5340 atcccgacgt gtcgacgtct gagcccgatg ttcgtcgtat caagcccaac caaaagtggt 5400 ttgtcaacca catcaaccag cacatgtttg ttcctttcct gtacggactg ctggcgttca 5460 aggtgcgcat tcaggacatc aacattttgt actttgtcaa gaccaatgac gctattcgtg 5520 tcaatcccat ctcgacatgg cacactgtga tgttctgggg cggcaaggct ttctttgtct 5580 ggtatcgcct gattgttccc ctgcagtatc tgcccctggg caaggtgctg ctcttgttca 5640 cggtcgcgga catggtgtcg tcttactggc tggcgctgac cttccaggcg aaccacgttg 5700 ttgaggaagt tcagtggccg ttgcctgacg agaacgggat catccaaaag gactgggcag 5760 ctatgcaggt cgagactacg caggattacg cacacgattc gcacctctgg accagcatca 5820 ctggcagctt gaactaccag gctgtgcacc atctgttccc caacgtgtcg cagcaccatt 5880 atcccgatat tctggccatc atcaagaaca cctgcagcga gtacaaggtt ccataccttg 5940 tcaaggatac gttttggcaa gcatttgctt cacatttgga gcacttgcgt gttcttggac 6000 tccgtcccaa ggaagagtag gcggccgcga cacaagtgtg agagtactaa ataaatgctt 6060 tggttgtacg aaatcattac actaaataaa ataatcaaag cttatatatg ccttccgcta 6120 aggccgaatg caaagaaatt ggttctttct cgttatcttt tgccactttt actagtacgt 6180 attaattact acttaatcat ctttgtttac ggctcattat atccggtcta gaggatccaa 6240 ggccgcgaag ttaaaagcaa tgttgtcact tgtcgtacta acacatgatg tgatagttta 6300 tgctagctag ctataacata agctgtctct gagtgtgttg tatattaata aagatcatca 6360 ctggtgaatg gtgatcgtgt acgtacccta cttagtaggc aatggaagca cttagagtgt 6420 gctttgtgca tggccttgcc tctgttttga gacttttgta atgttttcga gtttaaatct 6480 ttgcctttgc gtac 6494 <210> 6 <211> 4771 <212> DNA <213> Seqüência Artificial <220> <223> plasmideo pKR953 <400> 6 ggccgcgaag ttaaaagcaa tgttgtcact tgtcgtacta acacatgatg tgatagttta 60 tgctagctag ctataacata agctgtctct gagtgtgttg tatattaata aagatcatca 120 ctggtgaatg gtgatcgtgt acgtacccta cttagtaggc aatggaagca cttagagtgt 180 gctttgtgca tggccttgcc tctgttttga gacttttgta atgttttcga gtttaaatct 240 ttgcctttgc gtacgtctag agtcgacctg caggcatgca agcttggcgt aatcatggtc 300 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 360 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 420
gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 480
ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 540
ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 600
acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 660
aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 720
tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 780
aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 840
gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 900
acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 960
accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 1020
ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 1080
gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 1140
gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 1200
ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 1260
gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 1320
cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 1380
cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 1440
gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 1500
tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 1560
gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 1620
agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 1680
tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 1740
agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 1800
gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 1860
catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 1920
ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 1980
atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 2040
tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 2100
cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 2160
cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 2220 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 2280
aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 2340
ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 2400
aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga 2460
aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct 2520
cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac 2580
agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt 2640
tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca 2700
ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca 2760
ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt 2820
acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt 2880
ttcccagtca cgacgttgta aaacgacggc cagtgaattc gagctcggta cccggggatc 2940
ctctagacct gcaggccaac tgcgtttggg gctccagatt aaacgacgcc gtttcgttcc 3000
tttcgcttca cggcttaacg atgtcgtttc tgtctgtgcc caaaaaataa aggcatttgt 3060
tatttgcacc agatatttac taagtgcacc ctagtttgac aagtaggcga taattacaaa 3120
tagatgcggt gcaaataata aattttgaag gaaataatta caaaagaaca gaacttatat 3180
ttactttatt ttaaaaaact aaaatgaaag aacaaaaaaa gtaaaaaata caaaaaatgt 3240
gctttaacca ctttcattat ttgttacaga aagtatgatt ctactcaaat tgatctgttg 3300
tatctggtgc tgccttgtca cactggcgat ttcaatcccc taaagatatg gtgcaaactg 3360
cgaagtgatc aatatctgct cggttaattt agattaatta ataatattca acgtgatgta 3420
ccaaaaaaag acaatttttt gctccattga caaattaaac ctcatcaagg taatttccaa 3480
acctataagc aaaaaaattt cacattaatt ggcccgcaat cctattagtc ttattatact 3540
agagtaggaa aaaaaacaat tacacaactt gtcttattat tctctatgct aatgaatatt 3600
tttccctttt gttagaaatc agtgtttcct aatttattga gtattaattc cactcaccgc 3660
atatatttac cgttgaataa gaaaatttta cacataattc tttttaagat aaataatttt 3720
tttatactag atcttatatg attacgtgaa gccaagtggg ttatactaat gatatataat 3780
gtttgatagt aatcagttta taaaccaaat gcatggaaat gttacgtgga agcacgtaaa 3840
ttaacaagca ttgaagcaaa tgcagccacc gcaccaaaac caccccactt cacttccacg 3900
taccatattc catgcaacta caacacccta aaacttcaat aaatgccccc accttcactt 3960
cacttcaccc atcaatagca agcggccgca ccatggaggt ggtgaatgaa atagtctcaa 4020
ttgggcagga agttttaccc aaagttgatt atgcccaact ctggagtgat gccagtcact 4080 gtgaggtgct ttacttgtcc atcgcatttg tcatcttgaa gttcactctt ggcccccttg 4140
gtccaaaagg tcagtctcgt atgaagtttg ttttcaccaa ttacaacctt ctcatgtcca 4200
tttattcgtt gggatcattc ctctcaatgg catatgccat gtacaccatc ggtgttatgt 4260
ctgacaactg cgagaaggct tttgacaaca acgtcttcag gatcaccacg cagttgttct 4320
atttgagcaa gttcctggag tatattgact ccttctattt gccactgatg ggcaagcctc 4380
tgacctggtt gcaattcttc catcatttgg gggcaccgat ggatatgtgg ctgttctata 4440
attaccgaaa tgaagctgtt tggatttttg tgctgttgaa tggtttcatc cactggatca 4500
tgtacggtta ttattggacc agattgatca agctgaagtt ccccatgcca aaatccctga 4560
ttacatcaat gcagatcatt caattcaatg ttggtttcta cattgtctgg aagtacagga 4620
acattccctg ttatcgccaa gatgggatga ggatgtttgg ctggttcttc aattactttt 4680
atgttggcac agtcttgtgt ttgttcttga atttctatgt gcaaacgtat atcgtcagga 4740
agcacaaggg agccaaaaag attcagtgag c 4771
<210> 7 <211> 8584 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmídeo pKR1084 <220>
<221> misc_feature <222> (5299)..(5299) <223> η é a, c, g, ou t
<400> 7
ggagatccaa gcttggcgcg ccgttctata gtgtcaccta aatcgtatgt gtatgataca 60
taaggttatg tattaattgt agccgcgttc taacgacaat atgtccatat ggtgcactct 120
cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc 180
tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt 240
ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa 300
gggcctcgtg atacgcctat ttttataggt taatgtcatg accaaaatcc cttaacgtga 360
gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc 420
tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt 480
ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc 540
gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc 600
tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 660 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 720
gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga 780
actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc 840
ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 900
gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg 960
atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt 1020
tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc 1080
tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg 1140
aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc 1200
gcctctcccc gcgcgttggc cgattcatta atgcaggttg atcagatctc gatcccgcga 1260
aattaatacg actcactata gggagaccac aacggtttcc ctctagaaat aattttgttt 1320
aactttaaga aggagatata cccatggaaa agcctgaact caccgcgacg tctgtcgaga 1380
agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag 1440
aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct 1500
gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc 1560
cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc 1620
gccgtgcaca aggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc 1680
agccggtcgc ggaggctatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt 1740
tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg 1800
cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt 1860
ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc 1920
acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag 1980
cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct 2040
tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc 2100
atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc 2160
aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat 2220
gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa 2280
gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc 2340
ccagcactcg tccgagggca aaggaatagt gaggtacagc ttggatcgat ccggctgcta 2400
acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac 24 60
cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg 2520 gatgatcggg cgcgccgtcg acggatccgt acgagatccg gccgggtacg caaaggcaaa 2580
gatttaaact cgaaaacatt acaaaagtct caaaacagag gcaaggccat gcacaaagca 2640
cactctaagt gcttccattg cctactaagt agggtacgta cacgatcacc attcaccagt 2700
gatgatcttt attaatatac aacacactca gagacagctt atgttatagc tagctagcat 2760
aaactatcac atcatgtgtt agtacgacaa gtgacaacat tgcttttaac ttcgcggcct 2820
tggatcctct agaccggata taatgagccg taaacaaaga tgattaagta gtaattaata 2880
cgtactagta aaagtggcaa aagataacga gaaagaacca atttctttgc attcggcctt 2940
agcggaaggc atatataagc tttgattatt ttatttagtg taatgatttc gtacaaccaa 3000
agcatttatt tagtactctc acacttgtgt cgcggccgcc tactcttcct tgggacggag 3060
tccaagaaca cgcaagtgct ccaaatgtga agcaaatgct tgccaaaacg tatccttgac 3120
aaggtatgga accttgtact cgctgcaggt gttcttgatg atggccagaa tatcgggata 3180
atggtgctgc gacacgttgg ggaacagatg gtgcacagcc tggtagttca agctgccagt 3240
gatgctggtc cagaggtgcg aatcgtgtgc gtaatcctgc gtagtctcga cctgcatagc 3300
tgcccagtcc ttttggatga tcccgttctc gtcaggcaac ggccactgaa cttcctcaac 3360
aacgtggttc gcctggaagg tcagcgccag ccagtaagac gacaccatgt ccgcgaccgt 3420
gaacaagagc agcaccttgc ccaggggcag atactgcagg ggaacaatca ggcgatacca 3480
gacaaagaaa gccttgccgc cccagaacat cacagtgtgc catgtcgaga tgggattgac 3540
acgaatagcg tcattggtct tgacaaagta caaaatgttg atgtcctgaa tgcgcacctt 3600
gaacgccagc agtccgtaca ggaaaggaac aaacatgtgc tggttgatgt ggttgacaaa 3660
ccacttttgg ttgggcttga tacgacgaac atcgggctca gacgtcgaca cgtcgggatc 3720
tgctccagca atgttggtgt aggggtgatg gccgagcata tgttggtaca tccacaccag 3780
gtacgatgct ccgttgaaaa agtcgtgcgt ggctcccaga atcttccaga cagtggggtt 3840
gtgggtcact gaaaagtgag acgcatcatg aagagggttg agtccgactt gtgcgcacgc 3900
aaatcccatg atgattgcaa acaccacctg aagccatgtg cgttcgacaa cgaaaggcac 3960
aaagagctgc gcgtagtagg aagcgatcaa ggatccaaag ataagagcgt atcgtcccca 4020
gatctctggt ctattcttgg gatcaatgtt ccgatccgta aagtagccct cgactctcgt 4080
cttgatggtt ttgtggaaca ccgttggctc cgggaagatg ggcagctcat tcgagaccag 4140
tgtaccgaca tagtacttct tcataatggc atctgcagcc ccaaacgcgt gatacatctc 4200
aaagaccgga gtaacatctc ggccagctcc gagcaggaga gtgtccactc caccaggatg 4260
gcggctcaag aactttgtga catcgtacac cctgccgcgg atggccaaga gtaggtcgtc 4320
cttggtgtta tgggccgcca gctcttccca ggtgaaggtt tttccttggt ccgttcccat 4380 gcggccgctt ggggggctat ggaagacttt cttagttagt tgtgtgaata agcaatgttg 4440
ggagaatcgg gactacttat aggataggaa taaaacagaa aagtattaag tgctaatgaa 4500
atatttagac tgataattaa aatcttcacg tatgtccact tgatataaaa acgtcaggaa 4560
taaaggaagt acagtagaat ttaaaggtac tctttttata tatacccgtg ttctcttttt 4620
ggctagctag ttgcataaaa aataatctat atttttatca ttattttaaa tatcttatga 4680
gatggtaaat atttatcata atttttttta ctattattta ttatttgtgt gtgtaataca 4740
tatagaagtt aattacaaat tttatttact ttttcattat tttgatatga ttcaccatta 4800
atttagtgtt attatttata atagttcatt ttaatctttt tgtatatatt atgcgtgcag 4860
tacttttttc ctacatataa ctactattac attttattta tataatattt ttattaatga 4920
attttcgtga taatatgtaa tattgttcat tattatttca gattttttaa aaatatttgt 4980
gttattattt atgaaatatg taattttttt agtatttgat tttatgatga taaagtgttc 5040
taaattcaaa agaaggggga aagcgtaaac attaaaaaac gtcatcaaac aaaaacaaaa 5100
tcttgttaat aaagataaaa ctgtttgttt tgatcactgt tatttcgtaa tataaaaaca 5160
ttatttatat ttatattgtt gacaaccaaa tttgcctatc aaatctaacc aatataatgc 5220
atgcgtggca ggtaatgtac taccatgaac ttaagtcatg acataataaa ccgtgaatct 5280
gaccaatgca tgtacctanc taaattgtat ttgtgacacg aagcaaatga ttcaattcac 5340
aatggagatg ggaaacaaat aatgaagaac ccagaactaa gaaagctttt ctgaaaaata 5400
aaataaaggc aatgtcaaaa gtatactgca tcatcagtcc agaaagcaca tgatattttt 5460
ttatcagtat caatgcagct agttttattt tacaatatcg atatagctag tttaaatata 5520
ttgcagctag atttataaat atttgtgtta ttatttatca tttgtgtaat cctgttttta 5580
gtattttagt ttatatatga tgataatgta ttccaaattt aaaagaaggg aaataaattt 5640
aaacaagaaa aaaagtcatc aaacaaaaaa caaatgaaag ggtggaaaga tgttaccatg 5700
taatgtgaat gttacagtat ttcttttatt atagagttaa caaattaact aatatgattt 5760
tgttaataat gataaaatat tttttttatt attatttcat aatataaaaa tagtttactt 5820
aatataaaaa aaattctatc gttcacaaca aagttggcca cctaatttaa ccatgcatgt 5880
acccatggac catattaggt aaccatcaaa cctgatgaag agataaagag atgaagactt 5940
aagtcataac acaaaaccat aaaaaacaaa aatacaatca accgtcaatc tgaccaatgc 6000
atgaaaaagc tgcaatagtg agtggcgaca caaagcacat gattttctta caacggagat 6060
aaaaccaaaa aaatatttca tgaacaacct agaacaaata aagcttttat ataataaata 6120
tataaataaa taaaggctat ggaataatat acttcaatat atttggatta aataaattgt 6180
tggcggggtt gatatattta tacacaccta aagtcacttc aatctcattt tcacttaact 6240 tttatttttt ttttcttttt atttatcata aagagaatat tgataatata ctttttaaca 6300
tatttttatg acatttttta ttggtgaaaa cttattaaaa atcataaatt ttgtaagtta 6360
gatttattta aagagttcct cttcttattt taaatttttt aataaatttt taaataacta 6420
aaatttgtgt taaaaatgtt aaaaaatgtg ttattaaccc ttctcttcga ggacgtaccc 6480
ggccagatcc tgcaggccaa ctgcgtttgg ggctccagat taaacgacgc cgtttcgttc 6540
ctttcgcttc acggcttaac gatgtcgttt ctgtctgtgc ccaaaaaata aaggcatttg 6600
ttatttgcac cagatattta ctaagtgcac cctagtttga caagtaggcg ataattacaa 6660
atagatgcgg tgcaaataat aaattttgaa ggaaataatt acaaaagaac agaacttata 6720
tttactttat tttaaaaaac taaaatgaaa gaacaaaaaa agtaaaaaat acaaaaaatg 6780
tgctttaacc actttcatta tttgttacag aaagtatgat tctactcaaa ttgatctgtt 6840
gtatctggtg ctgccttgtc acactggcga tttcaatccc ctaaagatat ggtgcaaact 6900
gcgaagtgat caatatctgc tcggttaatt tagattaatt aataatattc aacgtgatgt 6960
accaaaaaaa gacaattttt tgctccattg acaaattaaa cctcatcaag gtaatttcca 7020
aacctataag caaaaaaatt tcacattaat tggcccgcaa tcctattagt cttattatac 7080
tagagtagga aaaaaaacaa ttacacaact tgtcttatta ttctctatgc taatgaatat 7140
ttttcccttt tgttagaaat cagtgtttcc taatttattg agtattaatt ccactcaccg 7200
catatattta ccgttgaata agaaaatttt acacataatt ctttttaaga taaataattt 7260
ttttatacta gatcttatat gattacgtga agccaagtgg gttatactaa tgatatataa 7320
tgtttgatag taatcagttt ataaaccaaa tgcatggaaa tgttacgtgg aagcacgtaa 7380
attaacaagc attgaagcaa atgcagccac cgcaccaaaa ccaccccact tcacttccac 7440
gtaccatatt ccatgcaact acaacaccct aaaacttcaa taaatgcccc caccttcact 7500
tcacttcacc catcaatagc aagcggccgc accatggagg tggtgaatga aatagtctca 7560
attgggcagg aagttttacc caaagttgat tatgcccaac tctggagtga tgccagtcac 7620
tgtgaggtgc tttacttgtc catcgcattt gtcatcttga agttcactct tggccccctt 7680
ggtccaaaag gtcagtctcg tatgaagttt gttttcacca attacaacct tctcatgtcc 7740
atttattcgt tgggatcatt cctctcaatg gcatatgcca tgtacaccat cggtgttatg 7800
tctgacaact gcgagaaggc ttttgacaac aacgtcttca ggatcaccac gcagttgttc 7860
tatttgagca agttcctgga gtatattgac tccttctatt tgccactgat gggcaagcct 7920
ctgacctggt tgcaattctt ccatcatttg ggggcaccga tggatatgtg gctgttctat 7980
aattaccgaa atgaagctgt ttggattttt gtgctgttga atggtttcat ccactggatc 8040
atgtacggtt attattggac cagattgatc aagctgaagt tccccatgcc aaaatccctg 8100 attacatcaa tgcagatcat tcaattcaat gttggtttct acattgtctg gaagtacagg 8160 aacattccct gttatcgcca agatgggatg aggatgtttg gctggttctt caattacttt 8220 tatgttggca cagtcttgtg tttgttcttg aatttctatg tgcaaacgta tatcgtcagg 8280 aagcacaagg gagccaaaaa gattcagtga gcggccgcga agttaaaagc aatgttgtca 8340 cttgtcgtac taacacatga tgtgatagtt tatgctagct agctataaca taagctgtct 8400 ctgagtgtgt tgtatattaa taaagatcat cactggtgaa tggtgatcgt gtacgtaccc 8460 tacttagtag gcaatggaag cacttagagt gtgctttgtg catggccttg cctctgtttt 8520 gagacttttg taatgttttc gagtttaaat ctttgccttt gcgtacgtct agagtcgacc 8580 tgca 8584 <210> 8 <211> 5754 <212> DNA <213> Seqüência Artificial <220> <223> plasmideo pKR1002 <400> 8 ggccgcaagt atgaactaaa atgcatgtag gtgtaagagc tcatggagag catggaatat 60 tgtatccgac catgtaacag tataataact gagctccatc tcacttcttc tatgaataaa 12 0 caaaggatgt tatgatatat taacactcta tctatgcacc ttattgttct atgataaatt 180 tcctcttatt attataaatc atctgaatcg tgacggctta tggaatgctt caaatagtac 240 aaaaacaaat gtgtactata agactttcta aacaattcta accttagcat tgtgaacgag 300 acataagtgt taagaagaca taacaattat aatggaagaa gtttgtctcc atttatatat 360 tatatattac ccacttatgt attatattag gatgttaagg agacataaca attataaaga 420 gagaagtttg tatccattta tatattatat actacccatt tatatattat acttatccac 480 ttatttaatg tctttataag gtttgatcca tgatatttct aatattttag ttgatatgta 540 tatgaaaggg tactatttga actctcttac tctgtataaa ggttggatca tccttaaagt 600 gggtctattt aattttattg cttcttacag ataaaaaaaa aattatgagt tggtttgata 660 aaatattgaa ggatttaaaa taataataaa taacatataa tatatgtata taaatttatt 720 ataatataac atttatctat aaaaaagtaa atattgtcat aaatctatac aatcgtttag 780 ccttgctgga cgaatctcaa ttatttaaac gagagtaaac atatttgact ttttggttat 840 ttaacaaatt attatttaac actatatgaa attttttttt ttatcagcaa agaataaaat 900 taaattaaga aggacaatgg tgtcccaatc cttatacaac caacttccac aagaaagtca 960 agtcagagac aacaaaaaaa caagcaaagg aaatttttta atttgagttg tcttgtttgc 1020 tgcataattt atgcagtaaa acactacaca taaccctttt agcagtagag caatggttga 1080
ccgtgtgctt agcttctttt attttatttt tttatcagca aagaataaat aaaataaaat 1140
gagacacttc agggatgttt caacctgcag gtctagagga tccccgggta ccgagctcga 1200
attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta 1260
atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg 1320
atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc 1380
tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct 1440
ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac 1500
gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca 1560
tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac 1620
gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt 1680
ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt 1740
atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta 1800
tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg 1860
tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac 1920
gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg 1980
aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc 2040
gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg 2100
ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat 2160
gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg 2220
gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg 2280
atcgttggga accggagctg aatgaagcca tacçaaacga cgagcgtgac accacgatgc 234 0
ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt 2400
cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct 2460
cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc 2520
gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca 2580
cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct 2640
cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt 2700
taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga 2760
ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca 2820
aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac 2880 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg 2940
taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag 3000
gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac 3060
cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt 3120
taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg 3180
agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc 3240
ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc 3300
gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc 3360
acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa 3420
acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt 3480
tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg 3540
ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag 3600
agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 3660
acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 3720
tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 3780
ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg 3840
catgcctgca ggtcgactct agacctgcag gatccatgcc cttcatttgc cgcttattaa 3900
ttaatttggt aacagtccgt actaatcagt tacttatcct tcccccatca taattaatct 3960
tggtagtctc gaatgccaca acactgacta gtctcttgga tcataagaaa aagccaagga 4020
acaaaagaag acaaaacaca atgagagtat cctttgcata gcaatgtcta agttcataaa 4080
attcaaacaa aaacgcaatc acacacagtg gacatcactt atccactagc tgatcaggat 4140
cgccgcgtca agaaaaaaaa actggacccc aaaagccatg cacaacaaca cgtactcaca 4200
aaggtgtcaa tcgagcagcc caaaacattc accaactcaa cccatcatga gccctcacat 4260
ttgttgtttc taacccaacc tcaaactcgt attctcttcc gccacctcat ttttgtttat 4320
ttcaacaccc gtcaaactgc atgccacccc gtggccaaat gtccatgcat gttaacaaga 4380
cctatgacta taaatagctg caatctcggc ccaggttttc atcatcaaga accagttcaa 4440
tatcctagta caccgtatta aagaatttaa gatatactgc ggccgcacca tgtctcctaa 4500
gcggcaagct ctgccaatca caattgatgg cgcaacttat gatgtgtctg cttgggtcaa 4560
tcaccaccct ggaggagctg acattatcga gaactatcgc aaccgcgatg cgaccgatgt 4620
cttcatggtg atgcactctc aagaagccgt cgccaagttg aagagaatgc ctgttatgga 4680
gccttcctct cctgacacac ctgttgcacc caagcctaag cgtgatgagc cccaggagga 4740 tttccgcaag ttgcgggagg aattcatctc caagggtatg ttcgagacga gtttcctttg 4800
gtatttttac aagacttcaa ctaccgtcgg tttgatggtc ctttccatct tgatgaccgt 4860
gtacacgaat tggtatttca ccgctgcttt ggttcttggc gtgtgctacc aacagctagg 4920
ctggttgtcc cacgactatt gccatcacca ggttttcaca aaccgcaaga ttaacgacgc 4980
tttcggtctc tttttcggta acgtgatgca gggatactca cagacttggt ggaaggatag 5040
gcacaatggt caccatgccg ccaccaatgt ggtcggccat gacccagata ttgataacct 5100
ccccatcctg gcttggtctc ccgaagatgt caagagggct actccttcga ctcggaatct 5160
catcaagtac cagcagtact acttcattcc caccattgca tcccttaggt tcatctggtg 5220
cctccaatcc atcggcggcg tcatgtccta caagagcgag gagaggaacc tgtactacaa 5280
gcgccagtac actaaggagg cgattggtct ggccctccac tgggtgctca aggccacttt 5340
ctattgcagt gccatgccta gctttgccac cggtttggga tgcttcttga tctccgagct 5400
gctcggagga tttggcattg ccatcgttgt gtttctgaat cactatcctt tggacaaggt 5460
tgaggagact gtctgggatg agcacgggtt cagcgccagc cagatccacg agacgttgaa 5520
cattaagccc ggccttctca ccgattgggt ctttggtggt ctcaactacc agattgagca 5580
ccacttgtgg cccaacatgc ccaggcacaa cctcacggca gcttccctgg aggtgcagaa 5640
gttgtgcgcc aagcacaacc tgccctacag ggccccagcc atcatccccg gggttcagaa 5700
attggtcagc ttcttaggcg agattgccca gctggctgct gtccctgaat gagc 5754
<210> 9 <211> 6524 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmídeo pKR226 <400> 9
gtacgtctag aggatccgtc gacggcgcgc ccgatcatcc ggatatagtt cctcctttca 60
gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt tattgctcag 120
cggtggcagc agccaactca gcttcctttc gggctttgtt agcagccgga tcgatccaag 180
ctgtacctca ctattccttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg 240
cgagtacttc tacacagcca tcggtccaga cggccgcgct tctgcgggcg atttgtgtac 300
gcccgacagt cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg 360
catcatcgaa attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca 420
tatacgcccg gagccgcggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg 480
tctgctgctc catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt 540 gggaatcccc gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg 600
tcaggacatt gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct 660
cggcccaaag catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca 720
tcacagtttg ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg 780
tagtgtattg accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat 840
cggccgcagc gatcgcatcc atagcctccg cgaccggctg cagaacagcg ggcagttcgg 900
tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc 960
tctcgctgaa ttccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt 1020
gccgataaac ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat 1080
atccacgccc tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca 1140
tcaggtcgga gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga 1200
gttcaggctt ttccatgggt atatctcctt cttaaagtta aacaaaatta tttctagagg 1260
gaaaccgttg tggtctccct atagtgagtc gtattaattt cgcgggatcg agatctgatc 1320
aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 1380
tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 1440
gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 1500
atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 1560
ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 1620
cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 1680
tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 1740
gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 1800
aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 1860
tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 1920
aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 1980
aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 2040
ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 2100
ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 2160
atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 2220
atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt 2280
gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa 2340
gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg 2400 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatggacata 2460
ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca catacgattt 2520
aggtgacact atagaacggc gcgccaagct gggtctagaa ctagaaacgt gatgccactt 2580
gttattgaag tcgattacag catctattct gttttactat ttataacttt gccatttctg 2640
acttttgaaa actatctctg gatttcggta tcgctttgtg aagatcgagc aaaagagacg 2700
ttttgtggac gcaatggtcc aaatccgttc tacatgaaca aattggtcac aatttccact 2760
aaaagtaaat aaatggcaag ttaaaaaagg aatatgcatt ttactgattg cctaggtgag 2820
ctccaagaga agttgaatct acacgtctac caaccgctaa aaaaagaaaa acattgatat 2880
gtaacctgat tccattagct tttgacttct tcaacagatt ctctacttag atttctaaca 2940
gaaatattat tactagcaca tcattttcag tctcactaca gcaaaaaatc caacggcaca 3000
atacagacaa caggagatat cagactacag agatagatag atgctactgc atgtagtaag 3060
ttaaataaaa ggaaaataaa atgtcttgct accaaaacta ctacagacta tgatgctcac 3120
cacaggccaa atcctgcaac taggacagca ttatcttata tatattgtac aaaacaagca 3180
tcaaggaaca tttggtctag gcaatcagta cctcgttcta ccatcaccct cagttatcac 3240
atccttgaag gatccattac tgggaatcat cggcaacaca tgctcctgat ggggcacaat 3300
gacatcaaga aggtaggggc caggggtgtc caacattctc tgaattgccg ctctaagctc 3360
ttccttcttc gtcactcgcg ctgccggtat cccacaagca tcagcaaact tgagcatgtt 3420
tgggaatatc tcgctctcgc tagacggatc tccaagatag gtgtgagctc tattggactt 3480
gtagaaccta tcctccaact gaaccaccat acccaaatgc tgattgttca acaacaatat 3540
cttaactggg agattctcca ctcttatagt ggccaactcc tgaacattca tgatgaaact 3600
accatcccca tcaatgtcaa ccacaacagc cccagggtta gcaacagcag caccaatagc 3660
cgcaggcaat ccaaaaccca tggctccaag accccctgag gtcaaccact gcctcggtct 3720
cttgtacttg taaaactgcg cagcccacat ttgatgctgc ccaaccccag tactaacaat 3780
agcatctcca ttagtcaact catcaagaac ctcgatagca tgctgcggag aaatcgcgtc 3840
ctggaatgtc ttgtaaccca atggaaactt gtgtttctgc acattaatct cttctctcca 3900
acctccaaga tcaaacttac cctccactcc tttctcctcc aaaatcatat taattccctt 3960
caaggccaac ttcaaatccg cgcaaaccga cacgtgcgcc tgcttgttct tcccaatctc 4020
ggcagaatca atatcaatgt gaacaatctt agccctacta gcaaaagcct caagcttccc 4080
agtaacacgg tcatcaaacc ttaccccaaa ggcaagcaac aaatcactat tgtcaacagc 4140
atagttagca taaacagtac catgcatacc cagcatctga agggaatatt catcaccaat 4200
aggaaaagtt ccaagaccca ttaaagtgct agcaacggga ataccagtga gttcaacaaa 4260 gcgcctcaat tcagcactgg aattcaaact gccaccgccg acgtagagaa cgggcttttg 4320 ggcctccatg atgagtctga caatgtgttc caattgggcc tcggcggggg gcctgggcag 4380
cctggcgagg taaccgggga ggttaacggg ctcgtcccaa ttaggcacgg cgagttgctg 4440
ctgaacgtct ttgggaatgt cgatgaggac cggaccgggg cggccggagg tggcgacgaa 4500
gaaagcctcg gcgacgacgc gggggatgtc gtcgacgtcg aggatgaggt agttgtgctt 4560
cgtgatggat ctgctcacct ccacgatcgg ggtttcttgg aaggcgtcgg tgccgatcat 4620
ccggcgggcg acctggccgg tgatggcgac gactgggacg ctgtccatta aagcgtcggc 4 680
gaggccgctc acgaggttgg tggcgccggg gccggaggtg gcaatgcaga cgccggggag 4740
gccggaggaa cgcgcgtagc cttcggcggc gaagacgccg ccctgctcgt ggcgcgggag 4800
cacgttgcgg atggcggcgg agcgcgtgag cgcctggtgg atctccatcg acgcaccgcc 4860
ggggtacgcg aacaccgtcg tcacgccctg cctctccagc gcctccacaa ggatgtccgc 4920
gcccttgcga ggttcgccgg aggcgaaccg tgacacgaag ggctccgtgg tcggcgcttc 4980
cttggtgaag ggcgccgccg tggggggttt ggagatggaa catttgattt tgagagcgtg 5040
gttgggtttg gtgagggttt gatgagagag agggagggtg gatctagtaa tgcgtttggg 5100
gaaggtgggg tgtgaagagg aagaagagaa tcgggtggtt ctggaagcgg tggccgccat 5160
tgtgttgtgt ggcatggtta tacttcaaaa actgcacaac aagcctagag ttagtaccta 5220
aacagtaaat ttacaacaga gagcaaagac acatgcaaaa atttcagcca taaaaaaagt 5280
tataatagaa tttaaagcaa aagtttcatt ttttaaacat atatacaaac aaactggatt 5340
tgaaggaagg gattaattcc cctgctcaaa gtttgaattc ctattgtgac ctatactcga 5400
ataaaattga agcctaagga atgtatgaga aacaagaaaa caaaacaaaa ctacagacaa 54 60
acaagtacaa ttacaaaatt cgctaaaatt ctgtaatcac caaaccccat ctcagtcagc 5520
acaaggccca aggtttattt tgaaataaaa aaaaagtgat tttatttctc ataagctaaa 5580
agaaagaaag gcaattatga aatgatttcg actagatctg aaagtccaac gcgtattccg 5640
cagatattaa agaaagagta gagtttcaca tggatcctag atggacccag ttgaggaaaa 5700
agcaaggcaa agcaaaccag aagtgcaaga tccgaaattg aaccacggaa tctaggattt 5760
ggtagaggga gaagaaaagt accttgagag gtagaagaga agagaagagc agagagatat 5820
atgaacgagt gtgtcttggt ctcaactctg aagcgatacg agtttagagg ggagcattga 5880
gttccaattt atagggaaac cgggtggcag gggtgagtta atgacggaaa agcccctaag 5940
taacgagatt ggattgtggg ttagattcaa ccgtttgcat ccgcggctta gattggggaa 6000
gtcagagtga atctcaaccg ttgactgagt tgaaaattga atgtagcaac caattgagcc 6060
aaccccagcc tttgcccttt gattttgatt tgtttgttgc atacttttta tttgtcttct 6120 ggttctgact ctctttctct cgtttcaatg ccaggttgcc tactcccaca ccactcacaa 6180
gaagattcta ctgttagtat taaatatttt ttaatgtatt aaatgatgaa tgcttttgta 6240
aacagaacaa gactatgtct aataagtgtc ttgcaacatt ttttaagaaa ttaaaaaaaa 6300
tatatttatt atcaaaatca aatgtatgaa aaatcatgaa taatataatt ttatacattt 6360
ttttaaaaaa tcttttaatt tcttaattaa tatcttaaaa ataatgatta atatttaacc 6420
caaaataatt agtatgattg gtaaggaaga tatccatgtt atgtttggat gtgagtttga 6480
tctagagcaa agcttactag agtcgacctg caggtcgact cgac 6524
<210> 10 <211> 9578 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmídeo BC-Tpom_ALS <400> 10
gtacgtctag aggatccgtc gacggcgcgc ccgatcatcc ggatatagtt cctcctttca 60
gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt tattgctcag 120
cggtggcagc agccaactca gcttcctttc gggctttgtt agcagccgga tcgatccaag 180
ctgtacctca ctattccttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg 240
cgagtacttc tacacagcca tcggtccaga cggccgcgct tctgcgggcg atttgtgtac 300
gcccgacagt cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg 360
catcatcgaa attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca 420
tatacgcccg gagccgcggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg 480
tctgctgctc catacaagcc ,aaccacggcc tccagaagaa gatgttggcg acctcgtatt 540
gggaatcccc gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg 600
tcaggacatt gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct 660
cggcccaaag catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca 720
tcacagtttg ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg 780
tagtgtattg accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat 840
cggccgcagc gatcgcatcc atagcctccg cgaccggctg cagaacagcg ggcagttcgg 900
tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc 960
tctcgctgaa ttccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt 1020
gccgataaac ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat 1080
atccacgccc tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca 1140 tcaggtcgga gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga 1200
gttcaggctt ttccatgggt atatctcctt cttaaagtta aacaaaatta tttctagagg 1260
gaaaccgttg tggtctccct atagtgagtc gtattaattt cgcgggatcg agatctgatc 1320
aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 1380
tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 1440
gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 1500
atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 1560
ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 1620
cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 1680
tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 1740
gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 1800
aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 1860
tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 1920
aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 1980
aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 2040
ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 2100
ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 2160
atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 2220
atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt 2280
gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa 2340
gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg 2400
ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatggacata 2460
ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca catacgattt 2520
aggtgacact atagaacggc gcgccaagct gggtctagaa ctagaaacgt gatgccactt 2580
gttattgaag tcgattacag catctattct gttttactat ttataacttt gccatttctg 2640
acttttgaaa actatctctg gatttcggta tcgctttgtg aagatcgagc aaaagagacg 2700
ttttgtggac gcaatggtcc aaatccgttc tacatgaaca aattggtcac aatttccact 2760
aaaagtaaat aaatggcaag ttaaaaaagg aatatgcatt ttactgattg cctaggtgag 2820
ctccaagaga agttgaatct acacgtctac caaccgctaa aaaaagaaaa acattgatat 2880
gtaacctgat tccattagct tttgacttct tcaacagatt ctctacttag atttctaaca 2940
gaaatattat tactagcaca tcattttcag tctcactaca gcaaaaaatc caacggcaca 3000 atacagacaa caggagatat cagactacag agatagatag atgctactgc atgtagtaag 3060
ttaaataaaa ggaaaataaa atgtcttgct accaaaacta ctacagacta tgatgctcac 3120
cacaggccaa atcctgcaac taggacagca ttatcttata tatattgtac aaaacaagca 3180
tcaaggaaca tttggtctag gcaatcagta cctcgttcta ccatcaccct cagttatcac 3240
atccttgaag gatccattac tgggaatcat cggcaacaca tgctcctgat ggggcacaat 3300
gacatcaaga aggtaggggc caggggtgtc caacattctc tgaattgccg ctctaagctc 3360
ttccttcttc gtcactcgcg ctgccggtat cccacaagca tcagcaaact tgagcatgtt 3420
tgggaatatc tcgctctcgc tagacggatc tccaagatag gtgtgagctc tattggactt 3480
gtagaaccta tcctccaact gaaccaccat acccaaatgc tgattgttca acaacaatat 3540
cttaactggg agattctcca ctcttatagt ggccaactcc tgaacattca tgatgaaact 3600
accatcccca tcaatgtcaa ccacaacagc cccagggtta gcaacagcag caccaatagc 3660
cgcaggcaat ccaaaaccca tggctccaag accccctgag gtcaaccact gcctcggtct 3720
cttgtacttg taaaactgcg cagcccacat ttgatgctgc ccaaccccag tactaacaat 3780
agcatctcca ttagtcaact catcaagaac ctcgatagca tgctgcggag aaatcgcgtc 3840
ctggaatgtc ttgtaaccca atggaaactt gtgtttctgc acattaatct cttctctcca 3900
acctccaaga tcaaacttac cctccactcc tttctcctcc aaaatcatat taattccctt 3960
caaggccaac ttcaaatccg cgcaaaccga cacgtgcgcc tgcttgttct tcccaatctc 4020
ggcagaatca atatcaatgt gaacaatctt agccctacta gcaaaagcct caagcttccc 4080
agtaacacgg tcatcaaacc ttaccccaaa ggcaagcaac aaatcactat tgtcaacagc 4140
atagttagca taaacagtac catgcatacc cagcatctga agggaatatt catcaccaat 4200
aggaaaagtt ccaagaccca ttaaagtgct agcaacggga ataccagtga gttcaacaaa 4260
gcgcctcaat tcagcactgg aattcaaact gccaccgccg acgtagagaa cgggcttttg 4320
ggcctccatg atgagtctga caatgtgttc caattgggcc tcggcggggg gcctgggcag 4380
cctggcgagg taaccgggga ggttaacggg ctcgtcccaa ttaggcacgg cgagttgctg 4440
ctgaacgtct ttgggaatgt cgatgaggac cggaccgggg cggccggagg tggcgacgaa 4500
gaaagcctcg gcgacgacgc gggggatgtc gtcgacgtcg aggatgaggt agttgtgctt 4560
cgtgatggat ctgctcacct ccacgatcgg ggtttcttgg aaggcgtcgg tgccgatcat 4 620
ccggcgggcg acctggccgg tgatggcgac gactgggacg ctgtccatta aagcgtcggc 4680
gaggccgctc acgaggttgg tggcgccggg gccggaggtg gcaatgcaga cgccggggag 4740
gccggaggaa cgcgcgtagc cttcggcggc gaagacgccg ccctgctcgt ggcgcgggag 4800
cacgttgcgg atggcggcgg agcgcgtgag cgcctggtgg atctccatcg acgcaccgcc 4 860 ggggtacgcg aacaccgtcg tcacgccctg cctctccagc gcctccacaa ggatgtccgc 4920
gcccttgcga ggttcgccgg aggcgaaccg tgacacgaag ggctccgtgg tcggcgcttc 4980
cttggtgaag ggcgccgccg tggggggttt ggagatggaa catttgattt tgagagcgtg 5040
gttgggtttg gtgagggttt gatgagagag agggagggtg gatctagtaa tgcgtttggg 5100
gaaggtgggg tgtgaagagg aagaagagaa tcgggtggtt ctggaagcgg tggccgccat 5160
tgtgttgtgt ggcatggtta tacttcaaaa actgcacaac aagcctagag ttagtaccta 5220
aacagtaaat ttacaacaga gagcaaagac acatgcaaaa atttcagcca taaaaaaagt 5280
tataatagaa tttaaagcaa aagtttcatt ttttaaacat atatacaaac aaactggatt 5340
tgaaggaagg gattaattcc cctgctcaaa gtttgaattc ctattgtgac ctatactcga 5400
ataaaattga agcctaagga atgtatgaga aacaagaaaa caaaacaaaa ctacagacaa 5460
acaagtacaa ttacaaaatt cgctaaaatt ctgtaatcac caaaccccat ctcagtcagc 5520
acaaggccca aggtttattt tgaaataaaa aaaaagtgat tttatttctc ataagctaaa 5580
agaaagaaag gcaattatga aatgatttcg actagatctg aaagtccaac gcgtattccg 5640
cagatattaa agaaagagta gagtttcaca tggatcctag atggacccag ttgaggaaaa 5700
agcaaggcaa agcaaaccag aagtgcaaga tccgaaattg aaccacggaa tctaggattt 5760
ggtagaggga gaagaaaagt accttgagag gtagaagaga agagaagagc agagagatat 5820
atgaacgagt gtgtcttggt ctcaactctg aagcgatacg agtttagagg ggagcattga 5880
gttccaattt atagggaaac cgggtggcag gggtgagtta atgacggaaa agcccctaag 5940
taacgagatt ggattgtggg ttagattcaa ccgtttgcat ccgcggctta gattggggaa 6000
gtcagagtga atctcaaccg ttgactgagt tgaaaattga atgtagcaac caattgagcc 6060
aaccccagcc tttgcccttt gattttgatt tgtttgttgc atacttttta tttgtcttct 6120
ggttctgact ctctttctct cgtttcaatg ccaggttgcc tactcccaca ccactcacaa 6180
gaagattcta ctgttagtat taaatatttt ttaatgtatt aaatgatgaa tgcttttgta 6240
aacagaacaa gactatgtct aataagtgtc ttgcaacatt ttttaagaaa ttaaaaaaaa 6300
tatatttatt atcaaaatca aatgtatgaa aaatcatgaa taatataatt ttatacattt 6360
ttttaaaaaa tcttttaatt tcttaattaa tatcttaaaa ataatgatta atatttaacc 6420
caaaataatt agtatgattg gtaaggaaga tatccatgtt atgtttggat gtgagtttga 6480
tctagagcaa agcttactag agtcgacctg caggatccat gcccttcatt tgccgcttat 6540
taattaattt ggtaacagtc cgtactaatc agttacttat ccttccccca tcataattaa 6600
tcttggtagt ctcgaatgcc acaacactga ctagtctctt ggatcataag aaaaagccaa 6660
ggaacaaaag aagacaaaac acaatgagag tatcctttgc atagcaatgt ctaagttcat 6720 aaaattcaaa caaaaacgca atcacacaca gtggacatca cttatccact agctgatcag 6780 gatcgccgcg tcaagaaaaa aaaactggac cccaaaagcc atgcacaaca acacgtactc 6840 acaaaggtgt caatcgagca gcccaaaaca ttcaccaact caacccatca tgagccctca 6900 catttgttgt ttctaaccca acctcaaact cgtattctct tccgccacct catttttgtt 6960 tatttcaaca cccgtcaaac tgcatgccac cccgtggcca aatgtccatg catgttaaca 7020 agacctatga ctataaatag ctgcaatctc ggcccaggtt ttcatcatca agaaccagtt 7080 caatatccta gtacaccgta ttaaagaatt taagatatac tgcggccgca ccatgtctcc 7140 taagcggcaa gctctgccaa tcacaattga tggcgcaact tatgatgtgt ctgcttgggt 7200 caatcaccac cctggaggag ctgacattat cgagaactat cgcaaccgcg atgcgaccga 7260 tgtcttcatg gtgatgcact ctcaagaagc cgtcgccaag ttgaagagaa tgcctgttat 7320 ggagccttcc tctcctgaca cacctgttgc acccaagcct aagcgtgatg agccccagga 7380 ggatttccgc aagttgcggg aggaattcat ctccaagggt atgttcgaga cgagtttcct 7440 ttggtatttt tacaagactt caactaccgt cggtttgatg gtcctttcca tcttgatgac 7500 cgtgtacacg aattggtatt tcaccgctgc tttggttctt ggcgtgtgct accaacagct 7560 aggctggttg tcccacgact attgccatca ccaggttttc acaaaccgca agattaacga 7620 cgctttcggt ctctttttcg gtaacgtgat gcagggatac tcacagactt ggtggaagga 7680 taggcacaat ggtcaccatg ccgccaccaa tgtggtcggc catgacccag atattgataa 7740 cctccccatc ctggcttggt ctcccgaaga tgtcaagagg gctactcctt cgactcggaa 7800 tctcatcaag taccagcagt actacttcat tcccaccatt gcatccctta ggttcatctg 7860 gtgcctccaa tccatcggcg gcgtcatgtc ctacaagagc gaggagagga acctgtacta 7920 caagcgccag tacactaagg aggcgattgg tctggccctc cactgggtgc tcaaggccac 7980 tttctattgc agtgccatgc ctagctttgc caccggtttg ggatgcttct tgatctccga 8040 gctgctcgga ggatttggca ttgccatcgt tgtgtttctg aatcactatc ctttggacaa 8100 ggttgaggag actgtctggg atgagcacgg gttcagcgcc agccagatcc acgagacgtt 8160 gaacattaag cccggccttc tcaccgattg ggtctttggt ggtctcaact accagattga 8220 gcaccacttg tggcccaaca tgcccaggca caacctcacg gcagcttccc tggaggtgca 8280 gaagttgtgc gccaagcaca acctgcccta cagggcccca gccatcatcc ccggggttca 8340 gaaattggtc agcttcttag gcgagattgc ccagctggct gctgtccctg aatgagcggc 8400 cgcaagtatg aactaaaatg catgtaggtg taagagctca tggagagcat ggaatattgt 8460 atccgaccat gtaacagtat aataactgag ctccatctca cttcttctat gaataaacaa 8520 aggatgttat gatatattaa cactctatct atgcacctta ttgttctatg ataaatttcc 8580 tcttattatt ataaatcatc tgaatcgtga cggcttatgg aatgcttcaa atagtacaaa 8640
aacaaatgtg tactataaga ctttctaaac aattctaacc ttagcattgt gaacgagaca 8700
taagtgttaa gaagacataa caattataat ggaagaagtt tgtctccatt tatatattat 8760
atattaccca cttatgtatt atattaggat gttaaggaga cataacaatt ataaagagag 8820
aagtttgtat ccatttatat attatatact acccatttat atattatact tatccactta 8880
tttaatgtct ttataaggtt tgatccatga tatttctaat attttagttg atatgtatat 8940
gaaagggtac tatttgaact ctcttactct gtataaaggt tggatcatcc ttaaagtggg 9000
tctatttaat tttattgctt cttacagata aaaaaaaaat tatgagttgg tttgataaaa 9060
tattgaagga tttaaaataa taataaataa catataatat atgtatataa atttattata 9120
atataacatt tatctataaa aaagtaaata ttgtcataaa tctatacaat cgtttagcct 9180
tgctggacga atctcaatta tttaaacgag agtaaacata tttgactttt tggttattta 9240
acaaattatt atttaacact atatgaaatt ttttttttta tcagcaaaga ataaaattaa 9300
attaagaagg acaatggtgt cccaatcctt atacaaccaa cttccacaag aaagtcaagt 9360
cagagacaac aaaaaaacaa gcaaaggaaa ttttttaatt tgagttgtct tgtttgctgc 9420
ataatttatg cagtaaaaca ctacacataa cccttttagc agtagagcaa tggttgaccg 9480
tgtgcttagc ttcttttatt ttattttttt atcagcaaag aataaataaa ataaaatgag 9540
acacttcagg gatgtttcaa cctgcaggtc gactcgac 9578
<210> 11 <211> 9578 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmídeo BC-Tpom_ALSrev <400> 11
gtacgtctag aggatccgtc gacggcgcgc ccgatcatcc ggatatagtt cctcctttca 60
gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt tattgctcag 120
cggtggcagc agccaactca gcttcctttc gggctttgtt agcagccgga tcgatccaag 180
ctgtacctca ctattccttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg 240
cgagtacttc tacacagcca tcggtccaga cggccgcgct tctgcgggcg atttgtgtac 300
gcccgacagt cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg 360
catcatcgaa attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca 420
tatacgcccg gagccgcggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg 480
tctgctgctc catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt 540 gggaatcccc gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg 600
tcaggacatt gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct 660
cggcccaaag catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca 720
tcacagtttg ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg 780
tagtgtattg accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat 840
cggccgcagc gatcgcatcc atagcctccg cgaccggctg cagaacagcg ggcagttcgg 900
tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc 960
tctcgctgaa ttccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt 1020
gccgataaac ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat 1080
atccacgccc tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca 1140
tcaggtcgga gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga 1200
gttcaggctt ttccatgggt atatctcctt cttaaagtta aacaaaatta tttctagagg 1260
gaaaccgttg tggtctccct atagtgagtc gtattaattt cgcgggatcg agatctgatc 1320
aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 1380
tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 1440
gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 1500
atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 1560
ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 1620
cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 1680
tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 1740
gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 1800
aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 1860
tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 1920
aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 1980
aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 2040
ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 2100
ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 2160
atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 2220
atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt 2280
gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa 2340
gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg 2400 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatggacata 2460 ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca catacgattt 2520 aggtgacact atagaacggc gcgccaagct gggtctagaa ctagaaacgt gatgccactt 2580 gttattgaag tcgattacag catctattct gttttactat ttataacttt gccatttctg 2640 acttttgaaa actatctctg gatttcggta tcgctttgtg aagatcgagc aaaagagacg 2700 ttttgtggac gcaatggtcc aaatccgttc tacatgaaca aattggtcac aatttccact 2760 aaaagtaaat aaatggcaag ttaaaaaagg aatatgcatt ttactgattg cctaggtgag 2820 ctccaagaga agttgaatct acacgtctac caaccgctaa aaaaagaaaa acattgatat 2880 gtaacctgat tccattagct tttgacttct tcaacagatt ctctacttag atttctaaca 2940 gaaatattat tactagcaca tcattttcag tctcactaca gcaaaaaatc caacggcaca 3000 atacagacaa caggagatat cagactacag agatagatag atgctactgc atgtagtaag 3060 ttaaataaaa ggaaaataaa atgtcttgct accaaaacta ctacagacta tgatgctcac 3120 cacaggccaa atcctgcaac taggacagca ttatcttata tatattgtac aaaacaagca 3180 tcaaggaaca tttggtctag gcaatcagta cctcgttcta ccatcaccct cagttatcac 3240 atccttgaag gatccattac tgggaatcat cggcaacaca tgctcctgat ggggcacaat 3300 Ί π gacaccaaga ttccttcttc gtcactcgcg ctgccggtat cccacaagca tcagcaaact tgagcatgtt 3420 tgggaatatc tcgctctcgc tagacggatc tccaagatag gtgtgagctc tattggactt 3480 gtagaaccta tcctccaact gaaccaccat acccaaatgc tgattgttca acaacaatat 3540 cttaactggg agattctcca ctcttatagt ggccaactcc tgaacattca tgatgaaact 3600 accatcccca tcaatgtcaa ccacaacagc cccagggtta gcaacagcag caccaatagc 3660 cgcaggcaat ccaaaaccca tggctccaag accccctgag gtcaaccact gcctcggtct 3720 cttgtacttg taaaactgcg cagcccacat ttgatgctgc ccaaccccag tactaacaat 3780 agcatctcca ttagtcaact catcaagaac ctcgatagca tgctgcggag aaatcgcgtc 3840 ctggaatgtc ttgtaaccca atggaaactt gtgtttctgc acattaatct cttctctcca 3900 acctccaaga tcaaacttac cctccactcc tttctcctcc aaaatcatat taattccctt 3960 caaggccaac ttcaaatccg cgcaaaccga cacgtgcgcc tgcttgttct tcccaatctc 4020 ggcagaatca atatcaatgt gaacaatctt agccctacta gcaaaagcct caagcttccc 4080 agtaacacgg tcatcaaacc ttaccccaaa ggcaagcaac aaatcactat tgtcaacagc 4140 atagttagca taaacagtac catgcatacc cagcatctga agggaatatt catcaccaat 4200 aggaaaagtt ccaagaccca ttaaagtgct agcaacggga ataccagtga gttcaacaaa 4260 gcgcctcaat tcagcactgg aattcaaact gccaccgccg acgtagagaa cgggcttttg 4320
ggcctccatg atgagtctga caatgtgttc caattgggcc tcggcggggg gcctgggcag 4380
cctggcgagg taaccgggga ggttaacggg ctcgtcccaa ttaggcacgg cgagttgctg 4440
ctgaacgtct ttgggaatgt cgatgaggac cggaccgggg cggccggagg tggcgacgaa 4500
gaaagcctcg gcgacgacgc gggggatgtc gtcgacgtcg aggatgaggt agttgtgctt 4560
cgtgatggat ctgctcacct ccacgatcgg ggtttcttgg aaggcgtcgg tgccgatcat 4 620
ccggcgggcg acctggccgg tgatggcgac gactgggacg ctgtccatta aagcgtcggc 4 680
gaggccgctc acgaggttgg tggcgccggg gccggaggtg gcaatgcaga cgccggggag 4740
gccggaggaa cgcgcgtagc cttcggcggc gaagacgccg ccctgctcgt ggcgcgggag 4800
cacgttgcgg atggcggcgg agcgcgtgag cgcctggtgg atctccatcg acgcaccgcc 4860
ggggtacgcg aacaccgtcg tcacgccctg cctctccagc gcctccacaa ggatgtccgc 4920
gcccttgcga ggttcgccgg aggcgaaccg tgacacgaag ggctccgtgg tcggcgcttc 4980
cttggtgaag ggcgccgccg tggggggttt ggagatggaa catttgattt tgagagcgtg 5040
gttgggtttg gtgagggttt gatgagagag agggagggtg gatctagtaa tgcgtttggg 5100
gaaggtgggg tgtgaagagg aagaagagaa tcgggtggtt ctggaagcgg tggccgccat 5160
tgtgttgtgt ggcatggtta tacttcaaaa actgcacaac aagcctagag ttagtaccta 522υ
aacagtaaat ttacaacaga gagcaaagac acatgcaaaa atttcagcca taaaaaaagt 5280
tataatagaa tttaaagcaa aagtttcatt ttttaaacat atatacaaac aaactggatt 5340
tgaaggaagg gattaattcc cctgctcaaa gtttgaattc ctattgtgac ctatactcga 5400
ataaaattga agcctaagga atgtatgaga aacaagaaaa caaaacaaaa ctacagacaa 5460
acaagtacaa ttacaaaatt cgctaaaatt ctgtaatcac caaaccccat ctcagtcagc 5520
acaaggccca aggtttattt tgaaataaaa aaaaagtgat tttatttctc ataagctaaa 5580
agaaagaaag gcaattatga aatgatttcg actagatctg aaagtccaac gcgtattccg 5640
cagatattaa agaaagagta gagtttcaca tggatcctag atggacccag ttgaggaaaa 5700
agcaaggcaa agcaaaccag aagtgcaaga tccgaaattg aaccacggaa tctaggattt 5760
ggtagaggga gaagaaaagt accttgagag gtagaagaga agagaagagc agagagatat 5820
atgaacgagt gtgtcttggt ctcaactctg aagcgatacg agtttagagg ggagcattga 5880
gttccaattt atagggaaac cgggtggcag gggtgagtta atgacggaaa agcccctaag 5940
taacgagatt ggattgtggg ttagattcaa ccgtttgcat ccgcggctta gattggggaa 6000
gtcagagtga atctcaaccg ttgactgagt tgaaaattga atgtagcaac caattgagcc 6060
aaccccagcc tttgcccttt gattttgatt tgtttgttgc atacttttta tttgtcttct 6120 ggttctgact ctctttctct cgtttcaatg ccaggttgcc tactcccaca ccactcacaa 6180
gaagattcta ctgttagtat taaatatttt ttaatgtatt aaatgatgaa tgcttttgta 6240
aacagaacaa gactatgtct aataagtgtc ttgcaacatt ttttaagaaa ttaaaaaaaa 6300
tatatttatt atcaaaatca aatgtatgaa aaatcatgaa taatataatt ttatacattt 6360
ttttaaaaaa tcttttaatt tcttaattaa tatcttaaaa ataatgatta atatttaacc 6420
caaaataatt agtatgattg gtaaggaaga tatccatgtt atgtttggat gtgagtttga 6480
tctagagcaa agcttactag agtcgacctg caggttgaaa catccctgaa gtgtctcatt 6540
ttattttatt tattctttgc tgataaaaaa ataaaataaa agaagctaag cacacggtca 6600
accattgctc tactgctaaa agggttatgt gtagtgtttt actgcataaa ttatgcagca 6660
aacaagacaa ctcaaattaa aaaatttcct ttgcttgttt ttttgttgtc tctgacttga 6720
ctttcttgtg gaagttggtt gtataaggat tgggacacca ttgtccttct taatttaatt 6780
ttattctttg ctgataaaaa aaaaaatttc atatagtgtt aaataataat ttgttaaata 6840
accaaaaagt caaatatgtt tactctcgtt taaataattg agattcgtcc agcaaggcta 6900
aacgattgta tagatttatg acaatattta cttttttata gataaatgtt atattataat 6960
aaatttatat acatatatta tatgttattt attattattt taaatccttc aatattttat 7020
caaaccaact cataattttt tttttatctg taagaagcaa taaaattaaa tagacccact 7080
ttaaggatga tccaaccttt atacagagta agagagttca aatagtaccc tttcatatac 7140
atatcaacta aaatattaga aatatcatgg atcaaacctt ataaagacat taaataagtg 7200
gataagtata atatataaat gggtagtata taatatataa atggatacaa acttctctct 7260
ttataattgt tatgtctcct taacatccta atataataca taagtgggta atatataata 7320
tataaatgga gacaaacttc ttccattata attgttatgt cttcttaaca cttatgtctc 7380
gttcacaatg ctaaggttag aattgtttag aaagtcttat agtacacatt tgtttttgta 7440
ctatttgaag cattccataa gccgtcacga ttcagatgat ttataataat aagaggaaat 7500
ttatcataga acaataaggt gcatagatag agtgttaata tatcataaca tcctttgttt 7560
attcatagaa gaagtgagat ggagctcagt tattatactg ttacatggtc ggatacaata 7620
ttccatgctc tccatgagct cttacaccta catgcatttt agttcatact tgcggccgct 7680
cattcaggga cagcagccag ctgggcaatc tcgcctaaga agctgaccaa tttctgaacc 7740
ccggggatga tggctggggc cctgtagggc aggttgtgct tggcgcacaa cttctgcacc 7800
tccagggaag ctgccgtgag gttgtgcctg ggcatgttgg gccacaagtg gtgctcaatc 7860
tggtagttga gaccaccaaa gacccaatcg gtgagaaggc cgggcttaat gttcaacgtc 7920
tcgtggatct ggctggcgct gaacccgtgc tcatcccaga cagtctcctc aaccttgtcc 7980 aaaggatagt gattcagaaa cacaacgatg gcaatgccaa atcctccgag cagctcggag 8040 atcaagaagc atcccaaacc ggtggcaaag ctaggcatgg cactgcaata gaaagtggcc 8100 ttgagcaccc agtggagggc cagaccaatc gcctccttag tgtactggcg cttgtagtac 8160 aggttcctct cctcgctctt gtaggacatg acgccgccga tggattggag gcaccagatg 8220 aacctaaggg atgcaatggt gggaatgaag tagtactgct ggtacttgat gagattccga 8280 gtcgaaggag tagccctctt gacatcttcg ggagaccaag ccaggatggg gaggttatca 8340 atatctgggt catggccgac cacattggtg gcggcatggt gaccattgtg cctatccttc 8400 caccaagtct gtgagtatcc ctgcatcacg ttaccgaaaa agagaccgaa agcgtcgtta 8460 atcttgcggt ttgtgaaaac ctggtgatgg caatagtcgt gggacaacca gcctagctgt 8520 tggtagcaca cgccaagaac caaagcagcg gtgaaatacc aattcgtgta cacggtcatc 8580 aagatggaaa ggaccatcaa accgacggta gttgaagtct tgtaaaaata ccaaaggaaa 8640 ctcgtctcga acataccctt ggagatgaat tcctcccgca acttgcggaa atcctcctgg 8700 ggctcatcac gcttaggctt gggtgcaaca ggtgtgtcag gagaggaagg ctccataaca 8760 ggcattctct tcaacttggc gacggcttct tgagagtgca tcaccatgaa gacatcggtc 8820 gcatcgcggt tgcgatagtt ctcgataatg tcagctcctc cagggtggtg attgacccaa 8880 gcagacacat cataagttgc gccatcaatt gtgattggca gagcttgccg cttaggagac 8940 atggtgcggc cgcagtatat cttaaattct ttaatacggt gtactaggat attgaactgg 9000 ttcttgatga tgaaaacctg ggccgagatt gcagctattt atagtcatag gtcttgttaa 9060 catgcatgga catttggcca cggggtggca tgcagtttga cgggtgttga aataaacaaa 9120 aatgaggtgg cggaagagaa tacgagtttg aggttgggtt agaaacaaca aatgtgaggg 9180 ctcatgatgg gttgagttgg tgaatgtttt gggctgctcg attgacacct ttgtgagtac 9240 gtgttgttgt gcatggcttt tggggtccag tttttttttc ttgacgcggc gatcctgatc 9300 agctagtgga taagtgatgt ccactgtgtg tgattgcgtt tttgtttgaa ttttatgaac 9360 ttagacattg ctatgcaaag gatactctca ttgtgttttg tcttcttttg ttccttggct 9420 ttttcttatg atccaagaga ctagtcagtg ttgtggcatt cgagactacc aagattaatt 9480 atgatggggg aaggataagt aactgattag tacggactgt taccaaatta attaataagc 9540 ggcaaatgaa gggcatggat cctgcaggtc gactcgac 9578
<210> 12
<211> 3983
<212> DNA
<213> Seqüência Artificial <220> <223> plasmídeo pKR132
<400> 12
ctagagtcga cctgcaggca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg 60
tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 120
gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 180
ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 240
ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 300
gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 360
tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 420
aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 480
aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 540
ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 600
tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 660
agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 720
gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 780
tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 840
acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc 900
tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 960
caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 1020
aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 1080
aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 1140
ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 1200
agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 1260
atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 1320
cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 1380
aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 1440
cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 1500
aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 1560
ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 1620
gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 1680
ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 1740
tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 1800 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 1860
ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 1920
tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 1980
agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 2040
acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 2100
ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 2160
gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg 2220
acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat 2280
gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg 2340
gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc 2400
tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa 2460
ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg 2520
cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa 2580
gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt 2640
tgtaaaacga cggccagtga attcgagctc ggtacccggg gatcctctag acctgcaggc 2700
caactgcgtt tggggctcca gattaaacga cgccgtttcg ttcctttcgc ttcacggctt 2760
aacgatgtcg tttctgtctg tgcccaaaaa ataaaggcat ttgttatttg caccagatat 2820
ttactaagtg caccctagtt tgacaagtag gcgataatta caaatagatg cggtgcaaat 2880
aataaatttt gaaggaaata attacaaaag aacagaactt atatttactt tattttaaaa 2940
aactaaaatg aaagaacaaa aaaagtaaaa aatacaaaaa atgtgcttta accactttca 3000
ttatttgtta cagaaagtat gattctactc aaattgatct gttgtatctg gtgctgcctt 3060
gtcacactgg cgatttcaat cccctaaaga tatggtgcaa actgcgaagt gatcaatatc 3120
tgctcggtta atttagatta attaataata ttcaacgtga tgtaccaaaa aaagacaatt 3180
ttttgctcca ttgacaaatt aaacctcatc aaggtaattt ccaaacctat aagcaaaaaa 3240
atttcacatt aattggcccg caatcctatt agtcttatta tactagagta ggaaaaaaaa 3300
caattacaca acttgtctta ttattctcta tgctaatgaa tatttttccc ttttgttaga 3360
aatcagtgtt tcctaattta ttgagtatta attccactca ccgcatatat ttaccgttga 3420
ataagaaaat tttacacata attcttttta agataaataa tttttttata ctagatctta 3480
tatgattacg tgaagccaag tgggttatac taatgatata taatgtttga tagtaatcag 3540
tttataaacc aaatgcatgg aaatgttacg tggaagcacg taaattaaca agcattgaag 3600
caaatgcagc caccgcacca aaaccacccc acttcacttc cacgtaccat attccatgca 3660 actacaacac cctaaaactt caataaatgc ccccaccttc acttcacttc acccatcaat 3720
agcaagcggc cgcgaagtta aaagcaatgt tgtcacttgt cgtactaaca catgatgtga 3780
tagtttatgc tagctagcta taacataagc tgtctctgag tgtgttgtat attaataaag 3840
atcatcactg gtgaatggtg atcgtgtacg taccctactt agtaggcaat ggaagcactt 3900
agagtgtgct ttgtgcatgg ccttgcctct gttttgagac ttttgtaatg ttttcgagtt 3960
taaatctttg cctttgcgta cgt 3983
<210> 13
<211> 29
<212> DNA
<213> Seqüência Artificial <220>
<223> primer EgD5-5Not
<400> 13
gcggccgcac catggctctc agtcttacc 29
<210> 14
<211> 27
<212> DNA
<213> Seqüência Artificial <220>
<223> primer EgD5-3Not
<4 00> 14
gcggccgctc aggaatcctg tgcgtcc 27
<210> 15 <211> 4906 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo pKR268 <400> 15
ggccgcatga gccgtaaagg ttcaatacaa cgagtgcttg ttttcttagg gacaagcatt 60
gtacttatgt atgattctgt gtaaccatga gtcttccacg ttgtactaat gtgaagggca 120 aaaataaaac acagaacaag ttcgtttttc tcaaataatg tgaaggtaga aaatggaacc 180 atgcctcctc tcttgcatgt gatttaaaat attagcagat ggtaccgtac gtgggcggat 24 0 cccccgggct gcaggaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct 300 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc 360 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc 420 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact 480 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc 540
gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc 600
gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga 660
aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag 720
acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa 780
atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat 840
tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg 900
gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa 960
gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt 1020
gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt 1080
ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat 1140
tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg 1200
acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta 1260
cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat 1320
catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag 1380
cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa 1440
ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca 1500
ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc 1560
ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt 1620
atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc 1680
gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat 1740
atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt 1800
tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac 18 60
cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 1920
ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 1980
actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta 2040
gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 2100
ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 2160
gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 2220
acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 2280
tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 2340 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 2400
cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 2460
cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 2520
ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc 2580
gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg 2640
agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt 2700
cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca 2760
attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct 2820
cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat 2880
gattacgcca agcttgcatg cctgcaggtc gactcgacgt acgatcccac atgcaagttt 2940
ttatttcaat cccttttcct ttgaataact gaccaagaac aacaagaaaa aaaaaaaaaa 3000
agaaaaggat cattttgaaa ggatattttt cgctcctatt caaatactgt atttttacca 3060
aaaaaactgt atttttccta cactctcaag ctttgttttt cgcttcgact ctcatgattt 3120
ccttcatatg ccaatcactc tatttataaa tggcataagg tagtgtgaac aattgcaaag 3180
cttgtcatca aaagcttgca atgtacaaat taatgttttt catgcctttc aaaattatct 3240
tgcagtgcat taatttcccc gtgactattt tggctttgac tccaacactg gccccgtaca 3360
tccgtccctc attacatgaa aagaaatatt gtttatattc ttaattaaaa atattgtccc 3420
ttctaaattt tcatatagtt aattattata ttactttttt ctctattcta ttagttctat 3480
tttcaaatta ttatttatgc atatgtaaag tacattatat ttttgctata tacttaaata 3540
tttctaaatt attaaaaaaa gactgatatg aaaaatttat tctttttaaa gctatatcat 3600
tttatatata ctttttcttt tcttttcttt cattttctat tcaatttaat aagaaataaa 3660
ttttgtaaat ttttatttat caatttataa aaatatttta ctttatatgt tttttcacat 3720
ttttgttaaa caaatcatat cattatgatt gaaagagagg aaattgacag tgagtaataa 3780
gtgatgagaa aaaaatgtgt tatttcctaa aaaaaaccta aacaaacatg tatctactct 3840
ctatttcatc tatctctcat ttcatttttc tctttatctc tttctttatt tttttatcat 3900
atcatttcac attaattatt tttactctct ttattttttc tctctatccc tctcttattt 3960
ccactcatat atacactcca aaattggggc atgcctttat cactactcta tctcctccac 4020
taaatcattt aaatgaaact gaaaagcatt ggcaagtctc ctcccctcct caagtgattt 4080
ccaactcagc attggcatct aattgattca gtatatctat tgcatgtgta aaagtctttc 4140
cacaatacat aactattaat taatcttaaa taaataaagg ataaaatatt tttttttctt 4200 cataaaatta aaatatgtta ttttttgttt agatgtatat tcgaataaat ctaaatatat 4260 gataatgatt ttttatattg attaaacata taatcaatat taaatatgat atttttttat 4320 ataggttgta cacataattt tataaggata aaaaatatga taaaaataaa ttttaaatat 4380 ttttatattt acgagaaaaa aaaatatttt agccataaat aaatgaccag catattttac 4440 aaccttagta attcataaat tcctatatgt atatttgaaa ttaaaaacag ataatcgtta 4500 agggaaggaa tcctacgtca tctcttgcca tttgtttttc atgcaaacag aaagggacga 4560 aaaaccacct caccatgaat cactcttcac accattttta ctagcaaaca agtctcaaca 4620 actgaagcca gctctctttc cgtttctttt tacaacactt tctttgaaat agtagtattt 4680 ttttttcaca tgatttatta acgtgccaaa agatgcttat tgaatagagt gcacatttgt 4740 aatgtactac taattagaac atgaaaaagc attgttctaa cacgataatc ctgtgaaggc 4800 gttaactcca aagatccaat ttcactatat aaattgtgac gaaagcaaaa tgaattcaca 4860 tagctgagag agaaaggaaa ggttaactaa gaagcaatac ttcagc 4906 <210> 16 <211> 6267 <212> DNA <213> Seqüência Artificial <220> <223> plasmideo Ann-EgD5 <400> 16 ggccgcatga gccgtaaagg ttcaatacaa cgagtgcttg ttttcttagg gacaagcatt 60 gtacttatgt atgattctgt gtaaccatga gtcttccacg ttgtactaat gtgaagggca 120 aaaataaaac acagaacaag ttcgtttttc tcaaataatg tgaaggtaga aaatggaacc 180 atgcctcctc tcttgcatgt gatttaaaat attagcagat ggtaccgtac gtgggcggat 240 cccccgggct gcaggaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct 300 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc 360 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc 420 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact 480 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc 540 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc 600 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga 660 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag 720 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa 780 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat 840 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg 900 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa 960 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt 1020 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt 1080 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat 1140 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg 1200 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta 1260 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat 1320 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag 1380 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa 1440 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca 1500 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc 1560 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt 1620 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc 1680 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat 1740 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaayatcctt 1800 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac 1860 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 1920 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 1980 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta 2040 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 2100 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 2160 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 2220 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 2280 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 2340 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 2400 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 2460 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 2520 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc 2580 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg 2640 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt 2700 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca 2760
attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct 2820
cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat 2880
gattacgcca agcttgcatg cctgcaggtc gactcgacgt acgatcccac atgcaagttt 2940
ttatttcaat cccttttcct ttgaataact gaccaagaac aacaagaaaa aaaaaaaaaa 3000
agaaaaggat cattttgaaa ggatattttt cgctcctatt caaatactgt atttttacca 3060
aaaaaactgt atttttccta cactctcaag ctttgttttt cgcttcgact ctcatgattt 3120
ccttcatatg ccaatcactc tatttataaa tggcataagg tagtgtgaac aattgcaaag 3180
cttgtcatca aaagcttgca atgtacaaat taatgttttt catgcctttc aaaattatct 3240
gcacccccta gctattaatc taacatctaa gtaaggctag tgaatttttt cgaatagtca 3300
tgcagtgcat taatttcccc gtgactattt tggctttgac tccaacactg gccccgtaca 3360
tccgtccctc attacatgaa aagaaatatt gtttatattc ttaattaaaa atattgtccc 3420
ttctaaattt tcatatagtt aattattata ttactttttt ctctattcta ttagttctat 3480
tttcaaatta ttatttatgc atatgtaaag tacattatat ttttgctata tacttaaata 3540
tttctaaatt attaaaaaaa gactgatatg aaaaatttat tctttttaaa gctatatcat 3600
tttatatata ctttttcttt tcttttcttt cattttctat tcaatttaat aagaaataaa 3660
ttttgtaaat ttttatttat caatttataa aaatatttta ctttatatgt tttttcacat 3720
ttttgttaaa caaatcatat cattatgatt gaaagagagg aaattgacag tgagtaataa 3780
gtgatgagaa aaaaatgtgt tatttcctaa aaaaaaccta aacaaacatg tatctactct 3840
ctatttcatc tatctctcat ttcatttttc tctttatctc tttctttatt tttttatcat 3900
atcatttcac attaattatt tttactctct ttattttttc tctctatccc tctcttattt 3960
ccactcatat atacactcca aaattggggc atgcctttat cactactcta tctcctccac 4020
taaatcattt aaatgaaact gaaaagcatt ggcaagtctc ctcccctcct caagtgattt 4080
ccaactcagc attggcatct aattgattca gtatatctat tgcatgtgta aaagtctttc 4140
cacaatacat aactattaat taatcttaaa taaataaagg ataaaatatt tttttttctt 4200
cataaaatta aaatatgtta ttttttgttt agatgtatat tcgaataaat ctaaatatat 4260
gataatgatt ttttatattg attaaacata taatcaatat taaatatgat atttttttat 4320
ataggttgta cacataattt tataaggata aaaaatatga taaaaataaa ttttaaatat 4380
ttttatattt acgagaaaaa aaaatatttt agccataaat aaatgaccag catattttac 4440
aaccttagta attcataaat tcctatatgt atatttgaaa ttaaaaacag ataatcgtta 4500
agggaaggaa tcctacgtca tctcttgcca tttgtttttc atgcaaacag aaagggacga 4560 aaaaccacct caccatgaat cactcttcac accattttta ctagcaaaca agtctcaaca 4620
actgaagcca gctctctttc cgtttctttt tacaacactt tctttgaaat agtagtattt 4680
ttttttcaca tgatttatta acgtgccaaa agatgcttat tgaatagagt gcacatttgt 4740
aatgtactac taattagaac atgaaaaagc attgttctaa cacgataatc ctgtgaaggc 4800
gttaactcca aagatccaat ttcactatat aaattgtgac gaaagcaaaa tgaattcaca 4860
tagctgagag agaaaggaaa ggttaactaa gaagcaatac ttcagcggcc gcaccatggc 4920
tctcagtctt accacagaac agctgttaga acgccctgat ttggttgcga ttgatggcat 4980
cctctacgac cttgaagggc ttgccaaagt tcatccagga ggagatttga ttctcgcttc 5040
tggtgcctct gatgcctccc ctctctttta ttcgatgcat ccatacgtca aaccggagaa 5100
ttccaaattg cttcaacagt tcgtccgagg gaagcatgac cgcacctcga aggacattgt 5160
ctacacgtat gattctccct tcgcacaaga cgttaagcgg acaatgcgcg aggtgatgaa 5220
agggaggaac tggtacgcaa cccctggctt ctggctgcgc accgttggga tcatcgccgt 5280
gacggccttt tgcgagtggc actgggctac cacggggatg gtgctgtggg gcctgctgac 5340
tggattcatg cacatgcaga tcggcttatc catccagcat gatgcgtccc acggggccat 5400
cagcaagaag ccttgggtca acgccctctt cgcctacggc attgacgtca tcggatcgtc 5460
ccggtggatt tggctgcagt cgcacatcat gcggcaccac acctacacca accagcacgg 5520
cctcgacctg gatgcggagt cggcagagcc gttcctggtg ttccacaact accccgccgc 5580
aaacaccgcc cgaaagtggt tccaccgctt ccaagcttgg tacatgtacc ttgtgctggg 5640
ggcatacggg gtatcgctgg tgtacaaccc gctctacatt ttccggatgc agcacaatga 5700
caccatccca gagtctgtca cggccatgcg ggagaatggc tttctgcggc gctaccgcac 5760
acttgcattc gtgatgcgag ctttcttcat cttccggacc gcattcttgc cctggtacct 5820
cactgggacc tcattgctga tcaccattcc tctggtgccc actgcaactg gtgccttctt 5880
gacgttcttc ttcattttgt cccacaattt tgatggctcc gaacggatcc ccgacaagaa 5940
ctgcaaggtt aagagctctg agaaggacgt tgaggctgac caaattgact ggtatcgggc 6000
gcaggtggag acgtcctcca catacggtgg ccccatcgcc atgttcttca ctggcggtct 6060
caatttccag atcgagcacc acctctttcc ccggatgtcg tcttggcact accccttcgt 6120
ccagcaggcg gtccgggagt gttgcgaacg ccatggagtg cgatatgttt tctatcctac 6180
catcgtcggc aacatcatct ccaccctgaa gtacatgcat aaggtgggtg tcgtccactg 6240
cgtgaaggac gcacaggatt cctgagc 6267
<210> 17 <211> 13153 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo BC-Tpom_Ann-EgD5_ALS <400> 17
gtacgtctag aggatccgtc gacggcgcgc ccgatcatcc ggatatagtt cctcctttca 60
gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt tattgctcag 120
cggtggcagc agccaactca gcttcctttc gggctttgtt agcagccgga tcgatccaag 180
ctgtacctca ctattccttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg 240
cgagtacttc tacacagcca tcggtccaga cggccgcgct tctgcgggcg atttgtgtac 300
gcccgacagt cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg 360
catcatcgaa attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca 420
tatacgcccg gagccgcggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg 480
tctgctgctc catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt 540
gggaatcccc gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg 600
tcaggacatt gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct 660
cggcccaaag catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca 720
tcacagtttg ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg 780
tagtgtattg accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat 840
cggccgcagc gatcgcatcc atagcctccg cgaccggctg cagaacagcg ggcagttcgg 900
tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc 960
tctcgctgaa ttccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt 1020
gccgataaac ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat 1080
atccacgccc tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca 1140
tcaggtcgga gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga 1200
gttcaggctt ttccatgggt atatctcctt cttaaagtta aacaaaatta tttctagagg 1260
gaaaccgttg tggtctccct atagtgagtc gtattaattt cgcgggatcg agatctgatc 1320
aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 1380
tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 1440
gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 1500
atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 1560
ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 1620
cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 1680 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 1740
gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 1800
aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 1860
tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 1920
aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 1980
aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 2040
ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 2100
ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 2160
atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 2220
atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt 2280
gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa 2340
gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg 24 00
ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatggacata 2460
ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca catacgattt 2520
aggtgacact atagaacggc gcgccaagct gggtctagaa ctagaaacgt gatgccactt 2580
gttattgaag tcgattacag catctattct gttttactat ttataacttt gccatttctg 2640
acttttgaaa actatctctg gatttcggta tcgctttgtg aagatcgagc aaaagagacg 2700
ttttgtggac gcaatggtcc aaatccgttc tacatgaaca aattggtcac aatttccact 2760
aaaagtaaat aaatggcaag ttaaaaaagg aatatgcatt ttactgattg cctaggtgag 2820
ctccaagaga agttgaatct acacgtctac caaccgctaa aaaaagaaaa acattgatat 2880
gtaacctgat tccattagct tttgacttct tcaacagatt ctctacttag atttctaaca 2940
gaaatattat tactagcaca tcattttcag tctcactaca gcaaaaaatc caacggcaca 3000
atacagacaa caggagatat cagactacag agatagatag atgctactgc atgtagtaag 3060
ttaaataaaa ggaaaataaa atgtcttgct accaaaacta ctacagacta tgatgctcac 3120
cacaggccaa atcctgcaac taggacagca ttatcttata tatattgtac aaaacaagca 3180
tcaaggaaca tttggtctag gcaatcagta cctcgttcta ccatcaccct cagttatcac 3240
atccttgaag gatccattac tgggaatcat cggcaacaca tgctcctgat ggggcacaat 3300
gacatcaaga aggtaggggc caggggtgtc caacattctc tgaattgccg ctctaagctc 3360
ttccttcttc gtcactcgcg ctgccggtat cccacaagca tcagcaaact tgagcatgtt 3420
tgggaatatc tcgctctcgc tagacggatc tccaagatag gtgtgagctc tattggactt 3480
gtagaaccta tcctccaact gaaccaccat acccaaatgc tgattgttca acaacaatat 3540 cttaactggg agattctcca ctcttatagt ggccaactcc tgaacattca tgatgaaact 3600 accatcccca tcaatgtcaa ccacaacagc cccagggtta gcaacagcag caccaatagc 3660 cgcaggcaat ccaaaaccca tggctccaag accccctgag gtcaaccact gcctcggtct 3720 cttgtacttg taaaactgcg cagcccacat ttgatgctgc ccaaccccag tactaacaat 3780 agcatctcca ttagtcaact catcaagaac ctcgatagca tgctgcggag aaatcgcgtc 3840 ctggaatgtc ttgtaaccca atggaaactt gtgtttctgc acattaatct cttctctcca 3900 acctccaaga tcaaacttac cctccactcc tttctcctcc aaaatcatat taattccctt 3960 caaggccaac ttcaaatccg cgcaaaccga cacgtgcgcc tgcttgttct tcccaatctc 4020 ggcagaatca atatcaatgt gaacaatctt agccctacta gcaaaagcct caagcttccc 4080 agtaacacgg tcatcaaacc ttaccccaaa ggcaagcaac aaatcactat tgtcaacagc 4140 atagttagca taaacagtac catgcatacc cagcatctga agggaatatt catcaccaat 4200 aggaaaagtt ccaagaccca ttaaagtgct agcaacggga ataccagtga gttcaacaaa 4260 gcgcctcaat tcagcactgg aattcaaact gccaccgccg acgtagagaa cgggcttttg 4320 ggcctccatg atgagtctga caatgtgttc caattgggcc tcggcggggg gcctgggcag 4380 cctggcgagg taaccgggga ggttaacggg ctcgtcccaa ttaggcacgg cgagttgctg 4440 ctgaacgtct ttgggaatgt cgatgaggac cggaccgggg cggccggagg tggcgacgaa 4500 gaaagcctcg gcgacgacgc gggggatgtc gtcgacgtcg aggatgaggt agttgtgctt 4560 cgtgatggat ctgctcacct ccacgatcgg ggtttcttgg aaggcgtcgg tgccgatcat 4620 ccggcgggcg acctggccgg tgatggcgac gactgggacg ctgtccatta aagcgtcggc 4680 gaggccgctc acgaggttgg tggcgccggg gccggaggtg gcaatgcaga cgccggggag 4740 gccggaggaa cgcgcgtagc cttcggcggc gaagacgccg ccctgctcgt ggcgcgggag 4800 cacgttgcgg atggcggcgg agcgcgtgag cgcctggtgg atctccatcg acgcaccgcc 4860 ggggtacgcg aacaccgtcg tcacgccctg cctctccagc gcctccacaa ggatgtccgc 4920 gcccttgcga ggttcgccgg aggcgaaccg tgacacgaag ggctccgtgg tcggcgcttc 4980 cttggtgaag ggcgccgccg tggggggttt ggagatggaa catttgattt tgagagcgtg 5040 gttgggtttg gtgagggttt gatgagagag agggagggtg gatctagtaa tgcgtttggg 5100 gaaggtgggg tgtgaagagg aagaagagaa tcgggtggtt ctggaagcgg tggccgccat 5160 tgtgttgtgt ggcatggtta tacttcaaaa actgcacaac aagcctagag ttagtaccta 5220 aacagtaaat ttacaacaga gagcaaagac acatgcaaaa atttcagcca taaaaaaagt 5280 tataatagaa tttaaagcaa aagtttcatt ttttaaacat atatacaaac aaactggatt 5340 tgaaggaagg gattaattcc cctgctcaaa gtttgaattc ctattgtgac ctatactcga 5400 ataaaattga agcctaagga atgtatgaga aacaagaaaa caaaacaaaa ctacagacaa 5460
acaagtacaa ttacaaaatt cgctaaaatt ctgtaatcac caaaccccat ctcagtcagc 5520
acaaggccca aggtttattt tgaaataaaa aaaaagtgat tttatttctc ataagctaaa 5580
agaaagaaag gcaattatga aatgatttcg actagatctg aaagtccaac gcgtattccg 5640
cagatattaa agaaagagta gagtttcaca tggatcctag atggacccag ttgaggaaaa 5700
agcaaggcaa agcaaaccag aagtgcaaga tccgaaattg aaccacggaa tctaggattt 5760
ggtagaggga gaagaaaagt accttgagag gtagaagaga agagaagagc agagagatat 5820
atgaacgagt gtgtcttggt ctcaactctg aagcgatacg agtttagagg ggagcattga 5880
gttccaattt atagggaaac cgggtggcag gggtgagtta atgacggaaa agcccctaag 5940
taacgagatt ggattgtggg ttagattcaa ccgtttgcat ccgcggctta gattggggaa 6000
gtcagagtga atctcaaccg ttgactgagt tgaaaattga atgtagcaac caattgagcc 6060
aaccccagcc tttgcccttt gattttgatt tgtttgttgc atacttttta tttgtcttct 6120
ggttctgact ctctttctct cgtttcaatg ccaggttgcc tactcccaca ccactcacaa 6180
gaagattcta ctgttagtat taaatatttt ttaatgtatt aaatgatgaa tgcttttgta 6240
aacagaacaa gactatgtct aataagtgtc ttgcaacatt ttttaagaaa ttaaaaaaaa 6300
tatatttatt atcaaaatca aatgtatgaa aaatcatgaa taatataatt ttatacattt 6360
ttttaaaaaa tcttttaatt tcttaattaa tatcttaaaa ataatgatta atatttaacc 6420
caaaataatt agtatgattg gtaaggaaga tatccatgtt atgtttggat gtgagtttga 6480
tctagagcaa agcttactag agtcgacctg caggatccat gcccttcatt tgccgcttat 6540
taattaattt ggtaacagtc cgtactaatc agttacttat ccttccccca tcataattaa 6600
tcttggtagt ctcgaatgcc acaacactga ctagtctctt ggatcataag aaaaagccaa 6660
ggaacaaaag aagacaaaac acaatgagag tatcctttgc atagcaatgt ctaagttcat 6720
aaaattcaaa caaaaacgca atcacacaca gtggacatca cttatccact agctgatcag 6780
gatcgccgcg tcaagaaaaa aaaactggac cccaaaagcc atgcacaaca acacgtactc 6840
acaaaggtgt caatcgagca gcccaaaaca ttcaccaact caacccatca tgagccctca 6900
catttgttgt ttctaaccca acctcaaact cgtattctct tccgccacct catttttgtt 6960
tatttcaaca cccgtcaaac tgcatgccac cccgtggcca aatgtccatg catgttaaca 7020
agacctatga ctataaatag ctgcaatctc ggcccaggtt ttcatcatca agaaccagtt 7080
caatatccta gtacaccgta ttaaagaatt taagatatac tgcggccgca ccatgtctcc 7140
taagcggcaa gctctgccaa tcacaattga tggcgcaact tatgatgtgt ctgcttgggt 7200
caatcaccac cctggaggag ctgacattat cgagaactat cgcaaccgcg atgcgaccga 7260 tgtcttcatg gtgatgcact ctcaagaagc cgtcgccaag ttgaagagaa tgcctgttat 7320
ggagccttcc tctcctgaca cacctgttgc acccaagcct aagcgtgatg agccccagga 7380
ggatttccgc aagttgcggg aggaattcat ctccaagggt atgttcgaga cgagtttcct 7440
ttggtatttt tacaagactt caactaccgt cggtttgatg gtcctttcca tcttgatgac 7500
cgtgtacacg aattggtatt tcaccgctgc tttggttctt ggcgtgtgct accaacagct 7560
aggctggttg tcccacgact attgccatca ccaggttttc acaaaccgca agattaacga 7620
cgctttcggt ctctttttcg gtaacgtgat gcagggatac tcacagactt ggtggaagga 7680
taggcacaat ggtcaccatg ccgccaccaa tgtggtcggc catgacccag atattgataa 7740
cctccccatc ctggcttggt ctcccgaaga tgtcaagagg gctactcctt cgactcggaa 7800
tctcatcaag taccagcagt actacttcat tcccaccatt gcatccctta ggttcatctg 7860
gtgcctccaa tccatcggcg gcgtcatgtc ctacaagagc gaggagagga acctgtacta 7920
caagcgccag tacactaagg aggcgattgg tctggccctc cactgggtgc tcaaggccac 7980
tttctattgc agtgccatgc ctagctttgc caccggtttg ggatgcttct tgatctccga 8040
gctgctcgga ggatttggca ttgccatcgt tgtgtttctg aatcactatc ctttggacaa 8100
ggttgaggag actgtctggg atgagcacgg gttcagcgcc agccagatcc acgagacgtt 8160
gaacattaag cccggccttc tcaccgattg ggtctttggt ggtctcaact accagattga 8220
gcaccacttg tggcccaaca tgcccaggca caacctcacg gcagcttccc tggaggtgca 8280
gaagttgtgc gccaagcaca acctgcccta cagggcccca gccatcatcc ccggggttca 8340
gaaattggtc agcttcttag gcgagattgc ccagctggct gctgtccctg aatgagcggc 8400
cgcaagtatg aactaaaatg catgtaggtg taagagctca tggagagcat ggaatattgt 8460
atccgaccat gtaacagtat aataactgag ctccatctca cttcttctat gaataaacaa 8520
aggatgttat gatatattaa cactctatct atgcacctta ttgttctatg ataaatttcc 8580
tcttattatt ataaatcatc tgaatcgtga cggcttatgg aatgcttcaa atagtacaaa 8640
aacaaatgtg tactataaga ctttctaaac aattctaacc ttagcattgt gaacgagaca 8700
taagtgttaa gaagacataa caattataat ggaagaagtt tgtctccatt tatatattat 8760
atattaccca cttatgtatt atattaggat gttaaggaga cataacaatt ataaagagag 8820
aagtttgtat ccatttatat attatatact acccatttat atattatact tatccactta 8880
tttaatgtct ttataaggtt tgatccatga tatttctaat attttagttg atatgtatat 8940
gaaagggtac tatttgaact ctcttactct gtataaaggt tggatcatcc ttaaagtggg 9000
tctatttaat tttattgctt cttacagata aaaaaaaaat tatgagttgg tttgataaaa 9060
tattgaagga tttaaaataa taataaataa catataatat atgtatataa atttattata 9120 atataacatt tatctataaa aaagtaaata ttgtcataaa tctatacaat cgtttagcct 9180
tgctggacga atctcaatta tttaaacgag agtaaacata tttgactttt tggttattta 9240
acaaattatt atttaacact atatgaaatt ttttttttta tcagcaaaga ataaaattaa 9300
attaagaagg acaatggtgt cccaatcctt atacaaccaa cttccacaag aaagtcaagt 9360
cagagacaac aaaaaaacaa gcaaaggaaa ttttttaatt tgagttgtct tgtttgctgc 9420
ataatttatg cagtaaaaca ctacacataa cccttttagc agtagagcaa tggttgaccg 9480
tgtgcttagc ttcttttatt ttattttttt atcagcaaag aataaataaa ataaaatgag 9540
acacttcagg gatgtttcaa cctgcaggtc gactcgacgt acgatcccac atgcaagttt 9600
ttatttcaat cccttttcct ttgaataact gaccaagaac aacaagaaaa aaaaaaaaaa 9660
agaaaaggat cattttgaaa ggatattttt cgctcctatt caaatactgt atttttacca 9720
aaaaaactgt atttttccta cactctcaag ctttgttttt cgcttcgact ctcatgattt 9780
ccttcatatg ccaatcactc tatttataaa tggcataagg tagtgtgaac aattgcaaag 9840
cttgtcatca aaagcttgca atgtacaaat taatgttttt catgcctttc aaaattatct 9900
gcacccccta gctattaatc taacatctaa gtaaggctag tgaatttttt cgaatagtca 9960
tgcagtgcat taatttcccc gtgàctattt tggctttgac tccaacactg gccccgtaca 10020
tccgtccctc attacatgaa aagaaatatt gtttatattc ttaattaaaa atattgtccc 10080
ttctaaattt tcatatagtt aattattata ttactttttt ctctattcta ttagttctat 10140
tttcaaatta ttatttatgc atatgtaaag tacattatat ttttgctata tacttaaata 10200
tttctaaatt attaaaaaaa gactgatatg aaaaatttat tctttttaaa gctatatcat 10260
tttatatata ctttttcttt tcttttcttt cattttctat tcaatttaat aagaaataaa 10320
ttttgtaaat ttttatttat caatttataa aaatatttta ctttatatgt tttttcacat 10380
ttttgttaaa caaatcatat cattatgatt gaaagagagg aaattgacag tgagtaataa 10440
gtgatgagaa aaaaatgtgt tatttcctaa aaaaaaccta aacaaacatg tatctactct 10500
ctatttcatc tatctctcat ttcatttttc tctttatctc tttctttatt tttttatcat 10560
atcatttcac attaattatt tttactctct ttattttttc tctctatccc tctcttattt 10620
ccactcatat atacactcca aaattggggc atgcctttat cactactcta tctcctccac 10680
taaatcattt aaatgaaact gaaaagcatt ggcaagtctc ctcccctcct caagtgattt 10740
ccaactcagc attggcatct aattgattca gtatatctat tgcatgtgta aaagtctttc 10800
cacaatacat aactattaat taatcttaaa taaataaagg ataaaatatt tttttttctt 10860
cataaaatta aaatatgtta ttttttgttt agatgtatat tcgaataaat ctaaatatat 10920
gataatgatt ttttatattg attaaacata taatcaatat taaatatgat atttttttat 10980 ataggttgta cacataattt tataaggata aaaaatatga taaaaataaa ttttaaatat 11040 ttttatattt acgagaaaaa aaaatatttt agccataaat aaatgaccag catattttac 11100 aaccttagta attcataaat tcctatatgt atatttgaaa ttaaaaacag ataatcgtta 11160 agggaaggaa tcctacgtca tctcttgcca tttgtttttc atgcaaacag aaagggacga 11220 aaaaccacct caccatgaat cactcttcac accattttta ctagcaaaca agtctcaaca 11280 actgaagcca gctctctttc cgtttctttt tacaacactt tctttgaaat agtagtattt 11340 ttttttcaca tgatttatta acgtgccaaa agatgcttat tgaatagagt gcacatttgt 11400 aatgtactac taattagaac atgaaaaagc attgttctaa cacgataatc ctgtgaaggc 11460 gttaactcca aagatccaat ttcactatat aaattgtgac gaaagcaaaa tgaattcaca 11520 tagctgagag agaaaggaaa ggttaactaa gaagcaatac ttcagcggcc gcaccatggc 11580 tctcagtctt accacagaac agctgttaga acgccctgat ttggttgcga ttgatggcat 11640 cctctacgac cttgaagggc ttgccaaagt tcatccagga ggagatttga ttctcgcttc 11700 tggtgcctct gatgcctccc ctctctttta ttcgatgcat ccatacgtca aaccggagaa 11760 ttccaaattg cttcaacagt tcgtccgagg gaagcatgac cgcacctcga aggacattgt 11820 ctacacgtat gattctccct tcgcacaaga cgttaagcgg acaatgcgcg aggtgatgaa 11880 agggaggaac tggtacgcaa cccctggctt ctggctgcgc accgttggga tcatcgccgt 1 T Ω Λ Λ gacggccttt tgcgagtggc actgggctac cacggggatg gtgctgtggg gcctgctgac 12000 tggattcatg cacatgcaga tcggcttatc catccagcat gatgcgtccc acggggccat 12060 cagcaagaag ccttgggtca acgccctctt cgcctacggc attgacgtca tcggatcgtc 12120 ccggtggatt tggctgcagt cgcacatcat gcggcaccac acctacacca accagcacgg 12180 cctcgacctg gatgcggagt cggcagagcc gttcctggtg ttccacaact accccgccgc 12240 aaacaccgcc cgaaagtggt tccaccgctt ccaagcttgg tacatgtacc ttgtgctggg 12300 ggcatacggg gtatcgctgg tgtacaaccc gctctacatt ttccggatgc agcacaatga 12360 caccatccca gagtctgtca cggccatgcg ggagaatggc tttctgcggc gctaccgcac 12420 acttgcattc gtgatgcgag ctttcttcat cttccggacc gcattcttgc cctggtacct 12480 cactgggacc tcattgctga tcaccattcc tctggtgccc actgcaactg gtgccttctt 12540 gacgttcttc ttcattttgt cccacaattt tgatggctcc gaacggatcc ccgacaagaa 12600 ctgcaaggtt aagagctctg agaaggacgt tgaggctgac caaattgact ggtatcgggc 12660 gcaggtggag acgtcctcca catacggtgg ccccatcgcc atgttcttca ctggcggtct 12720 caatttccag atcgagcacc acctctttcc ccggatgtcg tcttggcact accccttcgt 12780 ccagcaggcg gtccgggagt gttgcgaacg ccatggagtg cgatatgttt tctatcctac 12840 catcgtcggc aacatcatct ccaccctgaa gtacatgcat aaggtgggtg tcgtccactg 12900
cgtgaaggac gcacaggatt cctgagcggc cgcatgagcc gtaaaggttc aatacaacga 12960
gtgcttgttt tcttagggac aagcattgta cttatgtatg attctgtgta accatgagtc 13020
ttccacgttg tactaatgtg aagggcaaaa ataaaacaca gaacaagttc gtttttctca 13080
aataatgtga aggtagaaaa tggaaccatg cctcctctct tgcatgtgat ttaaaatatt 13140
agcagatggt acc 13153
<210> 18 <211> 13153 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo BC-Tpom Ann-EgD5 ALSrev <4 00> 18 gtacgtctag aggatccgtc gacggcgcgc ccgatcatcc ggatatagtt cctcctttca 60 gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt tattgctcag 120 cggtggcagc agccaactca gcttcctttc gggctttgtt agcagccgga tcgatccaag 180 ctgtacctca ctattccttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg 240 cgagtacttc tacacagcca tcggtccaga cggccgcgct tctycgggcg atttgtgtac o nn W gcccgacagt cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg 360 catcatcgaa attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca 420 tatacgcccg gagccgcggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg 480 tctgctgctc catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt 540 gggaatcccc gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg 600 tcaggacatt gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct 660 cggcccaaag catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca 720 tcacagtttg ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg 780 tagtgtattg accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat 840 cggccgcagc gatcgcatcc atagcctccg cgaccggctg cagaacagcg ggcagttcgg 900 tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc 960 tctcgctgaa ttccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt 1020 gccgataaac ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat 1080 atccacgccc tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca 1140 tcaggtcgga gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga 1200 gttcaggctt ttccatgggt atatctcctt cttaaagtta aacaaaatta tttctagagg 1260
gaaaccgttg tggtctccct atagtgagtc gtattaattt cgcgggatcg agatctgatc 1320
aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 1380
tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 1440
gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 1500
atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 1560
ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 1620
cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 1680
tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 1740
gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 1800
aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 1860
tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 1920
aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 1980
aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 2040
ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 2100
ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 2160
atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 2220
atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt 2280
gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa 2340
gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg 2400
ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatggacata 2460
ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca catacgattt 2520
aggtgacact atagaacggc gcgccaagct gggtctagaa ctagaaacgt gatgccactt 2580
gttattgaag tcgattacag catctattct gttttactat ttataacttt gccatttctg 2640
acttttgaaa actatctctg gatttcggta tcgctttgtg aagatcgagc aaaagagacg 2700
ttttgtggac gcaatggtcc aaatccgttc tacatgaaca aattggtcac aatttccact 2760
aaaagtaaat aaatggcaag ttaaaaaagg aatatgcatt ttactgattg cctaggtgag 2820
ctccaagaga agttgaatct acacgtctac caaccgctaa aaaaagaaaa acattgatat 2880
gtaacctgat tccattagct tttgacttct tcaacagatt ctctacttag atttctaaca 2940
gaaatattat tactagcaca tcattttcag tctcactaca gcaaaaaatc caacggcaca 3000
atacagacaa caggagatat cagactacag agatagatag atgctactgc atgtagtaag 3060 ttaaataaaa ggaaaataaa atgtcttgct accaaaacta ctacagacta tgatgctcac 3120
cacaggccaa atcctgcaac taggacagca ttatcttata tatattgtac aaaacaagca 3180
tcaaggaaca tttggtctag gcaatcagta cctcgttcta ccatcaccct cagttatcac 3240
atccttgaag gatccattac tgggaatcat cggcaacaca tgctcctgat ggggcacaat 3300
gacatcaaga aggtaggggc caggggtgtc caacattctc tgaattgccg ctctaagctc 3360
ttccttcttc gtcactcgcg ctgccggtat cccacaagca tcagcaaact tgagcatgtt 3420
tgggaatatc tcgctctcgc tagacggatc tccaagatag gtgtgagctc tattggactt 3480
gtagaaccta tcctccaact gaaccaccat acccaaatgc tgattgttca acaacaatat 3540
cttaactggg agattctcca ctcttatagt ggccaactcc tgaacattca tgatgaaact 3600
accatcccca tcaatgtcaa ccacaacagc cccagggtta gcaacagcag caccaatagc 3660
cgcaggcaat ccaaaaccca tggctccaag accccctgag gtcaaccact gcctcggtct 3720
cttgtacttg taaaactgcg cagcccacat ttgatgctgc ccaaccccag tactaacaat 3780
agcatctcca ttagtcaact catcaagaac ctcgatagca tgctgcggag aaatcgcgtc 3840
ctggaatgtc ttgtaaccca atggaaactt gtgtttctgc acattaatct cttctctcca 3900
acctccaaga tcaaacttac cctccactcc tttctcctcc aaaatcatat taattccctt 3960
caaggccaac ttcaaatccg cgcaaaecga cacgtgcgcc tgcttgttct tcccaatctc 4 020
ggcagaatca atatcaatgt gaacaatctt agccctacta gcaaaagcct caagcttccc 4080
agtaacacgg tcatcaaacc ttaccccaaa ggcaagcaac aaatcactat tgtcaacagc 4140
atagttagca taaacagtac catgcatacc cagcatctga agggaatatt catcaccaat 4200
aggaaaagtt ccaagaccca ttaaagtgct agcaacggga ataccagtga gttcaacaaa 4260
gcgcctcaat tcagcactgg aattcaaact gccaccgccg acgtagagaa cgggcttttg 4320
ggcctccatg atgagtctga caatgtgttc caattgggcc tcggcggggg gcctgggcag 4380
cctggcgagg taaccgggga ggttaacggg ctcgtcccaa ttaggcacgg cgagttgctg 4440
ctgaacgtct ttgggaatgt cgatgaggac cggaccgggg cggccggagg tggcgacgaa 4500
gaaagcctcg gcgacgacgc gggggatgtc gtcgacgtcg aggatgaggt agttgtgctt 4560
cgtgatggat ctgctcacct ccacgatcgg ggtttcttgg aaggcgtcgg tgccgatcat 4620
ccggcgggcg acctggccgg tgatggcgac gactgggacg ctgtccatta aagcgtcggc 4680
gaggccgctc acgaggttgg tggcgccggg gccggaggtg gcaatgcaga cgccggggag 4740
gccggaggaa cgcgcgtagc cttcggcggc gaagacgccg ccctgctcgt ggcgcgggag 4800
cacgttgcgg atggcggcgg agcgcgtgag cgcctggtgg atctccatcg acgcaccgcc 4860
ggggtacgcg aacaccgtcg tcacgccctg cctctccagc gcctccacaa ggatgtccgc 4920 gcccttgcga ggttcgccgg aggcgaaccg tgacacgaag ggctccgtgg tcggcgcttc 4980
cttggtgaag ggcgccgccg tggggggttt ggagatggaa catttgattt tgagagcgtg 5040
gttgggtttg gtgagggttt gatgagagag agggagggtg gatctagtaa tgcgtttggg 5100
gaaggtgggg tgtgaagagg aagaagagaa tcgggtggtt ctggaagcgg tggccgccat 5160
tgtgttgtgt ggcatggtta tacttcaaaa actgcacaac aagcctagag ttagtaccta 5220
aacagtaaat ttacaacaga gagcaaagac acatgcaaaa atttcagcca taaaaaaagt 5280
tataatagaa tttaaagcaa aagtttcatt ttttaaacat atatacaaac aaactggatt 5340
tgaaggaagg gattaattcc cctgctcaaa gtttgaattc ctattgtgac ctatactcga 5400
ataaaattga agcctaagga atgtatgaga aacaagaaaa caaaacaaaa ctacagacaa 5460
acaagtacaa ttacaaaatt cgctaaaatt ctgtaatcac caaaccccat ctcagtcagc 5520
acaaggccca aggtttattt tgaaataaaa aaaaagtgat tttatttctc ataagctaaa 5580
agaaagaaag gcaattatga aatgatttcg actagatctg aaagtccaac gcgtattccg 5640
cagatattaa agaaagagta gagtttcaca tggatcctag atggacccag ttgaggaaaa 5700
agcaaggcaa agcaaaccag aagtgcaaga tccgaaattg aaccacggaa tctaggattt 5760
ggtagaggga gaagaaaagt accttgagag gtagaagaga agagaagagc agagagatat 5820
atgaacgagt gtgtcttggt ctcaactctg aagcgatacg agtttagagg ggagcattga 5880
gttccaattt atagggaaac cgggtggcag gggtgagtta atgacggaaa agcccctaag 5940
taacgagatt ggattgtggg ttagattcaa ccgtttgcat ccgcggctta gattggggaa 6000
gtcagagtga atctcaaccg ttgactgagt tgaaaattga atgtagcaac caattgagcc 6060
aaccccagcc tttgcccttt gattttgatt tgtttgttgc atacttttta tttgtcttct 6120
ggttctgact ctctttctct cgtttcaatg ccaggttgcc tactcccaca ccactcacaa 6180
gaagattcta ctgttagtat taaatatttt ttaatgtatt aaatgatgaa tgcttttgta 6240
aacagaacaa gactatgtct aataagtgtc ttgcaacatt ttttaagaaa ttaaaaaaaa 6300
tatatttatt atcaaaatca aatgtatgaa aaatcatgaa taatataatt ttatacattt 6360
ttttaaaaaa tcttttaatt tcttaattaa tatcttaaaa ataatgatta atatttaacc 6420
caaaataatt agtatgattg gtaaggaaga tatccatgtt atgtttggat gtgagtttga 6480
tctagagcaa agcttactag agtcgacctg caggttgaaa catccctgaa gtgtctcatt 6540
ttattttatt tattctttgc tgataaaaaa ataaaataaa agaagctaag cacacggtca 6600
accattgctc tactgctaaa agggttatgt gtagtgtttt actgcataaa ttatgcagca 6660
aacaagacaa ctcaaattaa aaaatttcct ttgcttgttt ttttgttgtc tctgacttga 6720
ctttcttgtg gaagttggtt gtataaggat tgggacacca ttgtccttct taatttaatt 6780 ttattctttg ctgataaaaa aaaaaatttc atatagtgtt aaataataat ttgttaaata 6840 accaaaaagt caaatatgtt tactctcgtt taaataattg agattcgtcc agcaaggcta 6900
aacgattgta tagatttatg acaatattta cttttttata gataaatgtt atattataat 6960
aaatttatat acatatatta tatgttattt attattattt taaatccttc aatattttat 7020
caaaccaact cataattttt tttttatctg taagaagcaa taaaattaaa tagacccact 7080
ttaaggatga tccaaccttt atacagagta agagagttca aatagtaccc tttcatatac 7140
atatcaacta aaatattaga aatatcatgg atcaaacctt ataaagacat taaataagtg 7200
gataagtata atatataaat gggtagtata taatatataa atggatacaa acttctctct 7260
ttataattgt tatgtctcct taacatccta atataataca taagtgggta atatataata 7320
tataaatgga gacaaacttc ttccattata attgttatgt cttcttaaca cttatgtctc 7380
gttcacaatg ctaaggttag aattgtttag aaagtcttat agtacacatt tgtttttgta 7440
ctatttgaag cattccataa gccgtcacga ttcagatgat ttataataat aagaggaaat 7500
ttatcataga acaataaggt gcatagatag agtgttaata tatcataaca tcctttgttt 7560
attcatagaa gaagtgagat ggagctcagt tattatactg ttacatggtc ggatacaata 7620
ttccatgctc tccatgagct cttacaccta catgcatttt agttcatact tgcggccgct 7680
cattcaggga cagcagccag ctgggcaatc tcgcctaaga agctgaccaa tttctgaacc 7740
ccggggatga tggctggggc cctgtagggc aggttgtgct tggcgcacaa cttctgcacc 7800
tccagggaag ctgccgtgag gttgtgcctg ggcatgttgg gccacaagtg gtgctcaatc 7860
tggtagttga gaccaccaaa gacccaatcg gtgagaaggc cgggcttaat gttcaacgtc 7920
tcgtggatct ggctggcgct gaacccgtgc tcatcccaga cagtctcctc aaccttgtcc 7 980
aaaggatagt gattcagaaa cacaacgatg gcaatgccaa atcctccgag cagctcggag 8040
atcaagaagc atcccaaacc ggtggcaaag ctaggcatgg cactgcaata gaaagtggcc 8100
ttgagcaccc agtggagggc cagaccaatc gcctccttag tgtactggcg cttgtagtac 8160
aggttcctct cctcgctctt gtaggacatg acgccgccga tggattggag gcaccagatg 8220
aacctaaggg atgcaatggt gggaatgaag tagtactgct ggtacttgat gagattccga 8280
gtcgaaggag tagccctctt gacatcttcg ggagaccaag ccaggatggg gaggttatca 834 0
atatctgggt catggccgac cacattggtg gcggcatggt gaccattgtg cctatccttc 8400
caccaagtct gtgagtatcc ctgcatcacg ttaccgaaaa agagaccgaa agcgtcgtta 8460
atcttgcggt ttgtgaaaac ctggtgatgg caatagtcgt gggacaacca gcctagctgt 8520
tggtagcaca cgccaagaac caaagcagcg gtgaaatacc aattcgtgta cacggtcatc 8580
aagatggaaa ggaccatcaa accgacggta gttgaagtct tgtaaaaata ccaaaggaaa 8640 ctcgtctcga acataccctt ggagatgaat tcctcccgca acttgcggaa atcctcctgg 8700 ggctcatcac gcttaggctt gggtgcaaca ggtgtgtcag gagaggaagg ctccataaca 8760 ggcattctct tcaacttggc gacggcttct tgagagtgca tcaccatgaa gacatcggtc 8820 gcatcgcggt tgcgatagtt ctcgataatg tcagctcctc cagggtggtg attgacccaa 8880 gcagacacat cataagttgc gccatcaatt gtgattggca gagcttgccg cttaggagac 8940 atggtgcggc cgcagtatat cttaaattct ttaatacggt gtactaggat attgaactgg 9000 ttcttgatga tgaaaacctg ggccgagatt gcagctattt atagtcatag gtcttgttaa 9060 catgcatgga catttggcca cggggtggca tgcagtttga cgggtgttga aataaacaaa 9120 aatgaggtgg cggaagagaa tacgagtttg aggttgggtt agaaacaaca aatgtgaggg 9180 ctcatgatgg gttgagttgg tgaatgtttt gggctgctcg attgacacct ttgtgagtac 9240 gtgttgttgt gcatggcttt tggggtccag tttttttttc ttgacgcggc gatcctgatc 9300 agctagtgga taagtgatgt ccactgtgtg tgattgcgtt tttgtttgaa ttttatgaac 9360 ttagacattg ctatgcaaag gatactctca ttgtgttttg tcttcttttg ttccttggct 9420 ttttcttatg atccaagaga ctagtcagtg ttgtggcatt cgagactacc aagattaatt 9480 atgatggggg aaggataagt aactgattag tacggactgt taccaaatta attaataagc 9540 ggcaaatgaa gggcatggat cctgcaggtc gactcgacgt acgatcccac atgcaagttt 9600 ttatttcaat cccttttcct ttgaataact gaccaagaac aacaagaaaa aaaaaaaaaa 9660 agaaaaggat cattttgaaa ggatattttt cgctcctatt caaatactgt atttttacca 9720 aaaaaactgt atttttccta cactctcaag ctttgttttt cgcttcgact ctcatgattt 9780 ccttcatatg ccaatcactc tatttataaa tggcataagg tagtgtgaac aattgcaaag 9840 cttgtcatca aaagcttgca atgtacaaat taatgttttt catgcctttc aaaattatct 9900 gcacccccta gctattaatc taacatctaa gtaaggctag tgaatttttt cgaatagtca 9960 tgcagtgcat taatttcccc gtgactattt tggctttgac tccaacactg gccccgtaca 10020 tccgtccctc attacatgaa aagaaatatt gtttatattc ttaattaaaa atattgtccc 10080 ttctaaattt tcatatagtt aattattata ttactttttt ctctattcta ttagttctat 10140 tttcaaatta ttatttatgc atatgtaaag tacattatat ttttgctata tacttaaata 10200 tttctaaatt attaaaaaaa gactgatatg aaaaatttat tctttttaaa gctatatcat 10260 tttatatata ctttttcttt tcttttcttt cattttctat tcaatttaat aagaaataaa 10320 ttttgtaaat ttttatttat caatttataa aaatatttta ctttatatgt tttttcacat 10380 ttttgttaaa caaatcatat cattatgatt gaaagagagg aaattgacag tgagtaataa 10440 gtgatgagaa aaaaatgtgt tatttcctaa aaaaaaccta aacaaacatg tatctactct 10500 ctatttcatc tatctctcat ttcatttttc tctttatctc tttctttatt tttttatcat 10560 atcatttcac attaattatt tttactctct ttattttttc tctctatccc tctcttattt 10620 ccactcatat atacactcca aaattggggc atgcctttat cactactcta tctcctccac 10680 taaatcattt aaatgaaact gaaaagcatt ggcaagtctc ctcccctcct caagtgattt 10740 ccaactcagc attggcatct aattgattca gtatatctat tgcatgtgta aaagtctttc 10800 cacaatacat aactattaat taatcttaaa taaataaagg ataaaatatt tttttttctt 10860 cataaaatta aaatatgtta ttttttgttt agatgtatat tcgaataaat ctaaatatat 10920 gataatgatt ttttatattg attaaacata taatcaatat taaatatgat atttttttat 10980 ataggttgta cacataattt tataaggata aaaaatatga taaaaataaa ttttaaatat 11040 ttttatattt acgagaaaaa aaaatatttt agccataaat aaatgaccag catattttac 11100 aaccttagta attcataaat tcctatatgt atatttgaaa ttaaaaacag ataatcgtta 11160 agggaaggaa tcctacgtca tctcttgcca tttgtttttc atgcaaacag aaagggacga 11220 aaaaccacct caccatgaat cactcttcac accattttta ctagcaaaca agtctcaaca 11280 actgaagcca gctctctttc cgtttctttt tacaacactt tctttgaaat agtagtattt 11340 ttttttcaca tgatttatta acgtgccaaa agatgcttat tgaatagagt gcacatttgt 11400 aatgtactac taattagaac atgaaaaagc attgttctaa cacgataatc ctgtgaaggc 11460 gttaactcca aagatccaat ttcactatat aaattgtgac gaaagcaaaa tgaattcaca 11520 tagctgagag agaaaggaaa ggttaactaa gaagcaatac ttcagcggcc gcaccatggc 11580 tctcagtctt accacagaac agctgttaga acgccctgat ttggttgcga ttgatggcat 11640 cctctacgac cttgaagggc ttgccaaagt tcatccagga ggagatttga ttctcgcttc 11700 tggtgcctct gatgcctccc ctctctttta ttcgatgcat ccatacgtca aaccggagaa 11760 ttccaaattg cttcaacagt tcgtccgagg gaagcatgac cgcacctcga aggacattgt 11820 ctacacgtat gattctccct tcgcacaaga cgttaagcgg acaatgcgcg aggtgatgaa 11880 agggaggaac tggtacgcaa cccctggctt ctggctgcgc accgttggga tcatcgccgt 11940 gacggccttt tgcgagtggc actgggctac cacggggatg gtgctgtggg gcctgctgac 12000 tggattcatg cacatgcaga tcggcttatc catccagcat gatgcgtccc acggggccat 12060 cagcaagaag ccttgggtca acgccctctt cgcctacggc attgacgtca tcggatcgtc 12120 ccggtggatt tggctgcagt cgcacatcat gcggcaccac acctacacca accagcacgg 12180 cctcgacctg gatgcggagt cggcagagcc gttcctggtg ttccacaact accccgccgc 12240 aaacaccgcc cgaaagtggt tccaccgctt ccaagcttgg tacatgtacc ttgtgctggg 12300 ggcatacggg gtatcgctgg tgtacaaccc gctctacatt ttccggatgc agcacaatga 12360 caccatccca gagtctgtca cggccatgcg ggagaatggc tttctgcggc gctaccgcac 12420 acttgcattc gtgatgcgag ctttcttcat cttccggacc gcattcttgc cctggtacct 12480 cactgggacc tcattgctga tcaccattcc tctggtgccc actgcaactg gtgccttctt 12540 gacgttcttc ttcattttgt cccacaattt tgatggctcc gaacggatcc ccgacaagaa 12600 ctgcaaggtt aagagctctg agaaggacgt tgaggctgac caaattgact ggtatcgggc 12660 gcaggtggag acgtcctcca catacggtgg ccccatcgcc atgttcttca ctggcggtct 12720 caatttccag atcgagcacc acctctttcc ccggatgtcg tcttggcact accccttcgt 12780 ccagcaggcg gtccgggagt gttgcgaacg ccatggagtg cgatatgttt tctatcctac 12840 catcgtcggc aacatcatct ccaccctgaa gtacatgcat aaggtgggtg tcgtccactg 12900 cgtgaaggac gcacaggatt cctgagcggc cgcatgagcc gtaaaggttc aatacaacga 12960 gtgcttgttt tcttagggac aagcattgta cttatgtatg attctgtgta accatgagtc 13020 ttccacgttg tactaatgtg aagggcaaaa ataaaacaca gaacaagttc gtttttctca 13080 aataatgtga aggtagaaaa tggaaccatg cctcctctct tgcatgtgat ttaaaatatt 13140 agcagatggt acc 13153 <210> 19 <2 ii> 4975 <212> DNA <213> Seqüência Artificial <220> <223> plasmideo KS133 <220> <221> misc feature <222> (3952)..(3952) <223> η é a, c, g, ou t <400> 19 ggccgccgac tcgacgatga gcgagatgac cagctccggc cgcgacacaa gtgtgagagt 60 actaaataaa tgctttggtt gtacgaaatc attacactaa ataaaataat caaagcttat 120 atatgccttc cgctaaggcc gaatgcaaag aaattggttc tttctcgtta tcttttgcca 180 cttttactag tacgtattaa ttactactta atcatctttg tttacggctc attatatccg 240 tcgacggcgc gcccgatcat ccggatatag ttcctccttt cagcaaaaaa cccctcaaga 300 cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtggca gcagccaact 360 cagcttcctt tcgggctttg ttagcagccg gatcgatcca agctgtacct cactattcct 420 ttgccctcgg acgagtgctg gggcgtcggt ttccactatc ggcgagtact tctacacagc 480 catcggtcca gacggccgcg cttctgcggg cgatttgtgt acgcccgaca gtcccggctc 540 cggatcggac gattgcgtcg catcgaccct gcgcccaagc tgcatcatcg aaattgccgt 600
caaccaagct ctgatagagt tggtcaagac caatgcggag catatacgcc cggagccgcg 660
gcgatcctgc aagctccgga tgcctccgct cgaagtagcg cgtctgctgc tccatacaag 720
ccaaccacgg cctccagaag aagatgttgg cgacctcgta ttgggaatcc ccgaacatcg 780
cctcgctcca gtcaatgacc gctgttatgc ggccattgtc cgtcaggaca ttgttggagc 840
cgaaatccgc gtgcacgagg tgccggactt cggggcagtc ctcggcccaa agcatcagct 900
catcgagagc ctgcgcgacg gacgcactga cggtgtcgtc catcacagtt tgccagtgat 960
acacatgggg atcagcaatc gcgcatatga aatcacgcca tgtagtgtat tgaccgattc 1020
cttgcggtcc gaatgggccg aacccgctcg tctggctaag atcggccgca gcgatcgcat 1080
ccatagcctc cgcgaccggc tgcagaacag cgggcagttc ggtttcaggc aggtcttgca 1140
acgtgacacc ctgtgcacgg cgggagatgc aataggtcag gctctcgctg aattccccaa 1200
tgtcaagcac ttccggaatc gggagcgcgg ccgatgcaaa gtgccgataa acataacgat 1260
ctttgtagaa accatcggcg cagctattta cccgcaggac atatccacgc cctcctacat 1320
cgaagctgaa agcacgagat tcttcgccct ccgagagctg catcaggtcg gagacgctgt 1380
cgaacttttc gatcagaaac ttctcgacag acgtcgcggt gagttcaggc ttttccatgg 1440
gtatatctcc ttcttaaagt taaacaaaat tatttctaga gggaaaccgt tgtggtctcc 1500
ctatagtgag tcgtattaat ttcgcgggat cgagatctga tcaacctgca ttaatgaatc 1560
ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 1620
gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 1680
atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 174 0
caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 1800
cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 1860
taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 1920
ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc 1980
tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 2040
gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 2100
ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 2160
aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 2220
aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 2280
agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 2340
cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 2400 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgacatt aacctataaa 2460 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc 2520
tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga 2580
caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg 2640
gcatcagagc agattgtact gagagtgcac catatggaca tattgtcgtt agaacgcggc 2700
tacaattaat acataacctt atgtatcata cacatacgat ttaggtgaca ctatagaacg 2760
gcgcgccaag cttggatcct cgaagagaag ggttaataac acactttttt aacattttta 2820
acacaaattt tagttattta aaaatttatt aaaaaattta aaataagaag aggaactctt 2880
taaataaatc taacttacaa aatttatgat ttttaataag ttttcaccaa taaaaaatgt 2940
cataaaaata tgttaaaaag tatattatca atattctctt tatgataaat aaaaagaaaa 3000
aaaaaataaa agttaagtga aaatgagatt gaagtgactt taggtgtgta taaatatatc 3060
aaccccgcca acaatttatt taatccaaat atattgaagt atattattcc atagccttta 3120
tttatttata tatttattat ataaaagctt tatttgttct aggttgttca tgaaatattt 3180
ttttggtttt atctccgttg taagaaaatc atgtgctttg tgtcgccact cactattgca 3240
gctttttcat gcattggtca gattgacggt tgattgtatt tttgtttttt atggttttgt 3300
gttatgactt aagtcttcat ctctttatct cttcatcagg tttgatggtt acctaatatg 3360
gtccatgggt acatgcatgg ttaaattagg tggccaactt tgttgtgaac gatagaattt 3420
tttttatatt aagtaaacta tttttatatt atgaaataat aataaaaaaa atattttatc 3480
attattaaca aaatcatatt agttaatttg ttaactctat aataaaagaa atactgtaac 3540
attcacatta catggtaaca tctttccacc ctttcatttg ttttttgttt gatgactttt 3600
tttcttgttt aaatttattt cccttctttt aaatttggaa tacattatca tcatatataa 3660
actaaaatac taaaaacagg attacacaaa tgataaataa taacacaaat atttataaat 3720
ctagctgcaa tatatttaaa ctagctatat cgatattgta aaataaaact agctgcattg 3780
atactgataa aaaaatatca tgtgctttct ggactgatga tgcagtatac ttttgacatt 3840
gcctttattt tatttttcag aaaagctttc ttagttctgg gttcttcatt atttgtttcc 3900
catctccatt gtgaattgaa tcatttgctt cgtgtcacaa atacaattta gntaggtaca 3960
tgcattggtc agattcacgg tttattatgt catgacttaa gttcatggta gtacattacc 4020
tgccacgcat gcattatatt ggttagattt gataggcaaa tttggttgtc aacaatataa 4080
atataaataa tgtttttata ttacgaaata acagtgatca aaacaaacag ttttatcttt 4140
attaacaaga ttttgttttt gtttgatgac gttttttaat gtttacgctt tcccccttct 4200
tttgaattta gaacacttta tcatcataaa atcaaatact aaaaaaatta catatttcat 4260 aaataataac acaaatattt ttaaaaaatc tgaaataata atgaacaata ttacatatta 4320 tcacgaaaat tcattaataa aaatattata taaataaaat gtaatagtag ttatatgtag 4380 gaaaaaagta ctgcacgcat aatatataca aaaagattaa aatgaactat tataaataat 4440 aacactaaat taatggtgaa tcatatcaaa ataatgaaaa agtaaataaa atttgtaatt 4500 aacttctata tgtattacac acacaaataa taaataatag taaaaaaaat tatgataaat 4560 atttaccatc tcataagata tttaaaataa tgataaaaat atagattatt ttttatgcaa 4620 ctagctagcc aaaaagagaa cacgggtata tataaaaaga gtacctttaa attctactgt 4680 acttccttta ttcctgacgt ttttatatca agtggacata cgtgaagatt ttaattatca 4740 gtctaaatat ttcattagca cttaatactt ttctgtttta ttcctatcct ataagtagtc 4800 ccgattctcc caacattgct tattcacaca actaactaag aaagtcttcc atagcccccc 4860 aagcggccgg agctggtcat ctcgctcatc gtcgagtcgg cggccggagc tggtcatctc 4920 gctcatcgtc gagtcggcgg ccgccgactc gacgatgagc gagatgacca gctcc 4975 <210> 20 <211> 5626 <212> DNA <213> Seqüência Artificial <220> <223> plasmideo KS263 <220> <221> misc feature <222> (3989)..(3989) <223> η é a, c, g, ou t <400> 20 ggccgccgac tcgacgatga gcgagatgac cagctccggc cgccgactcg acgatgagcg 60 agatgaccag ctccggccgc gacacaagtg tgagagtact aaataaatgc tttggttgta 120 cgaaatcatt acactaaata aaataatcaa agcttatata tgccttccgc taaggccgaa 180 tgcaaagaaa ttggttcttt ctcgttatct tttgccactt ttactagtac gtattaatta 240 ctacttaatc atctttgttt acggctcatt atatccgtcg acggcgcgcc cgatcatccg 300 gatatagttc ctcctttcag caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt 360 tatgctagtt attgctcagc ggtggcagca gccaactcag cttcctttcg ggctttgtta 420 gcagccggat cgatccaagc tgtacctcac tattcctttg ccctcggacg agtgctgggg 480 cgtcggtttc cactatcggc gagtacttct acacagccat cggtccagac ggccgcgctt 540 ctgcgggcga tttgtgtacg cccgacagtc ccggctccgg atcggacgat tgcgtcgcat 600 cgaccctgcg cccaagctgc atcatcgaaa ttgccgtcaa ccaagctctg atagagttgg 660 tcaagaccaa tgcggagcat atacgcccgg agccgcggcg atcctgcaag ctccggatgc 720 ctccgctcga agtagcgcgt ctgctgctcc atacaagcca accacggcct ccagaagaag 780 atgttggcga cctcgtattg ggaatccccg aacatcgcct cgctccagtc aatgaccgct 840 gttatgcggc cattgtccgt caggacattg ttggagccga aatccgcgtg cacgaggtgc 900 cggacttcgg ggcagtcctc ggcccaaagc atcagctcat cgagagcctg cgcgacggac 960 gcactgacgg tgtcgtccat cacagtttgc cagtgataca catggggatc agcaatcgcg 1020 catatgaaat cacgccatgt agtgtattga ccgattcctt gcggtccgaa tgggccgaac 1080 ccgctcgtct ggctaagatc ggccgcagcg atcgcatcca tagcctccgc gaccggctgc 1140 agaacagcgg gcagttcggt ttcaggcagg tcttgcaacg tgacaccctg tgcacggcgg 1200 gagatgcaat aggtcaggct ctcgctgaat tccccaatgt caagcacttc cggaatcggg 1260 agcgcggccg atgcaaagtg ccgataaaca taacgatctt tgtagaaacc atcggcgcag 1320 ctatttaccc gcaggacata tccacgccct cctacatcga agctgaaagc acgagattct 1380 tcgccctccg agagctgcat caggtcggag acgctgtcga acttttcgat cagaaacttc 1440 tcgacagacg tcgcggtgag ttcaggcttt tccatgggta tatctccttc ttaaagttaa 1500 acaaaattat ttctagaggg aaaccgttgt ggtctcccta tagtgagtcg tattaatttc 1560 gagctgatca acctgcatta atgaatcggc caacgcgcgg ggagaggcgg 1620 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 1680 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 1740 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 1800 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 1860 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 1920 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 1980 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc 2040 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 2100 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 2160 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 2220 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc 2280 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 2340 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 2400 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 2460 acgttaaggg attttggtca tgacattaac ctataaaaat aggcgtatca cgaggccctt 2520 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac 2580
ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc 2640
gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag 2700
agtgcaccat atggacatat tgtcgttaga acgcggctac aattaataca taaccttatg 2760
tatcatacac atacgattta ggtgacacta tagaacggcg cgccaagctt ggatcctcga 2820
agagaagggt taataacaca cttttttaac atttttaaca caaattttag ttatttaaaa 2880
atttattaaa aaatttaaaa taagaagagg aactctttaa ataaatctaa cttacaaaat 2940
ttatgatttt taataagttt tcaccaataa aaaatgtcat aaaaatatgt taaaaagtat 3000
attatcaata ttctctttat gataaataaa aagaaaaaaa aaataaaagt taagtgaaaa 3060
tgagattgaa gtgactttag gtgtgtataa atatatcaac cccgccaaca atttatttaa 3120
tccaaatata ttgaagtata ttattccata gcctttattt atttatatat ttattatata 3180
aaagctttat ttgttctagg ttgttcatga aatatttttt tggttttatc tccgttgtaa 3240
gaaaatcatg tgctttgtgt cgccactcac tattgcagct ttttcatgca ttggtcagat 3300
tgacggttga ttgtattttt gttttttatg gttttgtgtt atgacttaag tcttcatctc 3360
tttatctctt catcaggttt gatggttacc taatatggtc catgggtaca tgcatggtta 3420
aattaggtgg ccaactttgt tgtgaacgat agaatttttt ttatattaag taaactattt 3480
ttatattatg aaataataat aaaaaaaata ttttatcatt attaacaaaa tcatattagt 3540
taatttgtta actctataat aaaagaaata ctgtaacatt cacattacat ggtaacatct 3600
ttccaccctt tcatttgttt tttgtttgat gacttttttt cttgtttaaa tttatttccc 3660
ttcttttaaa tttggaatac attatcatca tatataaact aaaatactaa aaacaggatt 3720
acacaaatga taaataataa cacaaatatt tataaatcta gctgcaatat atttaaacta 3780
gctatatcga tattgtaaaa taaaactagc tgcattgata ctgataaaaa aatatcatgt 3840
gctttctgga ctgatgatgc agtatacttt tgacattgcc tttattttat ttttcagaaa 3900
agctttctta gttctgggtt cttcattatt tgtttcccat ctccattgtg aattgaatca 3960
tttgcttcgt gtcacaaata caatttagnt aggtacatgc attggtcaga ttcacggttt 4020
attatgtcat gacttaagtt catggtagta cattacctgc cacgcatgca ttatattggt 4080
tagatttgat aggcaaattt ggttgtcaac aatataaata taaataatgt ttttatatta 4140
cgaaataaca gtgatcaaaa caaacagttt tatctttatt aacaagattt tgtttttgtt 4200
tgatgacgtt ttttaatgtt tacgctttcc cccttctttt gaatttagaa cactttatca 4260
tcataaaatc aaatactaaa aaaattacat atttcataaa taataacaca aatattttta 4320
aaaaatctga aataataatg aacaatatta catattatca cgaaaattca ttaataaaaa 4380 tattatataa ataaaatgta atagtagtta tatgtaggaa aaaagtactg cacgcataat 4440 atatacaaaa agattaaaat gaactattat aaataataac actaaattaa tggtgaatca 4500 tatcaaaata atgaaaaagt aaataaaatt tgtaattaac ttctatatgt attacacaca 4560 caaataataa ataatagtaa aaaaaattat gataaatatt taccatctca taagatattt 4620 aaaataatga taaaaatata gattattttt tatgcaacta gctagccaaa aagagaacac 4680 gggtatatat aaaaagagta cctttaaatt ctactgtact tcctttattc ctgacgtttt 4740 tatatcaagt ggacatacgt gaagatttta attatcagtc taaatatttc attagcactt 4800 aatacttttc tgttttattc ctatcctata agtagtcccg attctcccaa cattgcttat 4860 tcacacaact aactaagaaa gtcttccata gccccccaag cggccggagc tggtcatctc 4920 gctcatcgtc gagtcggcgg ccggagctgg tcatctcgct catcgtcgag tcggcggccc 4980 tcctccaccg tttaagattg cagaaatcag agcttcaata ccaaaacatt gctgggtcaa 5040 gaatccatgg agatccctca gttatgttct cagggatgtg cttgtaattg ctgcattggt 5100 ggctgcagca attcacttcg acaactggct tctctggcta atctattgcc ccattcaagg 5160 cacaatgttc tgggctctct ttgttcttgg acatgattgt ggccatggaa gcttttcaga 5220 tagccctttg ctgaatagcc tggtgggaca catcttgcat tcctcaattc ttgtgccata 5280 ccatggatgg agaattagcc acagaactca ccatcaaaac catggacaca ttgagaagga 5340 tgagtcatgg gttccattaa cagagaagat ttacaagaat ctagacagca tgacaagact 5400 cattagattc actgtgccat ttccattgtt tgtgtatcca atttatttgt tttcaagaag 5460 ccccggaaag gaaggctctc acttcaatcc ctacagcaat ctgttcccac ccagtgagag 5520 aaaaggaata gcaatatcaa cactgtgttg ggctaccatg ttttctctgc ttatctatct 5580 ctcattcata actagtccac ttctagtgct caagctctat ggaatt 5626 <210> 21 <211> 17 <212> DNA <213> Seqüência Artificial
<220>
<223> primer ELfad3-5Not
<400> 21
ccccaagcgg ccgcagc 17
<210> 22
<211> 20
<212> DNA
<213> Seqüência Artificial <220> <223> primer ELfad3-3Not
<400> 22
acttgtgtcg cggccgcagc 20
<210> 23
<211> 807
<212> DNA
<213> Seqüência Artificial
<220>
<223> ELVISLIVES/fad3 cassette ELfad3Not
<400> 23
gcggccggag ctggtcatct cgctcatcgt cgagtcggcg gccggagctg gtcatctcgc 60 tcatcgtcga gtcggcggcc ctcctccacc gtttaagatt gcagaaatca gagcttcaat 120 accaaaacat tgctgggtca agaatccatg gagatccctc agttatgttc tcagggatgt 180 gcttgtaatt gctgcattgg tggctgcagc aattcacttc gacaactggc ttctctggct 240 aatctattgc cccattcaag gcacaatgtt ctgggctctc tttgttcttg gacatgattg 300 tggccatgga agcttttcag atagcccttt gctgaatagc ctggtgggac acatcttgca 360 ttcctcaatt cttgtgccat accatggatg gagaattagc cacagaactc accatcaaaa 420 ccatggacac attgagaagg atgagtcatg ggttccatta acagagaaga tttacaagaa 480 tctagacagc atgacaagac tcattagatt cactgtgcca tttccattgt ttgtgtatcc 540 aatttatttg ttttcaagaa gccccggaaa ggaaggctct cacttcaatc cctacagcaa 600 tctgttccca cccagtgaga gaaaaggaat agcaatatca acactgtgtt gggctaccat 660 gttttctctg cttatctatc tctcattcat aactagtcca cttctagtgc tcaagctcta 720 tggaattggc cgccgactcg acgatgagcg agatgaccag ctccggccgc cgactcgacg 780 atgagcgaga tgaccagctc cggccgc 807
<210> 24
<211> 4480
<212> DNA
<213> Seqüência Artificial
<220>
<223> primer pKR179
<400> 24
ctagacctgc aggatccatg cccttcattt gccgcttatt aattaatttg gtaacagtcc 60 gtactaatca gttacttatc cttcccccat cataattaat cttggtagtc tcgaatgcca 120 caacactgac tagtctcttg gatcataaga aaaagccaag gaacaaaaga agacaaaaca 180 caatgagagt atcctttgca tagcaatgtc taagttcata aaattcaaac aaaaacgcaa 240 tcacacacag tggacatcac ttatccacta gctgatcagg atcgccgcgt caagaaaaaa 300 aaactggacc ccaaaagcca tgcacaacaa cacgtactca caaaggtgtc aatcgagcag 360
cccaaaacat tcaccaactc aacccatcat gagccctcac atttgttgtt tctaacccaa 420
cctcaaactc gtattctctt ccgccacctc atttttgttt atttcaacac ccgtcaaact 480
gcatgccacc ccgtggccaa atgtccatgc atgttaacaa gacctatgac tataaatagc 540
tgcaatctcg gcccaggttt tcatcatcaa gaaccagttc aatatcctag tacaccgtat 600
taaagaattt aagatatact gcggccgcaa gtatgaacta aaatgcatgt aggtgtaaga 660
gctcatggag agcatggaat attgtatccg accatgtaac agtataataa ctgagctcca 720
tctcacttct tctatgaata aacaaaggat gttatgatat attaacactc tatctatgca 780
ccttattgtt ctatgataaa tttcctctta ttattataaa tcatctgaat cgtgacggct 840
tatggaatgc ttcaaatagt acaaaaacaa atgtgtacta taagactttc taaacaattc 900
taaccttagc attgtgaacg agacataagt gttaagaaga cataacaatt ataatggaag 960
aagtttgtct ccatttatat attatatatt acccacttat gtattatatt aggatgttaa 1020
ggagacataa caattataaa gagagaagtt tgtatccatt tatatattat atactaccca 1080
tttatatatt atacttatcc acttatttaa tgtctttata aggtttgatc catgatattt 1140
ctaatatttt agttgatatg tatatgaaag ggtactattt gaactctctt actctgtata 1200
aaggttggat catccttaaa gtgggtctat ttaattttat tgcttcttac agataaaaaa 1260
aaaattatga gttggtttga taaaatattg aaggatttaa aataataata aataacatat 1320
aatatatgta tataaattta ttataatata acatttatct ataaaaaagt aaatattgtc 1380
ataaatctat acaatcgttt agccttgctg gacgaatctc aattatttaa acgagagtaa 1440
acatatttga ctttttggtt atttaacaaa ttattattta acactatatg aaattttttt 1500
ttttatcagc aaagaataaa attaaattaa gaaggacaat ggtgtcccaa tccttataca 1560
accaacttcc acaagaaagt caagtcagag acaacaaaaa aacaagcaaa ggaaattttt 1620
taatttgagt tgtcttgttt gctgcataat ttatgcagta aaacactaca cataaccctt 1680
ttagcagtag agcaatggtt gaccgtgtgc ttagcttctt ttattttatt tttttatcag 1740
caaagaataa ataaaataaa atgagacact tcagggatgt ttcaacctgc aggtctagag 1800
gatccccggg taccgagctc gaattcactg gccgtcgttt tacaacgtcg tgactgggaa 1860
aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt 1920
aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa 1980
tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg 2040
tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca 2100
acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct 2160 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg 2220
agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt 2280
tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt 2340
ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa 2400
taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt 2460
tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat 2520
gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag 2580
atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg 2640
ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata 2700
cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat 2760
ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc 2820
aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg 2880
ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac 2940
gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact 3000
ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa 3060
gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct 3120
ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc 3180
tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga 3240
cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac 3300
tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag 3360
atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg 3420
tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc 3480
tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag 3540
ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc 3600
cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac 3660
ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc 3720
gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt 3780
tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt 3840
gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc 3900
ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt 3960
tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca 4020 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt 4080 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt 4140 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag 4200 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg 4260 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc 4320 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt 4380 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat 4440 gaccatgatt acgccaagct tgcatgcctg caggtcgact 4480 <210> 25 <211> 5279 <212> DNA <213> Seqüência Artificial <220> <223> primer BC-ELfad3 <400> 25 ctagacctgc aggatccatg cccttcattt gccgcttatt aattaatttg gtaacagtcc 60 gtactaatca gttacttatc cttcccccat cataattaat cttggtagtc tcgaatgcca 120 caacactgac tagtctcttg aatcataaaa aaaagccaag gaacaaaaga agacaaaaca 180 caatgagagt atcctttgca tagcaatgtc taagttcata aaattcaaac aaaaacgcaa 240 tcacacacag tggacatcac ttatccacta gctgatcagg atcgccgcgt caagaaaaaa 300 aaactggacc ccaaaagcca tgcacaacaa cacgtactca caaaggtgtc aatcgagcag 360 cccaaaacat tcaccaactc aacccatcat gagccctcac atttgttgtt tctaacccaa 420 cctcaaactc gtattctctt ccgccacctc atttttgttt atttcaacac ccgtcaaact 480 gcatgccacc ccgtggccaa atgtccatgc atgttaacaa gacctatgac tataaatagc 540 tgcaatctcg gcccaggttt tcatcatcaa gaaccagttc aatatcctag tacaccgtat 600 taaagaattt aagatatact gcggccgcag ctggtcatct cgctcatcgt cgagtcggcg 660 gccggagctg gtcatctcgc tcatcgtcga gtcggcggcc ctcctccacc gtttaagatt 720 gcagaaatca gagcttcaat accaaaacat tgctgggtca agaatccatg gagatccctc 780 agttatgttc tcagggatgt gcttgtaatt gctgcattgg tggctgcagc aattcacttc 840 gacaactggc ttctctggct aatctattgc cccattcaag gcacaatgtt ctgggctctc 900 tttgttcttg gacatgattg tggccatgga agcttttcag atagcccttt gctgaatagc 960 ctggtgggac acatcttgca ttcctcaatt cttgtgccat accatggatg gagaattagc 1020 cacagaactc accatcaaaa ccatggacac attgagaagg atgagtcatg ggttccatta 1080 acagagaaga tttacaagaa tctagacagc atgacaagac tcattagatt cactgtgcca 1140
tttccattgt ttgtgtatcc aatttatttg ttttcaagaa gccccggaaa ggaaggctct 1200
cacttcaatc cctacagcaa tctgttccca cccagtgaga gaaaaggaat agcaatatca 1260
acactgtgtt gggctaccat gttttctctg cttatctatc tctcattcat aactagtcca 1320
cttctagtgc tcaagctcta tggaattggc cgccgactcg acgatgagcg agatgaccag 1380
ctccggccgc cgactcgacg atgagcgaga tgaccagctg cggccgcaag tatgaactaa 1440
aatgcatgta ggtgtaagag ctcatggaga gcatggaata ttgtatccga ccatgtaaca 1500
gtataataac tgagctccat ctcacttctt ctatgaataa acaaaggatg ttatgatata 1560
ttaacactct atctatgcac cttattgttc tatgataaat ttcctcttat tattataaat 1620
catctgaatc gtgacggctt atggaatgct tcaaatagta caaaaacaaa tgtgtactat 1680
aagactttct aaacaattct aaccttagca ttgtgaacga gacataagtg ttaagaagac 1740
ataacaatta taatggaaga agtttgtctc catttatata ttatatatta cccacttatg 1800
tattatatta ggatgttaag gagacataac aattataaag agagaagttt gtatccattt 1860
atatattata tactacccat ttatatatta tacttatcca cttatttaat gtctttataa 1920
ggtttgatcc atgatatttc taatatttta gttgatatgt atatgaaagg gtactatttg 1980
aactctctta ctctgtataa aggttggatc atccttaaag tgggtctatt taattttatt 2040
gcttcttaca gataaaaaaa aaattatgag ttggtttgat aaaatattga aggatttaaa 2100
ataataataa ataacatata atatatgtat ataaatttat tataatataa catttatcta 2160
taaaaaagta aatattgtca taaatctata caatcgttta gccttgctgg acgaatctca 2220
attatttaaa cgagagtaaa catatttgac tttttggtta tttaacaaat tattatttaa 2280
cactatatga aatttttttt tttatcagca aagaataaaa ttaaattaag aaggacaatg 2340
gtgtcccaat ccttatacaa ccaacttcca caagaaagtc aagtcagaga caacaaaaaa 2400
acaagcaaag gaaatttttt aatttgagtt gtcttgtttg ctgcataatt tatgcagtaa 2460
aacactacac ataacccttt tagcagtaga gcaatggttg accgtgtgct tagcttcttt 2520
tattttattt ttttatcagc aaagaataaa taaaataaaa tgagacactt cagggatgtt 2580
tcaacctgca ggtctagagg atccccgggt accgagctcg aattcactgg ccgtcgtttt 2640
acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc 2700
ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt 2760
gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg 2820
tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag 2880
ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc 2940 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc 3000 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa 3060 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg 3120 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata 3180 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg 3240 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac 3300 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact 3360 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat 3420 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga 3480 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac 3540 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat 3600 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac 3660 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct 3720 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac 3730 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga 3840 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg 3900 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact 3960 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac 4020 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta 4080 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt 4140 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga 4200 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc 4260 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt 4320 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc 4380 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc 4 4 4 0 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 4500 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 4560 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga 4620 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc 4680 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 4 7 4 0 gçgaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg 4800 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt 4860
tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc 4920
tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg 4980
aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc 5040
gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg 5100
gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca 5160
ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt 5220
tcacacagga aacagctatg accatgatta cgccaagctt gcatgcctgc aggtcgact 5279
<210> 26 <211> 12582 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmídeo BC-Tpom_Ann-HPfad3ABA'_ALS <400> 26
gtacgtctag aggatccgtc gacggcgcgc ccgatcatcc ggatatagtt cctcctttca 60
gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt tattgctcag 120
cggtggcagc agccaactca gcttcctttc gggctttgtt agcagccgga tcgatccaag 180
ctgtacctca ctattccttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg 240
cgagtacttc tacacagcca tcggtccaga cggccgcgct tctgcgggcg atttgtgtac 300
gcccgacagt cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg 360
catcatcgaa attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca 420
tatacgcccg gagccgcggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg 480
tctgctgctc catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt 540
gggaatcccc gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg 600
tcaggacatt gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct 660
cggcccaaag catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca 720
tcacagtttg ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg 780
tagtgtattg accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat 840
cggccgcagc gatcgcatcc atagcctccg cgaccggctg cagaacagcg ggcagttcgg 900
tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc 960
tctcgctgaa ttccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt 1020
gccgataaac ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat 1080 atccacgccc tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca 1140
tcaggtcgga gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga 1200
gttcaggctt ttccatgggt atatctcctt cttaaagtta aacaaaatta tttctagagg 1260
gaaaccgttg tggtctccct atagtgagtc gtattaattt cgcgggatcg agatctgatc 1320
aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 1380
tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 1440
gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 1500
atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 1560
ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 1620
cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 1680
tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 1740
gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 1800
aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 1860
tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 1920
aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 1980
aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 2040
ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 2100
ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 2160
atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 2220
atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt 2280
gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa 2340
gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg 2400
ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatggacata 2460
ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca catacgattt 2520
aggtgacact atagaacggc gcgccaagct gggtctagaa ctagaaacgt gatgccactt 2580
gttattgaag tcgattacag catctattct gttttactat ttataacttt gccatttctg 2640
acttttgaaa actatctctg gatttcggta tcgctttgtg aagatcgagc aaaagagacg 2700
ttttgtggac gcaatggtcc aaatccgttc tacatgaaca aattggtcac aatttccact 2760
aaaagtaaat aaatggcaag ttaaaaaagg aatatgcatt ttactgattg cctaggtgag 2820
ctccaagaga agttgaatct acacgtctac caaccgctaa aaaaagaaaa acattgatat 2880
gtaacctgat tccattagct tttgacttct tcaacagatt ctctacttag atttctaaca 2940 gaaatattat tactagcaca tcattttcag tctcactaca gcaaaaaatc caacggcaca 3000
atacagacaa caggagatat cagactacag agatagatag atgctactgc atgtagtaag 3060
ttaaataaaa ggaaaataaa atgtcttgct accaaaacta ctacagacta tgatgctcac 3120
cacaggccaa atcctgcaac taggacagca ttatcttata tatattgtac aaaacaagca 3180
tcaaggaaca tttggtctag gcaatcagta cctcgttcta ccatcaccct cagttatcac 3240
atccttgaag gatccattac tgggaatcat cggcaacaca tgctcctgat ggggcacaat 3300
gacatcaaga aggtaggggc caggggtgtc caacattctc tgaattgccg ctctaagctc 3360
ttccttcttc gtcactcgcg ctgccggtat cccacaagca tcagcaaact tgagcatgtt 3420
tgggaatatc tcgctctcgc tagacggatc tccaagatag gtgtgagctc tattggactt 3480
gtagaaccta tcctccaact gaaccaccat acccaaatgc tgattgttca acaacaatat 3540
cttaactggg agattctcca ctcttatagt ggccaactcc tgaacattca tgatgaaact 3600
accatcccca tcaatgtcaa ccacaacagc cccagggtta gcaacagcag caccaatagc 3660
cgcaggcaat ccaaaaccca tggctccaag accccctgag gtcaaccact gcctcggtct 3720
cttgtacttg taaaactgcg cagcccacat ttgatgctgc ccaaccccag tactaacaat 3780
agcatctcca ttagtcaact catcaagaac ctcgatagca tgctgcggag aaatcgcgtc 3840
nt nna a rti-η t-trrhasnrrs Λ t- rrrra ^ 3 Ct-1- rrt- rrt- f- c t" ^C aca^^aa^c1" pt-fpi-rí-pna 39Q0
---J-J----j----w — — \J
acctccaaga tcaaacttac cctccactcc tttctcctcc aaaatcatat taattccctt 3960
caaggccaac ttcaaatccg cgcaaaccga cacgtgcgcc tgcttgttct tcccaatctc 4020
ggcagaatca atatcaatgt gaacaatctt agccctacta gcaaaagcct caagcttccc 4080
agtaacacgg tcatcaaacc ttaccccaaa ggcaagcaac aaatcactat tgtcaacagc 4140
atagttagca taaacagtac catgcatacc cagcatctga agggaatatt catcaccaat 4200
aggaaaagtt ccaagaccca ttaaagtgct agcaacggga ataccagtga gt.tcaacaaa 4260
gcgcctcaat tcagcactgg aattcaaact gccaccgccg acgtagagaa cgggcttttg 4320
ggcctccatg atgagtctga caatgtgttc caattgggcc tcggcggggg gcctgggcag 4380
cctggcgagg taaccgggga ggttaacggg ctcgtcccaa ttaggcacgg cgagttgctg 4440
ctgaacgtct ttgggaatgt cgatgaggac cggaccgggg cggccggagg tggcgacgaa 4 500
gaaagcctcg gcgacgacgc gggggatgtc gtcgacgtcg aggatgaggt agttgtgctt 4560
cgtgatggat ctgctcacct ccacgatcgg ggtttcttgg aaggcgtcgg tgccgatcat 4 620
ccggcgggcg acctggccgg tgatggcgac gactgggacg ctgtccatta aagcgtcggc 4 680
gaggccgctc acgaggttgg tggcgccggg gccggaggtg gcaatgcaga cgccggggag 4740
gccggaggaa cgcgcgtagc cttcggcggc gaagacgccg ccctgctcgt ggcgcgggag 4800 cacgttgcgg atggcggcgg agcgcgtgag cgcctggtgg atctccatcg acgcaccgcc 4860
ggggtacgcg aacaccgtcg tcacgccctg cctctccagc gcctccacaa ggatgtccgc 4920
gcccttgcga ggttcgccgg aggcgaaccg tgacacgaag ggctccgtgg tcggcgcttc 4980
cttggtgaag ggcgccgccg tggggggttt ggagatggaa catttgattt tgagagcgtg 5040
gttgggtttg gtgagggttt gatgagagag agggagggtg gatctagtaa tgcgtttggg 5100
gaaggtgggg tgtgaagagg aagaagagaa tcgggtggtt ctggaagcgg tggccgccat 5160
tgtgttgtgt ggcatggtta tacttcaaaa actgcacaac aagcctagag ttagtaccta 5220
aacagtaaat ttacaacaga gagcaaagac acatgcaaaa atttcagcca taaaaaaagt 5280
tataatagaa tttaaagcaa aagtttcatt ttttaaacat atatacaaac aaactggatt 5340
tgaaggaagg gattaattcc cctgctcaaa gtttgaattc ctattgtgac ctatactcga 5400
ataaaattga agcctaagga atgtatgaga aacaagaaaa caaaacaaaa ctacagacaa 5460
acaagtacaa ttacaaaatt cgctaaaatt ctgtaatcac caaaccccat ctcagtcagc 5520
acaaggccca aggtttattt tgaaataaaa aaaaagtgat tttatttctc ataagctaaa 5580
agaaagaaag gcaattatga aatgatttcg actagatctg aaagtccaac gcgtattccg 5640
cagatattaa agaaagagta gagtttcaca tggatcctag atggacccag ttgaggaaaa 5700
agcaaggcaa agcaaaccag aagtgcaaga tccgaaattg aaccacggaa tctaggattt 5760
ggtagaggga gaagaaaagt accttgagag gtagaagaga agagaagagc agagagatat 5820
atgaacgagt gtgtcttggt ctcaactctg aagcgatacg agtttagagg ggagcattga 5880
gttccaattt atagggaaac cgggtggcag gggtgagtta atgacggaaa agcccctaag 5940
taacgagatt ggattgtggg ttagattcaa ccgtttgcat ccgcggctta gattggggaa 6000
gtcagagtga atctcaaccg ttgactgagt tgaaaattga atgtagcaac caattgagcc 6060
aaccccagcc tttgcccttt gattttgatt tgtttgttgc atacttttta tttgtcttct 6120
ggttctgact ctctttctct cgtttcaatg ccaggttgcc tactcccaca ccactcacaa 6180
gaagattcta ctgttagtat taaatatttt ttaatgtatt aaatgatgaa tgcttttgta 6240
aacagaacaa gactatgtct aataagtgtc ttgcaacatt ttttaagaaa ttaaaaaaaa 6300
tatatttatt atcaaaatca aatgtatgaa aaatcatgaa taatataatt ttatacattt 6360
ttttaaaaaa tcttttaatt tcttaattaa tatcttaaaa ataatgatta atatttaacc 6420
caaaataatt agtatgattg gtaaggaaga tatccatgtt atgtttggat gtgagtttga 6480
tctagagcaa agcttactag agtcgacctg caggttgaaa catccctgaa gtgtctcatt 6540
ttattttatt tattctttgc tgataaaaaa ataaaataaa agaagctaag cacacggtca 6600
accattgctc tactgctaaa agggttatgt gtagtgtttt actgcataaa ttatgcagca 6660 aacaagacaa ctcaaattaa aaaatttcct ttgcttgttt ttttgttgtc tctgacttga 6720
ctttcttgtg gaagttggtt gtataaggat tgggacacca ttgtccttct taatttaatt 6780
ttattctttg ctgataaaaa aaaaaatttc atatagtgtt aaataataat ttgttaaata 6840
accaaaaagt caaatatgtt tactctcgtt taaataattg agattcgtcc agcaaggcta 6900
aacgattgta tagatttatg acaatattta cttttttata gataaatgtt atattataat 6960
aaatttatat acatatatta tatgttattt attattattt taaatccttc aatattttat 7020
caaaccaact cataattttt tttttatctg taagaagcaa taaaattaaa tagacccact 7080
ttaaggatga tccaaccttt atacagagta agagagttca aatagtaccc tttcatatac 7140
atatcaacta aaatattaga aatatcatgg atcaaacctt ataaagacat taaataagtg 7200
gataagtata atatataaat gggtagtata taatatataa atggatacaa acttctctct 7260
ttataattgt tatgtctcct taacatccta atataataca taagtgggta atatataata 7320
tataaatgga gacaaacttc ttccattata attgttatgt cttcttaaca cttatgtctc 7380
gttcacaatg ctaaggttag aattgtttag aaagtcttat agtacacatt tgtttttgta 7440
ctatttgaag cattccataa gccgtcacga ttcagatgat ttataataat aagaggaaat 7500
ttatcataga acaataaggt gcatagatag agtgttaata tatcataaca tcctttgttt 7560
attcatagaa gaagtgagat ggagctcagt tattatactg ttacatggtc ggatacaata 7620
ttccatgctc tccatgagct cttacaccta catgcatttt agttcatact tgcggccgct 7680
cattcaggga cagcagccag ctgggcaatc tcgcctaaga agctgaccaa tttctgaacc 7740
ccggggatga tggctggggc cctgtagggc aggttgtgct tggcgcacaa cttctgcacc 7800
tccagggaag ctgccgtgag gttgtgcctg ggcatgttgg gccacaagtg gtgctcaatc 7 8 60
tggtagttga gaccaccaaa gacccaatcg gtgagaaggc cgggcttaat gttcaacgtc 7920
tcgtggatct ggctggcgct gaacccgtgc tcatcccaga cagtctcctc aaccttgtcc 7980
aaaggatagt gattcagaaa cacaacgatg gcaatgccaa atcctccgag cagctcggag 8040
atcaagaagc atcccaaacc ggtggcaaag ctaggcatgg cactgcaata gaaagtggcc 8100
ttgagcaccc agtggagggc cagaccaatc gcctccttag tgtactggcg cttgtagtac 8160
aggttcctct cctcgctctt gtaggacatg acgccgccga tggattggag gcaccagatg 8220
aacctaaggg atgcaatggt gggaatgaag tagtactgct ggtacttgat gagattccga 8280
gtcgaaggag tagccctctt gacatcttcg ggagaccaag ccaggatggg gaggttatca 8340
atatctgggt catggccgac cacattggtg gcggcatggt gaccattgtg cctatccttc 8400
caccaagtct gtgagtatcc ctgcatcacg ttaccgaaaa agagaccgaa agcgtcgtta 8460
atcttgcggt ttgtgaaaac ctggtgatgg caatagtcgt gggacaacca gcctagctgt 8520 tggtagcaca cgccaagaac caaagcagcg gtgaaatacc aattcgtgta cacggtcatc 8580 aagatggaaa ggaccatcaa accgacggta gttgaagtct tgtaaaaata ccaaaggaaa 8640 ctcgtctcga acataccctt ggagatgaat tcctcccgca acttgcggaa atcctcctgg 8700 ggctcatcac gcttaggctt gggtgcaaca ggtgtgtcag gagaggaagg ctccataaca 8760 ggcattctct tcaacttggc gacggcttct tgagagtgca tcaccatgaa gacatcggtc 8820 gcatcgcggt tgcgatagtt ctcgataatg tcagctcctc cagggtggtg attgacccaa 8880 gcagacacat cataagttgc gccatcaatt gtgattggca gagcttgccg cttaggagac 8940 atggtgcggc cgcagtatat cttaaattct ttaatacggt gtactaggat attgaactgg 9000 ttcttgatga tgaaaacctg ggccgagatt gcagctattt atagtcatag gtcttgttaa 9060 catgcatgga catttggcca cggggtggca tgcagtttga cgggtgttga aataaacaaa 9120 aatgaggtgg cggaagagaa tacgagtttg aggttgggtt agaaacaaca aatgtgaggg 9180 ctcatgatgg gttgagttgg tgaatgtttt gggctgctcg attgacacct ttgtgagtac 9240 gtgttgttgt gcatggcttt tggggtccag tttttttttc ttgacgcggc gatcctgatc 9300 agctagtgga taagtgatgt ccactgtgtg tgattgcgtt tttgtttgaa ttttatgaac 9360 ttagacattg ctatgcaaag gatactctca ttgtgttttg tcttcttttg ttccttggct 9420 ttttcttatg atccaagaga ctagtcagtg ttgtggcatt cgagactacc aagattaatt 9480 atgatggggg aaggataagt aactgattag tacggactgt taccaaatta attaataagc 9540 ggcaaatgaa gggcatggat cctgcaggtc gactcgacgt acgatcccac atgcaagttt 9600 ttatttcaat cccttttcct ttgaataact gaccaagaac aacaagaaaa aaaaaaaaaa 9660 agaaaaggat cattttgaaa ggatattttt cgctcctatt caaatactgt atttttacca 9720 aaaaaactgt atttttccta cactctcaag ctttgttttt cgcttcgact ctcatgattt 9780 ccttcatatg ccaatcactc tatttataaa tggcataagg tagtgtgaac aattgcaaag 9840 cttgtcatca aaagcttgca atgtacaaat taatgttttt catgcctttc aaaattatct 9900 gcacccccta gctattaatc taacatctaa gtaaggctag tgaatttttt cgaatagtca 9960 tgcagtgcat taatttcccc gtgactattt tggctttgac tccaacactg gccccgtaca 10020 tccgtccctc attacatgaa aagaaatatt gtttatattc ttaattaaaa atattgtccc 10080 ttctaaattt tcatatagtt aattattata ttactttttt ctctattcta ttagttctat 10140 tttcaaatta ttatttatgc atatgtaaag tacattatat ttttgctata tacttaaata 10200 tttctaaatt attaaaaaaa gactgatatg aaaaatttat tctttttaaa gctatatcat 10260 tttatatata CtttttCttt tcttttcttt cattttctat tcaatttaat aagaaataaa 10320 ttttgtaaat ttttatttat caatttataa aaatatttta ctttatatgt tttttcacat 10380 ttttgttaaa caaatcatat cattatgatt gaaagagagg aaattgacag tgagtaataa 10440
gtgatgagaa aaaaatgtgt tatttcctaa aaaaaaccta aacaaacatg tatctactct 10500
ctatttcatc tatctctcat ttcatttttc tctttatctc tttctttatt tttttatcat 10560
atcatttcac attaattatt tttactctct ttattttttc tctctatccc tctcttattt 10620
ccactcatat atacactcca aaattggggc atgcctttat cactactcta tctcctccac 10680
taaatcattt aaatgaaact gaaaagcatt ggcaagtctc ctcccctcct caagtgattt 10740
ccaactcagc attggcatct aattgattca gtatatctat tgcatgtgta aaagtctttc 10800
cacaatacat aactattaat taatcttaaa taaataaagg ataaaatatt tttttttctt 10860
cataaaatta aaatatgtta ttttttgttt agatgtatat tcgaataaat ctaaatatat 10920
gataatgatt ttttatattg attaaacata taatcaatat taaatatgat atttttttat 10980
ataggttgta cacataattt tataaggata aaaaatatga taaaaataaa ttttaaatat 11040
ttttatattt acgagaaaaa aaaatatttt agccataaat aaatgaccag catattttac 11100
aaccttagta attcataaat tcctatatgt atatttgaaa ttaaaaacag ataatcgtta 11160
agggaaggaa tcctacgtca tctcttgcca tttgtttttc atgcaaacag aaagggacga 11220
aaaaccacct caccatgaat cactcttcac accattttta ctagcaaaca agtctcaaca 11280
actgaagcca gctctctttc cgtttctttt tacaacactt tctttgaaat agtagtattt 11340
ttttttcaca tgatttatta acgtgccaaa agatgcttat tgaatagagt gcacatttgt 11400
aatgtactac taattagaac atgaaaaagc attgttctaa cacgataatc ctgtgaaggc 11460
gttaactcca aagatccaat ttcactatat aaattgtgac gaaagcaaaa tgaattcaca 11520
tagctgagag agaaaggaaa ggttaactaa gaagcaatac ttcagcggcc gcgttaaaga 11580
cacaaagcct ttagcctatg ctgccaataa tggataccaa caaaagggtt cttcttttga 11640
ttttgatcct agcgctcctc caccgtttaa gattgcagaa atcagagctt caataccaaa 11700
acattgctgg gtcaagaatc catggagatc cctcagttat gttctcaggg atgtgcttgt 11760
aattgctgca ttggtggctg cagcaattca cttcgacaac tggcttctct ggctaatcta 11820
ttgccccatt caaggcacaa tgttctgggc tctctttgtt cttggacatg attgtggcca 11880
tggaagcttt tcagatagcc ctttgctgaa tagcctggtg ggacacatct tgcattcctc 11940
aattcttgtg ccataccatg gatggagaat tagccacaga actcaccatc aaaaccatgg 12000
acacattgag aaggatgagt catgggttcc attaacagag aagatttaca agaatctaga 12060
cagcatgaca agactcatta gattcactgt gccatttcca ttgtttgtgt atccaattta 12120
tttgttttca agaagccccg gaaaggaagg ctctcacttc aatccctaca gcaatctgtt 12180
cccacccagt gagagaaaag gaatagcaat atcaacactg tgttgggcta ccatgttttc 12240 tctgcttatc tatctccttc catggccaca atcatgtcca agaacaaaga gagcccagaa
cattgtgcct tgaatggggc aatagattag ccagagaagc cagttgtcga agtgaattgc
tgcagccacc aatgcagcaa ttacaagcac atccctgaga acataactga gggatctcca
tggattcttg acccagcaat gttttggtat tgaagctctg atttctgcaa tcttaaacgg
tggaggagcg ctaggatcaa aatcaaaaga agaacccttt tgttggtatc cattattggc
agcataggct aaaggctttg tgtctttaac cctgcaggta cc
<210> 27
<211> 9103
<212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo BC-ELfad3_ALS
<400> 27
gtacgtctag aggatccgtc gacggcgcgc ccgatcatcc ggatatagtt cctcctttca 60
gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt tattgctcag 120 cggtggcagc agccaactca gcttcctttc gggctttgtt agcagccgga tcgatccaag 180 ctgtacctca ctattccttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg 240 tclC3CclCfCCcl tcggtccaga cggccgcgct tctgcgggcg 300 gcccgacagt cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg 360 catcatcgaa attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca 420 tatacgcccg gagccgcggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg 480 tctgctgctc catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt 540 gggaatcccc gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg 600 tcaggacatt gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct 660 cggcccaaag catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca 720 tcacagtttg ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg 780 tagtgtattg accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat 840 cggccgcagc gatcgcatcc atagcctccg cgaccggctg cagaacagcg ggcagttcgg 900 tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc 960 tctcgctgaa ttccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt 1020 gccgataaac ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat 1080 atccacgccc tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca 1140 tcaggtcgga gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga 1200
12300 12360 12420 12480 12540 12582 gttcaggctt ttccatgggt atatctcctt cttaaagtta aacaaaatta tttctagagg 1260
gaaaccgttg tggtctccct atagtgagtc gtattaattt cgcgggatcg agatctgatc 1320
aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 1380
tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 1440
gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 1500
atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 1560
ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 1620
cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 1680
tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 1740
gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 1800
aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 1860
tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 1920
aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 1980
aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 2040
ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 2100
ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 2160
atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 2220
atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt 2280
gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa 2340
gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg 2400
ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatggacata 2460
ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca catacgattt 2520
aggtgacact atagaacggc gcgccaagct gggtctagaa ctagaaacgt gatgccactt 2580
gttattgaag tcgattacag catctattct gttttactat ttataacttt gccatttctg 2640
acttttgaaa actatctctg gatttcggta tcgctttgtg aagatcgagc aaaagagacg 2700
ttttgtggac gcaatggtcc aaatccgttc tacatgaaca aattggtcac aatttccact 2760
aaaagtaaat aaatggcaag ttaaaaaagg aatatgcatt ttactgattg cctaggtgag 2820
ctccaagaga agttgaatct acacgtctac caaccgctaa aaaaagaaaa acattgatat 2880
gtaacctgat tccattagct tttgacttct tcaacagatt ctctacttag atttctaaca 2940
gaaatattat tactagcaca tcattttcag tctcactaca gcaaaaaatc caacggcaca 3000
atacagacaa caggagatat cagactacag agatagatag atgctactgc atgtagtaag 3060 ttaaataaaa ggaaaataaa atgtcttgct accaaaacta ctacagacta tgatgctcac 3120
cacaggccaa atcctgcaac taggacagca ttatcttata tatattgtac aaaacaagca 3180
tcaaggaaca tttggtctag gcaatcagta cctcgttcta ccatcaccct cagttatcac 3240
atccttgaag gatccattac tgggaatcat cggcaacaca tgctcctgat ggggcacaat 3300
gacatcaaga aggtaggggc caggggtgtc caacattctc tgaattgccg ctctaagctc 3360
ttccttcttc gtcactcgcg ctgccggtat cccacaagca tcagcaaact tgagcatgtt 3420
tgggaatatc tcgctctcgc tagacggatc tccaagatag gtgtgagctc tattggactt 3480
gtagaaccta tcctccaact gaaccaccat acccaaatgc tgattgttca acaacaatat 3540
cttaactggg agattctcca ctcttatagt ggccaactcc tgaacattca tgatgaaact 3600
accatcccca tcaatgtcaa ccacaacagc cccagggtta gcaacagcag caccaatagc 3660
cgcaggcaat ccaaaaccca tggctccaag accccctgag gtcaaccact gcctcggtct 3720
cttgtacttg taaaactgcg cagcccacat ttgatgctgc ccaaccccag tactaacaat 3780
agcatctcca ttagtcaact catcaagaac ctcgatagca tgctgcggag aaatcgcgtc 3840
ctggaatgtc ttgtaaccca atggaaactt gtgtttctgc acattaatct cttctctcca 3900
acctccaaga tcaaacttac cctccactcc tttctcctcc aaaatcatat taattccctt 3960
caaggccaac ttcaaatccg cgcaaaccga cacgtgcgcc tgcttgttct tcccaatctc 4020
ggcagaatca atatcaatgt gaacaatctt agccctacta gcaaaagcct caagcttccc 4080
agtaacacgg tcatcaaacc ttaccccaaa ggcaagcaac aaatcactat tgtcaacagc 4140
atagttagca taaacagtac catgcatacc cagcatctga agggaatatt catcaccaat 4200
aggaaaagtt ccaagaccca ttaaagtgct agcaacggga ataccagtga gttcaacaaa 4260
gcgcctcaat tcagcactgg aattcaaact gccaccgccg acgtagagaa cgggcttttg 4320
ggcctccatg atgagtctga caatgtgttc caattgggcc tcggcggggg gcctgggcag 4380
cctggcgagg taaccgggga ggttaacggg ctcgtcccaa ttaggcacgg cgagttgctg 4440
ctgaacgtct ttgggaatgt cgatgaggac cggaccgggg cggccggagg tggcgacgaa 4500
gaaagcctcg gcgacgacgc gggggatgtc gtcgacgtcg aggatgaggt agttgtgctt 4560
cgtgatggat ctgctcacct ccacgatcgg ggtttcttgg aaggcgtcgg tgccgatcat 4 620
ccggcgggcg acctggccgg tgatggcgac gactgggacg ctgtccatta aagcgtcggc 4680
gaggccgctc acgaggttgg tggcgccggg gccggaggtg gcaatgcaga cgccggggag 4740
gccggaggaa cgcgcgtagc cttcggcggc gaagacgccg ccctgctcgt ggcgcgggag 4800
cacgttgcgg atggcggcgg agcgcgtgag cgcctggtgg atctccatcg acgcaccgcc 4 860
ggggtacgcg aacaccgtcg tcacgccctg cctctccagc gcctccacaa ggatgtccgc 4920 gcccttgcga ggttcgccgg aggcgaaccg tgacacgaag ggctccgtgg tcggcgcttc 4980
cttggtgaag ggcgccgccg tggggggttt ggagatggaa catttgattt tgagagcgtg 5040
gttgggtttg gtgagggttt gatgagagag agggagggtg gatctagtaa tgcgtttggg 5100
gaaggtgggg tgtgaagagg aagaagagaa tcgggtggtt ctggaagcgg tggccgccat 5160
tgtgttgtgt ggcatggtta tacttcaaaa actgcacaac aagcctagag ttagtaccta 5220
aacagtaaat ttacaacaga gagcaaagac acatgcaaaa atttcagcca taaaaaaagt 5280
tataatagaa tttaaagcaa aagtttcatt ttttaaacat atatacaaac aaactggatt 5340
tgaaggaagg gattaattcc cctgctcaaa gtttgaattc ctattgtgac ctatactcga 5400
ataaaattga agcctaagga atgtatgaga aacaagaaaa caaaacaaaa ctacagacaa 5460
acaagtacaa ttacaaaatt cgctaaaatt ctgtaatcac caaaccccat ctcagtcagc 5520
acaaggccca aggtttattt tgaaataaaa aaaaagtgat tttatttctc ataagctaaa 5580
agaaagaaag gcaattatga aatgatttcg actagatctg aaagtccaac gcgtattccg 5640
cagatattaa agaaagagta gagtttcaca tggatcctag atggacccag ttgaggaaaa 5700
agcaaggcaa agcaaaccag aagtgcaaga tccgaaattg aaccacggaa tctaggattt 5760
ggtagaggga gaagaaaagt accttgagag gtagaagaga agagaagagc agagagatat 5820
atgaacgagt gtgtcttggt ctcaactctg aagcgatacg agtttagagg ggagcattga 5880
gttccaattt atagggaaac cgggtggcag gggtgagtta atgacggaaa agcccctaag 5940
taacgagatt ggattgtggg ttagattcaa ccgtttgcat ccgcggctta gattggggaa 6000
gtcagagtga atctcaaccg ttgactgagt tgaaaattga atgtagcaac caattgagcc 6060
aaccccagcc tttgcccttt gattttgatt tgtttgttgc atacttttta tttgtcttct 6120
ggttctgact ctctttctct cgtttcaatg ccaggttgcc tactcccaca ccactcacaa 6180
gaagattcta ctgttagtat taaatatttt ttaatgtatt aaatgatgaa tgcttttgta 6240
aacagaacaa gactatgtct aataagtgtc ttgcaacatt ttttaagaaa ttaaaaaaaa 6300
tatatttatt atcaaaatca aatgtatgaa aaatcatgaa taatataatt ttatacattt 6360
ttttaaaaaa tcttttaatt tcttaattaa tatcttaaaa ataatgatta atatttaacc 6420
caaaataatt agtatgattg gtaaggaaga tatccatgtt atgtttggat gtgagtttga 6480
tctagagcaa agcttactag agtcgacctg caggttgaaa catccctgaa gtgtctcatt 6540
ttattttatt tattctttgc tgataaaaaa ataaaataaa agaagctaag cacacggtca 6600
accattgctc tactgctaaa agggttatgt gtagtgtttt actgcataaa ttatgcagca 6660
aacaagacaa ctcaaattaa aaaatttcct ttgcttgttt ttttgttgtc tctgacttga 6720
ctttcttgtg gaagttggtt gtataaggat tgggacacca ttgtccttct taatttaatt 6780 ttattctttg ctgataaaaa aaaaaatttc atatagtgtt aaataataat ttgttaaata 6840
accaaaaagt caaatatgtt tactctcgtt taaataattg agattcgtcc agcaaggcta 6900
aacgattgta tagatttatg acaatattta cttttttata gataaatgtt atattataat 6960
aaatttatat acatatatta tatgttattt attattattt taaatccttc aatattttat 7020
caaaccaact cataattttt tttttatctg taagaagcaa taaaattaaa tagacccact 7080
ttaaggatga tccaaccttt atacagagta agagagttca aatagtaccc tttcatatac 7140
atatcaacta aaatattaga aatatcatgg atcaaacctt ataaagacat taaataagtg 7200
gataagtata atatataaat gggtagtata taatatataa atggatacaa acttctctct 7260
ttataattgt tatgtctcct taacatccta atataataca taagtgggta atatataata 7320
tataaatgga gacaaacttc ttccattata attgttatgt cttcttaaca cttatgtctc 7380
gttcacaatg ctaaggttag aattgtttag aaagtcttat agtacacatt tgtttttgta 7440
ctatttgaag cattccataa gccgtcacga ttcagatgat ttataataat aagaggaaat 7500
ttatcataga acaataaggt gcatagatag agtgttaata tatcataaca tcctttgttt 7560
attcatagaa gaagtgagat ggagctcagt tattatactg ttacatggtc ggatacaata 7620
ttccatgctc tccatgagct cttacaccta catgcatttt agttcatact tgcggccgca 7680
gctggtcatc tcgctcatcg tcgagtcggc ggccggagct ggtcatctcg ctcatcgtcg 7740
agtcggcggc caattccata gagcttgagc actagaagtg gactagttat gaatgagaga 7800
tagataagca gagaaaacat ggtagcccaa cacagtgttg atattgctat tccttttctc 7860
tcactgggtg ggaacagatt gctgtaggga ttgaagtgag agccttcctt tccggggctt 7920
cttgaaaaca aataaattgg atacacaaac aatggaaatg gcacagtgaa tctaatgagt 7980
cttgtcatgc tgtctagatt cttgtaaatc ttctctgtta atggaaccca tgactcatcc 8040
ttctcaatgt gtccatggtt ttgatggtga gttctgtggc taattctcca tccatggtat 8100
ggcacaagaa ttgaggaatg caagatgtgt cccaccaggc tattcagcaa agggctatct 8160
gaaaagcttc catggccaca atcatgtcca agaacaaaga gagcccagaa cattgtgcct 8220
tgaatggggc aatagattag ccagagaagc cagttgtcga agtgaattgc tgcagccacc 8280
aatgcagcaa ttacaagcac atccctgaga acataactga gggatctcca tggattcttg 8340
acccagcaat gttttggtat tgaagctctg atttctgcaa tcttaaacgg tggaggaggg 8400
ccgccgactc gacgatgagc gagatgacca gctccggccg ccgactcgac gatgagcgag 8460
atgaccagct gcggccgcag tatatcttaa attctttaat acggtgtact aggatattga 8520
actggttctt gatgatgaaa acctgggccg agattgcagc tatttatagt cataggtctt 8580
gttaacatgc atggacattt ggccacgggg tggcatgcag tttgacgggt gttgaaataa 8640 acaaaaatga ggtggcggaa gagaatacga gtttgaggtt gggttagaaa caacaaatgt 8700 gagggctcat gatgggttga gttggtgaat gttttgggct gctcgattga cacctttgtg 8760 agtacgtgtt gttgtgcatg gcttttgggg tccagttttt ttttcttgac gcggcgatcc 8820 tgatcagcta gtggataagt gatgtccact gtgtgtgatt gcgtttttgt ttgaatttta 8880 tgaacttaga cattgctatg caaaggatac tctcattgtg ttttgtcttc ttttgttcct 8940 tggctttttc ttatgatcca agagactagt cagtgttgtg gcattcgaga ctaccaagat 9000 taattatgat gggggaagga taagtaactg attagtacgg actgttacca aattaattaa 9060 taagcggcaa atgaagggca tggatcctgc aggtcgactc gac 9103 <210> 28 <211> 5705 <212> DNA <213> Seqüência Artificial <220> <223> plasmideo BC-ELfad3_ALS <4 00> 28 ggccgcatga gccgtaaagg ttcaatacaa cgagtgcttg ttttcttagg gacaagcatt 60 gtacttatgt atgattctgt gtaaccatga gtcttccacg ttgtactaat gtgaagggca 120 aaaataaaac acagaacaag ttcgtttttc tcaaataatg tgaaggtaga aaatggaacc 180 atgcctcctc tcttgcatgt gatttaaaat attagcagat ggtaccgtac gtgggcggat 240 cccccgggct gcaggaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct 300 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc 360 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc 420 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact 480 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc 540 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc 600 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga 660 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag 720 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa 780 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat 840 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg 900 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa 960 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt 1020 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt 1080 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat 1140 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg 1200 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta 1260 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat 1320 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag 1380 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa 1440 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca 1500 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc 1560 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt 1620 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc 1680 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat 1740 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt 1800 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac 1860 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 1920 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 1980 accccccctc cgaaggtaac tggcttcagc agagcgcaga tâCCaaâtaC 2040 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 2100 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 2160 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 2220 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 2280 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 2340 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 2400 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 2460 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 2520 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc 2580 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg 2640 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt 2700 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca 2760 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct 2820 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat 2880 gattacgcca agcttgcatg cctgcaggtc gactcgacgt acgatcccac atgcaagttt 2940 ttatttcaat cccttttcct ttgaataact gaccaagaac aacaagaaaa aaaaaaaaaa 3000
agaaaaggat cattttgaaa ggatattttt cgctcctatt caaatactgt atttttacca 3060
aaaaaactgt atttttccta cactctcaag ctttgttttt cgcttcgact ctcatgattt 3120
ccttcatatg ccaatcactc tatttataaa tggcataagg tagtgtgaac aattgcaaag 3180
cttgtcatca aaagcttgca atgtacaaat taatgttttt catgcctttc aaaattatct 3240
gcacccccta gctattaatc taacatctaa gtaaggctag tgaatttttt cgaatagtca 3300
tgcagtgcat taatttcccc gtgactattt tggctttgac tccaacactg gccccgtaca 3360
tccgtccctc attacatgaa aagaaatatt gtttatattc ttaattaaaa atattgtccc 3420
ttctaaattt tcatatagtt aattattata ttactttttt ctctattcta ttagttctat 3480
tttcaaatta ttatttatgc atatgtaaag tacattatat ttttgctata tacttaaata 3540
tttctaaatt attaaaaaaa gactgatatg aaaaatttat tctttttaaa gctatatcat 3600
tttatatata ctttttcttt tcttttcttt cattttctat tcaatttaat aagaaataaa 3660
ttttgtaaat ttttatttat caatttataa aaatatttta ctttatatgt tttttcacat 3720
ttttgttaaa caaatcatat cattatgatt gaaagagagg aaattgacag tgagtaataa 3780
gtgatgagaa aaaaatgtgt tatttcctaa aaaaaaccta aacaaacatg tatctactct 3840
ctatttcatc tatctctcat ttcatttttc tctttatctc tttctttatt tttttatcat 3900
atcatttcac attaattatt tttactctct ttattttttc tctctatccc tctcttattt 3960
ccactcatat atacactcca aaattggggc atgcctttat cactactcta tctcctccac 4020
taaatcattt aaatgaaact gaaaagcatt ggcaagtctc ctcccctcct caagtgattt 4080
ccaactcagc attggcatct aattgattca gtatatctat tgcatgtgta aaagtctttc 4140
cacaatacat aactattaat taatcttaaa taaataaagg ataaaatatt tttttttctt 4200
cataaaatta aaatatgtta ttttttgttt agatgtatat tcgaataaat ctaaatatat 4260
gataatgatt ttttatattg attaaacata taatcaatat taaatatgat atttttttat 4320
ataggttgta cacataattt tataaggata aaaaatatga taaaaataaa ttttaaatat 4380
ttttatattt acgagaaaaa aaaatatttt agccataaat aaatgaccag catattttac 4440
aaccttagta attcataaat tcctatatgt atatttgaaa ttaaaaacag ataatcgtta 4500
agggaaggaa tcctacgtca tctcttgcca tttgtttttc atgcaaacag aaagggacga 4560
aaaaccacct caccatgaat cactcttcac accattttta ctagcaaaca agtctcaaca 4 620
actgaagcca gctctctttc cgtttctttt tacaacactt tctttgaaat agtagtattt 4680
ttttttcaca tgatttatta acgtgccaaa agatgcttat tgaatagagt gcacatttgt 4740
aatgtactac taattagaac atgaaaaagc attgttctaa cacgataatc ctgtgaaggc 4 800 gttaactcca aagatccaat ttcactatat aaattgtgac gaaagcaaaa tgaattcaca 4860
tagctgagag agaaaggaaa ggttaactaa gaagcaatac ttcagcggcc gcagctggtc 4 920
atctcgctca tcgtcgagtc ggcggccgga gctggtcatc tcgctcatcg tcgagtcggc 4980
ggccctcctc caccgtttaa gattgcagaa atcagagctt caataccaaa acattgctgg 5040
gtcaagaatc catggagatc cctcagttat gttctcaggg atgtgcttgt aattgctgca 5100
ttggtggctg cagcaattca cttcgacaac tggcttctct ggctaatcta ttgccccatt 5160
caaggcacaa tgttctgggc tctctttgtt cttggacatg attgtggcca tggaagcttt 5220
tcagatagcc ctttgctgaa tagcctggtg ggacacatct tgcattcctc aattcttgtg 5280
ccataccatg gatggagaat tagccacaga actcaccatc aaaaccatgg acacattgag 5340
aaggatgagt catgggttcc attaacagag aagatttaca agaatctaga cagcatgaca 5400
agactcatta gattcactgt gccatttcca ttgtttgtgt atccaattta tttgttttca 5460
agaagccccg gaaaggaagg ctctcacttc aatccctaca gcaatctgtt cccacccagt 5520
gagagaaaag gaatagcaat atcaacactg tgttgggcta ccatgttttc tctgcttatc 5580
tatctctcat tcataactag tccacttcta gtgctcaagc tctatggaat tggccgccga 5640
ctcgacgatg agcgagatga ccagctccgg ccgccgactc gacgatgagc gagatgacca 5700
gctgc 5705
<210> 29 <211> 12591 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmídeo BC-Tpom_Ann-ELfad3_ALS <400> 29
gtacgtctag aggatccgtc gacggcgcgc ccgatcatcc ggatatagtt cctcctttca 60
gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt tattgctcag 120
cggtggcagc agccaactca gcttcctttc gggctttgtt agcagccgga tcgatccaag 180
ctgtacctca ctattccttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg 240
cgagtacttc tacacagcca tcggtccaga cggccgcgct tctgcgggcg atttgtgtac 300
gcccgacagt cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg 360
catcatcgaa attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca 420
tatacgcccg gagccgcggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg 480
tctgctgctc catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt 540
gggaatcccc gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg 600 tcaggacatt gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct 660
cggcccaaag catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca 720
tcacagtttg ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg 780
tagtgtattg accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat 840
cggccgcagc gatcgcatcc atagcctccg cgaccggctg cagaacagcg ggcagttcgg 900
tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc 960
tctcgctgaa ttccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt 1020
gccgataaac ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat 1080
atccacgccc tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca 1140
tcaggtcgga gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga 1200
gttcaggctt ttccatgggt atatctcctt cttaaagtta aacaaaatta tttctagagg 1260
gaaaccgttg tggtctccct atagtgagtc gtattaattt cgcgggatcg agatctgatc 1320
aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 1380
tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 1440
gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 1500
atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 1560
ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 1620
cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 1680
tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 1740
gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 1800
aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 1860
tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 1920
aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 1980
aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 2040
ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 2100
ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 2160
atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 2220
atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt 2280
gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa 2340
gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg 2400
ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatggacata 24 60 ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca catacgattt 2520
aggtgacact atagaacggc gcgccaagct gggtctagaa ctagaaacgt gatgccactt 2580
gttattgaag tcgattacag catctattct gttttactat ttataacttt gccatttctg 2640
acttttgaaa actatctctg gatttcggta tcgctttgtg aagatcgagc aaaagagacg 2700
ttttgtggac gcaatggtcc aaatccgttc tacatgaaca aattggtcac aatttccact 2760
aaaagtaaat aaatggcaag ttaaaaaagg aatatgcatt ttactgattg cctaggtgag 2820
ctccaagaga agttgaatct acacgtctac caaccgctaa aaaaagaaaa acattgatat 2880
gtaacctgat tccattagct tttgacttct tcaacagatt ctctacttag atttctaaca 2940
gaaatattat tactagcaca tcattttcag tctcactaca gcaaaaaatc caacggcaca 3000
atacagacaa caggagatat cagactacag agatagatag atgctactgc atgtagtaag 3060
ttaaataaaa ggaaaataaa atgtcttgct accaaaacta ctacagacta tgatgctcac 3120
cacaggccaa atcctgcaac taggacagca ttatcttata tatattgtac aaaacaagca 3180
tcaaggaaca tttggtctag gcaatcagta cctcgttcta ccatcaccct cagttatcac 3240
atccttgaag gatccattac tgggaatcat cggcaacaca tgctcctgat ggggcacaat 3300
gacatcaaga aggtaggggc caggggtgtc caacattctc tgaattgccg ctctaagctc 3360
ttccttcttc gtcactcgcg ctgccggtat cccacaagca tcagcaaact tgagcatgtt 3420
tgggaatatc tcgctctcgc tagacggatc tccaagatag gtgtgagctc tattggactt 3480
gtagaaccta tcctccaact gaaccaccat acccaaatgc tgattgttca acaacaatat 3540
cttaactggg agattctcca ctcttatagt ggccaactcc tgaacattca tgatgaaact 3600
accatcccca tcaatgtcaa ccacaacagc cccagggtta gcaacagcag caccaatagc 3660
cgcaggcaat ccaaaaccca tggctccaag accccctgag gtcaaccact gcctcggtct 3720
cttgtacttg taaaactgcg cagcccacat ttgatgctgc ccaaccccag tactaacaat 3780
agcatctcca ttagtcaact catcaagaac ctcgatagca tgctgcggag aaatcgcgtc 3840
ctggaatgtc ttgtaaccca atggaaactt gtgtttctgc acattaatct cttctctcca 3900
acctccaaga tcaaacttac cctccactcc tttctcctcc aaaatcatat taattccctt 3960
caaggccaac ttcaaatccg cgcaaaccga cacgtgcgcc tgcttgttct tcccaatctc 4020
ggcagaatca atatcaatgt gaacaatctt agccctacta gcaaaagcct caagcttccc 4080
agtaacacgg tcatcaaacc ttaccccaaa ggcaagcaac aaatcactat tgtcaacagc 4140
atagttagca taaacagtac catgcatacc cagcatctga agggaatatt catcaccaat 4200
aggaaaagtt ccaagaccca ttaaagtgct agcaacggga ataccagtga gttcaacaaa 4260
gcgcctcaat tcagcactgg aattcaaact gccaccgccg acgtagagaa cgggcttttg 4320 ggcctccatg atgagtctga caatgtgttc caattgggcc tcggcggggg gcctgggcag 4380
cctggcgagg taaccgggga ggttaacggg ctcgtcccaa ttaggcacgg cgagttgctg 4440
ctgaacgtct ttgggaatgt cgatgaggac cggaccgggg cggccggagg tggcgacgaa 4500
gaaagcctcg gcgacgacgc gggggatgtc gtcgacgtcg aggatgaggt agttgtgctt 4560
cgtgatggat ctgctcacct ccacgatcgg ggtttcttgg aaggcgtcgg tgccgatcat 4620
ccggcgggcg acctggccgg tgatggcgac gactgggacg ctgtccatta aagcgtcggc 4680
gaggccgctc acgaggttgg tggcgccggg gccggaggtg gcaatgcaga cgccggggag 4740
gccggaggaa cgcgcgtagc cttcggcggc gaagacgccg ccctgctcgt ggcgcgggag 4800
cacgttgcgg atggcggcgg agcgcgtgag cgcctggtgg atctccatcg acgcaccgcc 4860
ggggtacgcg aacaccgtcg tcacgccctg cctctccagc gcctccacaa ggatgtccgc 4920
gcccttgcga ggttcgccgg aggcgaaccg tgacacgaag ggctccgtgg tcggcgcttc 4980
cttggtgaag ggcgccgccg tggggggttt ggagatggaa catttgattt tgagagcgtg 5040
gttgggtttg gtgagggttt gatgagagag agggagggtg gatctagtaa tgcgtttggg 5100
gaaggtgggg tgtgaagagg aagaagagaa tcgggtggtt ctggaagcgg tggccgccat 5160
tgtgttgtgt ggcatggtta tacttcaaaa actgcacaac aagcctagag ttagtaccta 5220
aacagtaaat ttacaacaga gagcaaagac acatgcaaaa atttcagcca taaaaaaagt 5280
tataatagaa tttaaagcaa aagtttcatt ttttaaacat atatacaaac aaactggatt 5340
tgaaggaagg gattaattcc cctgctcaaa gtttgaattc ctattgtgac ctatactcga 5400
ataaaattga agcctaagga atgtatgaga aacaagaaaa caaaacaaaa ctacagacaa 5460
acaagtacaa ttacaaaatt cgctaaaatt ctgtaatcac caaaccccat ctcagtcagc 5520
acaaggccca aggtttattt tgaaataaaa aaaaagtgat tttatttctc ataagctaaa 5580
agaaagaaag gcaattatga aatgatttcg actagatctg aaagtccaac gcgtattccg 5640
cagatattaa agaaagagta gagtttcaca tggatcctag atggacccag ttgaggaaaa 5700
agcaaggcaa agcaaaccag aagtgcaaga tccgaaattg aaccacggaa tctaggattt 5760
ggtagaggga gaagaaaagt accttgagag gtagaagaga agagaagagc agagagatat 5820
atgaacgagt gtgtcttggt ctcaactctg aagcgatacg agtttagagg ggagcattga 5880
gttccaattt atagggaaac cgggtggcag gggtgagtta atgacggaaa agcccctaag 5940
taacgagatt ggattgtggg ttagattcaa ccgtttgcat ccgcggctta gattggggaa 6000
gtcagagtga atctcaaccg ttgactgagt tgaaaattga atgtagcaac caattgagcc 6060
aaccccagcc tttgcccttt gattttgatt tgtttgttgc atacttttta tttgtcttct 6120
ggttctgact ctctttctct cgtttcaatg ccaggttgcc tactcccaca ccactcacaa 6180 gaagattcta ctgttagtat taaatatttt ttaatgtatt aaatgatgaa tgcttttgta 6240
aacagaacaa gactatgtct aataagtgtc ttgcaacatt ttttaagaaa ttaaaaaaaa 6300
tatatttatt atcaaaatca aatgtatgaa aaatcatgaa taatataatt ttatacattt 6360
ttttaaaaaa tcttttaatt tcttaattaa tatcttaaaa ataatgatta atatttaacc 6420
caaaataatt agtatgattg gtaaggaaga tatccatgtt atgtttggat gtgagtttga 6480
tctagagcaa agcttactag agtcgacctg caggttgaaa catccctgaa gtgtctcatt 6540
ttattttatt tattctttgc tgataaaaaa ataaaataaa agaagctaag cacacggtca 6600
accattgctc tactgctaaa agggttatgt gtagtgtttt actgcataaa ttatgcagca 6660
aacaagacaa ctcaaattaa aaaatttcct ttgcttgttt ttttgttgtc tctgacttga 6720
ctttcttgtg gaagttggtt gtataaggat tgggacacca ttgtccttct taatttaatt 6780
ttattctttg ctgataaaaa aaaaaatttc atatagtgtt aaataataat ttgttaaata 6840
accaaaaagt caaatatgtt tactctcgtt taaataattg agattcgtcc agcaaggcta 6900
aacgattgta tagatttatg acaatattta cttttttata gataaatgtt atattataat 6960
aaatttatat acatatatta tatgttattt attattattt taaatccttc aatattttat 7020
caaaccaact cataattttt tttttatctg taagaagcaa taaaattaaa tagacccact 7080
ttaaggatga tccaaccttt atacagagta agagagttca aatagtaccc tttcatatac 7140
atatcaacta aaatattaga aatatcatgg atcaaacctt ataaagacat taaataagtg 7200
gataagtata atatataaat gggtagtata taatatataa atggatacaa acttctctct 7260
ttataattgt tatgtctcct taacatccta atataataca taagtgggta atatataata 7320
tataaatgga gacaaacttc ttccattata attgttatgt cttcttaaca cttatgtctc 7380
gttcacaatg ctaaggttag aattgtttag aaagtcttat agtacacatt tgtttttgta 7440
ctatttgaag cattccataa gccgtcacga ttcagatgat ttataataat aagaggaaat 7500
ttatcataga acaataaggt gcatagatag agtgttaata tatcataaca tcctttgttt 7560
attcatagaa gaagtgagat ggagctcagt tattatactg ttacatggtc ggatacaata 7620
ttccatgctc tccatgagct cttacaccta catgcatttt agttcatact tgcggccgct 7680
cattcaggga cagcagccag ctgggcaatc tcgcctaaga agctgaccaa tttctgaacc 7740
ccggggatga tggctggggc cctgtagggc aggttgtgct tggcgcacaa cttctgcacc 7800
tccagggaag ctgccgtgag gttgtgcctg ggcatgttgg gccacaagtg gtgctcaatc 7860
tggtagttga gaccaccaaa gacccaatcg gtgagaaggc cgggcttaat gttcaacgtc 7 920
tcgtggatct ggctggcgct gaacccgtgc tcatcccaga cagtctcctc aaccttgtcc 7980
aaaggatagt gattcagaaa cacaacgatg gcaatgccaa atcctccgag cagctcggag 8040 atcaagaagc atcccaaacc ggtggcaaag ctaggcatgg cactgcaata gaaagtggcc 8100 ttgagcaccc agtggagggc cagaccaatc gcctccttag tgtactggcg cttgtagtac 8160 aggttcctct cctcgctctt gtaggacatg acgccgccga tggattggag gcaccagatg 8220 aacctaaggg atgcaatggt gggaatgaag tagtactgct ggtacttgat gagattccga 8280 gtcgaaggag tagccctctt gacatcttcg ggagaccaag ccaggatggg gaggttatca 8340 atatctgggt catggccgac cacattggtg gcggcatggt gaccattgtg cctatccttc 8400 caccaagtct gtgagtatcc ctgcatcacg ttaccgaaaa agagaccgaa agcgtcgtta 8460 atcttgcggt ttgtgaaaac ctggtgatgg caatagtcgt gggacaacca gcctagctgt 8520 tggtagcaca cgccaagaac caaagcagcg gtgaaatacc aattcgtgta cacggtcatc 8580 aagatggaaa ggaccatcaa accgacggta gttgaagtct tgtaaaaata ccaaaggaaa 8640 ctcgtctcga acataccctt ggagatgaat tcctcccgca acttgcggaa atcctcctgg 8700 ggctcatcac gcttaggctt gggtgcaaca ggtgtgtcag gagaggaagg ctccataaca 8760 ggcattctct tcaacttggc gacggcttct tgagagtgca tcaccatgaa gacatcggtc 8820 gcatcgcggt tgcgatagtt ctcgataatg tcagctcctc cagggtggtg attgacccaa 8880 gcagacacat cataagttgc gccatcaatt gtgattggca gagcttgccg cttaggagac 8940 atggtgcggc cgcagtatat cttaaattct ttaatacggt gtactaggat attgaactgg 9000 ttcttgatga tgaaaacctg ggccgagatt gcagctattt atagtcatag gtcttgttaa 9060 catgcatgga catttggcca cggggtggca tgcagtttga cgggtgttga aataaacaaa 9120 aatgaggtgg cggaagagaa tacgagtttg aggttgggtt agaaacaaca aatgtgaggg 9180 ctcatgatgg gttgagttgg tgaatgtttt gggctgctcg attgacacct ttgtgagtac 9240 gtgttgttgt gcatggcttt tggggtccag tttttttttc ttgacgcggc gatcctgatc 9300 agctagtgga taagtgatgt ccactgtgtg tgattgcgtt tttgtttgaa ttttatgaac 9360 ttagacattg ctatgcaaag gatactctca ttgtgttttg tcttcttttg ttccttggct 9420 ttttcttatg atccaagaga ctagtcagtg ttgtggcatt cgagactacc aagattaatt 9480 atgatggggg aaggataagt aactgattag tacggactgt taccaaatta attaataagc 9540 ggcaaatgaa gggcatggat cctgcaggtc gactcgacgt acgatcccac atgcaagttt 9600 ttatttcaat cccttttcct ttgaataact gaccaagaac aacaagaaaa aaaaaaaaaa 9660 agaaaaggat cattttgaaa ggatattttt cgctcctatt caaatactgt atttttacca 9720 aaaaaactgt atttttccta cactctcaag ctttgttttt cgcttcgact ctcatgattt 9780 ccttcatatg ccaatcactc tatttataaa tggcataagg tagtgtgaac aattgcaaag 9840 cttgtcatca aaagcttgca atgtacaaat taatgttttt catgcctttc aaaattatct 9900 gcacccccta gctattaatc taacatctaa gtaaggctag tgaatttttt cgaatagtca 9960
tgcagtgcat taatttcccc gtgactattt tggctttgac tccaacactg gccccgtaca 10020
tccgtccctc attacatgaa aagaaatatt gtttatattc ttaattaaaa atattgtccc 10080
ttctaaattt tcatatagtt aattattata ttactttttt ctctattcta ttagttctat 10140
tttcaaatta ttatttatgc atatgtaaag tacattatat ttttgctata tacttaaata 10200
tttctaaatt attaaaaaaa gactgatatg aaaaatttat tctttttaaa gctatatcat 10260
tttatatata ctttttcttt tcttttcttt cattttctat tcaatttaat aagaaataaa 10320
ttttgtaaat ttttatttat caatttataa aaatatttta ctttatatgt tttttcacat 10380
ttttgttaaa caaatcatat cattatgatt gaaagagagg aaattgacag tgagtaataa 10440
gtgatgagaa aaaaatgtgt tatttcctaa aaaaaaccta aacaaacatg tatctactct 10500
ctatttcatc tatctctcat ttcatttttc tctttatctc tttctttatt tttttatcat 10560
atcatttcac attaattatt tttactctct ttattttttc tctctatccc tctcttattt 10620
ccactcatat atacactcca aaattggggc atgcctttat cactactcta tctcctccac 10680
taaatcattt aaatgaaact gaaaagcatt ggcaagtctc ctcccctcct caagtgattt 10740
ccaactcagc attggcatct aattgattca gtatatctat tgcatgtgta aaagtctttc 10800
cacaatacat aactattaat taatcttaaa taaataaagg ataaaatatt tttttttctt 10860
cataaaatta aaatatgtta ttttttgttt agatgtatat tcgaataaat ctaaatatat 10920
gataatgatt ttttatattg attaaacata taatcaatat taaatatgat atttttttat 10980
ataggttgta cacataattt tataaggata aaaaatatga taaaaataaa ttttaaatat 11040
ttttatattt acgagaaaaa aaaatatttt agccataaat aaatgaccag catattttac 11100
aaccttagta attcataaat tcctatatgt atatttgaaa ttaaaaacag ataatcgtta 11160
agggaaggaa tcctacgtca tctcttgcca tttgtttttc atgcaaacag aaagggacga 11220
aaaaccacct caccatgaat cactcttcac accattttta ctagcaaaca agtctcaaca 11280
actgaagcca gctctctttc cgtttctttt tacaacactt tctttgaaat agtagtattt 11340
ttttttcaca tgatttatta acgtgccaaa agatgcttat tgaatagagt gcacatttgt 11400
aatgtactac taattagaac atgaaaaagc attgttctaa cacgataatc ctgtgaaggc 114 60
gttaactcca aagatccaat ttcactatat aaattgtgac gaaagcaaaa tgaattcaca 11520
tagctgagag agaaaggaaa ggttaactaa gaagcaatac ttcagcggcc gcagctggtc 11580
atctcgctca tcgtcgagtc ggcggccgga gctggtcatc tcgctcatcg tcgagtcggc 11640
ggccctcctc caccgtttaa gattgcagaa atcagagctt caataccaaa acattgctgg 11700
gtcaagaatc catggagatc cctcagttat gttctcaggg atgtgcttgt aattgctgca 11760 ttggtggctg cagcaattca cttcgacaac tggcttctct ggctaatcta ttgccccatt 11820
caaggcacaa tgttctgggc tctctttgtt cttggacatg attgtggcca tggaagcttt 11880
tcagatagcc ctttgctgaa tagcctggtg ggacacatct tgcattcctc aattcttgtg 11940
ccataccatg gatggagaat tagccacaga actcaccatc aaaaccatgg acacattgag 12000
aaggatgagt catgggttcc attaacagag aagatttaca agaatctaga cagcatgaca 12060
agactcatta gattcactgt gccatttcca ttgtttgtgt atccaattta tttgttttca 12120
agaagccccg gaaaggaagg ctctcacttc aatccctaca gcaatctgtt cccacccagt 12180
gagagaaaag gaatagcaat atcaacactg tgttgggcta ccatgttttc tctgcttatc 12240
tatctctcat tcataactag tccacttcta gtgctcaagc tctatggaat tggccgccga 12300
ctcgacgatg agcgagatga ccagctccgg ccgccgactc gacgatgagc gagatgacca 12360
gctgcggccg catgagccgt aaaggttcaa tacaacgagt gcttgttttc ttagggacaa 12420
gcattgtact tatgtatgat tctgtgtaac catgagtctt ccacgttgta ctaatgtgaa 12480
gggcaaaaat aaaacacaga acaagttcgt ttttctcaaa taatgtgaag gtagaaaatg 12540
gaaccatgcc tcctctcttg catgtgattt aaaatattag cagatggtac c 12591
<210> 30
<211> 32
<212> DNA
<213> Seqüência Artificial <220>
<223> primer HPfad3-l
<400> 30
gcggccgcgt taaagacaca aagcctttag cc
<210> 31
<211> 34
<212> DNA
<213> Seqüência Artificial
<220>
<223> primer HPfad3-2
<4 00> 31
gattgtggcc atggaaggag atagataagc agag
<210> 32
<211> 709
<212> DNA
<213> Seqüência Artificial
32
34
<220>
<223> fragmento HPfad3AB <400> 32
gcggccgcgt taaagacaca aagcctttag cctatgctgc caataatgga taccaacaaa
60
agggttcttc ttttgatttt gatcctagcg ctcctccacc gtttaagatt gcagaaatca 120
gagcttcaat accaaaacat tgctgggtca agaatccatg gagatccctc agttatgttc 180
tcagggatgt gcttgtaatt gctgcattgg tggctgcagc aattcacttc gacaactggc 240
ttctctggct aatctattgc cccattcaag gcacaatgtt ctgggctctc tttgttcttg 300
gacatgattg tggccatgga agcttttcag atagcccttt gctgaatagc ctggtgggac 360
acatcttgca ttcctcaatt cttgtgccat accatggatg gagaattagc cacagaactc 420
accatcaaaa ccatggacac attgagaagg atgagtcatg ggttccatta acagagaaga
480
tttacaagaa tctagacagc atgacaagac tcattagatt cactgtgcca tttccattgt 540
ttgtgtatcc aatttatttg ttttcaagaa gccccggaaa ggaaggctct cacttcaatc 600
cctacagcaa tctgttccca cccagtgaga gaaaaggaat agcaatatca acactgtgtt 660
gggctaccat gttttctctg cttatctatc tccttccatg gccacaatc
709
<210> 33
<211> 34
<212> DNA
<213> Seqüência Artificial <22Ü>
<223> primer HPfad3-3
<400> 33
ctctgcttat ctatctcctt ccatggccac aatc 34
<210> 34
<211> 36
<212> DNA
<213> Seqüência Artificial <220>
<223> primer HPfad3-4
<210> 35 <211> 343 <212> DNA
<213> Seqüência Artificial <220>
<223> fragmento HPfad3A' <400> 35
ctctgcttat ctatctcctt ccatggccac aatcatgtcc aagaacaaag agagcccaga 60
acattgtgcc ttgaatgggg caatagatta gccagagaag ccagttgtcg aagtgaattg 120
<400> 34
ggtacctgca gggttaaaga cacaaagcct ttagcc
36 ctgcagccac caatgcagca attacaagca catccctgag aacataactg agggatctcc 180 atggattctt gacccagcaa tgttttggta ttgaagctct gatttctgca atcttaaacg 240 gtggaggagc gctaggatca aaatcaaaag aagaaccctt ttgttggtat ccattattgg 300 cagcataggc taaaggcttt gtgtctttaa ccctgcaggt acc 343
<210> 36 <211> 1018 <212> DNA
<213> Seqüência Artificial <220>
<223> fragmento HPfad3ABA1 <400> 36
gcggccgcgt taaagacaca aagcctttag cctatgctgc caataatgga taccaacaaa 60
agggttcttc ttttgatttt gatcctagcg ctcctccacc gtttaagatt gcagaaatca 120
gagcttcaat accaaaacat tgctgggtca agaatccatg gagatccctc agttatgttc 180
tcagggatgt gcttgtaatt gctgcattgg tggctgcagc aattcacttc gacaactggc 240
ttctctggct aatctattgc cccattcaag gcacaatgtt ctgggctctc tttgttcttg 300
gacatgattg tggccatgga agcttttcag atagcccttt gctgaatagc ctggtgggac 360
acatcttgca ttcctcaatt cttgtgccat accatggatg gagaattagc cacagaactc 420
accatcaaaa ccatggacac attgagaagg atgagtcatg ggttccatta acagagaaga 480
tttacaagaa tctagacagc atgacaagac tcattagatt cactgtgcca tttccattgt 540
ttgtgtatcc aatttatttg ttttcaagaa gccccggaaa ggaaggctct cacttcaatc 600
cctacagcaa tctgttccca cccagtgaga gaaaaggaat agcaatatca acactgtgtt 660
gggctaccat gttttctctg cttatctatc tccttccatg gccacaatca tgtccaagaa 720
caaagagagc ccagaacatt gtgccttgaa tggggcaata gattagccag agaagccagt 780
tgtcgaagtg aattgctgca gccaccaatg cagcaattac aagcacatcc ctgagaacat 840
aactgaggga tctccatgga ttcttgaccc agcaatgttt tggtattgaa gctctgattt 900
ctgcaatctt aaacggtgga ggagcgctag gatcaaaatc aaaagaagaa cccttttgtt 960
ggtatccatt attggcagca taggctaaag gctttgtgtc tttaaccctg caggtacc 1018
<210> 37 <211> 4304 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo BC-HPfad3ABA' <400> 37
ctagacctgc aggatccatg cccttcattt gccgcttatt aattaatttg gtaacagtcc 60
gtactaatca gttacttatc cttcccccat cataattaat cttggtagtc tcgaatgcca 120
caacactgac tagtctcttg gatcataaga aaaagccaag gaacaaaaga agacaaaaca 180
caatgagagt atcctttgca tagcaatgtc taagttcata aaattcaaac aaaaacgcaa 240
tcacacacag tggacatcac ttatccacta gctgatcagg atcgccgcgt caagaaaaaa 300
aaactggacc ccaaaagcca tgcacaacaa cacgtactca caaaggtgtc aatcgagcag 360
cccaaaacat tcaccaactc aacccatcat gagccctcac atttgttgtt tctaacccaa 420
cctcaaactc gtattctctt ccgccacctc atttttgttt atttcaacac ccgtcaaact 480
gcatgccacc ccgtggccaa atgtccatgc atgttaacaa gacctatgac tataaatagc 540
tgcaatctcg gcccaggttt tcatcatcaa gaaccagttc aatatcctag tacaccgtat 600
taaagaattt aagatatact gcggccgcgt taaagacaca aagcctttag cctatgctgc 660
caataatgga taccaacaaa agggttcttc ttttgatttt gatcctagcg ctcctccacc 720
gtttaagatt gcagaaatca gagcttcaat accaaaacat tgctgggtca agaatccatg 780
gagatccctc agttatgttc tcagggatgt gcttgtaatt gctgcattgg tggctgcagc 840
aattcacttc gacaactggc ttctctggct aatctattgc cccattcaag gcacaatgtt 900
ctgggctctc tttgttcttg gacatgattg tggccatgga agcttttcag atagcccttt 960
gctgaatagc ctggtgggac acatcttgca ttcctcaatt cttgtgccat accatggatg 1020
gagaattagc cacagaactc accatcaaaa ccatggacac attgagaagg atgagtcatg 1080
ggttccatta acagagaaga tttacaagaa tctagacagc atgacaagac tcattagatt 1140
cactgtgcca tttccattgt ttgtgtatcc aatttatttg ttttcaagaa gccccggaaa 1200
ggaaggctct cacttcaatc cctacagcaa tctgttccca cccagtgaga gaaaaggaat 1260
agcaatatca acactgtgtt gggctaccat gttttctctg cttatctatc tccttccatg 1320
gccacaatca tgtccaagaa caaagagagc ccagaacatt gtgccttgaa tggggcaata 1380
gattagccag agaagccagt tgtcgaagtg aattgctgca gccaccaatg cagcaattac 1440
aagcacatcc ctgagaacat aactgaggga tctccatgga ttcttgaccc agcaatgttt 1500
tggtattgaa gctctgattt ctgcaatctt aaacggtgga ggagcgctag gatcaaaatc 1560
aaaagaagaa cccttttgtt ggtatccatt attggcagca taggctaaag gctttgtgtc 1620
tttaaccctg caggtaccga gctcgaattc actggccgtc gttttacaac gtcgtgactg 1680
ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg 1740
gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg 1800
cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat 1860 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc 1920
gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca 1980
agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg 2040
cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat 2100
ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt 2160
atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct 2220
tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc 2280
cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa 2340
agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg 2400
taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt 2460
tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg 2520
catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac 2580
ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc 2640
ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa 2700
catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc 2760
aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt 2820
aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga 2880
taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa 2940
atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa 3000
gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa 3060
tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt 3120
ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt 3180
gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg 3240
agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt 3300
aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca 3360
agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac 3420
tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac 3480
atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct 3540
taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg 3600
gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca 3660
gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt 3720 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta 3780 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc 3840 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc 3900 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa 3960 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag 4020 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg 4080 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga 4140 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat 4200 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag 4260 ctatgaccat gattacgcca agcttgcatg cctgcaggtc gact 4304
<210> 38
<211> 5696
<212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo Ann-HPfad3ABA'
<400> 38
ttgctgcagc caccaatgca gcaattacaa gcacatccct gagaacataa ctgagggatc 60 tccatggatt cttgacccag caatgttttg gtattgaagc tctgatttct gcaatcttaa 120 acggtggagg agcgctagga tcaaaatcaa aagaagaacc cttttgttgg tatccattat 180 tggcagcata ggctaaaggc tttgtgtctt taaccctgca ggtaccgtac gtgggcggat 240 cccccgggct gcaggaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct 300 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc 360 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc 420 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact 480 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc 540 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc 600 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga 660 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag 720 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa 780 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat 840 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg 900 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa 960 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt 1020
gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt 1080
ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat 1140
tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg 1200
acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta 1260
cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat 1320
catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag 1380
cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa 1440
ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca 1500
ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc 1560
ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt 1620
atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc 1680
gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat 1740
atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt 1800
tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac 1860
cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 1920
ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 1980
actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta 2040
gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 2100
ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 2160
gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 2220
acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 2280
tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 2340
gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 2400
cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 2460
cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 2520
ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc 2580
gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg 2640
agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt 2700
cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca 2760
attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct 2820 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat 2880
gattacgcca agcttgcatg cctgcaggtc gactcgacgt acgatcccac atgcaagttt 2940
ttatttcaat cccttttcct ttgaataact gaccaagaac aacaagaaaa aaaaaaaaaa 3000
agaaaaggat cattttgaaa ggatattttt cgctcctatt caaatactgt atttttacca 3060
aaaaaactgt atttttccta cactctcaag ctttgttttt cgcttcgact ctcatgattt 3120
ccttcatatg ccaatcactc tatttataaa tggcataagg tagtgtgaac aattgcaaag 3180
cttgtcatca aaagcttgca atgtacaaat taatgttttt catgcctttc aaaattatct 3240
gcacccccta gctattaatc taacatctaa gtaaggctag tgaatttttt cgaatagtca 3300
tgcagtgcat taatttcccc gtgactattt tggctttgac tccaacactg gccccgtaca 3360
tccgtccctc attacatgaa aagaaatatt gtttatattc ttaattaaaa atattgtccc 3420
ttctaaattt tcatatagtt aattattata ttactttttt ctctattcta ttagttctat 3480
tttcaaatta ttatttatgc atatgtaaag tacattatat ttttgctata tacttaaata 3540
tttctaaatt attaaaaaaa gactgatatg aaaaatttat tctttttaaa gctatatcat 3600
tttatatata ctttttcttt tcttttcttt cattttctat tcaatttaat aagaaataaa 3660
ttttgtaaat ttttatttat caatttataa aaatatttta ctttatatgt tttttcacat 3720
ttttgttaaa caaatcatat cattatgatt gaaagagagg aaattgacag tgagtaataa 3780
gtgatgagaa aaaaatgtgt tatttcctaa aaaaaaccta aacaaacatg tatctactct 3840
ctatttcatc tatctctcat ttcatttttc tctttatctc tttctttatt tttttatcat 3900
atcatttcac attaattatt tttactctct ttattttttc tctctatccc tctcttattt 3960
ccactcatat atacactcca aaattggggc atgcctttat cactactcta tctcctccac 4020
taaatcattt aaatgaaact gaaaagcatt ggcaagtctc ctcccctcct caagtgattt 4080
ccaactcagc attggcatct aattgattca gtatatctat tgcatgtgta aaagtctttc 4140
cacaatacat aactattaat taatcttaaa taaataaagg ataaaatatt tttttttctt 4200
cataaaatta aaatatgtta ttttttgttt agatgtatat tcgaataaat ctaaatatat 4260
gataatgatt ttttatattg attaaacata taatcaatat taaatatgat atttttttat 4320
ataggttgta cacataattt tataaggata aaaaatatga taaaaataaa ttttaaatat 4380
ttttatattt acgagaaaaa aaaatatttt agccataaat aaatgaccag catattttac 4440
aaccttagta attcataaat tcctatatgt atatttgaaa ttaaaaacag ataatcgtta 4500
agggaaggaa tcctacgtca tctcttgcca tttgtttttc atgcaaacag aaagggacga 4560
aaaaccacct caccatgaat cactcttcac accattttta ctagcaaaca agtctcaaca 4620
actgaagcca gctctctttc cgtttctttt tacaacactt tctttgaaat agtagtattt 4680 ttttttcaca tgatttatta acgtgccaaa agatgcttat tgaatagagt gcacatttgt 4740 aatgtactac taattagaac atgaaaaagc attgttctaa cacgataatc ctgtgaaggc 4800 gttaactcca aagatccaat ttcactatat aaattgtgac gaaagcaaaa tgaattcaca 4860 tagctgagag agaaaggaaa ggttaactaa gaagcaatac ttcagcggcc gcgttaaaga 4920 cacaaagcct ttagcctatg ctgccaataa tggataccaa caaaagggtt cttcttttga 4980 ttttgatcct agcgctcctc caccgtttaa gattgcagaa atcagagctt caataccaaa 5040 acattgctgg gtcaagaatc catggagatc cctcagttat gttctcaggg atgtgcttgt 5100 aattgctgca ttggtggctg cagcaattca cttcgacaac tggcttctct ggctaatcta 5160 ttgccccatt caaggcacaa tgttctgggc tctctttgtt cttggacatg attgtggcca 5220 tggaagcttt tcagatagcc ctttgctgaa tagcctggtg ggacacatct tgcattcctc 5280 aattcttgtg ccataccatg gatggagaat tagccacaga actcaccatc aaaaccatgg 5340 acacattgag aaggatgagt catgggttcc attaacagag aagatttaca agaatctaga 5400 cagcatgaca agactcatta gattcactgt gccatttcca ttgtttgtgt atccaattta 5460 tttgttttca agaagccccg gaaaggaagg ctctcacttc aatccctaca gcaatctgtt 5520 cccacccagt gagagaaaag gaatagcaat atcaacactg tgttgggcta ccatgttttc 5580 tctgcttatc tatctccttc catggccaca atcatgtcca agaacaaaga gagcccagaa 5640 cattgtgcct tgaatggggc aatagattag ccagagaagc cagttgtcga agtgaa 5696
<210> 39 <211> 8145 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo HPfad3ABA'_ALS <400> 39
gtacgtctag aggatccgtc gacggcgcgc ccgatcatcc ggatatagtt cctcctttca 60
gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt tattgctcag 120
cggtggcagc agccaactca gcttcctttc gggctttgtt agcagccgga tcgatccaag 180
ctgtacctca ctattccttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg 240
cgagtacttc tacacagcca tcggtccaga cggccgcgct tctgcgggcg atttgtgtac 300
gcccgacagt cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg 360
catcatcgaa attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca 420
tatacgcccg gagccgcggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg 480
tctgctgctc catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt 540 gggaatcccc gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg 600
tcaggacatt gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct 660
cggcccaaag catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca 720
tcacagtttg ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg 780
tagtgtattg accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat 840
cggccgcagc gatcgcatcc atagcctccg cgaccggctg cagaacagcg ggcagttcgg 900
tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc 960
tctcgctgaa ttccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt 1020
gccgataaac ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat 1080
j atccacgccc tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca 1140
tcaggtcgga gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga 1200
gttcaggctt ttccatgggt atatctcctt cttaaagtta aacaaaatta tttctagagg 1260
gaaaccgttg tggtctccct atagtgagtc gtattaattt cgcgggatcg agatctgatc 1320
aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 1380
tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 1440
gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 1500
atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 1560
ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 1620
cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 1680
(tl·1 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 1740
gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 1800
aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 1860
tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 1920
aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 1980
aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 2040
ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 2100
ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 2160
ãtcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 2220
atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt 2280
gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa 2340
gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg 2400 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatggacata ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca catacgattt aggtgacact atagaacggc gcgccaagct gggtctagaa ctagaaacgt gatgccactt gttattgaag tcgattacag catctattct gttttactat ttataacttt gccatttctg acttttgaaa actatctctg gatttcggta tcgctttgtg aagatcgagc aaaagagacg ttttgtggac gcaatggtcc aaatccgttc tacatgaaca aattggtcac aatttccact aaaagtaaat aaatggcaag ttaaaaaagg aatatgcatt ttactgattg cctaggtgag ctccaagaga agttgaatct acacgtctac caaccgctaa aaaaagaaaa acattgatat gtaacctgat tccattagct tttgacttct tcaacagatt ctctacttag atttctaaca gaaatattat tactagcaca tcattttcag tctcactaca gcaaaaaatc caacggcaca atacagacaa caggagatat cagactacag agatagatag atgctactgc atgtagtaag ttaaataaaa ggaaaataaa atgtcttgct accaaaacta ctacagacta tgatgctcac cacaggccaa atcctgcaac taggacagca ttatcttata tatattgtac aaaacaagca tcaaggaaca tttggtctag gcaatcagta cctcgttcta ccatcaccct cagttatcac atccttgaag gatccattac tgggaatcat cggcaacaca tgctcctgat ggggcacaat gacatcaaga aggtaggggc caggggtgtc caacattctc tgaattgccg ctctaagctc ttccttcttc gtcactcgcg ctgccggtat cccacaagca tcagcaaact tgagcatgtt tgggaatatc tcgctctcgc tagacggatc tccaagatag gtgtgagctc tattggactt gtagaaccta tcctccaact gaaccaccat acccaaatgc tgattgttca acaacaatat cttaactggg agattctcca ctcttatagt ggccaactcc tgaacattca tgatgaaact accatcccca tcaatgtcaa ccacaacagc cccagggtta gcaacagcag caccaatagc cgcaggcaat ccaaaaccca tggctccaag accccctgag gtcaaccact gcctcggtct cttgtacttg taaaactgcg cagcccacat ttgatgctgc ccaaccccag tactaacaat agcatctcca ttagtcaact catcaagaac ctcgatagca tgctgcggag aaatcgcgtc ctggaatgtc ttgtaaccca atggaaactt gtgtttctgc acattaatct cttctctcca acctccaaga tcaaacttac cctccactcc tttctcctcc aaaatcatat taattccctt caaggccaac ttcaaatccg cgcaaaccga cacgtgcgcc tgcttgttct tcccaatctc ggcagaatca atatcaatgt gaacaatctt agccctacta gcaaaagcct caagcttccc agtaacacgg tcatcaaacc ttaccccaaa ggcaagcaac aaatcactat tgtcaacagc atagttagca taaacagtac catgcatacc cagcatctga agggaatatt catcaccaat aggaaaagtt ccaagaccca ttaaagtgct agcaacggga ataccagtga gttcaacaaa
2460
2520
2580
2640
2700
2760
2820
2880
2940
3000
3060
3120
3180
3240
3300
3360
3420
3480
3540
3600
3660
3720
3780
3840
3900
3960
4020
4080
4140
4200
4260 gcgcctcaat tcagcactgg aattcaaact gccaccgccg acgtagagaa cgggcttttg 4320 ggcctccatg atgagtctga caatgtgttc caattgggcc tcggcggggg gcctgggcag 4380
cctggcgagg taaccgggga ggttaacggg ctcgtcccaa ttaggcacgg cgagttgctg 4440
ctgaacgtct ttgggaatgt cgatgaggac cggaccgggg cggccggagg tggcgacgaa 4500
gaaagcctcg gcgacgacgc gggggatgtc gtcgacgtcg aggatgaggt agttgtgctt 4560
cgtgatggat ctgctcacct ccacgatcgg ggtttcttgg aaggcgtcgg tgccgatcat 4620
ccggcgggcg acctggccgg tgatggcgac gactgggacg ctgtccatta aagcgtcggc 4680
gaggccgctc acgaggttgg tggcgccggg gccggaggtg gcaatgcaga cgccggggag 4740
gccggaggaa cgcgcgtagc cttcggcggc gaagacgccg ccctgctcgt ggcgcgggag 4800
cacgttgcgg atggcggcgg agcgcgtgag cgcctggtgg atctccatcg acgcaccgcc 4860
ggggtacgcg aacaccgtcg tcacgccctg cctctccagc gcctccacaa ggatgtccgc 4920
gcccttgcga ggttcgccgg aggcgaaccg tgacacgaag ggctccgtgg tcggcgcttc 4980
cttggtgaag ggcgccgccg tggggggttt ggagatggaa catttgattt tgagagcgtg 5040
gttgggtttg gtgagggttt gatgagagag agggagggtg gatctagtaa tgcgtttggg 5100
gaaggtgggg tgtgaagagg aagaagagaa tcgggtggtt ctggaagcgg tggccgccat 5160
tgtgttgtgt ggcatggtta tacttcaaaa actgcacaac aagcctagag ttagtaccta 5220
aacagtaaat ttacaacaga gagcaaagac acatgcaaaa atttcagcca taaaaaaagt 5280
tataatagaa tttaaagcaa aagtttcatt ttttaaacat atatacaaac aaactggatt 5340
tgaaggaagg gattaattcc cctgctcaaa gtttgaattc ctattgtgac ctatactcga 5400
ataaaattga agcctaagga atgtatgaga aacaagaaaa caaaacaaaa ctacagacaa 54 60
acaagtacaa ttacaaaatt cgctaaaatt ctgtaatcac caaaccccat ctcagtcagc 5520
acaaggccca aggtttattt tgaaataaaa aaaaagtgat tttatttctc ataagctaaa 5580
agaaagaaag gcaattatga aatgatttcg actagatctg aaagtccaac gcgtattccg 5640
cagatattaa agaaagagta gagtttcaca tggatcctag atggacccag ttgaggaaaa 5700
agcaaggcaa agcaaaccag aagtgcaaga tccgaaattg aaccacggaa tctaggattt 5760
ggtagaggga gaagaaaagt accttgagag gtagaagaga agagaagagc agagagatat 5820
atgaacgagt gtgtcttggt ctcaactctg aagcgatacg agtttagagg ggagcattga 5880
gttccaattt atagggaaac cgggtggcag gggtgagtta atgacggaaa agcccctaag 5940
taacgagatt ggattgtggg ttagattcaa ccgtttgcat ccgcggctta gattggggaa 6000
gtcagagtga atctcaaccg ttgactgagt tgaaaattga atgtagcaac caattgagcc 6060
aaccccagcc tttgcccttt gattttgatt tgtttgttgc atacttttta tttgtcttct 6120 ggttctgact ctctttctct cgtttcaatg ccaggttgcc tactcccaca ccactcacaa 6180 gaagattcta ctgttagtat taaatatttt ttaatgtatt aaatgatgaa tgcttttgta 6240 aacagaacaa gactatgtct aataagtgtc ttgcaacatt ttttaagaaa ttaaaaaaaa 6300 tatatttatt atcaaaatca aatgtatgaa aaatcatgaa taatataatt ttatacattt 6360 ttttaaaaaa tcttttaatt tcttaattaa tatcttaaaa ataatgatta atatttaacc 6420 caaaataatt agtatgattg gtaaggaaga tatccatgtt atgtttggat gtgagtttga 6480 tctagagcaa agcttactag agtcgacctg cagggttaaa gacacaaagc ctttagccta 6540 tgctgccaat aatggatacc aacaaaaggg ttcttctttt gattttgatc ctagcgctcc 6600 tccaccgttt aagattgcag aaatcagagc ttcaatacca aaacattgct gggtcaagaa 6660 tccatggaga tccctcagtt atgttctcag ggatgtgctt gtaattgctg cattggtggc 6720 tgcagcaatt cacttcgaca actggcttct ctggctaatc tattgcccca ttcaaggcac 6780 aatgttctgg gctctctttg ttcttggaca tgattgtggc catggaagga gatagataag 6840 cagagaaaac atggtagccc aacacagtgt tgatattgct attccttttc tctcactggg 6900 tgggaacaga ttgctgtagg gattgaagtg agagccttcc tttccggggc ttcttgaaaa 6960 caaataaatt ggatacacaa acaatggaaa tggcacagtg aatctaatga gtcttgtcat 7020 gctgtctaga ttcttgtaaa tcttctctgt taatggaacc catgactcat ccttctcaat 7080 gtgtccatgg ttttgatggt gagttctgtg gctaattctc catccatggt atggcacaag 7140 aattgaggaa tgcaagatgt gtcccaccag gctattcagc aaagggctat ctgaaaagct 7200 tccatggcca caatcatgtc caagaacaaa gagagcccag aacattgtgc cttgaatggg 7260 gcaatagatt agccagagaa gccagttgtc gaagtgaatt gctgcagcca ccaatgcagc 7320 aattacaagc acatccctga gaacataact gagggatctc catggattct tgacccagca 7380 atgttttggt attgaagctc tgatttctgc aatcttaaac ggtggaggag cgctaggatc 7440 aaaatcaaaa gaagaaccct tttgttggta tccattattg gcagcatagg ctaaaggctt 7500 tgtgtcttta acgcggccgc agtatatctt aaattcttta atacggtgta ctaggatatt 7560 gaactggttc ttgatgatga aaacctgggc cgagattgca gctatttata gtcataggtc 7620 ttgttaacat gcatggacat ttggccacgg ggtggcatgc agtttgacgg gtgttgaaat 7680 aaacaaaaat gaggtggcgg aagagaatac gagtttgagg ttgggttaga aacaacaaat 7740 gtgagggctc atgatgggtt gagttggtga atgttttggg ctgctcgatt gacacctttg 7800 tgagtacgtg ttgttgtgca tggcttttgg ggtccagttt ttttttcttg acgcggcgat 7860 cctgatcagc tagtggataa gtgatgtcca ctgtgtgtga ttgcgttttt gtttgaattt 7920 tatgaactta gacattgcta tgcaaaggat actctcattg tgttttgtct tcttttgttc 7980 cttggctttt tcttatgatc caagagacta gtcagtgttg tggcattcga gactaccaag 8040 attaattatg atgggggaag gataagtaac tgattagtac ggactgttac caaattaatt 8100 aataagcggc aaatgaaggg catggatcct gcaggtcgac tcgac 8145
<210> 40 <211> 11781 <212> DNA <213> Seqüência Artificial <220> <223> plasmídeo ρKKE2 <220> <221> misc feature <222> (1180)..(1180) <223> η é a, c, g, ou t <400> 40
gtacgtcctc gaagagaagg gttaataaca cattttttaa catttttaac acaaatttta 60 gttatttaaa aatttattaa aaaatttaaa ataagaagag gaactcttta aataaatcta 120 acttacaaaa tttatgattt ttaataagtt ttcaccaata aaaaatgtca taaaaatatg 180 ttaaaaagta tattatcaat attctcttta tgataaataa aaagaaaaaa aaaataaaag 240 ttaagtgaaa atgagattga agtgacttta ggtgtgtata aatatatcaa ccccgccaac 300 aatttattta atccaaatat attgaagtat attattccat agcctttatt tatttatata 360 tttattatat aaaagcttta tttgttctag gttgttcatg aaatattttt ttggttttat 420 ctccgttgta agaaaatcat gtgctttgtg tcgccactca ctattgcagc tttttcatgc 480 attggtcaga ttgacggttg attgtatttt tgttttttat ggttttgtgt tatgacttaa 540 gtcttcatct ctttatctct tcatcaggtt tgatggttac ctaatatggt ccatgggtac 600 atgcatggtt aaattaggtg gccaactttg ttgtgaacga tagaattttt tttatattaa 660 gtaaactatt tttatattat gaaataataa taaaaaaaat attttatcat tattaacaaa 720 atcatattag ttaatttgtt aactctataa taaaagaaat actgtaacat tcacattaca 780 tggtaacatc tttccaccct ttcatttgtt ttttgtttga tgactttttt tcttgtttaa 840 atttatttcc cttcttttaa atttggaata cattatcatc atatataaac taaaatacta 900 aaaacaggat tacacaaatg ataaataata acacaaatat ttataaatct agctgcaata 960 tatttaaact agctatatcg atattgtaaa ataaaactag ctgcattgat actgataaaa 1020 aaatatcatg tgctttctgg actgatgatg cagtatactt ttgacattgc ctttatttta 1080 tttttcagaa aagctttctt agttctgggt tcttcattat ttgtttccca tctccattgt 1140 gaattgaatc atttgcttcg tgtcacaaat acaatttagn taggtacatg cattggtcag 1200 attcacggtt tattatgtca tgacttaagt tcatggtagt acattacctg ccacgcatgc 1260
attatattgg ttagatttga taggcaaatt tggttgtcaa caatataaat ataaataatg 1320
tttttatatt acgaaataac agtgatcaaa acaaacagtt ttatctttat taacaagatt 1380
ttgtttttgt ttgatgacgt tttttaatgt ttacgctttc ccccttcttt tgaatttaga 1440
acactttatc atcataaaat caaatactaa aaaaattaca tatttcataa ataataacac 1500
aaatattttt aaaaaatctg aaataataat gaacaatatt acatattatc acgaaaattc 1560
attaataaaa atattatata aataaaatgt aatagtagtt atatgtagga aaaaagtact 1620
gcacgcataa tatatacaaa aagattaaaa tgaactatta taaataataa cactaaatta 1680
atggtgaatc atatcaaaat aatgaaaaag taaataaaat ttgtaattaa cttctatatg 1740
tattacacac acaaataata aataatagta aaaaaaatta tgataaatat ttaccatctc 1800
ataagatatt taaaataatg ataaaaatat agattatttt ttatgcaact agctagccaa 1860
aaagagaaca cgggtatata taaaaagagt acctttaaat tctactgtac ttcctttatt 1920
cctgacgttt ttatatcaag tggacatacg tgaagatttt aattatcagt ctaaatattt 1980
cattagcact taatactttt ctgttttatt cctatcctat aagtagtccc gattctccca 2040
acattgctta ttcacacaac taactaagaa agtcttccat agccccccaa gcggccgcat 2100
gggaacggac caaggaaaaa ccttcacctg ggaagagctg gcggcccata acaccaagga 2160
cgacctactc ttggccatcc gcggcagggt gtacgatgtc acaaagttct tgagccgcca 2220
tcctggtgga gtggacactc tcctgctcgg agctggccga gatgttactc cggtctttga 2280
gatgtatcac gcgtttgggg ctgcagatgc cattatgaag aagtactatg tcggtacact 2340
ggtctcgaat gagctgccca tcttcccgga gccaacggtg ttccacaaaa ccatcaagac 2400
gagagtcgag ggctacttta cggatcggaa cattgatccc aagaatagac cagagatctg 2460
gggacgatac gctcttatct ttggatcctt gatcgcttcc tactacgcgc agctctttgt 2520
gcctttcgtt gtcgaacgca catggcttca ggtggtgttt gcaatcatca tgggatttgc 2580
gtgcgcacaa gtcggactca accctcttca tgatgcgtct cacttttcag tgacccacaa 2640
ccccactgtc tggaagattc tgggagccac gcacgacttt ttcaacggag catcgtacct 2700
ggtgtggatg taccaacata tgctcggcca tcacccctac accaacattg ctggagcaga 2760
tcccgacgtg tcgacgtctg agcccgatgt tcgtcgtatc aagcccaacc aaaagtggtt 2820
tgtcaaccac atcaaccagc acatgtttgt tcctttcctg tacggactgc tggcgttcaa 2880
ggtgcgcatt caggacatca acattttgta ctttgtcaag accaatgacg ctattcgtgt 2940
caatcccatc tcgacatggc acactgtgat gttctggggc ggcaaggctt tctttgtctg 3000
gtatcgcctg attgttcccc tgcagtatct gcccctgggc aaggtgctgc tcttgttcac 3060 ggtcgcggac atggtgtcgt cttactggct ggcgctgacc ttccaggcga accacgttgt 3120 tgaggaagtt cagtggccgt tgcctgacga gaacgggatc atccaaaagg actgggcagc 3180 tatgcaggtc gagactacgc aggattacgc acacgattcg cacctctgga ccagcatcac 3240 tggcagcttg aactaccagg ctgtgcacca tctgttcccc aacgtgtcgc agcaccatta 3300 tcccgatatt ctggccatca tcaagaacac ctgcagcgag tacaaggttc cataccttgt 3360 caaggatacg ttttggcaag catttgcttc acatttggag cacttgcgtg ttcttggact 3420 ccgtcccaag gaagagtagg cggccgcgac acaagtgtga gagtactaaa taaatgcttt 3480 ggttgtacga aatcattaca ctaaataaaa taatcaaagc ttatatatgc cttccgctaa 3540 ggccgaatgc aaagaaattg gttctttctc gttatctttt gccactttta ctagtacgta 3600 ttaattacta cttaatcatc tttgtttacg gctcattata tccgtacgga tccgtcgacg 3660 gcgcgcccga tcatccggat atagttcctc ctttcagcaa aaaacccctc aagacccgtt 3720 tagaggcccc aaggggttat gctagttatt gctcagcggt ggcagcagcc aactcagctt 3780 cctttcgggc tttgttagca gccggatcga tccaagctgt acctcactat tcctttgccc 3840 tcggacgagt gctggggcgt cggtttccac tatcggcgag tacttctaca cagccatcgg 3900 tccagacggc cgcgcttctg cgggcgattt gtgtacgccc gacagtcccg gctccggatc 3960 ggacgattgc gtcgcatcga ccctgcgccc aagctgcatc atcgaaattg ccgtcaacca 4020 agctctgata gagttggtca agaccaatgc ggagcatata cgcccggagc cgcggcgatc 4080 ctgcaagctc cggatgcctc cgctcgaagt agcgcgtctg ctgctccata caagccaacc 4140 acggcctcca gaagaagatg ttggcgacct cgtattggga atccccgaac atcgcctcgc 4200 tccagtcaat gaccgctgtt atgcggccat tgtccgtcag gacattgttg gagccgaaat 4260 ccgcgtgcac gaggtgccgg acttcggggc agtcctcggc ccaaagcatc agctcatcga 4320 gagcctgcgc gacggacgca ctgacggtgt cgtccatcac agtttgccag tgatacacat 4380 ggggatcagc aatcgcgcat atgaaatcac gccatgtagt gtattgaccg attccttgcg 4440 gtccgaatgg gccgaacccg ctcgtctggc taagatcggc cgcagcgatc gcatccatag 4500 cctccgcgac cggctgcaga acagcgggca gttcggtttc aggcaggtct tgcaacgtga 4560 caccctgtgc acggcgggag atgcaatagg tcaggctctc gctgaattcc ccaatgtcaa 4620 gcacttccgg aatcgggagc gcggccgatg caaagtgccg ataaacataa cgatctttgt 4680 agaaaccatc ggcgcagcta tttacccgca ggacatatcc acgccctcct acatcgaagc 4740 tgaaagcacg agattcttcg ccctccgaga gctgcatcag gtcggagacg ctgtcgaact 4800 tttcgatcag aaacttctcg acagacgtcg cggtgagttc aggcttttcc atgggtatat 4860 ctccttctta aagttaaaca aaattatttc tagagggaaa ccgttgtggt ctccctatag 4920 tgagtcgtat taatttcgcg ggatcgagat ctgatcaacc tgcattaatg aatcggccaa 4980
cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 5040 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 5100
ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 5160
gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 5220
gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 5280
taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 5340
accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc 5400
tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 5460
cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 5520
agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 5580
gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca 5640
gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 5700
tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 5760
acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 5820
cagtggaacg aaaactcacg ttaagggatt ttggtcatga cattaaccta taaaaatagg 5880
cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac 5940
atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc 6000
cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca 6060
gagcagattg tactgagagt gcaccatatg gacatattgt cgttagaacg cggctacaat 6120
taatacataa ccttatgtat catacacata cgatttaggt gacactatag aacggcgcgc 6180
caagcttgtt gaaacatccc tgaagtgtct cattttattt tatttattct ttgctgataa 6240
aaaaataaaa taaaagaagc taagcacacg gtcaaccatt gctctactgc taaaagggtt 6300
atgtgtagtg ttttactgca taaattatgc agcaaacaag acaactcaaa ttaaaaaatt 6360
tcctttgctt gtttttttgt tgtctctgac ttgactttct tgtggaagtt ggttgtataa 6420
ggattgggac accattgtcc ttcttaattt aattttattc tttgctgata aaaaaaaaaa 6480
tttcatatag tgttaaataa taatttgtta aataaccaaa aagtcaaata tgtttactct 6540
cgtttaaata attgagattc gtccagcaag gctaaacgat tgtatagatt tatgacaata 6600
tttacttttt tatagataaa tgttatatta taataaattt atatacatat attatatgtt 6660
atttattatt attttaaatc cttcaatatt ttatcaaacc aactcataat ttttttttta 6720
tctgtaagaa gcaataaaat taaatagacc cactttaagg atgatccaac ctttatacag 6780 agtaagagag ttcaaatagt accctttcat atacatatca actaaaatat tagaaatatc 6840
atggatcaaa ccttataaag acattaaata agtggataag tataatatat aaatgggtag 6900
tatataatat ataaatggat acaaacttct ctctttataa ttgttatgtc tccttaacat 6960
cctaatataa tacataagtg ggtaatatat aatatataaa tggagacaaa cttcttccat 7020
tataattgtt atgtcttctt aacacttatg tctcgttcac aatgctaagg ttagaattgt 7080
ttagaaagtc ttatagtaca catttgtttt tgtactattt gaagcattcc ataagccgtc 7140
acgattcaga tgatttataa taataagagg aaatttatca tagaacaata aggtgcatag 7200
atagagtgtt aatatatcat aacatccttt gtttattcat agaagaagtg agatggagct 7260
cagttattat actgttacat ggtcggatac aatattccat gctctccatg agctcttaca 7320
cctacatgca ttttagttca tacttgcggc cagcttttac atggcgggaa actccttgaa 7380
gaactcgatc gagatgcgct cgaggtgcac gacgacctcg gccatgcccg cgatgaagcc 7440
cgtctcgtgg tatgggatgt cgtactgctt gcagagcgac ttgacgagca cgttgagcgc 7500
cgggaggttg tgccggggca ccatcgggaa caagtggtgg tcgatctggt agttgaggcc 7560
gcccatgaac cagtcgatcc agagcgacga cgtcacgttg cgcgtcgaga gcacttgcag 7620
cttccaaaaa tcgggcttgc tgtccttgtc aaagacctcc atgccgttgt ggccgacgct 7680
aaagaccatc gcgaggaaga ggccgcacga cgcctggctc acaaagagga acgcagccgc 7740
ttggagcagc gacatgttgg ctgcgtacac aaggccgagg ttccagccgt agtagaggag 7800
gaggccggcg cgctcgagca gcgggtactg gaccttgtca aaggtgccgc cgggcccaac 7860
gttgtagaag gcgtacatgg ccgactggat cacccacgag atacgcgcaa agagcaagat 7920
gggaaagtac aggtacgctt ggtagcgcat gaagaagagc ccgacgggcg agtcgaccgc 7980
gtgctgcgcc atcttgagcg accacgcgag aatcggcatc gtgtcaatgt ccgggtcgcc 8040
gtggaaggcg atctcgggcg tcgcgtggag gttggggatc gcatggtgcg tgttgtgctt 8100
gttcttccac cactgcaccg agaagccctg ccagaggttg ccgaccatga cgccgacgag 8160
gtcgccaaac aagtggttct caaacacttg gtggtgcaga aagtcatggg cgagccagcc 8220
gcactgctgg taaaagaggc caaggatgac agccgcgacc atgtacatgg ccgtcgagtc 8280
aaagtggagg caaatggccg ccgacacaag cgcaatgctc agcgtcgagg cgcacttgta 8340
gaggtagtag agcttgctcg agtcgtacaa gccgaggcgc ttgacttcaa ggcgcagctt 8400
gcggtacgac gcaatgaagt ccgactggct cttcttgacc tcgtccgaga tcgacgtgtc 8460
gacggccgcc gtcgactggt cgacgtcgcc gacgtagtac tgctcgagga gcttgagcgc 8520
cgagctcggg tggaagacag cgaacgcatc ggtcgcgtct tcgccggcct gcgtgaacat 8580
gacgacgccg cccgggtggt cctcaaaggc cgagatgtcg tacaccttgt ggtggatcac 8640 gatccacgcg ttgtcttggc ggttgtgctc acggatggtc gcccacgaga tcttctcggc 8700 cttttgcccc tggaccatga attggccgca gtatatctta aattctttaa tacggtgtac 8760 taggatattg aactggttct tgatgatgaa aacctgggcc gagattgcag ctatttatag 8820 tcataggtct tgttaacatg catggacatt tggccacggg gtggcatgca gtttgacggg 8880 tgttgaaata aacaaaaatg aggtggcgga agagaatacg agtttgaggt tgggttagaa 8940 acaacaaatg tgagggctca tgatgggttg agttggtgaa tgttttgggc tgctcgattg 9000 acacctttgt gagtacgtgt tgttgtgcat ggcttttggg gtccagtttt tttttcttga 9060 cgcggcgatc ctgatcagct agtggataag tgatgtccac tgtgtgtgat tgcgtttttg 9120 tttgaatttt atgaacttag acattgctat gcaaaggata ctctcattgt gttttgtctt 9180 cttttgttcc ttggcttttt cttatgatcc aagagactag tcagtgttgt ggcattcgag 9240 actaccaaga ttaattatga tgggggaagg ataagtaact gattagtacg gactgttacc 9300 aaattaatta ataagcggca aatgaagggc atggatcaaa agcttggatc tcctgcaggc 9360 tagcctaagt acgtactcaa aatgccaaca aataaaaaaa aagttgcttt aataatgcca 9420 aaacaaatta ataaaacact tacaacaccg gatttttttt aattaaaatg tgccatttag 9480 gataaatagt taatattttt aataattatt taaaaagccg tatctactaa aatgattttt 9540 atttggttga aaatattaat atgtttaaat caacacaatc tatcaaaatt aaactaaaaa 9600 aaaaataagt gtacgtggtt aacattagta cagtaatata agaggaaaat gagaaattaa 9660 gaaattgaaa gcgagtctaa tttttaaatt atgaacctgc atatataaaa ggaaagaaag 9720 aatccaggaa gaaaagaaat gaaaccatgc atggtcccct cgtcatcacg agtttctgcc 9780 atttgcaata gaaacactga aacacctttc tctttgtcac ttaattgaga tgccgaagcc 9840 acctcacacc atgaacttca tgaggtgtag cacccaaggc ttccatagcc atgcatactg 9900 aagaatgtct caagctcagc accctacttc tgtgacgtgt ccctcattca ccttcctctc 9960 ttccctataa ataaccacgc ctcaggttct ccgcttcaca actcaaacat tctctccatt 10020 ggtccttaaa cactcatcag tcatcaccgc ggccgcatgg agtcgattgc gccattcctc 10080 ccatcaaaga tgccgcaaga tctgtttatg gaccttgcca ccgctatcgg tgtccgggcc 10140 gcgccctatg tcgatcctct cgaggccgcg ctggtggccc aggccgagaa gtacatcccc 10200 acgattgtcc atcacacgcg tgggttcctg gtcgcggtgg agtcgccttt ggcccgtgag 10260 ctgccgttga tgaacccgtt ccacgtgctg ttgatcgtgc tcgcttattt ggtcacggtc 10320 tttgtgggca tgcagatcat gaagaacttt gagcggttcg aggtcaagac gttttcgctc 10380 ctgcacaact tttgtctggt ctcgatcagc gcctacatgt gcggtgggat cctgtacgag 10440 gcttatcagg ccaactatgg actgtttgag aacgctgctg atcatacctt caagggtctt 10500 cctatggcca agatgatctg gctcttctac ttctccaaga tcatggagtt tgtcgacacc 10560 atgatcatgg tcctcaagaa gaacaaccgc cagatctcct tcttgcacgt ttaccaccac 10620 agctccatct tcaccatctg gtggttggtc acctttgttg cacccaacgg tgaagcctac 10680 ttctctgctg cgttgaactc gttcatccat gtgatcatgt acggctacta cttcttgtcg 10740 gccttgggct tcaagcaggt gtcgttcatc aagttctaca tcacgcgctc gcagatgaca 10800 cagttctgca tgatgtcggt ccagtcttcc tgggacatgt acgccatgaa ggtccttggc 10860 cgccccggat accccttctt catcacggct ctgctttggt tctacatgtg gaccatgctc 10920 ggtctcttct acaactttta cagaaagaac gccaagttgg ccaagcaggc caaggccgac 10980 gctgccaagg agaaggcaag gaagttgcag taagcggccg catttcgcac caaatcaatg 11040 aaagtaataa tgaaaagtct gaataagaat acttaggctt agatgccttt gttacttgtg 11100 taaaataact tgagtcatgt acctttggcg gaaacagaat aaataaaagg tgaaattcca 11160 atgctctatg tataagttag taatacttaa tgtgttctac ggttgtttca atatcatcaa 11220 actctaattg aaactttaga accacaaatc tcaatctttt cttaatgaaa tgaaaaatct 11280 taattgtacc atgtttatgt taaacacctt acaattggtt ggagaggagg accaaccgat 11340 gggacaacat tgggagaaag agattcaatg gagatttgga taggagaaca acattctttt 11400 tcacttcaat acaagatgag tgcaacacta aggatatgta tgagactttc agaagctacg 11460 acaacataga tgagtgaggt ggtgattcct agcaagaaag acattagagg aagccaaaat 11520 cgaacaagga agacatcaag ggcaagagac aggaccatcc atctcaggaa aaggagcttt 11580 gggatagtcc gagaagttgt acaagaaatt ttttggaggg tgagtgatgc attgctggtg 11640 actttaactc aatcaaaatt gagaaagaaa gaaaagggag ggggctcaca tgtgaataga 11700 agggaaacgg gagaatttta cagttttgat ctaatgggca tcccagctag tggtaacata 11760 ttcaccatgt ttaaccttca c 11781 <210> 41 <211> 11366 <212> DNA <213> Seqüência Artificial <220> <223> plasmideo pKR973 <220> <221> misc feature <222> (5237) .. (5237) <223> n é a, c, g, ou t <400> 41 ggagatccaa gcttggcgcg ccgttctata gtgtcaccta aatcgtatgt gtatgataca 60 taaggttatg tattaattgt agccgcgttc taacgacaat atgtccatat ggtgcactct 120 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc 180 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt 240 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa 300 gggcctcgtg atacgcctat ttttataggt taatgtcatg accaaaatcc cttaacgtga 360 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc 420 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt 480 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc 540 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc 600 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 660 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 720 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga 780 actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc 840 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 900 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg 960 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt 1020 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc 1080 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg 1140 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc 1200 gcctctcccc gcgcgttggc cgattcatta atgcaggttg atcagatctc gatcccgcga 1260 aattaatacg actcactata gggagaccac aacggtttcc ctctagaaat aattttgttt 1320 aactttaaga aggagatata cccatggaaa agcctgaact caccgcgacg tctgtcgaga 1380 agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag 1440 aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct 1500 gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc 1560 cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc 1620 gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc 1680 agccggtcgc ggaggctatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt 1740 tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg 1800 cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt 1860 ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc 1920 acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag 1980
cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct 2040
tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc 2100
atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc 2160
aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat 2220
gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa 2280
gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc 2340
ccagcactcg tccgagggca aaggaatagt gaggtacagc ttggatcgat ccggctgcta 2400
acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac 2460
cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg 2520
gatgatcggg cgcgccgtcg acggatccgt acccggggat cctctagacg tacgcaaagg 2580
caaagattta aactcgaaaa cattacaaaa gtctcaaaac agaggcaagg ccatgcacaa 2640
agcacactct aagtgcttcc attgcctact aagtagggta cgtacacgat caccattcac 2700
cagtgatgat ctttattaat atacaacaca ctcagagaca gcttatgtta tagctagcta 2760
gcataaacta tcacatcatg tgttagtacg acaagtgaca acattgcttt taacttcgcg 2820
gccttggatc ctctagaccg gatataatga gccgtaaaca aagatgatta agtagtaatt 2880
aatacgtact agtaaaagtg gcaaaagata acgagaaaga accaatttct ttgcattcgg 2940
ccttagcgga aggcatatat aagctttgat tattttattt agtgtaatga tttcgtacaa 3000
ccaaagcatt tatttagtac tctcacactt gtgtcgcggc cgctcagtgg aggaggtgct 3060
cgaaggcgag agccttgatg ttctggtgca cgcactggaa gaggttgccc tcgcggtact 3120
cgaggccatg cttcttgaag aagtcgcgta cctcgggctg gatagtcatt aggttgccgg 3180
tgggcatcat ggggaagagg tgatgctcgg tctgcaggga gatgaagccc gtgagcacgt 3240
tcactagcca gccgcccgtg atgttacggg aggtcttggc ggtctgctcg acgagcgtca 3300
tgccctcgac gtgctcgcgg tcgaggatgt cctcaccata gtgggttgca aagacaacga 3360
tgcccgtcgc gaagccgcgt gcaaccatca tgagagggat gaggttgaga tgcgctgcga 3420
agacccagca caggagcgcg tagtgagcgg caagagcggc ggcctgcatc cacatcttag 3480
gcaggcgcac agccacgtac gcgatggact ccaggcgcca gtagaggtcg aggatggcca 3540
tggtcggcac atagtagtac tgctgccagc gctggaagaa gttgagccgc ttggcaatgg 3600
acgggttctc tcgcacgtag atgagcaggg gcgccgtctt gatgtccggg tccgaaccat 3660
cctcgttggt gcacacgtgg tgcgtgttat ggcgcgcgcg ccaccaggcc acgtcgtagc 3720
cctggaggaa gccgagggcg tatcccatgg cgttgttcgc cttgacgagc gtgttgaccg 3780 agcccttgaa gactgcgtga tgcaggtagt cgtgagcaag ccagccgtcg aggtagaagc 3840
caaagcccat ggagagcgcg cccgcgaggg gcgagacgcc gctccgcacg agataggcgc 3900
ccacgaagat gggcgcaagg acgatggtga gcttgaacag ctcgtccaac gggcaagggt 3960
cgtagagacc agccctcttg tggcgctcaa tcatctcctt gaactccgtg cccatggtgg 4020
accgcgaggg caggggcacg gaagcgggcg catcctccga cttcttgagc atgccctcca 4080
tcatcttgat ggccttgggc gagtggtaag actcgaactg ctccgtcgcg tcgcgctcct 4140
ggaagatgcg gaacaccttg ctcccgcccg ggtgtgcctt ctggaacttt gtgacatcca 4200
cgcgcttgcc gtagagcacg acgtgcacgc tcttgctatc gacggagctc acctcagcga 4260
gagacgcgtc ggtcccgctc accgcctgcg cgccgccgtc tccacccttg cccatggtgc 4320
ggccgcttgg ggggctatgg aagactttct tagttagttg tgtgaataag caatgttggg 4380
agaatcggga ctacttatag gataggaata aaacagaaaa gtattaagtg ctaatgaaat 4440
atttagactg ataattaaaa tcttcacgta tgtccacttg atataaaaac gtcaggaata 4500
aaggaagtac agtagaattt aaaggtactc tttttatata tacccgtgtt ctctttttgg 4560
ctagctagtt gcataaaaaa taatctatat ttttatcatt attttaaata tcttatgaga 4620
tggtaaatat ttatcataat tttttttact attatttatt atttgtgtgt gtaatacata 4680
tagaagttaa ttacaaattt tatttacttt ttcattattt tgatatgatt caccattaat 4740
ttagtgttat tatttataat agttcatttt aatctttttg tatatattat gcgtgcagta 4800
cttttttcct acatataact actattacat tttatttata taatattttt attaatgaat 4860
tttcgtgata atatgtaata ttgttcatta ttatttcaga ttttttaaaa atatttgtgt 4920
tattatttat gaaatatgta atttttttag tatttgattt tatgatgata aagtgttcta 4980
aattcaaaag aagggggaaa gcgtaaacat taaaaaacgt catcaaacaa aaacaaaatc 5040
ttgttaataa agataaaact gtttgttttg atcactgtta tttcgtaata taaaaacatt 5100
atttatattt atattgttga caaccaaatt tgcctatcaa atctaaccaa tataatgcat 5160
gcgtggcagg taatgtacta ccatgaactt aagtcatgac ataataaacc gtgaatctga 5220
ccaatgcatg tacctancta aattgtattt gtgacacgaa gcaaatgatt caattcacaa 5280
tggagatggg aaacaaataa tgaagaaccc agaactaaga aagcttttct gaaaaataaa 5340
ataaaggcaa tgtcaaaagt atactgcatc atcagtccag aaagcacatg atattttttt 5400
atcagtatca atgcagctag ttttatttta caatatcgat atagctagtt taaatatatt 5460
gcagctagat ttataaatat ttgtgttatt atttatcatt tgtgtaatcc tgtttttagt 5520
attttagttt atatatgatg ataatgtatt ccaaatttaa aagaagggaa ataaatttaa 5580
acaagaaaaa aagtcatcaa acaaaaaaca aatgaaaggg tggaaagatg ttaccatgta 5640 atgtgaatgt tacagtattt cttttattat agagttaaca aattaactaa tatgattttg 5700
ttaataatga taaaatattt tttttattat tatttcataa tataaaaata gtttacttaa 5760
tataaaaaaa attctatcgt tcacaacaaa gttggccacc taatttaacc atgcatgtac 5820
ccatggacca tattaggtaa ccatcaaacc tgatgaagag ataaagagat gaagacttaa 5880
gtcataacac aaaaccataa aaaacaaaaa tacaatcaac cgtcaatctg accaatgcat 5940
gaaaaagctg caatagtgag tggcgacaca aagcacatga ttttcttaca acggagataa 6000
aaccaaaaaa atatttcatg aacaacctag aacaaataaa gcttttatat aataaatata 6060
taaataaata aaggctatgg aataatatac ttcaatatat ttggattaaa taaattgttg 6120
gcggggttga tatatttata cacacctaaa gtcacttcaa tctcattttc acttaacttt 6180
tatttttttt ttctttttat ttatcataaa gagaatattg ataatatact ttttaacata 6240
tttttatgac attttttatt ggtgaaaact tattaaaaat cataaatttt gtaagttaga 6300
tttatttaaa gagttcctct tcttatttta aattttttaa taaattttta aataactaaa 6360
atttgtgtta aaaatgttaa aaaatgtgtt attaaccctt ctcttcgagg acgtacgaag 6420
cttagggcgg atcccccggg cgacctcgag ggggggcccg gtacgtgaag gttaaacatg 6480
gtgaatatgt taccactagc tgggatgccc attagatcaa aactgtaaaa ttctcccgtt 6540
tcccttctat tcacatgtga gccccctccc ttttctttct ttctcaattt tgattgagtt 6600
aaagtcacca gcaatgcatc actcaccctc caaaaaattt cttgtacaac ttctcggact 6660
atcccaaagc tccttttcct gagatggatg gtcctgtctc ttgcccttga tgtcttcctt 6720
gttcgatttt ggcttcctct aatgtctttc ttgctaggaa tcaccacctc actcatctat 6780
gttgtcgtag cttctgaaag tctcatacat atccttagtg ttgcactcat cttgtattga 6840
agtgaaaaag aatgttgttc tcctatccaa atctccattg aatctctttc tcccaatgtt 6900
gtcccatcgg ttggtcctcc tctccaacca attgtaaggt gtttaacata aacatggtac 6960
aattaagatt tttcatttca ttaagaaaag attgagattt gtggttctaa agtttcaatt 7020
agagtttgat gatattgaaa caaccgtaga acacattaag tattactaac ttatacatag 7080
agcattggaa tttcaccttt tatttattct gtttccgcca aaggtacatg actcaagtta 7140
ttttacacaa gtaacaaagg catctaagcc taagtattct tattcagact tttcattatt 7200
actttcattg atttggtgcg aaatgcggcc gcctactctt ccttgggacg gagtccaaga 7260
acacgcaagt gctccaaatg tgaagcaaat gcttgccaaa acgtatcctt gacaaggtat 7320
ggaaccttgt actcgctgca ggtgttcttg atgatggcca gaatatcggg ataatggtgc 7380
tgcgacacgt tggggaacag atggtgcaca gcctggtagt tcaagctgcc agtgatgctg 7440
gtccagaggt gcgaatcgtg tgcgtaatcc tgcgtagtct cgacctgcat agctgcccag 7500 tccttttgga tgatcccgtt ctcgtcaggc aacggccact gaacttcctc aacaacgtgg 7560 ttcgcctgga aggtcagcgc cagccagtaa gacgacacca tgtccgcgac cgtgaacaag 7620
agcagcacct tgcccagggg cagatactgc aggggaacaa tcaggcgata ccagacaaag 7 680
aaagccttgc cgccccagaa catcacagtg tgccatgtcg agatgggatt gacacgaata 7740
gcgtcattgg tcttgacaaa gtacaaaatg ttgatgtcct gaatgcgcac cttgaacgcc 7800
agcagtccgt acaggaaagg aacaaacatg tgctggttga tgtggttgac aaaccacttt 7860
tggttgggct tgatacgacg aacatcgggc tcagacgtcg acacgtcggg atctgctcca 7920
gcaatgttgg tgtaggggtg atggccgagc atatgttggt acatccacac caggtacgat 7980
gctccgttga aaaagtcgtg cgtggctccc agaatcttcc agacagtggg gttgtgggtc 8040
actgaaaagt gagacgcatc atgaagaggg ttgagtccga cttgtgcgca cgcaaatccc 8100
atgatgattg caaacaccac ctgaagccat gtgcgttcga caacgaaagg cacaaagagc 8160
tgcgcgtagt aggaagcgat caaggatcca aagataagag cgtatcgtcc ccagatctct 8220
ggtctattct tgggatcaat gttccgatcc gtaaagtagc cctcgactct cgtcttgatg 8280
gttttgtgga acaccgttgg ctccgggaag atgggcagct cattcgagac cagtgtaccg 8340
acatagtact tcttcataat ggcatctgca gccccaaacg cgtgatacat ctcaaagacc 8400
ggagtaacat ctcggccagc tccgagcagg agagtgtcca ctccaccagg atggcggctc 8460
aagaactttg tgacatcgta caccctgccg cggatggcca agagtaggtc gtccttggtg 8520
ttatgggccg ccagctcttc ccaggtgaag gtttttcctt ggtccgttcc catgcggccg 8580
cggtgatgac tgatgagtgt ttaaggacca atggagagaa tgtttgagtt gtgaagcgga 8640
gaacctgagg cgtggttatt tatagggaag agaggaaggt gaatgaggga cacgtcacag 8700
aagtagggtg ctgagcttga gacattcttc agtatgcatg gctatggaag ccttgggtgc 8760
tacacctcat gaagttcatg gtgtgaggtg gcttcggcat ctcaattaag tgacaaagag 8820
aaaggtgttt cagtgtttct attgcaaatg gcagaaactc gtgatgacga ggggaccatg 8880
catggtttca tttcttttct tcctggattc tttctttcct tttatatatg caggttcata 8940
atttaaaaat tagactcgct ttcaatttct taatttctca ttttcctctt atattactgt 9000
actaatgtta accacgtaca cttatttttt ttttagttta attttgatag attgtgttga 9060
tttaaacata ttaatatttt caaccaaata aaaatcattt tagtagatac ggctttttaa 9120
ataattatta aaaatattaa ctatttatcc taaatggcac attttaatta aaaaaaatcc 9180
ggtgttgtaa gtgttttatt aatttgtttt ggcattatta aagcaacttt ttttttattt 9240
gttggcattt tgagtacgta cttaggctag cctgcaggcc aactgcgttt ggggctccag 9300
attaaacgac gccgtttcgt tcctttcgct tcacggctta acgatgtcgt ttctgtctgt 9360 gcccaaaaaa taaaggcatt tgttatttgc accagatatt tactaagtgc accctagttt 9420 gacaagtagg cgataattac aaatagatgc ggtgcaaata ataaattttg aaggaaataa 9480 ttacaaaaga acagaactta tatttacttt attttaaaaa actaaaatga aagaacaaaa 9540 aaagtaaaaa atacaaaaaa tgtgctttaa ccactttcat tatttgttac agaaagtatg 9600 attctactca aattgatctg ttgtatctgg tgctgccttg tcacactggc gatttcaatc 9660 ccctaaagat atggtgcaaa ctgcgaagtg atcaatatct gctcggttaa tttagattaa 9720 ttaataatat tcaacgtgat gtaccaaaaa aagacaattt tttgctccat tgacaaatta 9780 aacctcatca aggtaatttc caaacctata agcaaaaaaa tttcacatta attggcccgc 9840 aatcctatta gtcttattat actagagtag gaaaaaaaac aattacacaa cttgtcttat 9900 tattctctat gctaatgaat atttttccct tttgttagaa atcagtgttt cctaatttat 9960 tgagtattaa ttccactcac cgcatatatt taccgttgaa taagaaaatt ttacacataa 10020 ttctttttaa gataaataat ttttttatac tagatcttat atgattacgt gaagccaagt 10080 gggttatact aatgatatat aatgtttgat agtaatcagt ttataaacca aatgcatgga 10140 aatgttacgt ggaagcacgt aaattaacaa gcattgaagc aaatgcagcc accgcaccaa 10200 aaccacccca cttcacttcc acgtaccata ttccatgcaa ctacaacacc ctaaaacttc 10260 aataaatgcc cccaccttca cttcacttca cccatcaata gcaagcggcc gcaccatgga 10320 ggtggtgaat gaaatagtct caattgggca ggaagtttta cccaaagttg attatgccca 10380 actctggagt gatgccagtc actgtgaggt gctttacttg tccatcgcat ttgtcatctt 10440 gaagttcact cttggccccc ttggtccaaa aggtcagtct cgtatgaagt ttgttttcac 10500 caattacaac cttctcatgt ccatttattc gttgggatca ttcctctcaa tggcatatgc 10560 catgtacacc atcggtgtta tgtctgacaa ctgcgagaag gcttttgaca acaacgtctt 10620 caggatcacc acgcagttgt tctatttgag caagttcctg gagtatattg actccttcta 10680 tttgccactg atgggcaagc ctctgacctg gttgcaattc ttccatcatt tgggggcacc 10740 gatggatatg tggctgttct ataattaccg aaatgaagct gtttggattt ttgtgctgtt 10800 gaatggtttc atccactgga tcatgtacgg ttattattgg accagattga tcaagctgaa 10860 gttccccatg ccaaaatccc tgattacatc aatgcagatc attcaattca atgttggttt 10920 ctacattgtc tggaagtaca ggaacattcc ctgttatcgc caagatggga tgaggatgtt 10980 tggctggttc ttcaattact tttatgttgg cacagtcttg tgtttgttct tgaatttcta 11040 tgtgcaaacg tatatcgtca ggaagcacaa gggagccaaa aagattcagt gagcggccgc 11100 gaagttaaaa gcaatgttgt cacttgtcgt actaacacat gatgtgatag tttatgctag 11160 ctagctataa cataagctgt ctctgagtgt gttgtatatt aataaagatc atcactggtg 11220 aatggtgatc gtgtacgtac cctacttagt aggcaatgga agcacttaga gtgtgctttg 11280 tgcatggcct tgcctctgtt ttgagacttt tgtaatgttt tcgagtttaa atctttgcct 11340 ttgcgtacgt ctagagtcga cctgca 11366
<210> 42
<211> 30
<212> DNA
<213> Seqüência Artificial <220>
<223> primer oEugELl-1
<400> 42
agcggccgca ccatggaggt ggtgaatgaa
<210> 43
<211> 30
<212> DNA
<213> Seqüência Artificial
30
<220>
<223> primer oEugELl-2 <400> 43
tgcggccgct cactgaatct ttttggctcc 30
<210> 44 <211> 12947 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo pKR1005 <400> 44
ggtcgactcg acgtacgtct agaggatccg tcgacggcgc gcccgatcat ccggatatag 60
ttcctccttt cagcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta 120 gttattgctc agcggtggca gcagccaact cagcttcctt tcgggctttg ttagcagccg 180 gatcgatcca agctgtacct cactattcct ttgccctcgg acgagtgctg gggcgtcggt 240 ttccactatc ggcgagtact tctacacagc catcggtcca gacggccgcg cttctgcggg 300 cgatttgtgt acgcccgaca gtcccggctc cggatcggac gattgcgtcg catcgaccct 360 gcgcccaagc tgcatcatcg aaattgccgt caaccaagct ctgatagagt tggtcaagac 420 caatgcggag catatacgcc cggagccgcg gcgatcctgc aagctccgga tgcctccgct 480 cgaagtagcg cgtctgctgc tccatacaag ccaaccacgg cctccagaag aagatgttgg 540 cgacctcgta ttgggaatcc ccgaacatcg cctcgctcca gtcaatgacc gctgttatgc 600 ggccattgtc cgtcaggaca ttgttggagc cgaaatccgc gtgcacgagg tgccggactt 660 cggggcagtc ctcggcccaa agcatcagct catcgagagc ctgcgcgacg gacgcactga 720 cggtgtcgtc catcacagtt tgccagtgat acacatgggg atcagcaatc gcgcatatga 780 aatcacgcca tgtagtgtat tgaccgattc cttgcggtcc gaatgggccg aacccgctcg 840 tctggctaag atcggccgca gcgatcgcat ccatagcctc cgcgaccggc tgcagaacag 900 cgggcagttc ggtttcaggc aggtcttgca acgtgacacc ctgtgcacgg cgggagatgc 960 aataggtcag gctctcgctg aattccccaa tgtcaagcac ttccggaatc gggagcgcgg 1020 ccgatgcaaa gtgccgataa acataacgat ctttgtagaa accatcggcg cagctattta 1080 cccgcaggac atatccacgc cctcctacat cgaagctgaa agcacgagat tcttcgccct 1140 ccgagagctg catcaggtcg gagacgctgt cgaacttttc gatcagaaac ttctcgacag 1200 acgtcgcggt gagttcaggc ttttccatgg gtatatctcc ttcttaaagt taaacaaaat 1260 tatttctaga gggaaaccgt tgtggtctcc ctatagtgag tcgtattaat ttcgcgggat 1320 cgagatctga tcaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 1380 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 1440 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 1500 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 1560 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 1620 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 1680 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 1740 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag 1800 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 1860 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 1920 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 1980 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg 2040 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 2100 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 2160 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 2220 gggattttgg tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc 2280 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca 2340 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt 2400 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac 2460 catatggaca tattgtcgtt agaacgcggc tacaattaat acataacctt atgtatcata 2520 cacatacgat ttaggtgaca ctatagaacg gcgcgccaag ctgggtctag aactagaaac 2580 gtgatgccac ttgttattga agtcgattac agcatctatt ctgttttact atttataact 2640 ttgccatttc tgacttttga aaactatctc tggatttcgg tatcgctttg tgaagatcga 2700 gcaaaagaga cgttttgtgg acgcaatggt ccaaatccgt tctacatgaa caaattggtc 2760 acaatttcca ctaaaagtaa ataaatggca agttaaaaaa ggaatatgca ttttactgat 2820 tgcctaggtg agctccaaga gaagttgaat ctacacgtct accaaccgct aaaaaaagaa 2880 aaacattgaa tatgtaacct gattccatta gcttttgact tcttcaacag attctctact 2940 tagatttcta acagaaatat tattactagc acatcatttt cagtctcact acagcaaaaa 3000 atccaacggc acaatacaga caacaggaga tatcagacta cagagataga tagatgctac 3060 tgcatgtagt aagttaaata aaaggaaaat aaaatgtctt gctaccaaaa ctactacaga 3120 ctatgatgct caccacaggc caaatcctgc aactaggaca gcattatctt atatatattg 3180 tacaaaacaa gcatcaagga acatttggtc taggcaatca gtacctcgtt ctaccatcac 3240 cctcagttat cacatccttg aaggatccat tactgggaat catcggcaac acatgctcct 3300 gatggggcac aatgacatca agaaggtagg ggccaggggt gtccaacatt ctctgaattg 3360 ccgctctaag ctcttccttc ttcgtcactc gcgctgccgg tatcccacaa gcatcagcaa 3420 acttgagcat gtttgggaat atctcgctct cgctagacgg atctccaaga taggtgtgag 3480 ctctattgga cttgtagaac ctatcctcca actgaaccac catacccaaa tgctgattgt 3540 tcaacaacaa tatcttaact gggagattct ccactcttat agtggccaac tcctgaacat 3600 tcatgatgaa actaccatcc ccatcaatgt caaccacaac agccccaggg ttagcaacag 3660 cagcaccaat agccgcaggc aatccaaaac ccatggctcc aagaccccct gaggtcaacc 3720 actgcctcgg tctcttgtac ttgtaaaact gcgcagccca catttgatgc tgcccaaccc 3780 cagtactaac aatagcatct ccattagtca actcatcaag aacctcgata gcatgctgcg 3840 gagaaatcgc gtcctggaat gtcttgtaac ccaatggaaa cttgtgtttc tgcacattaa 3900 tctcttctct ccaacctcca agatcaaact taccctccac tcctttctcc tccaaaatca 3960 tattaattcc cttcaaggcc aacttcaaat ccgcgcaaac cgacacgtgc gcctgcttgt 4020 tcttcccaat ctcggcagaa tcaatatcaa tgtgaacaat cttagcccta ctagcaaaag 4080 cctcaagctt cccagtaaca cggtcatcaa accttacccc aaaggcaagc aacaaatcac 4140 tattgtcaac agcatagtta gcataaacag taccatgcat acccagcatc tgaagggaat 4200 attcatcacc aataggaaaa gttccaagac ccattaaagt gctagcaacg ggaataccag 4260 tgagttcaac aaagcgcctc aattcagcac tggaattcaa actgccaccg ccgacgtaga 4320 gaacgggctt ttgggcctcc atgatgagtc tgacaatgtg ttccaattgg gcctcggcgg 4380 ggggcctggg cagcctggcg aggtaaccgg ggaggttaac gggctcgtcc caattaggca 4440
cggcgagttg ctgctgaacg tctttgggaa tgtcgatgag gaccggaccg gggcggccgg 4500
aggtggcgac gaagaaagcc tcggcgacga cgcgggggat gtcgtcgacg tcgaggatga 4560
ggtagttgtg cttcgtgatg gatctgctca cctccacgat cggggtttct tggaaggcgt 4620
cggtgccgat catccggcgg gcgacctggc cggtgatggc gacgactggg acgctgtcca 4680
ttaaagcgtc ggcgaggccg ctcacgaggt tggtggcgcc ggggccggag gtggcaatgc 4740
agacgccggg gaggccggag gaacgcgcgt agccttcggc ggcgaagacg ccgccctgct 4800
cgtggcgcgg gagcacgttg cggatggcgg cggagcgcgt gagcgcctgg tggatctcca 4860
tcgacgcacc gccggggtac gcgaacaccg tcgtcacgcc ctgcctctcc agcgcctcca 4920
caaggatgtc cgcgcccttg cgaggttcgc cggaggcgaa ccgtgacacg aagggctccg 4980
tggtcggcgc ttccttggtg aagggcgccg ccgtgggggg tttggagatg gaacatttga 5040
ttttgagagc gtggttgggt ttggtgaggg tttgatgaga gagagggagg gtggatctag 5100
taatgcgttt ggggaaggtg gggtgtgaag aggaagaaga gaatcgggtg gttctggaag 5160
cggtggccgc cattgtgttg tgtggcatgg ttatacttca aaaactgcac aacaagccta 5220
gagttagtac ctaaacagta aatttacaac agagagcaaa gacacatgca aaaatttcag 5280
ccataaaaaa agttataata gaatttaaag caaaagtttc attttttaaa catatataca 5340
aacaaactgg atttgaagga agggattaat tcccctgctc aaagtttgaa ttcctattgt 5400
gacctatact cgaataaaat tgaagcctaa ggaatgtatg agaaacaaga aaacaaaaca 5460
aaactacaga caaacaagta caattacaaa attcgctaaa attctgtaat caccaaaccc 5520
catctcagtc agcacaaggc ccaaggttta ttttgaaata aaaaaaaagt gattttattt 5580
ctcataagct aaaagaaaga aaggcaatta tgaaatgatt tcgactagat ctgaaagtca 5640
aacgcgtatt ccgcagatat taaagaaaga gtagagtttc acatggatcc tagatggacc 5700
cagttgagga aaaagcaagg caaagcaaac cagaagtgca agatccgaaa ttgaaccacg 5760
gaatctagga tttggtagag ggagaagaaa agtaccttga gaggtagaag agaagagaag 5820
agcagagaga tatatgaacg agtgtgtctt ggtctcaact ctgaagcgat acgagtttag 5880
aggggagcat tgagttccaa tttataggga aaccgggtgg caggggtgag ttaatgacgg 5940
aaaagcccct aagtaacgag attggattgt gggttagatt caaccgtttg catccgcggc 6000
ttagattggg gaagtcagag tgaatctcaà ccgttgactg agttgaaaat tgaatgtagc 6060
aaccaattga gccaacccca gcctttgccc tttgattttg atttgtttgt tgcatacttt 6120
ttatttgtct tctggttctg actctctttc tctcgtttca atgccaggtt gcctactccc 6180
acaccactca caagaagatt ctactgttag tattaaatat tttttaatgt attaaatgat 6240 gaatgctttt gtaaacagaa caagactatg tctaataagt gtcttgcaac attttttaag 6300
aaattaaaaa aaatatattt attatcaaaa tcaaatgtat gaaaaatcat gaataatata 6360
attttataca tttttttaaa aaatctttta atttcttaat taatatctta aaaataatga 6420
ttaatattta acccaaaata attagtatga ttggtaagga agatatccat gttatgtttg 6480
gatgtgagtt tgatctagag caaagcttac tagagtcgac ctgcagcccg ggggatccgc 654 0
ccacgtacgg taccatctgc taatatttta aatcacatgc aagagaggag gcatggttcc 6600
attttctacc ttcacattat ttgagaaaaa cgaacttgtt ctgtgtttta tttttgccct 6660
tcacattagt acaacgtgga agactcatgg ttacacagaa tcatacataa gtacaatgct 6720
tgtccctaag aaaacaagca ctcgttgtat tgaaccttta cggctcatgc ggccgcgaat 6780
tcactagtga ttgaattcgc ggccgcttag tccgacttgg ccttggcggc cgcggccgac 6840
tctttgagcg tgaagatctg cgccgtctcg ggcacagcgc cgtagttgac aaagaggtgc 6900
gcggtcttga agaaggccgt gatgatgggc tcgtcgttcc tgcgcacgag gtgcgggtac 6960
gcggccgcaa agtgcttggt ggcttcgttg agcttgtagt gcggaatgat cgggaacaag 7020
tggtggacct ggtgcgtgcc aatgtggtgg ctcaggttgt ccacgaacgc gccgtacgag 7 080
cggtcgacgc tcgagaggtt gcccttgacg tacgtccact ccgagtcgcc gtaccacggc 7140
gtcgcttcgt cgttgtggtg caagaaggtc gtaatgacga ggaacgaagc aaagacaaag 7200
agcggcgcat agtagtagag gcccatgacg gcaaagccga gcgagtatgt gaggtacgcg 7260
tacgcggcga agaaggcggc ccagacgccg agcgacacga tgacggccga cgcgcggcga 7320
aggaggagcg ggtcccacgg gtcaaagtgg ctcatcgtgc gcggggcata cccgaccttc 7380
aagtagacaa accacgcacc gccgagcgtg tagacccatt ggcgcacgtc ctggaggtcc 7440
ttgaccgacc ggtgcgggta aaagatctcg tccttatcaa tgttgcccgt gttcttgtgg 7500
tggtggcggt gcgtcacgcg ccagctctcg aacggcgtca aaatcgcaga gtgcatgatg 7560
cagccgatga taaagttgac gctgtggtag cgcgagaagg ccgagtggcc gcagtcgtgg 7 620
ccgaccgtga agaagcccca gaagatgacg ccctgcacgt agatgtaggt ggcgcaaacg 7 68 0
agcgcgtgga gcagaacgtt atcggcaatg aacggcgtcg agcgcgccgc gtagagcagc 7740
gccgccgagg ccgacgcgtt gaagatcgcg cgggccgtgt agtagagcga gaggccgagg 7800
ttcgactcaa agcacgcgtt cgggatcgag tgcttgagct ccgtgagcgt cgggaactcg 7860
accttcgtct tatcctcagt catgcggccg ctgaagtatt gcttcttagt taacctttcc 7920
tttctctctc agctatgtga attcattttg ctttcgtcac aatttatata gtgaaattgg 7980
atctttggag ttaacgcctt cacaggatta tcgtgttaga acaatgcttt ttcatgttct 8040
aattagtagt acattacaaa tgtgcactct attcaataag catcttttgg cacgttaata 8100 aatcatgtga aaaaaaaata ctactatttc aaagaaagtg ttgtaaaaag aaacggaaag 8160 agagctggct tcagttgttg agacttgttt gctagtaaaa atggtgtgaa gagtgattca 8220 tggtgaggtg gtttttcgtc cctttctgtt tgcatgaaaa acaaatggca agagatgacg 8280 taggattcct tcccttaacg attatctgtt tttaatttca aatatacata taggaattta 8340 tgaattacta aggttgtaaa atatgctggt catttattta tggctaaaat attttttttt 8400 ctcgtaaata taaaaatatt taaaatttat ttttatcata ttttttatcc ttataaaatt 8460 atgtgtacaa cctatataaa aaaatatcat atttaatatt gattatatgt ttaatcaata 8520 taaaaaatca ttatcatata tttagattta ttcgaatata catctaaaca aaaaataaca 8580 tattttaatt ttatgaagaa aaaaaaatat tttatccttt atttatttaa gattaattaa 8640 tagttatgta ttgtggaaag acttttacac atgcaataga tatactgaat caattagatg 8700 ccaatgctga gttggaaatc acttgaggag gggaggagac ttgccaatgc ttttcagttt 8760 catttaaatg atttagtgga ggagatagag tagtgataaa ggcatgcccc aattttggag 8820 tgtatatatg agtggaaata agagagggat agagagaaaa aataaagaga gtaaaaataa 8880 ttaatgtgaa atgatatgat aaaaaaataa agaaagagat aaagagaaaa atgaaatgag 8940 agatagatga aatagagagt agatacatgt ttgtttaggt tttttttagg aaataacaca 9000 tttttttctc atcacttatt actcactgtc aatttcctct ctttcaatca taatgatatg 9060 atttgtttaa caaaaatgtg aaaaaacata taaagtaaaa tatttttata aattgataaa 9120 taaaaattta caaaatttat ttcttattaa attgaataga aaatgaaaga aaagaaaaga 9180 aaaagtatat ataaaatgat atagctttaa aaagaataaa tttttcatat cagtcttttt 9240 ttaataattt agaaatattt aagtatatag caaaaatata atgtacttta catatgcata 9300 aataataatt tgaaaataga actaatagaa tagagaaaaa agtaatataa taattaacta 9360 tatgaaaatt tagaagggac aatattttta attaagaata taaacaatat ttcttttcat 9420 gtaatgaggg acggatgtac ggggccagtg ttggagtcaa agccaaaata gtcacgggga 9480 aattaatgca ctgcatgact attcgaaaaa attcactagc cttacttaga tgttagatta 9540 atagctaggg ggtgcagata attttgaaag gcatgaaaaa cattaatttg tacattgcaa 9600 gcttttgatg acaagctttg caattgttca cactacctta tgccatttat aaatagagtg 9660 attggcatat gaaggaaatc atgagagtcg aagcgaaaaa çaaagcttga gagtgtagga 9720 aaaatacagt ttttttggta aaaatacagt atttgaatag gagcgaaaaa tatcctttca 9780 aaatgatcct tttctttttt tttttttttc ttgttgttct tggtcagtta ttcaaaggaa 9840 aagggattga aataaaaact tgcatgtggg atcgtacgtc gagtcgacct gcaggatcca 9900 tgcccttcat ttgccgctta ttaattaatt tggtaacagt ccgtactaat cagttactta 9960 tccttccccc atcataatta atcttggtag tctcgaatgc cacaacactg actagtctct 10020
tggatcataa gaaaaagcca aggaacaaaa gaagacaaaa cacaatgaga gtatcctttg 10080
catagcaatg tctaagttca taaaattcaa acaaaaacgc aatcacacac agtggacatc 10140
acttatccac tagctgatca ggatcgccgc gtcaagaaaa aaaaactgga ccccaaaagc 10200
catgcacaac aacacgtact cacaaaggtg tcaatcgagc agcccaaaac attcaccaac 10260
tcaacccatc atgagccctc acatttgttg tttctaaccc aacctcaaac tcgtattctc 10320
ttccgccacc tcatttttgt ttatttcaac acccgtcaaa ctgcatgcca ccccgtggcc 10380
aaatgtccat gcatgttaac aagacctatg actataaata gctgcaatct cggcccaggt 10440
tttcatcatc aagaaccagt tcaatatcct agtacaccgt attaaagaat ttaagatata 10500
ctgcggccgc accatgtctc ctaagcggca agctctgcca atcacaattg atggcgcaac 10560
ttatgatgtg tctgcttggg tcaatcacca ccctggagga gctgacatta tcgagaacta 10620
tcgcaaccgc gatgcgaccg atgtcttcat ggtgatgcac tctcaagaag ccgtcgccaa 10680
gttgaagaga atgcctgtta tggagccttc ctctcctgac acacctgttg cacccaagcc 10740
taagcgtgat gagccccagg aggatttccg caagttgcgg gaggaattca tctccaaggg 10800
tatgttcgag acgagtttcc tttggtattt ttacaagact tcaactaccg tcggtttgat 10860
ggtcctttcc atcttgatga ccgtgtacac gaattggtat ttcaccgctg ctttggttct 10920
tggcgtgtgc taccaacagc taggctggtt gtcccacgac tattgccatc accaggtttt 10980
cacaaaccgc aagattaacg acgctttcgg tctctttttc ggtaacgtga tgcagggata 11040
ctcacagact tggtggaagg ataggcacaa tggtcaccat gccgccacca atgtggtcgg 11100
ccatgaccca gatattgata acctccccat cctggcttgg tctcccgaag atgtcaagag 11160
ggctactcct tcgactcgga atctcatcaa gtaccagcag tactacttca ttcccaccat 11220
tgcatccctt aggttcatct ggtgcctcca atccatcggc ggcgtcatgt cctacaagag 11280
cgaggagagg aacctgtact acaagcgcca gtacactaag gaggcgattg gtctggccct 11340
ccactgggtg ctcaaggcca ctttctattg cagtgccatg cctagctttg ccaccggttt 11400
gggatgcttc ttgatctccg agctgctcgg aggatttggc attgccatcg ttgtgtttct 11460
gaatcactat cctttggaca aggttgagga gactgtctgg gatgagcacg ggttcagcgc 11520
cagccagatc cacgagacgt tgaacattaa gcccggcctt ctcaccgatt gggtctttgg 11580
tggtctcaac taccagattg agcaccactt gtggcccaac atgcccaggc acaacctcac 1164 0
ggcagcttcc ctggaggtgc agaagttgtg cgccaagcac aacctgccct acagggcccc 11700
agccatcatc cccggggttc agaaattggt cagcttctta ggcgagattg cccagctggc 11760
tgctgtccct gaatgagcgg ccgcaagtat gaactaaaat gcatgtaggt gtaagagctc 11820 atggagagca tggaatattg tatccgacca tgtaacagta taataactga gctccatctc 11880 acttcttcta tgaataaaca aaggatgtta tgatatatta acactctatc tatgcacctt 11940 attgttctat gataaatttc ctcttattat tataaatcat ctgaatcgtg acggcttatg 12000 gaatgcttca aatagtacaa aaacaaatgt gtactataag actttctaaa caattctaac 12060 cttagcattg tgaacgagac ataagtgtta agaagacata acaattataa tggaagaagt 12120 ttgtctccat ttatatatta tatattaccc acttatgtat tatattagga tgttaaggag 12180 acataacaat tataaagaga gaagtttgta tccatttata tattatatac tacccattta 12240 tatattatac ttatccactt atttaatgtc tttataaggt ttgatccatg atatttctaa 12300 tattttagtt gatatgtata tgaaagggta ctatttgaac tctcttactc tgtataaagg 12360 ttggatcatc cttaaagtgg gtctatttaa ttttattgct tcttacagat aaaaaaaaaa 12420 ttatgagttg gtttgataaa atattgaagg atttaaaata ataataaata acatataata 12480 tatgtatata aatttattat aatataacat ttatctataa aaaagtaaat attgtcataa 12540 atctatacaa tcgtttagcc ttgctggacg aatctcaatt atttaaacga gagtaaacat 12600 atttgacttt ttggttattt aacaaattat tatttaacac tatatgaaat tttttttttt 12660 atcagcaaag aataaaatta aattaagaag gacaatggtg tcccaatcct tatacaacca 12720 acttccacaa gaaagtcaag tcagagacaa caaaaaaaca agcaaaggaa attttttaat 12780 ttgagttgtc ttgtttgctg cataatttat gcagtaaaac actacacata acccttttag 12840 cagtagagca atggttgacc gtgtgcttag cttcttttat tttatttttt tatcagcaaa 12900 gaataaataa aataaaatga gacacttcag ggatgtttca acctgca 12947 <210> 45 <211> 4311 <212> DNA <213> Seqüência Artificial <220> <223> plasmideo pKR906 <400> 45 agcggccgca ccatggaggt ggtgaatgaa atagtctcaa ttgggcagga agttttaccc 60 aaagttgatt atgcccaact ctggagtgat gccagtcact gtgaggtgct ttacttgtcc 120 atcgcatttg tcatcttgaa gttcactctt ggcccccttg gtccaaaagg tcagtctcgt 180 atgaagtttg ttttcaccaa ttacaacctt ctcatgtcca tttattcgtt gggatcattc 240 ctctcaatgg catatgccat gtacaccatc ggtgttatgt ctgacaactg cgagaaggct 300 tttgacaaca acgtcttcag gatcaccacg cagttgttct atttgagcaa gttcctggag 360 tatattgact ccttctattt gccactgatg ggcaagcctc tgacctggtt gcaattcttc 420 catcatttgg gggcaccgat ggatatgtgg ctgttctata attaccgaaa tgaagctgtt 480 tggatttttg tgctgttgaa tggtttcatc cactggatca tgtacggtta ttattggacc 540 agattgatca agctgaagtt ccccatgcca aaatccctga ttacatcaat gcagatcatt 600 caattcaatg ttggtttcta cattgtctgg aagtacagga acattccctg ttatcgccaa 660 gatgggatga ggatgtttgg ctggttcttc aattactttt atgttggcac agtcttgtgt 720 ttgttcttga atttctatgt gcaaacgtat atcgtcagga agcacaaggg agccaaaaag 780 attcagtgag cggccgcacc tgaattccag cacactggcg gccgttacta gtggatccga 840 gctcggtacc aagcttgatg catagcttga gtattctaac gcgtcaccta aatagcttgg 900 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 960 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 1020 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 1080 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 1140 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 1200 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 1260 caaaaggcca gcaaaagccc aggaaccgta aaaaggccgc gttgctggcg tttttccata 1320 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 1380 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 1440 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 1500 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 1560 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 1620 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 1680 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 1740 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 1800 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 1860 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 1920 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 1980 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt agcacgtgtc 2040 agtcctgctc ctcggccacg aagtgcacgc agttgccggc cgggtcgcgc agggcgaact 2100 cccgccccca cggctgctcg ccgatctcgg tcatggccgg cccggaggcg tcccggaagt 2160 tcgtggacac gacctccgac cactcggcgt acagctcgtc caggccgcgc acccacaccc 2220 aggccagggt gttgtccggc accacctggt cctggaccgc gctgatgaac agggtcacgt 2280 cgtcccggac cacaccggcg aagtcgtcct ccacgaagtc ccgggagaac ccgagccggt 2340 cggtccagaa ctcgaccgct ccggcgacgt cgcgcgcggt gagcaccgga acggcactgg 2400 tcaacttggc catggtggcc ctcctcacgt gctattattg aagcatttat cagggttatt 2460 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 2520 gcacatttcc ccgaaaagtg ccacctgtat gcggtgtgaa ataccgcaca gatgcgtaag 2580 gagaaaatac cgcatcagga aattgtaagc gttaataatt cagaagaact cgtcaagaag 2640 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg 2700 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg 2760 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc 2820 caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg 2880 catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc 2940 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg 3000 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc 3060 atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc 3120 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc 3180 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cttgcagttc 324 0 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag 3300 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag 3360 cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa 3420 cgatcctcat cctgtctctt gatcagagct tgatcccctg cgccatcaga tccttggcgg 3480 cgagaaagcc atccagttta ctttgcaggg cttcccaacc ttaccagagg gcgccccagc 3540 tggcaattcc ggttcgcttg ctgtccataa aaccgcccag tctagctatc gccatgtaag 3600 cccactgcaa gctacctgct ttctctttgc gcttgcgttt tcccttgtcc agatagccca 3660 gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtgaaaag 3720 gatctaggtg aagatccttt ttgataatct catgcctgac atttatattc cccagaacat 3780 caggttaatg gcgtttttga tgtcattttc gcggtggctg agatcagcca cttcttcccc 3840 gataacggag accggcacac tggccatatc ggtggtcatc atgcgccagc tttcatcccc 3900 gatatgcacc accgggtaaa gttcacggga gactttatct gacagcagac gtgcactggc 3960 cagggggatc accatccgtc gccccggcgt gtcaataata tcactctgta catccacaaa 4020 cagacgataa cggctctctc ttttataggt gtaaacctta aactgccgta cgtataggct 4080 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa 4140 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg 4200 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tgggccctct 4260 agatgcatgc tcgagcggcc gccagtgtga tggatatctg cagaattcag g 4311
<210> 46
<211> 777
<212> DNA
<213> Euglena gracilis
<400> 46
atggaggtgg tgaatgaaat agtctcaatt gggcaggaag ttttacccaa agttgattat 60 gcccaactct ggagtgatgc cagtcactgt gaggtgcttt acttgtccat cgcatttgtc 120 atcttgaagt tcactcttgg cccccttggt ccaaaaggtc agtctcgtat gaagtttgtt 180 ttcaccaatt acaaccttct catgtccatt tattcgttgg gatcattcct ctcaatggca 240 tatgccatgt acaccatcgg tgttatgtct gacaactgcg agaaggcttt tgacaacaac 300 gtcttcagga tcaccacgca gttgttctat ttgagcaagt tcctggagta tattgactcc 360 ttctatttgc cactgatggg caagcctctg acctggttgc aattcttcca tcatttgggg 420 gcaccgatgg atatgtggct gttctataat taccgaaatg aagctgtttg gatttttgtg 480 ctgttgaatg gtttcatcca ctggatcatg tacggttatt attggaccag attgatcaag 540 ctgaagttcc ccatgccaaa atccctgatt acatcaatgc agatcattca attcaatgtt 600 ggtttctaca ttgtctggaa gtacaggaac attccctgtt atcgccaaga tgggatgagg 660 atgtttggct ggttcttcaa ttacttttat gttggcacag tcttgtgttt gttcttgaat 720 ttctatgtgc aaacgtatat cgtcaggaag cacaagggag ccaaaaagat tcagtga 777
<210> 47 <211> 258 <212> PRT <213> Euglena gracilis
<400> 47
Met Glu Val Val Asn Glu Ile Val Ser Ile Gly Gln Glu Val Leu Pro 1 5 10 15
Lys Val Asp Tyr Ala Gln Leu Trp Ser Asp Ala Ser His Cys Glu Val 20 25 30
Leu Tyr Leu Ser Ile Ala Phe Val Ile Leu Lys Phe Thr Leu Gly Pro 35 40 45
Leu Gly Pro Lys Gly Gln Ser Arg Met Lys Phe Val Phe Thr Asn Tyr 50 55 60 Asn Leu Leu Met Ser Ile Tyr Ser Leu Gly Ser Phe Leu Ser Met Ala 65 70 75 80
Tyr Ala Met Tyr Thr Ile Gly Val Met Ser Asp Asn Cys Glu Lys Ala 85 90 95
Phe Asp Asn Asn Val Phe Arg Ile Thr Thr Gln Leu Phe Tyr Leu Ser 100 105 110
Lys Phe Leu Glu Tyr Ile Asp Ser Phe Tyr Leu Pro Leu Met Gly Lys 115 120 125
Pro Leu Thr Trp Leu Gln Phe Phe His His Leu Gly Ala Pro Met Asp 130 135 140
Met Trp Leu Phe Tyr Asn Tyr Arg Asn Glu Ala Val Trp Ile Phe Val 145 150 155 160
Leu Leu Asn Gly Phe Ile His Trp Ile Met Tyr Gly Tyr Tyr Trp Thr 165 170 175
Arg Leu Ile Lys Leu Lys Phe Pro Met Pro Lys Ser Leu Ile Thr Ser 180 185 190
Met Gln Ile Ile Gln Phe Asn Val Gly Phe Tyr Ile Val Trp Lys Tyr 195 200 205
Arg Asn Ile Pro Cys Tyr Arg Gln Asp Gly Met Arg Met Phe Gly Trp 210 215 220
Phe Phe Asn Tyr Phe Tyr Val Gly Thr Val Leu Cys Leu Phe Leu Asn 225 230 235 240
Phe Tyr Val Gln Thr Tyr Ile Val Arg Lys His Lys Gly Ala Lys Lys 245 250 255
Ile Gln
<210> 48 <211> 1269 <212> DNA
<213> Pavlova lutheri <400> 48
atgggcaagg gtggagacgg cggcgcgcag gcggtgagcg ggaccgacgc gtctctcgct gaggtgagct ccgtcgatag caagagcgtg cacgtcgtgc tctacggcaa gcgcgtggat 120 gtcacaaagt tccagaaggc acacccgggc gggagcaagg tgttccgcat cttccaggag 180 cgcgacgcga cggagcagtt cgagtcttac cactcgccca aggccatcaa gatgatggag 240 ggcatgctca agaagtcgga ggatgcgccc gcttccgtgc ccctgccctc gcggtccacc 300 atgggcacgg agttcaagga gatgattgag cgccacaaga gggctggtct ctacgaccct 360 tgcccgttgg acgagctgtt caagctcacc atcgtccttg cgcccatctt cgtgggcgcc 420 tatctcgtgc ggagcggcgt ctcgcccctc gcgggcgcgc tctccatggg ctttggcttc 480 tacctcgacg gctggcttgc tcacgactac ctgcatcacg cagtcttcaa gggctcggtc 540 aacacgctcg tcaaggcgaa caacgccatg ggatacgccc tcggcttcct ccagggctac 600 gacgtggcct ggtggcgcgc gcgccataac acgcaccacg tgtgcaccaa cgaggatggt 660 tcggacccgg acatcaagac ggcgcccctg ctcatctacg tgcgagagaa cccgtccatt 720 gccaagcggc tcaacttctt ccagcgctgg cagcagtact actatgtgcc gaccatggcc 780 atcctcgacc tctactggcg cctggagtcc atcgcgtacg tggctgtgcg cctgcctaag 840 atgtggatgc aggccgccgc tcttgccgct cactacgcgc tcctgtgctg ggtcttcgca 900 gcgcatctca acctcatccc tctcatgatg gttgcacgcg gcttcgcgac gggcatcgtt 960 gtctttgcaa cccactatgg tgaggacatc ctcgaccgcg agcacgtcga gggcatgacg 1020 ctcgtcgagc agaccgccaa gacctcccgt aacatcacgg gcggctggct agtgaacgtg 1080 ctcacgggct tcatctccct gcagaccgag catcacctct tccccatgat gcccaccggc 1140 aacctaatga ctatccagcc cgaggtacgc gacttcttca agaagcatgg cctcgagtac 1200 cgcgagggca acctcttcca gtgcgtgcac cagaacatca aggctctcgc cttcgagcac 1260 ctcctccac 1269
<210> 49 <211> 423 <212> PRT
<213> Pavlova Iutheri <400> 49
Met Gly Lys Gly Gly Asp Gly Gly Ala Gln Ala Val Ser Gly Thr Asp 15 10 15
Ala Ser Leu Ala Glu Val Ser Ser Val Asp Ser Lys Ser Val His Val 20 25 30
Val Leu Tyr Gly Lys Arg Val Asp Val Thr Lys Phe Gln Lys Ala His 35 40 45 Pro Gly Gly Ser Lys Val Phe Arg Ile Phe Gln Glu Arg Asp Ala Thr 50 55 60
Glu Gln Phe Glu Ser Tyr His Ser Pro Lys Ala Ile Lys Met Met Glu 65 70 75 80
Gly Met Leu Lys Lys Ser Glu Asp Ala Pro Ala Ser Val Pro Leu Pro 85 90 95
Ser Arg Ser Thr Met Gly Thr Glu Phe Lys Glu Met Ile Glu Arg His 100 105 110
Lys Arg Ala Gly Leu Tyr Asp Pro Cys Pro Leu Asp Glu Leu Phe Lys 115 120 125
Leu Thr Ile Val Leu Ala Pro Ile Phe Val Gly Ala Tyr Leu Val Arg 130 135 140
Ser Gly Val Ser Pro Leu Ala Gly Ala Leu Ser Met Gly Phe Gly Phe 145 150 155 160
Tyr Leu Asp Gly Trp Leu Ala His Asp Tyr Leu His His Ala Val Phe 165 170 175
Lys Gly Ser Val Asn Thr Leu Val Lys Ala Asn Asn Ala Met Gly Tyr 180 185 190
Ala Leu Gly Phe Leu Gln Gly Tyr Asp Val Ala Trp Trp Arg Ala Arg 195 200 205
His Asn Thr His His Val Cys Thr Asn Glu Asp Gly Ser Asp Pro Asp 210 215 220
Ile Lys Thr Ala Pro Leu Leu Ile Tyr Val Arg Glu Asn Pro Ser Ile 225 230 235 240
Ala Lys Arg Leu Asn Phe Phe Gln Arg Trp Gln Gln Tyr Tyr Tyr Val 245 250 255
Pro Thr Met Ala Ile Leu Asp Leu Tyr Trp Arg Leu Glu Ser Ile Ala 260 265 270
Tyr Val Ala Val Arg Leu Pro Lys Met Trp Met Gln Ala Ala Ala Leu 275 280 285
Ala Ala His Tyr Ala Leu Leu Cys Trp Val Phe Ala Ala His Leu Asn 290 295 300 Leu Ile Pro Leu Met Met Val Ala Arg Gly Phe Ala Thr Gly Ile Val 305 310 315 320
Val Phe Ala Thr His Tyr Gly Glu Asp Ile Leu Asp Arg Glu His Val 325 330 335
Glu Gly Met Thr Leu Val Glu Gln Thr Ala Lys Thr Ser Arg Asn Ile 340 345 350
Thr Gly Gly Trp Leu Val Asn Val Leu Thr Gly Phe Ile Ser Leu Gln 355 360 365
Thr Glu His His Leu Phe Pro Met Met Pro Thr Gly Asn Leu Met Thr 370 375 380
Ile Gln Pro Glu Val Arg Asp Phe Phe Lys Lys His Gly Leu Glu Tyr 385 390 395 400
Arg Glu Gly Asn Leu Phe Gln Cys Val His Gln Asn Ile Lys Ala Leu 405 410 415
Ala Phe Glu His Leu Leu His 420
<210> 50
<211> 1260
<212> DNA
<213> Tetruetreptia pomquetensis CCMP1491
<400> 50
atgtctccta agcggcaagc tctgccaatc acaattgatg gcgcaactta tgatgtgtct 60 gcttgggtca atcaccaccc tggaggagct gacattatcg agaactatcg caaccgcgat 120 gcgaccgatg tcttcatggt gatgcactct caagaagccg tcgccaagtt gaagagaatg 180 cctgttatgg agccttcctc tcctgacaca cctgttgcac ccaagcctaa gcgtgatgag 240 ccccaggagg atttccgcaa gttgcgggag gaattcatct ccaagggtat gttcgagacg 300 agtttccttt ggtattttta caagacttca actaccgtcg gtttgatggt cctttccatc 360 ttgatgaccg tgtacacgaa ttggtatttc accgctgctt tggttcttgg cgtgtgctac 420 caacagctag gctggttgtc ccacgactat tgccatcacc aggttttcac aaaccgcaag 480 attaacgacg ctttcggtct ctttttcggt aacgtgatgc agggatactc acagacttgg 540 tggaaggata ggcacaatgg tcaccatgcc gccaccaatg tggtcggcca tgacccagat 600 attgataacc tccccatcct ggcttggtct cccgaagatg tcaagagggc tactccttcg 660 actcggaatc tcatcaagta ccagcagtac tacttcattc ccaccattgc atcccttagg 720
ttcatctggt gcctccaatc catcggcggc gtcatgtcct acaagagcga ggagaggaac 780
ctgtactaca agcgccagta cactaaggag gcgattggtc tggccctcca ctgggtgctc 840
aaggccactt tctattgcag tgccatgcct agctttgcca ccggtttggg atgcttcttg 900
atctccgagc tgctcggagg atttggcatt gccatcgttg tgtttctgaa tcactatcct 960
ttggacaagg ttgaggagac tgtctgggat gagcacgggt tcagcgccag ccagatccac 1020
gagacgttga acattaagcc cggccttctc accgattggg tctttggtgg tctcaactac 1080
cagattgagc accacttgtg gcccaacatg cccaggcaca acctcacggc agcttccctg 1140
gaggtgcaga agttgtgcgc caagcacaac ctgccctaca gggccccagc catcatcccc 1200
ggggttcaga aattggtcag cttcttaggc gagattgccc agctggctgc tgtccctgaa 1260
<210> 51 <211> 420 <212> PRT
<213> Tetruetreptia pomquetensis CCMP1491 <400> 51
Met Ser Pro Lys Arg Gln Ala Leu Pro Ile Thr Ile Asp Gly Ala Thr 1 5 10 15
Tyr Asp Val Ser Ala Trp Val Asn His His Pro Gly Gly Ala Asp Ile 20 25 30
Ile Glu Asn Tyr Arg Asn Arg Asp Ala Thr Asp Ala Phe Met Val Met 35 40 45
His Ser Gln Glu Ala Val Ala Lys Leu Lys Arg Met Pro Val Met Glu 50 55 60
Pro Ser Ser Pro Asp Thr Pro Val Ala Pro Lys Pro Lys Arg Asp Glu 65 70 75 80
Pro Gln Glu Asp Phe Arg Lys Leu Arg Glu Glu Phe Ile Ser Lys Gly 85 90 95
Met Phe Glu Thr Ser Phe Leu Trp Tyr Phe Tyr Lys Thr Ser Thr Thr 100 105 110
Val Gly Leu Met Val Leu Ser Ile Leu Met Thr Val Tyr Thr Asn Trp 115 120 125
Tyr Phe Thr Ala Ala Leu Val Leu Gly Val Cys Tyr Gln Gln Leu Gly 130 135 140 Trp Leu Ser His Asp Tyr Cys His His Gln Val Phe Thr Asn Arg Lys 145 150 155 160
Ile Asn Asp Ala Phe Gly Leu Phe Phe Gly Asn Val Met Gln Gly Tyr 165 170 175
Ser Gln Thr Trp Trp Lys Asp Arg His Asn Gly His His Ala Ala Thr 180 185 190
Asn Val Val Gly His Asp Pro Asp Ile Asp Asn Leu Pro Ile Leu Ala 195 200 205
Trp Ser Pro Glu Asp Val Lys Arg Ala Thr Pro Ser Thr Arg Asn Leu 210 215 220
Ile Lys Tyr Gln Gln Tyr Tyr Phe Ile Pro Thr Ile Ala Ser Leu Arg 225 230 235 240
Phe Ile Trp Cys Leu Gln Ser Ile Gly Gly Val Met Ser Tyr Lys Ser 245 250 255
Glu Glu Arg Asn Leu Tyr Tyr Lys Arg Arg Tyr Thr Lys Glu Ala Ile 260 265 270
Gly Leu Ala Leu Pro Trp Val Leu Lys Ala Thr Phe Tyr Cys Ser Ala 275 280 285
Met Pro Ser Phe Ala Thr Gly Leu Gly Cys Phe Leu Ile Ser Glu Leu 290 295 300
Leu Gly Gly Phe Gly Ile Ala Ile Val Val Phe Leu Asn His Tyr Pro 305 310 315 320
Leu Asp Lys Val Glu Glu Thr Val Trp Asp Glu His Gly Phe Ser Ala 325 330 335
Ser Gln Ile His Glu Thr Leu Asn Ile Lys Pro Gly Leu Leu Thr Asp 340 345 350
Trp Val Phe Gly Gly Leu Asn Tyr Gln Ile Glu His His Leu Trp Pro 355 360 365
Asn Met Pro Arg His Asn Leu Thr Ala Ala Ser Leu Glu Val Gln Lys 370 375 380 Leu Cys Ala Lys His Asn Leu Pro Tyr Arg Ala Pro Ala Ile Ile Pro 385 390 395 400
Gly Val Gln Lys Leu Val Ser Phe Leu Gly Glu Ile Ala Gln Leu Ala 405 410 415
Ala Val Pro Glu 420
<210> 52 <211> 1341 <212> DNA
<213> Mortierella alpina <400> 52
atgggaacgg accaaggaaa aaccttcacc tgggaagagc tggcggccca taacaccaag 60 gacgacctac tcttggccat ccgcggcagg gtgtacgatg tcacaaagtt cttgagccgc 120 catcctggtg gagtggacac tctcctgctc ggagctggcc gagatgttac tccggtcttt 180 gagatgtatc acgcgtttgg ggctgcagat gccattatga agaagtacta tgtcggtaca 240 ctggtctcga atgagctgcc catcttcccg gagccaacgg tgttccacaa aaccatcaag 300 acgagagtcg agggctactt tacggatcgg aacattgatc ccaagaatag accagagatc 360 tggggacgat acgctcttat ctttggatcc ttgatcgctt cctactacgc gcagctcttt 420 gtgcctttcg ttgtcgaacg cacatggctt caggtggtgt ttgcaatcat catgggattt 480 gcgtgcgcac aagtcggact caaccctctt catgatgcgt ctcacttttc agtgacccac 540 aaccccactg tctggaagat tctgggagcc acgcacgact ttttcaacgg agcatcgtac 600 ctggtgtgga tgtaccaaca tatgctcggc catcacccct acaccaacat tgctggagca 660 gatcccgacg tgtcgacgtc tgagcccgat gttcgtcgta tcaagcccaa ccaaaagtgg 720 tttgtcaacc acatcaacca gcacatgttt gttcctttcc tgtacggact gctggcgttc 780 aaggtgcgca ttcaggacat caacattttg tactttgtca agaccaatga cgctattcgt 840 gtcaatccca tctcgacatg gcacactgtg atgttctggg gcggcaaggc tttctttgtc 900 tggtatcgcc tgattgttcc cctgcagtat ctgcccctgg gcaaggtgct gctcttgttc 960 acggtcgcgg acatggtgtc gtcttactgg ctggcgctga ccttccaggc gaaccacgtt 1020 gttgaggaag ttcagtggcc gttgcctgac gagaacggga tcatccaaaa ggactgggca 1080 gctatgcagg tcgagactac gcaggattac gcacacgatt cgcacctctg gaccagcatc 1140 actggcagct tgaactacca ggctgtgcac catctgttcc ccaacgtgtc gcagcaccat 1200 tatcccgata ttctggccat catcaagaac acctgcagcg agtacaaggt tccatacctt 1260 gtcaaggata cgttttggca agcatttgct tcacatttgg agcacttgcg tgttcttgga 1320 ctccgtccca aggaagagta g 1341
<210> 53 <211> 446 <212> PRT
<213> Mortierella alpina <400> 53
Met Gly Thr Asp Gln Gly Lys Thr Phe Thr Trp Glu Glu Leu Ala Ala 1 5 10 15
His Asn Thr Lys Asp Asp Leu Leu Leu Ala Ile Arg Gly Arg Val Tyr 20 25 30
Asp Val Thr Lys Phe Leu Ser Arg His Pro Gly Gly Val Asp Thr Leu 35 40 45
Leu Leu Gly Ala Gly Arg Asp Val Thr Pro Val Phe Glu Met Tyr His 50 55 60
Ala Phe Gly Ala Ala Asp Ala Ile Met Lys Lys Tyr Tyr Val Gly Thr 65 70 75 80
Leu Val Ser Asn Glu Leu Pro Ile Phe Pro Glu Pro Thr Val Phe His 85 90 95
Lys Thr Ile Lys Thr Arg Val Glu Gly Tyr Phe Thr Asp Arg Asn Ile 100 105 110
Asp Pro Lys Asn Arg Pro Glu Ile Trp Gly Arg Tyr Ala Leu Ile Phe 115 120 125
Gly Ser Leu Ile Ala Ser Tyr Tyr Ala Gln Leu Phe Val Pro Phe Val 130 135 140
Val Glu Arg Thr Trp Leu Gln Val Val Phe Ala Ile Ile Met Gly Phe 145 150 155 160
Ala Cys Ala Gln Val Gly Leu Asn Pro Leu His Asp Ala Ser His Phe 165 170 175
Ser Val Thr His Asn Pro Thr Val Trp Lys Ile Leu Gly Ala Thr His 180 185 190
Asp Phe Phe Asn Gly Ala Ser Tyr Leu Val Trp Met Tyr Gln His Met 195 200 205 Leu Gly His His Pro Tyr Thr Asn Ile Ala Gly Ala Asp Pro Asp Val 210 215 220
Ser Thr Ser Glu Pro Asp Val Arg Arg Ile Lys Pro Asn Gln Lys Trp 225 230 235 240
Phe Val Asn His Ile Asn Gln His Met Phe Val Pro Phe Leu Tyr Gly 245 250 255
Leu Leu Ala Phe Lys Val Arg Ile Gln Asp Ile Asn Ile Leu Tyr Phe 260 265 270
Val Lys Thr Asn Asp Ala Ile Arg Val Asn Pro Ile Ser Thr Trp His 275 280 285
Thr Val Met Phe Trp Gly Gly Lys Ala Phe Phe Val Trp Tyr Arg Leu 290 295 300
Ile Val Pro Leu Gln Tyr Leu Pro Leu Gly Lys Val Leu Leu Leu Phe 305 310 315 320
Thr Val Ala Asp Met Val Ser Ser Tyr Trp Leu Ala Leu Thr Phe Gln 325 330 335
Ala Asn His Val Val Glu Glu Val Gln Trp Pro Leu Pro Asp Glu Asn 340 345 350
Gly Ile Ile Gln Lys Asp Trp Ala Ala Met Gln Val Glu Thr Thr Gln 355 360 365
Asp Tyr Ala His Asp Ser His Leu Trp Thr Ser Ile Thr Gly Ser Leu 370 375 380
Asn Tyr Gln Ala Val His His Leu Phe Pro Asn Val Ser Gln His His 385 390 395 400
Tyr Pro Asp Ile Leu Ala Ile Ile Lys Asn Thr Cys Ser Glu Tyr Lys 405 410 415
Val Pro Tyr Leu Val Lys Asp Thr Phe Trp Gln Ala Phe Ala Ser His 420 425 430
Leu Glu His Leu Arg Val Leu Gly Leu Arg Pro Lys Glu Glu 435 440 445
<210> 54 <211> 1350 <212> DNA
<213> Euglena gracilis <400> 54
atggctctca gtcttaccac agaacagctg ttagaacgcc ctgatttggt tgcgattgat 60 ggcatcctct acgaccttga agggcttgcc aaagttcatc caggaggaga tttgattctc 120 gcttctggtg cctctgatgc ctcccctctc ttttattcaa tgcatccata cgtcaaaccg 180 gagaattcca aattgcttca acagttcgtc cgagggaagc atgaccgcac ctcgaaggac 240
attgtctaca cgtatgattc tcccttcgca caagacgtta agcggacaat gcgcgaggtg 300
atgaaaggga ggaactggta cgcaacccct ggcttctggc tgcgcaccgt tgggatcatc 360
gccgtgacgg ccttttgcga gtggcactgg gctaccacgg ggatggtgct gtggggcctg 420
ttgactggat tcatgcacat gcagatcggc ttatccatcc agcatgatgc gtcccacggg 480
gccatcagca agaagccttg ggtcaacgcc ctcttcgcct acggcattga cgtcatcgga 540
tcgtcccggt ggatttggct gcagtcgcac atcatgcggc accacaccta caccaaccag 600
cacggcctcg acctggatgc ggagtcggca gagccgttcc tggtgttcca caactacccc 660
gccgcaaaca ccgcccgaaa gtggttccac cgcttccaag cttggtacat gtaccttgtg 720
ctgggggcat acggggtatc gctggtgtac aacccgctct acattttccg gatgcagcac 780
aatgacacca tcccagagtc tgtcacggcc atgcgggaga atggctttct gcggcgctac 840
cgcacacttg cattcgtgat gcgagctttc ttcatcttcc ggaccgcatt cttgccctgg 900
tacctcactg ggacctcatt gctgatcacc attcctctgg tgcccactgc aactggtgcc 960
ttcttgacgt tcttcttcat tttgtcccac aattttgatg gctccgaacg gatccccgac 1020
aagaactgca aggttaagag ctctgagaag gacgttgagg ctgaccaaat tgactggtat 1080
cgggcgcagg tggagacgtc ctccacatac ggtggcccca tcgccatgtt cttcactggc 1140
ggtctcaatt tccagatcga gcaccacctc tttccccgga tgtcgtcttg gcactacccc 1200
ttcgtccagc aggcggtccg ggagtgttgc gaacgccatg gagtgcgata tgttttctac 1260
cctaccatcg tcggcaacat catctccacc ctgaagtaca tgcataaggt gggtgtcgtc 1320
cactgcgtga aggacgcaca ggattcctga 1350
<210> 55 <211> 449 <212> PRT
<213> Euglena gracilis <400> 55
Met Ala Leu Ser Leu Thr Thr Glu Gln Leu Leu Glu Arg Pro Asp Leu 1 5 10 15 Val Ala Ile Asp Gly Ile Leu Tyr Asp Leu Glu Gly Leu Ala Lys Val 20 25 30
His Pro Gly Gly Asp Leu Ile Leu Ala Ser Gly Ala Ser Asp Ala Ser 35 40 45
Pro Leu Phe Tyr Ser Met His Pro Tyr Val Lys Pro Glu Asn Ser Lys 50 55 60
Leu Leu Gln Gln Phe Val Arg Gly Lys His Asp Arg Thr Ser Lys Asp 65 70 75 80
Ile Val Tyr Thr Tyr Asp Ser Pro Phe Ala Gln Asp Val Lys Arg Thr 85 90 95
Met Arg Glu Val Met Lys Gly Arg Asn Trp Tyr Ala Thr Pro Gly Phe 100 105 110
Trp Leu Arg Thr Val Gly Ile Ile Ala Val Thr Ala Phe Cys Glu Trp 115 120 125
His Trp Ala Thr Thr Gly Met Val Leu Trp Gly Leu Leu Thr Gly Phe 130 135 140
Met His Met Gln Ile Gly Leu Ser Ile Gln His Asp Ala Ser His Gly 145 150 155 160
Ala Ile Ser Lys Lys Pro Trp Val Asn Ala Leu Phe Ala Tyr Gly Ile 165 170 175
Asp Val Ile Gly Ser Ser Arg Trp Ile Trp Leu Gln Ser His Ile Met 180 185 190
Arg His His Thr Tyr Thr Asn Gln His Gly Leu Asp Leu Asp Ala Glu 195 200 205
Ser Ala Glu Pro Phe Leu Val Phe His Asn Tyr Pro Ala Ala Asn Thr 210 215 220
Ala Arg Lys Trp Phe His Arg Phe Gln Ala Trp Tyr Met Tyr Leu Val 225 230 235 240
Leu Gly Ala Tyr Gly Val Ser Leu Val Tyr Asn Pro Leu Tyr Ile Phe 245 250 255
Arg Met Gln His Asn Asp Thr Ile Pro Glu Ser Val Thr Ala Met Arg 260 265 270 Glu Asn Gly Phe Leu Arg Arg Tyr Arg Thr Leu Ala Phe Val Met Arg 275 280 285
Ala Phe Phe Ile Phe Arg Thr Ala Phe Leu Pro Trp Tyr Leu Thr Gly 290 295 300
Thr Ser Leu Leu Ile Thr Ile Pro Leu Val Pro Thr Ala Thr Gly Ala 305 310 315 320
Phe Leu Thr Phe Phe Phe Ile Leu Ser His Asn Phe Asp Gly Ser Glu 325 330 335
Arg Ile Pro Asp Lys Asn Cys Lys Val Lys Ser Ser Glu Lys Asp Val 340 345 350
Glu Ala Asp Gln Ile Asp Trp Tyr Arg Ala Gln Val Glu Thr Ser Ser 355 360 365
Thr Tyr Gly Gly Pro Ile Ala Met Phe Phe Thr Gly Gly Leu Asn Phe 370 375 380
Gln Ile Glu His His Leu Phe Pro Arg Met Ser Ser Trp His Tyr Pro 385 390 395 400
Phe Val Gln Gln Ala Val Arg Glu Cys Cys Glu Arg His Gly Val Arg 405 410 415
Tyr Val Phe Tyr Pro Thr Ile Val Gly Asn Ile Ile Ser Thr Leu Lys 420 425 430
Tyr Met His Lys Val Gly Val Val His Cys Val Lys Asp Ala Gln Asp 435 440 445
Ser
<210> 56 <211> 1077 <212> DNA
<213> Saprolegnia diclina <400> 56
atgactgagg ataagacgaa ggtcgagttc ccgacgctca cggagctcaa gcactcgatc 60
ccgaacgcgt gctttgagtc gaacctcggc ctctcgctct actacacggc ccgcgcgatc 120 ttcaacgcgt cggcctcggc ggcgctgctc tacgcggcgc gctcgacgcc gttcattgcc 180 gataacgttc tgctccacgc gctcgtttgc gccacctaca tctacgtgca gggcgtcatc 240
ttctggggct tcttcacggt cggccacgac tgcggccact cggccttctc gcgctaccac 300
agcgtcaact ttatcatcgg ctgcatcatg cactctgcga ttttgacgcc gttcgagagc 360
tggcgcgtga cgcaccgcca ccaccacaag aacacgggca acattgataa ggacgagatc 420
ttttacccgc accggtcggt caaggacctc caggacgtgc gccaatgggt ctacacgctc 480
ggcggtgcgt ggtttgtcta cttgaaggtc gggtatgccc cgcgcacgat gagccacttt 540
gacccgtggg acccgctcct ccttcgccgc gcgtcggccg tcatcgtgtc gctcggcgtc 600
tgggccgcct tcttcgccgc gtacgcgtac ctcacatact cgctcggctt tgccgtcatg 660
ggcctctact actatgcgcc gctctttgtc tttgcttcgt tcctcgtcat tacgaccttc 720
ttgcaccaca acgacgaagc gacgccgtgg tacggcgact cggagtggac gtacgtcaag 780
ggcaacctct cgagcgtcga ccgctcgtac ggcgcgttcg tggacaacct gagccaccac 840
attggcacgc accaggtcca ccacttgttc ccgatcattc cgcactacaa gctcaacgaa 900
gccaccaagc actttgcggc cgcgtacccg cacctcgtgc gcaggaacga cgagcccatc 960
atcacggcct tcttcaagac cgcgcacctc tttgtcaact acggcgctgt gcccgagacg 1020
gcgcagatct tcacgctcaa agagtcggcc gcggccgcca aggccaagtc ggactaa 1077
<210> 57 <211> 358 <212> PRT
<213> Saprolegnia diclina <400> 57
Met Thr Glu Asp Lys Thr Lys Val Glu Phe Pro Thr Leu Thr Glu Leu 15 10 15
Lys His Ser Ile Pro Asn Ala Cys Phe Glu Ser Asn Leu Gly Leu Ser 20 25 30
Leu Tyr Tyr Thr Ala Arg Ala Ile Phe Asn Ala Ser Ala Ser Ala Ala 35 40 45
Leu Leu Tyr Ala Ala Arg Ser Thr Pro Phe Ile Ala Asp Asn Val Leu 50 55 60
Leu His Ala Leu Val Cys Ala Thr Tyr Ile Tyr Val Gln Gly Val Ile 65 70 75 80
Phe Trp Gly Phe Phe Thr Val Gly His Asp Cys Gly His Ser Ala Phe 85 90 95 Ser Arg Tyr His Ser Val Asn Phe Ile Ile Gly Cys Ile Met His Ser 100 105 110
Ala Ile Leu Thr Pro Phe Glu Ser Trp Arg Val Thr His Arg His His 115 120 125
His Lys Asn Thr Gly Asn Ile Asp Lys Asp Glu Ile Phe Tyr Pro His 130 135 140
Arg Ser Val Lys Asp Leu Gln Asp Val Arg Gln Trp Val Tyr Thr Leu 145 150 155 160
Gly Gly Ala Trp Phe Val Tyr Leu Lys Val Gly Tyr Ala Pro Arg Thr 165 170 175
Met Ser His Phe Asp Pro Trp Asp Pro Leu Leu Leu Arg Arg Ala Ser 180 185 190
Ala Val Ile Val Ser Leu Gly Val Trp Ala Ala Phe Phe Ala Ala Tyr 195 200 205
Ala Tyr Leu Thr Tyr Ser Leu Gly Phe Ala Val Met Gly Leu Tyr Tyr 210 215 220
Tyr Ala Pro Leu Phe Val Phe Ala Ser Phe Leu Val lie Thr Thr Phe 225 230 235 240
Leu His His Asn Asp Glu Ala Thr Pro Trp Tyr Gly Asp Ser Glu Trp 245 250 255
Thr Tyr Val Lys Gly Asn Leu Ser Ser Val Asp Arg Ser Tyr Gly Ala 260 265 270
Phe Val Asp Asn Leu Ser His His Ile Gly Thr His Gln Val His His 275 280 285
Leu Phe Pro Ile Ile Pro His Tyr Lys Leu Asn Glu Ala Thr Lys His 290 295 300
Phe Ala Ala Ala Tyr Pro His Leu Val Arg Arg Asn Asp Glu Pro Ile 305 310 315 320
Ile Thr Ala Phe Phe Lys Thr Ala His Leu Phe Val Asn Tyr Gly Ala 325 330 335
Val Pro Glu Thr Ala Gln Ile Phe Thr Leu Lys Glu Ser Ala Ala Ala 340 345 350 Ala Lys Ala Lys Ser Asp 355
<210> 58 <211> 1209 <212> DNA
<213> Fusarium moniliforme <400> 58
atggcgactc gacagcgaac tgccaccact gttgtggtcg aggaccttcc caaggtcact 60
cttgaggcca agtctgaacc tgtgttcccc gatatcaaga ccatcaagga tgccattccc 120
gcgcactgct tccagccctc gctcgtcacc tcattctact acgtcttccg cgattttgcc 180
atggtctctg ccctcgtctg ggctgctctc acctacatcc ccagcatccc cgaccagacc 240
ctccgcgtcg cagcttggat ggtctacggc ttcgtccagg gtctgttctg caccggtgtc 300
tggattctcg gccatgagtg cggccacggt gctttctctc tccacggaaa ggtcaacaat 360
gtgaccggct ggttcctcca ctcgttcctc ctcgtcccct acttcagctg gaagtactct 420
caccaccgcc accaccgctt caccggccac atggatctcg acatggcttt cgtccccaag 480
actgagccca agccctccaa gtcgctcatg attgctggca ttgacgtcgc cgagcttgtt 540
gaggacaccc ccgctgctca gatggtcaag ctcatcttcc accagctttt cggatggcag 600
gcgtacctct tcttcaacgc tagctctggc aagggcagca agcagtggga gcccaagact 660
ggcctctcca agtggttccg agtcagtcac ttcgagccta ccagcgctgt cttccgcccc 720
aacgaggcca tcttcatcct catctccgat atcggtcttg ctctaatggg aactgctctg 780
tactttgctt ccaagcaagt tggtgtttcg accattctct tcctctacct tgttccctac 840
ctgtgggttc accactggct cgttgccatt acctacctcc accaccacca caccgagctc 900
cctcactaca ccgctgaggg ctggacctac gtcaagggag ctctcgccac tgtcgaccgt 960
gagtttggct tcatcggaaa gcacctcttc cacggtatca ttgagaagca cgttgttcac 1020
catctcttcc ctaagatccc cttctacaag gctgacgagg ccaccgaggc catcaagccc 1080
gtcattggcg accactactg ccacgacgac cgaagcttcc tgggccagct gtggaccatc 1140
ttcggcacgc tcaagtacgt cgagcacgac cctgcccgac ccggtgccat gcgatggaac 1200
aaggactag 1209
<210> 59
<211> 402
<212> PRT
<213> Fusarium monili forme
<400> 59 Met Ala Thr Arg Gln Arg Thr Ala Thr Thr Val Val Val Glu Asp Leu 1 5 10 15
Pro Lys Val Thr Leu Glu Ala Lys Ser Glu Pro Val Phe Pro Asp Ile 20 25 30
Lys Thr Ile Lys Asp Ala Ile Pro Ala His Cys Phe Gln Pro Ser Leu 35 40 45
Val Thr Ser Phe Tyr Tyr Val Phe Arg Asp Phe Ala Met Val Ser Ala 50 55 60
Leu Val Trp Ala Ala Leu Thr Tyr Ile Pro Ser Ile Pro Asp Gln Thr 65 70 75 80
Leu Arg Val Ala Ala Trp Met Val Tyr Gly Phe Val Gln Gly Leu Phe 85 90 95
Cys Thr Gly Val Trp Ile Leu Gly His Glu Cys Gly His Gly Ala Phe 100 105 110
Ser Leu His Gly Lys Val Asn Asn Val Thr Gly Trp Phe Leu His Ser 115 120 125
Phe Leu Leu Val Pro Tyr Phe Ser Trp Lys Tyr Ser His His Arg His 130 135 140
His Arg Phe Thr Gly His Met Asp Leu Asp Met Ala Phe Val Pro Lys 145 150 155 160
Thr Glu Pro Lys Pro Ser Lys Ser Leu Met Ile Ala Gly Ile Asp Val 165 170 175
Ala Glu Leu Val Glu Asp Thr Pro Ala Ala Gln Met Val Lys Leu Ile 180 185 190
Phe His Gln Leu Phe Gly Trp Gln Ala Tyr Leu Phe Phe Asn Ala Ser 195 200 205
Ser Gly Lys Gly Ser Lys Gln Trp Glu Pro Lys Thr Gly Leu Ser Lys 210 215 220
Trp Phe Arg Val Ser His Phe Glu Pro Thr Ser Ala Val Phe Arg Pro 225 230 235 240
Asn Glu Ala Ile Phe Ile Leu Ile Ser Asp Ile Gly Leu Ala Leu Met 245 250 255 Gly Thr Ala Leu Tyr Phe Ala Ser Lys Gln Val Gly Val Ser Thr Ile 260 265 270
Leu Phe Leu Tyr Leu Val Pro Tyr Leu Trp Val His His Trp Leu Val 275 280 285
Ala Ile Thr Tyr Leu His His His His Thr Glu Leu Pro His Tyr Thr 290 295 300
Ala Glu Gly Trp Thr Tyr Val Lys Gly Ala Leu Ala Thr Val Asp Arg 305 310 315 320
Glu Phe Gly Phe Ile Gly Lys His Leu Phe His Gly Ile Ile Glu Lys 325 330 335
His Val Val His His Leu Phe Pro Lys Ile Pro Phe Tyr Lys Ala Asp 340 345 350
Glu Ala Thr Glu Ala Ile Lys Pro Val Ile Gly Asp His Tyr Cys His 355 360 365
Asp Asp Arg Ser Phe Leu Gly Gln Leu Trp Thr Ile Phe Gly Thr Leu 370 375 380
Lys Tyr Val Glu His Asp Pro Ala Arg Pro Gly Ala Met Arg Trp Asn 385 390 395 400
Lys Asp
<210> 60
<211> 39
<212> DNA
<213> Seqüência Artificial <220>
<223> oligonucleotideo SMART IV
<400> 60
aagcagtggt atcaacgcag agtggccatt acggccggg
<210> 61 <211> 37 <212> DNA <213> Seqüência Artificial <220> <223> adaptor primer from Invitrogen <400> 61
ggccacgcgt cgactagtac tttttttttt ttttttt 37
<210> 62
<211> 31
<212> DNA
<213> Seqüência Artificial <220>
<223> primer TpomNot-5
<4 00> 62
gcggccgcac catgtctcct aagcggcaag c 31
<210> 63
<211> 27
<212> DNA
<213> Seqüência Artificial <220>
<223> primer tpomNot-3
<4 00> 63
gcggccgctc attcagggac agcagcc 27
<210> 64
<211> 4300
<212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo pLF114-10
<4 00> 64
taatacgact cactataggg cgaattgggc ccgacgtcgc atgctcccgg ccgccatggc ggccgcggga attcgattgg cggccgcacc atgtctccta agcggcaagc tctgccaatc acaattgatg gcgcaactta tgatgtgtct gcttgggtca atcaccaccc tggaggagct gacattatcg agaactatcg caaccgcgat gcgaccgatg tcttcatggt gatgcactct caagaagccg tcgccaagtt gaagagaatg cctgttatgg agccttcctc tcctgacaca cctgttgcac ccaagcctaa gcgtgatgag ccccaggagg atttccgcaa gttgcgggag gaattcatct ccaagggtat gttcgagacg agtttccttt ggtattttta caagacttca actaccgtcg gtttgatggt cctttccatc ttgatgaccg tgtacacgaa ttggtatttc accgctgctt tggttcttgg cgtgtgctac caacagctag gctggttgtc ccacgactat tgccatcacc aggttttcac aaaccgcaag attaacgacg ctttcggtct ctttttcggt aacgtgatgc agggatactc acagacttgg tggaaggata ggcacaatgg tcaccatgcc gccaccaatg tggtcggcca tgacccagat attgataacc tccccatcct ggcttggtct cccgaagatg tcaagagggc tactccttcg actcggaatc tcatcaagta ccagcagtac
60 120 180 240 300 360 420 480 540 600 660 720 780 tacttcattc ccaccattgc atcccttagg ttcatctggt gcctccaatc catcggcggc 840
gtcatgtcct acaagagcga ggagaggaac ctgtactaca agcgccagta cactaaggag 900
gcgattggtc tggccctcca ctgggtgctc aaggccactt tctattgcag tgccatgcct 960
agctttgcca ccggtttggg atgcttcttg atctccgagc tgctcggagg atttggcatt 1020
gccatcgttg tgtttctgaa tcactatcct ttggacaagg ttgaggagac tgtctgggat 1080
gagcacgggt tcagcgccag ccagatccac gagacgttga acattaagcc cggccttctc 1140
accgattggg tctttggtgg tctcaactac cagattgagc accacttgtg gcccaacatg 1200
cccaggcaca acctcacggc agcttccctg gaggtgcaga agttgtgcgc caagcacaac 1260
ctgccctaca gggccccagc catcatcccc ggggttcaga aattggtcag cttcttaggc 1320
gagattgccc agctggctgc tgtccctgaa tgagcggccg caatcactag tgaattcgcg 1380
gccgcctgca ggtcgaccat atgggagagc tcccaacgcg ttggatgcat agcttgagta 1440
ttctatagtg tcacctaaat agcttggcgt aatcatggtc atagctgttt cctgtgtgaa 1500
attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct 1560
ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc 1620
agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg 1680
gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 1740
ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 1800
gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 1860
aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 1920
gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 1980
ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 2040
cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 2100
cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 2160
gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 2220
cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 2280
agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg 2340
ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 2400
ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 2460
gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 2520
cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 2580
attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 2640 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 2700 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 2760 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 2820 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 2880 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 2940 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 3000 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 3060 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 3120 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 3180 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 3240 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 3300 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 3360 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 3420 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 3480 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 3540 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 3600 cgcgcacatt tccccgaaaa gtgccacctg atgcggtgtg aaataccgca cagatgcgta 3660 aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa 3720 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata 3780 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac 3840 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc 3900 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa 3960 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg 4020 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg 4080 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca 4140 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt 4200 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt 4260 ttcccagtca cgacgttgta aaacgacggc cagtgaattg 4300 <210> 65 <211> 6021 <212> DNA <213> Seqüência Artificial <220>
<223> plasmideo pKR271 <400> 65
ggccgcgaat tcaatcacta gtgaattcgc ggccgcatga gccgtaaagg ttcaatacaa 60
cgagtgcttg ttttcttagg gacaagcatt gtacttatgt atgattctgt gtaaccatga 120
gtcttccacg ttgtactaat gtgaagggca aaaataaaac acagaacaag ttcgtttttc 180
tcaaataatg tgaaggtaga aaatggaacc atgcctcctc tcttgcatgt gatttaaaat 240
attagcagat ggtaccgtac gtgggcggat cccccgggct gcaggaattc actggccgtc 300
gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca 360
catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa 420
cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg 480
tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag 540
ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc 600
ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt 660
tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag 720
gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg 780
cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga 840
caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat 900
ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca 960
gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc 1020
gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca 1080
atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg 1140
caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca 1200
gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata 1260
accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag 1320
ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg 1380
gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca 1440
acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta 1500
atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct 1560
ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca 1620
gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag 1680
gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat 1740 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt 1800
taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 1860
cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 1920
gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 1980
gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 2040
agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag 2100
aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 2160
agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 2220
cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 2280
accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 2340
aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 2400
ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 2460
cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 2520
gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 2580
tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 2640
agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc 2700
aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc 2760
gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca 2820
ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa 2880
caatttcaca caggaaacag ctatgaccat gattacgcca agcttgcatg cctgcaggtc 2940
gactcgacgt acgatcccac atgcaagttt ttatttcaat cccttttcct ttgaataact 3000
gaccaagaac aacaagaaaa aaaaaaaaaa agaaaaggat cattttgaaa ggatattttt 3060
cgctcctatt caaatactgt atttttacca aaaaaactgt atttttccta cactctcaag 3120
ctttgttttt cgcttcgact ctcatgattt ccttcatatg ccaatcactc tatttataaa 3180
tggcataagg tagtgtgaac aattgcaaag cttgtcatca aaagcttgca atgtacaaat 3240
taatgttttt catgcctttc aaaattatct gcacccccta gctattaatc taacatctaa 3300
gtaaggctag tgaatttttt cgaatagtca tgcagtgcat taatttcccc gtgactattt 3360
tggctttgac tccaacactg gccccgtaca tccgtccctc attacatgaa aagaaatatt 3420
gtttatattc ttaattaaaa atattgtccc ttctaaattt tcatatagtt aattattata 3480
ttactttttt ctctattcta ttagttctat tttcaaatta ttatttatgc atatgtaaag 3540
tacattatat ttttgctata tacttaaata tttctaaatt attaaaaaaa gactgatatg 3600 aaaaatttat tctttttaaa gctatatcat tttatatata ctttttcttt tcttttcttt 3660
cattttctat tcaatttaat aagaaataaa ttttgtaaat ttttatttat caatttataa 3720
aaatatttta ctttatatgt tttttcacat ttttgttaaa caaatcatat cattatgatt 3780
gaaagagagg aaattgacag tgagtaataa gtgatgagaa aaaaatgtgt tatttcctaa 3840
aaaaaaccta aacaaacatg tatctactct ctatttcatc tatctctcat ttcatttttc 3900
tctttatctc tttctttatt tttttatcat atcatttcac attaattatt tttactctct 3960
ttattttttc tctctatccc tctcttattt ccactcatat atacactcca aaattggggc 4020
atgcctttat cactactcta tctcctccac taaatcattt aaatgaaact gaaaagcatt 4080
ggcaagtctc ctcccctcct caagtgattt ccaactcagc attggcatct aattgattca 4140
gtatatctat tgcatgtgta aaagtctttc cacaatacat aactattaat taatcttaaa 4200
taaataaagg ataaaatatt tttttttctt cataaaatta aaatatgtta ttttttgttt 4260
agatgtatat tcgaataaat ctaaatatat gataatgatt ttttatattg attaaacata 4320
taatcaatat taaatatgat atttttttat ataggttgta cacataattt tataaggata 4380
aaaaatatga taaaaataaa ttttaaatat ttttatattt acgagaaaaa aaaatatttt 4440
agccataaat aaatgaccag catattttac aaccttagta attcataaat tcctatatgt 4500
atatttgaaa ttaaaaacag ataatcgtta agggaaggaa tcctacgtca tctcttgcca 4560
tttgtttttc atgcaaacag aaagggacga aaaaccacct caccatgaat cactcttcac 4 620
accattttta ctagcaaaca agtctcaaca actgaagcca gctctctttc cgtttctttt 4680
tacaacactt tctttgaaat agtagtattt ttttttcaca tgatttatta acgtgccaaa 4740
agatgcttat tgaatagagt gcacatttgt aatgtactac taattagaac atgaaaaagc 4800
attgttctaa cacgataatc ctgtgaaggc gttaactcca aagatccaat ttcactatat 4860
aaattgtgac gaaagcaaaa tgaattcaca tagctgagag agaaaggaaa ggttaactaa 4920
gaagcaatac ttcagcggcc gcatgactga ggataagacg aaggtcgagt tcccgacgct 4980
cacggagctc aagcactcga tcccgaacgc gtgctttgag tcgaacctcg gcctctcgct 5040
ctactacacg gcccgcgcga tcttcaacgc gtcggcctcg gcggcgctgc tctacgcggc 5100
gcgctcgacg ccgttcattg ccgataacgt tctgctccac gcgctcgttt gcgccaccta 5160
catctacgtg cagggcgtca tcttctgggg cttcttcacg gtcggccacg actgcggcca 5220
ctcggccttc tcgcgctacc acagcgtcaa ctttatcatc ggctgcatca tgcactctgc 5280
gattttgacg ccgttcgaga gctggcgcgt gacgcaccgc caccaccaca agaacacggg 5340
caacattgat aaggacgaga tcttttaccc gcaccggtcg gtcaaggacc tccaggacgt 5400
gcgccaatgg gtctacacgc tcggcggtgc gtggtttgtc tacttgaagg tcgggtatgc 5460 cccgcgcacg atgagccact ttgacccgtg ggacccgctc ctccttcgcc gcgcgtcggc 5520
cgtcatcgtg tcgctcggcg tctgggccgc cttcttcgcc gcgtacgcgt acctcacata 5580
ctcgctcggc tttgccgtca tgggcctcta ctactatgcg ccgctctttg tctttgcttc 5640
gttcctcgtc attacgacct tcttgcacca caacgacgaa gcgacgccgt ggtacggcga 5700
ctcggagtgg acgtacgtca agggcaacct ctcgagcgtc gaccgctcgt acggcgcgtt 5760
cgtggacaac ctgagccacc acattggcac gcaccaggtc caccacttgt tcccgatcat 5820
tccgcactac aagctcaacg aagccaccaa gcactttgcg gccgcgtacc cgcacctcgt 5880
gcgcaggaac gacgagccca tcatcacggc cttcttcaag accgcgcacc tctttgtcaa 5940
ctacggcgct gtgcccgaga cggcgcagat cttcacgctc aaagagtcgg ccgcggccgc 6000
caaggccaag tcggactaag c 6021
<210> 66 <211> 9892 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmídeo pKR886r <400> 66
ggtcgactcg acgtacgtct agaggatccg tcgacggcgc gcccgatcat ccggatatag 60
ttcctccttt cagcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta 120
gttattgctc agcggtggca gcagccaact cagcttcctt tcgggctttg ttagcagccg 180
gatcgatcca agctgtacct cactattcct ttgccctcgg acgagtgctg gggcgtcggt 240
ttccactatc ggcgagtact tctacacagc catcggtcca gacggccgcg cttctgcggg 300
cgatttgtgt acgcccgaca gtcccggctc cggatcggac gattgcgtcg catcgaccct 360
gcgcccaagc tgcatcatcg aaattgccgt caaccaagct ctgatagagt tggtcaagac 420
caatgcggag catatacgcc cggagccgcg gcgatcctgc aagctccgga tgcctccgct 480
cgaagtagcg cgtctgctgc tccatacaag ccaaccacgg cctccagaag aagatgttgg 540
cgacctcgta ttgggaatcc ccgaacatcg cctcgctcca gtcaatgacc gctgttatgc 600
ggccattgtc cgtcaggaca ttgttggagc cgaaatccgc gtgcacgagg tgccggactt 660
cggggcagtc ctcggcccaa agcatcagct catcgagagc ctgcgcgacg gacgcactga 720
cggtgtcgtc catcacagtt tgccagtgat acacatgggg atcagcaatc gcgcatatga 780
aatcacgcca tgtagtgtat tgaccgattc cttgcggtcc gaatgggccg aacccgctcg 840
tctggctaag atcggccgca gcgatcgcat ccatagcctc cgcgaccggc tgcagaacag 900
cgggcagttc ggtttcaggc aggtcttgca acgtgacacc ctgtgcacgg cgggagatgc 960 aataggtcag gctctcgctg aattccccaa tgtcaagcac ttccggaatc gggagcgcgg 1020
ccgatgcaaa gtgccgataa acataacgat ctttgtagaa accatcggcg cagctattta 1080
cccgcaggac atatccacgc cctcctacat cgaagctgaa agcacgagat tcttcgccct 1140
ccgagagctg catcaggtcg gagacgctgt cgaacttttc gatcagaaac ttctcgacag 1200
acgtcgcggt gagttcaggc ttttccatgg gtatatctcc ttcttaaagt taaacaaaat 1260
tatttctaga gggaaaccgt tgtggtctcc ctatagtgag tcgtattaat ttcgcgggat 1320
cgagatctga tcaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 1380
attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 1440
cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 1500
gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 1560
ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 1620
agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 1680
tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 1740
ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag 1800
gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 1860
ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 1920
gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 1980
aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg 2040
aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 2100
ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 2160
gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 2220
gggattttgg tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc 2280
gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca 2340
gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt 2400
ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac 2460
catatggaca tattgtcgtt agaacgcggc tacaattaat acataacctt atgtatcata 2520
cacatacgat ttaggtgaca ctatagaacg gcgcgccaag ctgggtctag aactagaaac 2580
gtgatgccac ttgttattga agtcgattac agcatctatt ctgttttact atttataact 2640
ttgccatttc tgacttttga aaactatctc tggatttcgg tatcgctttg tgaagatcga 2700
gcaaaagaga cgttttgtgg acgcaatggt ccaaatccgt tctacatgaa caaattggtc 2760
acaatttcca ctaaaagtaa ataaatggca agttaaaaaa ggaatatgca ttttactgat 2820 tgcctaggtg agctccaaga gaagttgaat ctacacgtct accaaccgct aaaaaaagaa 2880
aaacattgat atgtaacctg attccattag cttttgactt cttcaacaga ttctctactt 2940
agatttctaa cagaaatatt attactagca catcattttc agtctcacta cagcaaaaaa 3000
tccaacggca caatacagac aacaggagat atcagactac agagatagat agatgctact 3060
gcatgtagta agttaaataa aaggaaaata aaatgtcttg ctaccaaaac tactacagac 3120
tatgatgctc accacaggcc aaatcctgca actaggacag cattatctta tatatattgt 3180
acaaaacaag catcaaggaa catttggtct aggcaatcag tacctcgttc taccatcacc 3240
ctcagttatc acatccttga aggatccatt actgggaatc atcggcaaca catgctcctg 3300
atggggcaca atgacatcaa gaaggtaggg gccaggggtg tccaacattc tctgaattgc 3360
cgctctaagc tcttccttct tcgtcactcg cgctgccggt atcccacaag catcagcaaa 3420
cttgagcatg tttgggaata tctcgctctc gctagacgga tctccaagat aggtgtgagc 3480
tctattggac ttgtagaacc tatcctccaa ctgaaccacc atacccaaat gctgattgtt 3540
caacaacaat atcttaactg ggagattctc cactcttata gtggccaact cctgaacatt 3600
catgatgaaa ctaccatccc catcaatgtc aaccacaaca gccccagggt tagcaacagc 3660
agcaccaata gccgcaggca atccaaaacc catggctcca agaccccctg aggtcaacca 3720
ctgcctcggt ctcttgtact tgtaaaactg cgcagcccac atttgatgct gcccaacccc 3780
agtactaaca atagcatctc cattagtcaa ctcatcaaga acctcgatag catgctgcgg 3840
agaaatcgcg tcctggaatg tcttgtaacc caatggaaac ttgtgtttct gcacattaat 3900
ctcttctctc caacctccaa gatcaaactt accctccact cctttctcct ccaaaatcat 3960
attaattccc ttcaaggcca acttcaaatc cgcgcaaacc gacacgtgcg cctgcttgtt 4020
cttcccaatc tcggcagaat caatatcaat gtgaacaatc ttagccctac tagcaaaagc 4080
ctcaagcttc ccagtaacac ggtcatcaaa ccttacccca aaggcaagca acaaatcact 414 0
attgtcaaca gcatagttag cataaacagt accatgcata cccagcatct gaagggaata 4200
ttcatcacca ataggaaaag ttccaagacc cattaaagtg ctagcaacgg gaataccagt 4260
gagttcaaca aagcgcctca attcagcact ggaattcaaa ctgccaccgc cgacgtagag 4320
aacgggcttt tgggcctcca tgatgagtct gacaatgtgt tccaattggg cctcggcggg 4380
gggcctgggc agcctggcga ggtaaccggg gaggttaacg ggctcgtccc aattaggcac 4440
ggcgagttgc tgctgaacgt ctttgggaat gtcgatgagg accggaccgg ggcggccgga 4500
ggtggcgacg aagaaagcct cggcgacgac gcgggggatg tcgtcgacgt cgaggatgag 4560
gtagttgtgc ttcgtgatgg atctgctcac ctccacgatc ggggtttctt ggaaggcgtc 4620
ggtgccgatc atccggcggg cgacctggcc ggtgatggcg acgactggga cgctgtccat 4 680 taaagcgtcg gcgaggccgc tcacgaggtt ggtggcgccg gggccggagg tggcaatgca 4740
gacgccgggg aggccggagg aacgcgcgta gccttcggcg gcgaagacgc cgccctgctc 4800
gtggcgcggg agcacgttgc ggatggcggc ggagcgcgtg agcgcctggt ggatctccat 4860
cgacgcaccg ccggggtacg cgaacaccgt cgtcacgccc tgcctctcca gcgcctccac 4920
aaggatgtcc gcgcccttgc gaggttcgcc ggaggcgaac cgtgacacga agggctccgt 4980
ggtcggcgct tccttggtga agggcgccgc cgtggggggt ttggagatgg aacatttgat 5040
tttgagagcg tggttgggtt tggtgagggt ttgatgagag agagggaggg tggatctagt 5100
aatgcgtttg gggaaggtgg ggtgtgaaga ggaagaagag aatcgggtgg ttctggaagc 5160
ggtggccgcc attgtgttgt gtggcatggt tatacttcaa aaactgcaca acaagcctag 5220
agttagtacc taaacagtaa atttacaaca gagagcaaag acacatgcaa aaatttcagc 5280
cataaaaaaa gttataatag aatttaaagc aaaagtttca ttttttaaac atatatacaa 5340
acaaactgga tttgaaggaa gggattaatt cccctgctca aagtttgaat tcctattgtg 5400
acctatactc gaataaaatt gaagcctaag gaatgtatga gaaacaagaa aacaaaacaa 5460
aactacagac aaacaagtac aattacaaaa ttcgctaaaa ttctgtaatc accaaacccc 5520
atctcagtca gcacaaggcc caaggtttat tttgaaataa aaaaaaagtg attttatttc 5580
tcataagcta aaagaaagaa aggcaattat gaaatgattt cgactagatc tgaaagtcca 5640
acgcgtattc cgcagatatt aaagaaagag tagagtttca catggatcct agatggaccc 5700
agttgaggaa aaagcaaggc aaagcaaacc agaagtgcaa gatccgaaat tgaaccacgg 5760
aatctaggat ttggtagagg gagaagaaaa gtaccttgag aggtagaaga gaagagaaga 5820
gcagagagat atatgaacga gtgtgtcttg gtctcaactc tgaagcgata cgagtttaga 5880
ggggagcatt gagttccaat ttatagggaa accgggtggc aggggtgagt taatgacgga 5940
aaagccccta agtaacgaga ttggattgtg ggttagattc aaccgtttgc atccgcggct 6000
tagattgggg aagtcagagt gaatctcaac cgttgactga gttgaaaatt gaatgtagca 6060
accaattgag ccaaccccag cctttgccct ttgattttga tttgtttgtt gcatactttt 6120
tatttgtctt ctggttctga ctctctttct ctcgtttcaa tgccaggttg cctactccca 6180
caccactcac aagaagattc tactgttagt attaaatatt ttttaatgta ttaaatgatg 6240
aatgcttttg taaacagaac aagactatgt ctaataagtg tcttgcaaca ttttttaaga 6300
aattaaaaaa aatatattta ttatcaaaat caaatgtatg aaaaatcatg aataatataa 6360
ttttatacat ttttttaaaa aatcttttaa tttcttaatt aatatcttaa aaataatgat 6420
taatatttaa cccaaaataa ttagtatgat tggtaaggaa gatatccatg ttatgtttgg 6480
atgtgagttt gatctagagc aaagcttact agagtcgacc tgcagcccgg gggatccgcc 6540 cacgtacggt accatctgct aatattttaa atcacatgca agagaggagg catggttcca 6600 ttttctacct tcacattatt tgagaaaaac gaacttgttc tgtgttttat ttttgccctt 6660 cacattagta caacgtggaa gactcatggt tacacagaat catacataag tacaatgctt 6720 gtccctaaga aaacaagcac tcgttgtatt gaacctttac ggctcatgcg gccgcgaatt 6780 cactagtgat tgaattcgcg gccgcttagt ccgacttggc cttggcggcc gcggccgact 6840 ctttgagcgt gaagatctgc gccgtctcgg gcacagcgcc gtagttgaca aagaggtgcg 6900 cggtcttgaa gaaggccgtg atgatgggct cgtcgttcct gcgcacgagg tgcgggtacg 6960 cggccgcaaa gtgcttggtg gcttcgttga gcttgtagtg cggaatgatc gggaacaagt 7020 ggtggacctg gtgcgtgcca atgtggtggc tcaggttgtc cacgaacgcg ccgtacgagc 7080 ggtcgacgct cgagaggttg cccttgacgt acgtccactc cgagtcgccg taccacggcg 7140 tcgcttcgtc gttgtggtgc aagaaggtcg taatgacgag gaacgaagca aagacaaaga 7200 gcggcgcata gtagtagagg cccatgacgg caaagccgag cgagtatgtg aggtacgcgt 7260 acgcggcgaa gaaggcggcc cagacgccga gcgacacgat gacggccgac gcgcggcgaa 7320 ggaggagcgg gtcccacggg tcaaagtggc tcatcgtgcg cggggcatac ccgaccttca 7380 agtagacaaa ccacgcaccg ccgagcgtgt agacccattg gcgcacgtcc tggaggtcct 7440 tgaccgaccg gtgcgggtaa aagatctcgt ccttatcaat gttgcccgtg ttcttgtggt 7500 ggtggcggtg cgtcacgcgc cagctctcga acggcgtcaa aatcgcagag tgcatgatgc 7560 agccgatgat aaagttgacg ctgtggtagc gcgagaaggc cgagtggccg cagtcgtggc 7620 cgaccgtgaa gaagccccag aagatgacgc cctgcacgta gatgtaggtg gcgcaaacga 7680 gcgcgtggag cagaacgtta tcggcaatga acggcgtcga gcgcgccgcg tagagcagcg 7740 ccgccgaggc cgacgcgttg aagatcgcgc gggccgtgta gtagagcgag aggccgaggt 7800 tcgactcaaa gcacgcgttc gggatcgagt gcttgagctc cgtgagcgtc gggaactcga 7860 ccttcgtctt atcctcagtc atgcggccgc tgaagtattg cttcttagtt aacctttcct 7920 ttctctctca gctatgtgaa ttcattttgc tttcgtcaca atttatatag tgaaattgga 7980 tctttggagt taacgccttc acaggattat cgtgttagaa caatgctttt tcatgttcta 8040 attagtagta cattacaaat gtgcactcta ttcaataagc atcttttggc acgttaataa 8100 atcatgtgaa aaaaaaatac tactatttca aagaaagtgt tgtaaaaaga aacggaaaga 8160 gagctggctt cagttgttga gacttgtttg ctagtaaaaa tggtgtgaag agtgattcat 8220 ggtgaggtgg tttttcgtcc ctttctgttt gcatgaaaaa caaatggcaa gagatgacgt 8280 aggattcctt cccttaacga ttatctgttt ttaatttcaa atatacatat aggaatttat 8340 gaattactaa ggttgtaaaa tatgctggtc atttatttat ggctaaaata tttttttttc 8400 tcgtaaatat aaaaatattt aaaatttatt tttatcatat tttttatcct tataaaatta 8460
tgtgtacaac ctatataaaa aaatatcata tttaatattg attatatgtt taatcaatat 8520
aaaaaatcat tatcatatat ttagatttat tcgaatatac atctaaacaa aaaataacat 8580
attttaattt tatgaagaaa aaaaaatatt ttatccttta tttatttaag attaattaat 8640
agttatgtat tgtggaaaga cttttacaca tgcaatagat atactgaatc aattagatgc 8700
caatgctgag ttggaaatca cttgaggagg ggaggagact tgccaatgct tttcagtttc 8760
atttaaatga tttagtggag gagatagagt agtgataaag gcatgcccca attttggagt 8820
gtatatatga gtggaaataa gagagggata gagagaaaaa ataaagagag taaaaataat 8880
taatgtgaaa tgatatgata aaaaaataaa gaaagagata aagagaaaaa tgaaatgaga 8940
gatagatgaa atagagagta gatacatgtt tgtttaggtt ttttttagga aataacacat 9000
ttttttctca tcacttatta ctcactgtca atttcctctc tttcaatcat aatgatatga 9060
tttgtttaac aaaaatgtga aaaaacatat aaagtaaaat atttttataa attgataaat 9120
aaaaatttac aaaatttatt tcttattaaa ttgaatagaa aatgaaagaa aagaaaagaa 9180
aaagtatata taaaatgata tagctttaaa aagaataaat ttttcatatc agtctttttt 9240
taataattta gaaatattta agtatatagc aaaaatataa tgtactttac atatgcataa 9300
ataataattt gaaaatagaa ctaatagaat agagaaaaaa gtaatataat aattaactat 9360
atgaaaattt agaagggaca atatttttaa ttaagaatat aaacaatatt tcttttcatg 9420
taatgaggga cggatgtacg gggccagtgt tggagtcaaa gccaaaatag tcacggggaa 9480
attaatgcac tgcatgacta ttcgaaaaaa ttcactagcc ttacttagat gttagattaa 9540
tagctagggg gtgcagataa ttttgaaagg catgaaaaac attaatttgt acattgcaag 9600
cttttgatga caagctttgc aattgttcac actaccttat gccatttata aatagagtga 9660
ttggcatatg aaggaaatca tgagagtcga agcgaaaaac aaagcttgag agtgtaggaa 9720
aaatacagtt tttttggtaa aaatacagta tttgaatagg agcgaaaaat atcctttcaa 9780
aatgatcctt ttcttttttt ttttttttct tgttgttctt ggtcagttat tcaaaggaaa 9840
agggattgaa ataaaaactt gcatgtggga tcgtacgtcg agtcgacctg ca 9892
<210> 67 <211> 7085 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmídeo pKR72 <400> 67
gtacggatcc gtcgacggcg cgcccgatca tccggatata gttcctcctt tcagcaaaaa 60 acccctcaag acccgtttag aggccccaag gggttatgct agttattgct cagcggtggc 120 agcagccaac tcagcttcct ttcgggcttt gttagcagcc ggatcgatcc aagctgtacc 180 tcactattcc tttgccctcg gacgagtgct ggggcgtcgg tttccactat cggcgagtac 240 ttctacacag ccatcggtcc agacggccgc gcttctgcgg gcgatttgtg tacgcccgac 300 agtcccggct ccggatcgga cgattgcgtc gcatcgaccc tgcgcccaag ctgcatcatc 360 gaaattgccg tcaaccaagc tctgatagag ttggtcaaga ccaatgcgga gcatatacgc 420 ccggagccgc ggcgatcctg caagctccgg atgcctccgc tcgaagtagc gcgtctgctg 480 ctccatacaa gccaaccacg gcctccagaa gaagatgttg gcgacctcgt attgggaatc 540 cccgaacatc gcctcgctcc agtcaatgac cgctgttatg cggccattgt ccgtcaggac 600 attgttggag ccgaaatccg cgtgcacgag gtgccggact tcggggcagt cctcggccca 660 aagcatcagc tcatcgagag cctgcgcgac ggacgcactg acggtgtcgt ccatcacagt 720 ttgccagtga tacacatggg gatcagcaat cgcgcatatg aaatcacgcc atgtagtgta 780 ttgaccgatt ccttgcggtc cgaatgggcc gaacccgctc gtctggctaa gatcggccgc 840 agcgatcgca tccatagcct ccgcgaccgg ctgcagaaca gcgggcagtt cggtttcagg 900 caggtcttgc aacgtgacac cctgtgcacg gcgggagatg caataggtca ggctctcgct 960 gaattcccca atgtcaagca cttccggaat cgggagcgcg gccgatgcaa agtgccgata 1020 aacataacga tctttgtaga aaccatcggc gcagctattt acccgcagga catatccacg 1080 ccctcctaca tcgaagctga aagcacgaga ttcttcgccc tccgagagct gcatcaggtc 1140 ggagacgctg tcgaactttt cgatcagaaa cttctcgaca gacgtcgcgg tgagttcagg 1200 cttttccatg ggtatatctc cttcttaaag ttaaacaaaa ttatttctag agggaaaccg 1260 ttgtggtctc cctatagtga gtcgtattaa tttcgcggga tcgagatcga tccaattcca 1320 atcccacaaa aatctgagct taacagcaca gttgctcctc tcagagcaga atcgggtatt 1380 caacaccctc atatcaacta ctacgttgtg tataacggtc cacatgccgg tatatacgat 1440 gactggggtt gtacaaaggc ggcaacaaac ggcgttcccg gagttgcaca caagaaattt 1500 gccactatta cagaggcaag agcagcagct gacgcgtaca caacaagtca gcaaacagac 1560 aggttgaact tcatccccaa aggagaagct caactcaagc ccaagagctt tgctaaggcc 1620 ctaacaagcc caccaaagca aaaagcccac tggctcacgc taggaaccaa aaggcccagc 1680 agtgatccag ccccaaaaga gatctccttt gccccggaga ttacaatgga cgatttcctc 1740 tatctttacg atctaggaag gaagttcgaa ggtgaaggtg acgacactat gttcaccact 1800 gataatgaga aggttagcct cttcaatttc agaaagaatg ctgacccaca gatggttaga 1860 gaggcctacg cagcaggtct catcaagacg atctacccga gtaacaatct ccaggagatc 1920 aaataccttc ccaagaaggt taaagatgca gtcaaaagat tcaggactaa ttgcatcaag 1980 aacacagaga aagacatatt tctcaagatc agaagtacta ttccagtatg gacgattcaa 2040 ggcttgcttc ataaaccaag gcaagtaata gagattggag tctctaaaaa ggtagttcct 2100 actgaatcta aggccatgca tggagtctaa gattcaaatc gaggatctaa cagaactcgc 2160 cgtgaagact ggcgaacagt tcatacagag tcttttacga ctcaatgaca agaagaaaat 2220 cttcgtcaac atggtggagc acgacactct ggtctactcc aaaaatgtca aagatacagt 2280 ctcagaagac caaagggcta ttgagacttt tcaacaaagg ataatttcgg gaaacctcct 2340 cggattccat tgcccagcta tctgtcactt catcgaaagg acagtagaaa aggaaggtgg 2400 ctcctacaaa tgccatcatt gcgataaagg aaaggctatc attcaagatg cctctgccga 2460 cagtggtccc aaagatggac ccccacccac gaggagcatc gtggaaaaag aagacgttcc 2520 aaccacgtct tcaaagcaag tggattgatg tgacatctcc actgacgtaa gggatgacgc 2580 acaatcccac tatccttcgc aagacccttc ctctatataa ggaagttcat ttcatttgga 2640 gaggacacgc tcgagctcat ttctctatta cttcagccat aacaaaagaa ctcttttctc 2700 ttcttattaa accatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat 2760 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc 2820 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg 2880 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga 2940 agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca 3000 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc 3060 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt 3120 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga 3180 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca 3240 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca 3300 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga 3360 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag 3420 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct 3480 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca 3540 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat 3600 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt 3660 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg 3720 tccgagggca aaggaatagt gaggtaccta aagaaggagt gcgtcgaagc agatcgttca 3780 aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc gatgattatc 3840 atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg catgacgtta 3900 tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata cgcgatagaa 3960 aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta 4020 gatcgatgtc gaatcgatca acctgcatta atgaatcggc caacgcgcgg ggagaggcgg 4080 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 4140 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 4200 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 4260 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 4320 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 4380 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 4440 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc 4500 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 4560 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 4620 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 4680 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc 4740 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 4800 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 4860 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 4920 acgttaaggg attttggtca tgacattaac ctataaaaat aggcgtatca cgaggccctt 4980 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac 5040 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc 5100 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag 5160 agtgcaccat atggacatat tgtcgttaga acgcggctac aattaataca taaccttatg 5220 tatcatacac atacgattta ggtgacacta tagaacggcg cgccaagctt gttgaaacat 5280 ccctgaagtg tctcatttta ttttatttat tctttgctga taaaaaaata aaataaaaga 5340 agctaagcac acggtcaacc attgctctac tgctaaaagg gttatgtgta gtgttttact 5400 gcataaatta tgcagcaaac aagacaactc aaattaaaaa atttcctttg cttgtttttt 5460 tgttgtctct gacttgactt tcttgtggaa gttggttgta taaggattgg gacaccattg 5520 tccttcttaa tttaatttta ttctttgctg ataaaaaaaa aaatttcata tagtgttaaa 5580 taataatttg ttaaataacc aaaaagtcaa atatgtttac tctcgtttaa ataattgaga 5640 ttcgtccagc aaggctaaac gattgtatag atttatgaca atatttactt ttttatagat 5700
aaatgttata ttataataaa tttatataca tatattatat gttatttatt attattttaa 5760
atccttcaat attttatcaa accaactcat aatttttttt ttatctgtaa gaagcaataa 5820
aattaaatag acccacttta aggatgatcc aacctttata cagagtaaga gagttcaaat 5880
agtacccttt catatacata tcaactaaaa tattagaaat atcatggatc aaaccttata 5940
aagacattaa ataagtggat aagtataata tataaatggg tagtatataa tatataaatg 6000
gatacaaact tctctcttta taattgttat gtctccttaa catcctaata taatacataa 6060
gtgggtaata tataatatat aaatggagac aaacttcttc cattataatt gttatgtctt 6120
cttaacactt atgtctcgtt cacaatgcta aggttagaat tgtttagaaa gtcttatagt 6180
acacatttgt ttttgtacta tttgaagcat tccataagcc gtcacgattc agatgattta 6240
taataataag aggaaattta tcatagaaca ataaggtgca tagatagagt gttaatatat 6300
cataacatcc tttgtttatt catagaagaa gtgagatgga gctcagttat tatactgtta 6360
catggtcgga tacaatattc catgctctcc atgagctctt acacctacat gcattttagt 6420
tcatacttgc ggccgcagta tatcttaaat tctttaatac ggtgtactag gatattgaac 6480
tggttcttga tgatgaaaac ctgggccgag attgcagcta tttatagtca taggtcttgt 6540
taacatgcat ggacatttgg ccacggggtg gcatgcagtt tgacgggtgt tgaaataaac 6600
aaaaatgagg tggcggaaga gaatacgagt ttgaggttgg gttagaaaca acaaatgtga 6660
gggctcatga tgggttgagt tggtgaatgt tttgggctgc tcgattgaca cctttgtgag 6720
tacgtgttgt tgtgcatggc ttttggggtc cagttttttt ttcttgacgc ggcgatcctg 6780
atcagctagt ggataagtga tgtccactgt gtgtgattgc gtttttgttt gaattttatg 6840
aacttagaca ttgctatgca aaggatactc tcattgtgtt ttgtcttctt ttgttccttg 6900
gctttttctt atgatccaag agactagtca gtgttgtggc attcgagact accaagatta 6960
attatgatgg gggaaggata agtaactgat tagtacggac tgttaccaaa ttaattaata 7020
agcggcaaat gaagggcatg gatcaaaagc ttggatctcc tgcaggatct ggccggccgg 7080
atctc 7085
<210> 68
<211> 32
<212> DNA
<213> Seqüência Artificial <220>
<223> primer oCon-1
<400> 68
aatctagacc tgcaggatcc atgcccttca tt
32 <210> 69
<211> 33
<212> DNA
<213> Seqüência Artificial <220>
<223> primer oCon-2
<400> 69
tttctagacc tgcaggttga aacatccctg aag 33
<210> 70
<211> 2635
<212> DNA
<213> Seqüência Artificial <220>
<223> cassete Kti/NotI/Kti3'Salb3' modificado
<220>
<221> misc_feature <222> (1229)..(1229) <223> η é a, c, g, ou t
<400> 70
ggtaccgggc cccccctcga ggtcgcccgg gggatccgcc ctaagcttcg tacgtcctcg 60
aagagaaggg ttaataacac attttttaac atttttaaca caaattttag ttatttaaaa 120
atttattaaa aaatttaaaa taagaagagg aactctttaa ataaatctaa cttacaaaat 180
ttatgatttt taataagttt tcaccaataa aaaatgtcat aaaaatatgt taaaaagtat 240
attatcaata ttctctttat gataaataaa aagaaaaaaa aaataaaagt taagtgaaaa 300
tgagattgaa gtgactttag gtgtgtataa atatatcaac cccgccaaca atttatttaa 360
tccaaatata ttgaagtata ttattccata gcctttattt atttatatat ttattatata 420
aaagctttat ttgttctagg ttgttcatga aatatttttt tggttttatc tccgttgtaa 480
gaaaatcatg tgctttgtgt cgccactcac tattgcagct ttttcatgca ttggtcagat 540
tgacggttga ttgtattttt gttttttatg gttttgtgtt atgacttaag tcttcatctc 600
tttatctctt catcaggttt gatggttacc taatatggtc catgggtaca tgcatggtta 660
aattaggtgg ccaactttgt tgtgaacgat agaatttttt ttatattaag taaactattt 720
ttatattatg aaataataat aaaaaaaata ttttatcatt attaacaaaa tcatattagt 780
taatttgtta actctataat aaaagaaata ctgtaacatt cacattacat ggtaacatct 840
ttccaccctt tcatttgttt tttgtttgat gacttttttt cttgtttaaa tttatttccc 900
ttcttttaaa tttggaatac attatcatca tatataaact aaaatactaa aaacaggatt 960
acacaaatga taaataataa cacaaatatt tataaatcta gctgcaatat atttaaacta 1020 gctatatcga tattgtaaaa taaaactagc tgcattgata ctgataaaaa aatatcatgt 1080
gctttctgga ctgatgatgc agtatacttt tgacattgcc tttattttat ttttcagaaa 1140
agctttctta gttctgggtt cttcattatt tgtttcccat ctccattgtg aattgaatca 1200
tttgcttcgt gtcacaaata caatttagnt aggtacatgc attggtcaga ttcacggttt 1260
attatgtcat gacttaagtt catggtagta cattacctgc cacgcatgca ttatattggt 1320
tagatttgat aggcaaattt ggttgtcaac aatataaata taaataatgt ttttatatta 1380
cgaaataaca gtgatcaaaa caaacagttt tatctttatt aacaagattt tgtttttgtt 1440
tgatgacgtt ttttaatgtt tacgctttcc cccttctttt gaatttagaa cactttatca 1500
tcataaaatc aaatactaaa aaaattacat atttcataaa taataacaca aatattttta 1560
aaaaatctga aataataatg aacaatatta catattatca cgaaaattca ttaataaaaa 1620
tattatataa ataaaatgta atagtagtta tatgtaggaa aaaagtactg cacgcataat 1680
atatacaaaa agattaaaat gaactattat aaataataac actaaattaa tggtgaatca 1740
tatcaaaata atgaaaaagt aaataaaatt tgtaattaac ttctatatgt attacacaca 1800
caaataataa ataatagtaa aaaaaattat gataaatatt taccatctca taagatattt 1860
aaaataatga taaaaatata gattattttt tatgcaacta gctagccaaa aagagaacac 1920
gggtatatat aaaaagagta cctttaaatt ctactgtact tcctttattc ctgacgtttt 1980
tatatcaagt ggacatacgt gaagatttta attatcagtc taaatatttc attagcactt 2040
aatacttttc tgttttattc ctatcctata agtagtcccg attctcccaa cattgcttat 2100
tcacacaact aactaagaaa gtcttccata gccccccaag cggccgcgac acaagtgtga 2160
gagtactaaa taaatgcttt ggttgtacga aatcattaca ctaaataaaa taatcaaagc 2220
ttatatatgc cttccgctaa ggccgaatgc aaagaaattg gttctttctc gttatctttt 2280
gccactttta ctagtacgta ttaattacta cttaatcatc tttgtttacg gctcattata 2340
tccggtctag aggatccaag gccgcgaagt taaaagcaat gttgtcactt gtcgtactaa 2400
cacatgatgt gatagtttat gctagctagc tataacataa gctgtctctg agtgtgttgt 2460
atattaataa agatcatcac tggtgaatgg tgatcgtgta cgtaccctac ttagtaggca 2520
atggaagcac ttagagtgtg ctttgtgcat ggccttgcct ctgttttgag acttttgtaa 2580
tgttttcgag tttaaatctt tgcctttgcg tacgtctaga ggatccccgg gtacc 2635
<210> 71 <211> 9240 <212> DNA
<213> Seqüência Artificial
<220> <223> plasmídeo pKR970 <220>
<221> misc_feature <222> (6592)..(6592) <223> η é a, c, g, ou t
<400> 71
gtacggatcc gtcgacggcg cgcccgatca tccggatata gttcctcctt tcagcaaaaa 60
acccctcaag acccgtttag aggccccaag gggttatgct agttattgct cagcggtggc 120
agcagccaac tcagcttcct ttcgggcttt gttagcagcc ggatcgatcc aagctgtacc 180
tcactattcc tttgccctcg gacgagtgct ggggcgtcgg tttccactat cggcgagtac 240
ttctacacag ccatcggtcc agacggccgc gcttctgcgg gcgatttgtg tacgcccgac 300
agtcccggct ccggatcgga cgattgcgtc gcatcgaccc tgcgcccaag ctgcatcatc 360
gaaattgccg tcaaccaagc tctgatagag ttggtcaaga ccaatgcgga gcatatacgc 420
ccggagccgc ggcgatcctg caagctccgg atgcctccgc tcgaagtagc gcgtctgctg 480
ctccatacaa gccaaccacg gcctccagaa gaagatgttg gcgacctcgt attgggaatc 540
cccgaacatc gcctcgctcc agtcaatgac cgctgttatg cggccattgt ccgtcaggac 600
attgttggag ccgaaatccg cgtgcacgag gtgccggact tcggggcagt cctcggccca 660
aagcatcagc tcatcgagag cctgcgcgac ggacgcactg acggtgtcgt ccatcacagt 720
ttgccagtga tacacatggg gatcagcaat cgcgcatatg aaatcacgcc atgtagtgta 780
ttgaccgatt ccttgcggtc cgaatgggcc gaacccgctc gtctggctaa gatcggccgc 840
agcgatcgca tccatagcct ccgcgaccgg ctgcagaaca gcgggcagtt cggtttcagg 900
caggtcttgc aacgtgacac cctgtgcacg gcgggagatg caataggtca ggctctcgct 960
gaattcccca atgtcaagca cttccggaat cgggagcgcg gccgatgcaa agtgccgata 1020
aacataacga tctttgtaga aaccatcggc gcagctattt acccgcagga catatccacg 1080
ccctcctaca tcgaagctga aagcacgaga ttcttcgccc tccgagagct gcatcaggtc 1140
ggagacgctg tcgaactttt cgatcagaaa cttctcgaca gacgtcgcgg tgagttcagg 1200
cttttccatg ggtatatctc cttcttaaag ttaaacaaaa ttatttctag agggaaaccg 1260
ttgtggtctc cctatagtga gtcgtattaa tttcgcggga tcgagatctg atcaacctgc 1320
attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 1380
cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 1440
caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 1500
caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 1560
ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 1620 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 1680
ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 1740
tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 1800
gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 1860
ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 1920
ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 1980
gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 2040
aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 2100
tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 2160
ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgacat 2220
taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg 2280
gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg 2340
ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc 2400
ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatggac atattgtcgt 24 60
tagaacgcgg ctacaattaa tacataacct tatgtatcat acacatacga tttaggtgac 2520
actatagaac ggcgcgccaa gcttggatct cctgcaggct agcctaagta cgtactcaaa 2580
atgccaacaa ataaaaaaaa agttgcttta ataatgccaa aacaaattaa taaaacactt 2640
acaacaccgg atttttttta attaaaatgt gccatttagg ataaatagtt aatattttta 2700
ataattattt aaaaagccgt atctactaaa atgattttta tttggttgaa aatattaata 2760
tgtttaaatc aacacaatct atcaaaatta aactaaaaaa aaaataagtg tacgtggtta 2820
acattagtac agtaatataa gaggaaaatg agaaattaag aaattgaaag cgagtctaat 2880
ttttaaatta tgaacctgca tatataaaag gaaagaaaga atccaggaag aaaagaaatg 2940
aaaccatgca tggtcccctc gtcatcacga gtttctgcca tttgcaatag aaacactgaa 3000
acacctttct ctttgtcact taattgagat gccgaagcca cctcacacca tgaacttcat 3060
gaggtgtagc acccaaggct tccatagcca tgcatactga agaatgtctc aagctcagca 3120
ccctacttct gtgacgtgtc cctcattcac cttcctctct tccctataaa taaccacgcc 3180
tcaggttctc cgcttcacaa ctcaaacatt ctctccattg gtccttaaac actcatcagt 3240
catcaccgcg gccgcatggg aacggaccaa ggaaaaacct tcacctggga agagctggcg 3300
gcccataaca ccaaggacga cctactcttg gccatccgcg gcagggtgta cgatgtcaca 3360
aagttcttga gccgccatcc tggtggagtg gacactctcc tgctcggagc tggccgagat 3420
gttactccgg tctttgagat gtatcacgcg tttggggctg cagatgccat tatgaagaag 3480 tactatgtcg gtacactggt ctcgaatgag ctgcccatct tcccggagcc aacggtgttc 3540
cacaaaacca tcaagacgag agtcgagggc tactttacgg atcggaacat tgatcccaag 3600
aatagaccag agatctgggg acgatacgct cttatctttg gatccttgat cgcttcctac 3660
tacgcgcagc tctttgtgcc tttcgttgtc gaacgcacat ggcttcaggt ggtgtttgca 3720
atcatcatgg gatttgcgtg cgcacaagtc ggactcaacc ctcttcatga tgcgtctcac 3780
ttttcagtga cccacaaccc cactgtctgg aagattctgg gagccacgca cgactttttc 3840
aacggagcat cgtacctggt gtggatgtac caacatatgc tcggccatca cccctacacc 3900
aacattgctg gagcagatcc cgacgtgtcg acgtctgagc ccgatgttcg tcgtatcaag 3960
cccaaccaaa agtggtttgt caaccacatc aaccagcaca tgtttgttcc tttcctgtac 4020
ggactgctgg cgttcaaggt gcgcattcag gacatcaaca ttttgtactt tgtcaagacc 4080
aatgacgcta ttcgtgtcaa tcccatctcg acatggcaca ctgtgatgtt ctggggcggc 4140
aaggctttct ttgtctggta tcgcctgatt gttcccctgc agtatctgcc cctgggcaag 4200
gtgctgctct tgttcacggt cgcggacatg gtgtcgtctt actggctggc gctgaccttc 4260
caggcgaacc acgttgttga ggaagttcag tggccgttgc ctgacgagaa cgggatcatc 4320
caaaaggact gggcagctat gcaggtcgag actacgcagg attacgcaca cgattcgcac 4380
ctctggacca gcatcactgg cagcttgaac taccagactg tgcaccatct gttccccaac 4440
gtgtcgcagc accattatcc cgatattctg gccatcatca agaacacctg cagcgagtac 4500
aaggttccat accttgtcaa ggatacgttt tggcaagcat ttgcttcaca tttggagcac 4560
ttgcgtgttc ttggactccg tcccaaggaa gagtaggcgg ccgcatttcg caccaaatca 4620
atgaaagtaa taatgaaaag tctgaataag aatacttagg cttagatgcc tttgttactt 4680
gtgtaaaata acttgagtca tgtacctttg gcggaaacag aataaataaa aggtgaaatt 4740
ccaatgctct atgtataagt tagtaatact taatgtgttc tacggttgtt tcaatatcat 4800
caaactctaa ttgaaacttt agaaccacaa atctcaatct tttcttaatg aaatgaaaaa 4860
tcttaattgt accatgttta tgttaaacac cttacaattg gttggagagg aggaccaacc 4920
gatgggacaa cattgggaga aagagattca atggagattt ggataggaga acaacattct 4980
ttttcacttc aatacaagat gagtgcaaca ctaaggatat gtatgagact ttcagaagct 5040
acgacaacat agatgagtga ggtggtgatt cctagcaaga aagacattag aggaagccaa 5100
aatcgaacaa ggaagacatc aagggcaaga gacaggacca tccatctcag gaaaaggagc 5160
tttgggatag tccgagaagt tgtacaagaa attttttgga gggtgagtga tgcattgctg 5220
gtgactttaa ctcaatcaaa attgagaaag aaagaaaagg gagggggctc acatgtgaat 5280
agaagggaaa cgggagaatt ttacagtttt gatctaatgg gcatcccagc tagtggtaac 5340 atattcacca tgtttaacct tcacgtaccg ggccccccct cgaggtcgcc cgggggatcc 5400
gccctaagct tcgtacgtcc tcgaagagaa gggttaataa cacatttttt aacattttta 5460
acacaaattt tagttattta aaaatttatt aaaaaattta aaataagaag aggaactctt 5520
taaataaatc taacttacaa aatttatgat ttttaataag ttttcaccaa taaaaaatgt 5580
cataaaaata tgttaaaaag tatattatca atattctctt tatgataaat aaaaagaaaa 5640
aaaaaataaa agttaagtga aaatgagatt gaagtgactt taggtgtgta taaatatatc 5700
aaccccgcca acaatttatt taatccaaat atattgaagt atattattcc atagccttta 5760
tttatttata tatttattat ataaaagctt tatttgttct aggttgttca tgaaatattt 5820
ttttggtttt atctccgttg taagaaaatc atgtgctttg tgtcgccact cactattgca 5880
gctttttcat gcattggtca gattgacggt tgattgtatt tttgtttttt atggttttgt 5940
gttatgactt aagtcttcat ctctttatct cttcatcagg tttgatggtt acctaatatg 6000
gtccatgggt acatgcatgg ttaaattagg tggccaactt tgttgtgaac gatagaattt 6060
tttttatatt aagtaaacta tttttatatt atgaaataat aataaaaaaa atattttatc 6120
attattaaca aaatcatatt agttaatttg ttaactctat aataaaagaa atactgtaac 6180
attcacatta catggtaaca tctttccacc ctttcatttg ttttttgttt gatgactttt 6240
tttcttgttt aaatttattt cccttctttt aaatttggaa tacattatca tcatatataa 6300
actaaaatac taaaaacagg attacacaaa tgataaataa taacacaaat atttataaat 6360
ctagctgcaa tatatttaaa ctagctatat cgatattgta aaataaaact agctgcattg 6420
atactgataa aaaaatatca tgtgctttct ggactgatga tgcagtatac ttttgacatt 6480
gcctttattt tatttttcag aaaagctttc ttagttctgg gttcttcatt atttgtttcc 6540
catctccatt gtgaattgaa tcatttgctt cgtgtcacaa atacaattta gntaggtaca 6600
tgcattggtc agattcacgg tttattatgt catgacttaa gttcatggta gtacattacc 6660
tgccacgcat gcattatatt ggttagattt gataggcaaa tttggttgtc aacaatataa 6720
atataaataa tgtttttata ttacgaaata acagtgatca aaacaaacag ttttatcttt 6780
attaacaaga ttttgttttt gtttgatgac gttttttaat gtttacgctt tcccccttct 6840
tttgaattta gaacacttta tcatcataaa atcaaatact aaaaaaatta catatttcat 6900
aaataataac acaaatattt ttaaaaaatc tgaaataata atgaacaata ttacatatta 6960
tcacgaaaat tcattaataa aaatattata taaataaaat gtaatagtag ttatatgtag 7020
gaaaaaagta ctgcacgcat aatatataca aaaagattaa aatgaactat tataaataat 7080
aacactaaat taatggtgaa tcatatcaaa ataatgaaaa agtaaataaa atttgtaatt 7140
aacttctata tgtattacac acacaaataa taaataatag taaaaaaaat tatgataaat 7200 atttaccatc tcataagata tttaaaataa tgataaaaat atagattatt ttttatgcaa 7260
ctagctagcc aaaaagagaa cacgggtata tataaaaaga gtacctttaa attctactgt 7320
acttccttta ttcctgacgt ttttatatca agtggacata cgtgaagatt ttaattatca 7380
gtctaaatat ttcattagca cttaatactt ttctgtttta ttcctatcct ataagtagtc 7440
ccgattctcc caacattgct tattcacaca actaactaag aaagtcttcc atagcccccc 7500
aagcggccgc accatgggca agggtggaga cggcggcgcg caggcggtga gcgggaccga 7560
cgcgtctctc gctgaggtga gctccgtcga tagcaagagc gtgcacgtcg tgctctacgg 7620
caagcgcgtg gatgtcacaa agttccagaa ggcacacccg ggcgggagca aggtgttccg 7 680
catcttccag gagcgcgacg cgacggagca gttcgagtct taccactcgc ccaaggccat 7740
caagatgatg gagggcatgc tcaagaagtc ggaggatgcg cccgcttccg tgcccctgcc 7 800
ctcgcggtcc accatgggca cggagttcaa ggagatgatt gagcgccaca agagggctgg 7860
tctctacgac ccttgcccgt tggacgagct gttcaagctc accatcgtcc ttgcgcccat 7920
cttcgtgggc gcctatctcg tgcggagcgg cgtctcgccc ctcgcgggcg cgctctccat 7980
gggctttggc ttctacctcg acggctggct tgctcacgac tacctgcatc acgcagtctt 8040
caagggctcg gtcaacacgc tcgtcaaggc gaacaacgcc atgggatacg ccctcggctt 8100
cctccagggc tacgacgtgg cctggtggcg cgcgcgccat aacacgcacc acgtgtgcac 8160
caacgaggat ggttcggacc cggacatcaa gacggcgccc ctgctcatct acgtgcgaga 8220
gaacccgtcc attgccaagc ggctcaactt cttccagcgc tggcagcagt actactatgt 8280
gccgaccatg gccatcctcg acctctactg gcgcctggag tccatcgcgt acgtggctgt 8340
gcgcctgcct aagatgtgga tgcaggccgc cgctcttgcc gctcactacg cgctcctgtg 8400
ctgggtcttc gcagcgcatc tcaacctcat ccctctcatg atggttgcac gcggcttcgc 8460
gacgggcatc gttgtctttg caacccacta tggtgaggac atcctcgacc gcgagcacgt 8520
cgagggcatg acgctcgtcg agcagaccgc caagacctcc cgtaacatca cgggcggctg 8580
gctagtgaac gtgctcacgg gcttcatctc cctgcagacc gagcatcacc tcttccccat 8640
gatgcccacc ggcaacctaa tgactatcca gcccgaggta cgcgacttct tcaagaagca 8700
tggcctcgag taccgcgagg gcaacctctt ccagtgcgtg caccagaaca tcaaggctct 8760
cgccttcgag cacctcctcc actgagcggc cgcgacacaa gtgtgagagt actaaataaa 8820
tgctttggtt gtacgaaatc attacactaa ataaaataat caaagcttat atatgccttc 8880
cgctaaggcc gaatgcaaag aaattggttc tttctcgtta tcttttgcca cttttactag 8940
tacgtattaa ttactactta atcatctttg tttacggctc attatatccg gtctagagga 9000
tccaaggccg cgaagttaaa agcaatgttg tcacttgtcg tactaacaca tgatgtgata 9060 gtttatgcta gctagctata acataagctg tctctgagtg tgttgtatat taataaagat 9120 catcactggt gaatggtgat cgtgtacgta ccctacttag taggcaatgg aagcacttag 9180 agtgtgcttt gtgcatggcc ttgcctctgt tttgagactt ttgtaatgtt ttcgagttta 9240
<210> 72
<211> 30
<212> DNA
<213> Seqüência Artificial <220>
<223> primer PvDES5'Not-l
<400> 72
gcggccgcac catgggcaag ggtggagacg 30
<210> 73
<211> 27
<212> DNA
<213> Seqüência Artificial <220>
<223> PvDES3'Not-I
<400> 73
gcggccgctc agtggaggag gtgctcg 27
<210> 74 <211> 4308 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo pLF113 <400> 74
taatacgact cactataggg cgaattgggc ccgacgtcgc atgctcccgg ccgccatggc 60
ggccgcggga attcgattgc ggccgcacca tgggcaaggg tggagacggc ggcgcgcagg 12 0
cggtgagcgg gaccgacgcg tctctcgctg aggtgagctc cgtcgatagc aagagcgtgc 180
acgtcgtgct ctacggcaag cgcgtggatg tcacaaagtt ccagaaggca cacccgggcg 24 0
ggagcaaggt gttccgcatc ttccaggagc gcgacgcgac ggagcagttc gagtcttacc 300
actcgcccaa ggccatcaag atgatggagg gcatgctcaa gaagtcggag gatgcgcccg 360
cttccgtgcc cctgccctcg cggtccacca tgggcacgga gttcaaggag atgattgagc 420
gccacaagag ggctggtctc tacgaccctt gcccgttgga cgagctgttc aagctcacca 480
tcgtccttgc gcccatcttc gtgggcgcct atctcgtgcg gagcggcgtc tcgcccctcg 54 0
cgggcgcgct ctccatgggc tttggcttct acctcgacgg ctggcttgct cacgactacc 600
tgcatcacgc agtcttcaag ggctcggtca acacgctcgt caaggcgaac aacgccatgg 660 gatacgccct cggcttcctc cagggctacg acgtggcctg gtggcgcgcg cgccataaca 720
cgcaccacgt gtgcaccaac gaggatggtt cggacccgga catcaagacg gcgcccctgc 780
tcatctacgt gcgagagaac ccgtccattg ccaagcggct caacttcttc cagcgctggc 840
agcagtacta ctatgtgccg accatggcca tcctcgacct ctactggcgc ctggagtcca 900
tcgcgtacgt ggctgtgcgc ctgcctaaga tgtggatgca ggccgccgct cttgccgctc 960
actacgcgct cctgtgctgg gtcttcgcag cgcatctcaa cctcatccct ctcatgatgg 1020
ttgcacgcgg cttcgcgacg ggcatcgttg tctttgcaac ccactatggt gaggacatcc 1080
tcgaccgcga gcacgtcgag ggcatgacgc tcgtcgagca gaccgccaag acctcccgta 1140
acatcacggg cggctggcta gtgaacgtgc tcacgggctt catctccctg cagaccgagc 1200
atcacctctt ccccatgatg cccaccggca acctaatgac tatccagccc gaggtacgcg 1260
acttcttcaa gaagcatggc ctcgagtacc gcgagggcaa cctcttccag tgcgtgcacc 1320
agaacatcaa ggctctcgcc ttcgagcacc tcctccactg agcggccgca atcactagtg 1380
aattcgcggc cgcctgcagg tcgaccatat gggagagctc ccaacgcgtt ggatgcatag 1440
cttgagtatt ctatagtgtc acctaaatag cttggcgtaa tcatggtcat agctgtttcc 1500
tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 1560
taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 1620
cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 1680
gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 1740
ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 1800
agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 1860
ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 1920
caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 1980
gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 2040
cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 2100
tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 2160
gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 2220
cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 2280
tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg 2340
tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 2400
caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 2460
aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 2520 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 2580
ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 2640
tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 2700
atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 2760
tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 2820
aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 2880
catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 2940
gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 3000
ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 3060
aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 3120
atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 3180
cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 3240
gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 3300
agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 3360
gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 3420
caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 3480
ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 3540
tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 3600
aggggttccg cgcacatttc cccgaaaagt gccacctgat gcggtgtgaa ataccgcaca 3660
gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt 3720
cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat 3780
cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa 3840
gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg 3900
cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa 3960
agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc 4020
gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag 4080
tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg 4140
cgcgtccatt cgccattcag gc.tgcgcaac tgttgggaag ggcgatcggt gcgggcctct 4200
tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg 4260
ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattg 4308 9
<210> 75
<211> 5492
<212> DNA
<213> Artificial
<220>
<223> plasmideo pKR287
<400> 75 ggccgcattt cgcaccaaat caatgaaagt aataatgaaa agtctgaata agaatactta 60 ggcttagatg cctttgttac ttgtgtaaaa taacttgagt catgtacctt tggcggaaac 120 agaataaata aaaggtgaaa ttccaatgct ctatgtataa gttagtaata cttaatgtgt 180 tctacggttg tttcaatatc atcaaactct aattgaaact ttagaaccac aaatctcaat 240 cttttcttaa tgaaatgaaa aatcttaatt gtaccatgtt tatgttaaac accttacaat 300 tggttggaga ggaggaccaa ccgatgggac aacattggga gaaagagatt caatggagat 360 ttggatagga gaacaacatt ctttttcact tcaatacaag atgagtgcaa cactaaggat 420 atgtatgaga ctttcagaag ctacgacaac atagatgagt gaggtggtga ttcctagcaa 480 gaaagacatt agaggaagcc aaaatcgaac aaggaagaca tcaagggcaa gagacaggac 540 catccatctc aggaaaagga gctttgggat agtccgagaa gttgtacaag aaattttttg 600 gagggtgagt gatgcattgc tggtgacttt aactcaatca aaattgagaa agaaagaaaa 660 gggagggggc tcacatgtga atagaaggga aacgggagaa ttttacagtt ttgatctaat -ι o η ι ΐ- W gggcatccca gctagtggta acatattcac catgtttaac cttcacgtac gtctagagga 780 tccccgggta ccgagctcga attcactggc cgtcgtttta caacgtcgtg actgggaaaa 840 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa 900 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg 960 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg 1020 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac 1080 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt 1140 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag 1200 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc 1260 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt 1320 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata 1380 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt 1440 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc 1500 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat 1560 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct 1620 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca 1680 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg 1740 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa 1800 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg 1860 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga 1920 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg 1980 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt 2040 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg 2100 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc 2160 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca 2220 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc 2280 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat 2340 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc 2400 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg 2460 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct 2520 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct 2580 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 2640 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 2700 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 2760 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 2820 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 2880 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 2940 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 3000 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 3060 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat 3120 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc 3180 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc 3240 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa 3300 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc 3360 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga 3420 ccatgattac gccaagcttg catgcctgca ggctagccta agtacgtact caaaatgcca 3480 acaaataaaa aaaaagttgc tttaataatg ccaaaacaaa ttaataaaac acttacaaca 3540
ccggattttt tttaattaaa atgtgccatt taggataaat agttaatatt tttaataatt 3600
atttaaaaag ccgtatctac taaaatgatt tttatttggt tgaaaatatt aatatgttta 3660
aatcaacaca atctatcaaa attaaactaa aaaaaaaata agtgtacgtg gttaacatta 3720
gtacagtaat ataagaggaa aatgagaaat taagaaattg aaagcgagtc taatttttaa 3780
attatgaacc tgcatatata aaaggaaaga aagaatccag gaagaaaaga aatgaaacca 3840
tgcatggtcc cctcgtcatc acgagtttct gccatttgca atagaaacac tgaaacacct 3900
ttctctttgt cacttaattg agatgccgaa gccacctcac accatgaact tcatgaggtg 3960
tagcacccaa ggcttccata gccatgcata ctgaagaatg tctcaagctc agcaccctac 4020
ttctgtgacg tgtccctcat tcaccttcct ctcttcccta taaataacca cgcctcaggt 4080
tctccgcttc acaactcaaa cattctctcc attggtcctt aaacactcat cagtcatcac 4140
cgcggccgca tgggaacgga ccaaggaaaa accttcacct gggaagagct ggcggcccat 4200
aacaccaagg acgacctact cttggccatc cgcggcaggg tgtacgatgt cacaaagttc 4260
ttgagccgcc atcctggtgg agtggacact ctcctgctcg gagctggccg agatgttact 4320
ccggtctttg agatgtatca cgcgtttggg gctgcagatg ccattatgaa gaagtactat 4380
gtcggtacac tggtctcgaa tgagctgcec atcttcccgg agccaacggt gttccacaaa 4440
accatcaaga cgagagtcga gggctacttt acggatcgga acattgatcc caagaataga 4500
ccagagatct ggggacgata cgctcttatc tttggatcct tgatcgcttc ctactacgcg 4560
cagctctttg tgcctttcgt tgtcgaacgc acatggcttc aggtggtgtt tgcaatcatc 4620
atgggatttg cgtgcgcaca agtcggactc aaccctcttc atgatgcgtc tcacttttca 4680
gtgacccaca accccactgt ctggaagatt ctgggagcca cgcacgactt tttcaacgga 4740
gcatcgtacc tggtgtggat gtaccaacat atgctcggcc atcaccccta caccaacatt 4800
gctggagcag atcccgacgt gtcgacgtct gagcccgatg ttcgtcgtat caagcccaac 4860
caaaagtggt ttgtcaacca catcaaccag cacatgtttg ttcctttcct gtacggactg 4920
ctggcgttca aggtgcgcat tcaggacatc aacattttgt actttgtcaa gaccaatgac 4980
gctattcgtg tcaatcccat ctcgacatgg cacactgtga tgttctgggg cggcaaggct 5040
ttctttgtct ggtatcgcct gattgttccc ctgcagtatc tgcccctggg caaggtgctg 5100
ctcttgttca cggtcgcgga catggtgtcg tcttactggc tggcgctgac cttccaggcg 5160
aaccacgttg ttgaggaagt tcagtggccg ttgcctgacg agaacgggat catccaaaag 5220
gactgggcag ctatgcaggt cgagactacg caggattacg cacacgattc gcacctctgg 5280
accagcatca ctggcagctt gaactaccag gctgtgcacc atctgttccc caacgtgtcg 5340 cagcaccatt atcccgatat tctggccatc atcaagaaca cctgcagcga gtacaaggtt 5400 ccataccttg tcaaggatac gttttggcaa gcatttgctt cacatttgga gcacttgcgt 54 60 gttcttggac tccgtcccaa ggaagagtag gc 5492
<210> 76
<211> 13293
<212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo pKR1155 <220>
<221> misc_feature
<222> (2769)..(2769)
<223> n é a, c, g, ou t
<400> 76
ggatctggcc ggccggatct cgtacgcaaa ggcaaagatt taaactcgaa aacattacaa 60 aagtctcaaa acagaggcaa ggccatgcac aaagcacact ctaagtgctt ccattgccta 120 ctaagtaggg tacgtacacg atcaccattc accagtgatg atctttatta atatacaaca 180 cactcagaga cagcttatgt tatagctagc tagcataaac tatcacatca tgtgttagta 240 cgacaagtga caacattgct tttaacttcg cggccttgga tcctctagac cggatataat 300 gagccgtaaa caaagatgat taagtagtaa ttaatacgta ctagtaaaag tggcaaaaga 360 taacgagaaa gaaccaattt ctttgcattc ggccttagcg gaaggcatat ataagctttg 420 attattttat ttagtgtaat gatttcgtac aaccaaagca tttatttagt actctcacac 480 ttgtgtcgcg gccgcttagg aatcctgtgc gtccttcacg cagtggacga cacccacctt 540 atgcatgtac ttcagggtgg agatgatgtt gccgacgatg gtagggtaga aaacatatcg 600 cactccatgg cgttcgcaac actcccggac cgcctgctgg acgaaggggt agtgccaaga 660 cgacatccgg ggaaagaggt ggtgctcgat ctggaaattg agaccgccag tgaagaacat 720 ggcgatgggg ccaccgtatg tggaggacgt ctccacctgc gcccgatacc agtcaatttg 780 gtcagcctca acgtccttct cagatctctt aaccttgcag ttcttgtcgg ggatccgttc 840 ggagccatca aaattgtggg acaaaatgaa gaagaacgtc aagaaggcac cagttgcggt 900 gggcaccaga ggaatggtga tcagcaatga ggtcccagtg aggtaccagg gcaagaatgc 960 ggtccggaag atgaagaaag ctcgcatcac gaatgcaagt gtgcggtagc gccgcagaaa 1020 gccattttcc cgcatggccg tgacagactc tgggatggtg tcattgtgct gcatccggaa 1080 aatgtagagc gggttgtaca ccagcgatac cccgtatgcc cccagcacaa ggtacatgta 1140 ccaagcttgg aagcggtgga accactttcg ggcggtgttt gcggcggggt agttgtggaa 1200 caccaggaac ggctctgccg actccgcatc caggtcgagg ccgtgctggt tggtgtaggt 1260
gtggtgccgc atgatgtgcg actgcagcca aatccaccgg gacgatccga tgacgtcaat 1320
gccgtaggcg aagagggcgt tgacccaagg cttcttgctg atggccccgt gggacgcatc 1380
atgctggatg gataagccga tctgcatgtg catgaatcca gtcaacaggc cccacagcac 1440
catccccgtg gtagcccagt gccactcgca aaaggccgtc acggcgatga tcccaacggt 1500
gcgcagccag aagccagggg ttgcgtacca gttcctccct ttcatcacct cgcgcattgt 1560
ccgcttaacg tcttgtgcga agggagaatc atacgtgtag acaatgtcct tcgaggtgcg 1620
gtcatgcttc cctcggacga actgttgaag caatttggaa ttctccggtt tgacgtatgg 1680
atgcattgaa taaaagagag gggaggcatc agaggcacca gaagcgagaa tcaaatctcc 174 0
tcctggatga actttggcaa gcccttcaag gtcgtagagg atgccatcaa tcgcaaccaa 1800
atcagggcgt tctaacagct gttctgtggt aagactgaga gccatggttt gcggccgctt 1860
ggggggctat ggaagacttt cttagttagt tgtgtgaata agcaatgttg ggagaatcgg 1920
gactacttat aggataggaa taaaacagaa aagtattaag tgctaatgaa atatttagac 1980
tgataattaa aatcttcacg tatgtccact tgatataaaa acgtcaggaa taaaggaagt 2040
acagtagaat ttaaaggtac tctttttata tatacccgtg ttctcttttt ggctagctag 2100
ttgcataaaa aataatctat atttttatca ttattttaaa tatcttatga gatggtaaat 2160
atttatcata atttttttta ctattattta ttatttgtgt gtgtaataca tatagaagtt 2220
aattacaaat tttatttact ttttcattat tttgatatga ttcaccatta atttagtgtt 2280
attatttata atagttcatt ttaatctttt tgtatatatt atgcgtgcag tacttttttc 2340
ctacatataa ctactattac attttattta tataatattt ttattaatga attttcgtga 2400
taatatgtaa tattgttcat tattatttca gattttttaa aaatatttgt gttattattt 2460
atgaaatatg taattttttt agtatttgat tttatgatga taaagtgttc taaattcaaa 2520
agaaggggga aagcgtaaac attaaaaaac gtcatcaaac aaaaacaaaa tcttgttaat 2580
aaagataaaa ctgtttgttt tgatcactgt tatttcgtaa tataaaaaca ttatttatat 2640
ttatattgtt gacaaccaaa tttgcctatc aaatctaacc aatataatgc atgcgtggca 2700
ggtaatgtac taccatgaac ttaagtcatg acataataaa ccgtgaatct gaccaatgca 2760
tgtacctanc taaattgtat ttgtgacacg aagcaaatga ttcaattcac aatggagatg 2820
gçaaacaaat aatgaagaac ccagaactaa gaaagctttt ctgaaaaata aaataaaggc 2880
aatgtcaaaa gtatactgca tcatcagtcc agaaagcaca tgatattttt ttatcagtat 2940
caatgcagct agttttattt tacaatatcg atatagctag tttaaatata ttgcagctag 3000
atttataaat atttgtgtta ttatttatca tttgtgtaat cctgttttta gtattttagt 3060 ttatatatga tgataatgta ttccaaattt aaaagaaggg aaataaattt aaacaagaaa 3120
aaaagtcatc aaacaaaaaa caaatgaaag ggtggaaaga tgttaccatg taatgtgaat 3180
gttacagtat ttcttttatt atagagttaa caaattaact aatatgattt tgttaataat 3240
gataaaatat tttttttatt attatttcat aatataaaaa tagtttactt aatataaaaa 3300
aaattctatc gttcacaaca aagttggcca cctaatttaa ccatgcatgt acccatggac 3360
catattaggt aaccatcaaa cctgatgaag agataaagag atgaagactt aagtcataac 3420
acaaaaccat aaaaaacaaa aatacaatca accgtcaatc tgaccaatgc atgaaaaagc 3480
tgcaatagtg agtggcgaca caaagcacat gattttctta caacggagat aaaaccaaaa 3540
aaatatttca tgaacaacct agaacaaata aagcttttat ataataaata tataaataaa 3600
taaaggctat ggaataatat acttcaatat atttggatta aataaattgt tggcggggtt 3660
gatatattta tacacaccta aagtcacttc aatctcattt tcacttaact tttatttttt 3720
ttttcttttt atttatcata aagagaatat tgataatata ctttttaaca tatttttatg 3780
acatttttta ttggtgaaaa cttattaaaa atcataaatt ttgtaagtta gatttattta 3840
aagagttcct cttcttattt taaatttttt aataaatttt taaataacta aaatttgtgt 3900
taaaaatgtt aaaaaatgtg ttattaaccc ttctcttcga ggacgtacgg atccttcatc 3960
catgcccttc atttgccgct tattaattaa tttggtaaca gtccgtacta atcagttact 4020
tatccttccc ccatcataat taatcttggt agtctcgaat gccacaacac tgactagtct 4080
cttggatcat aagaaaaagc caaggaacaa aagaagacaa aacacaatga gagtatcctt 4140
tgcatagcaa tgtctaagtt cataaaattc aaacaaaaac gcaatcacac acagtggaca 4200
tcacttatcc actagctgat caggatcgcc gcgtcaagaa aaaaaaactg gaccccaaaa 4260
gccatgcaca acaacacgta ctcacaaagg tgtcaatcga gcagcccaaa acattcacca 4320
actcaaccca tcatgagccc tcacatttgt tgtttctaac ccaacctcaa actcgtattc 4380
tcttccgcca cctcattttt gtttatttca acacccgtca aactgcatgc caccccgtgg 4440
ccaaatgtcc atgcatgtta acaagaccta tgactataaa tagctgcaat ctcggcccag 4500
gttttcatca tcaagaacca gttcaatatc ctagtacacc gtattaaaga atttaagata 4560
tactccatgg aagcagccaa agaattggtt tccatcgtcc aagaggagct ccccaaggtg 4620
gactatgccc agctttggca ggatgccagc agctgtgagg tcctttacct ctcggtggca 4680
ttcgtggcga tcaagttcat gctgcgccca ctggacctga agcgccaggc caccttgaag 4740
aagctgttca cagcatacaa cttcctcatg tcgatctatt cctttggctc cttcctggcc 4800
atggcctatg ccctatcagt aactggcatc ctctccggcg actgtgagac ggcgttcaac 4860
aacgatgtgt tcaggatcac aactcagctg ttctacctca gcaagttcgt agagtacatc 4920 gactccttct accttcccct tatggacaag ccactgtcgt tccttcagtt cttccatcat 4980 ttgggggccc ccattgacat gtggctattc tacaaatacc gcaacgaagg agtctggatc 5040 tttgtcctgt tgaatgggtt cattcactgg atcatgtacg gttactattg gacgcggctc 5100 atcaagctga acttccccat gcccaagaac ctgatcacct ccatgcagat catccagttc 5160 aatgtcgggt tctacatcgt ctggaagtac cgcaatgtgc catgctaccg ccaggatggg 5220 atgcgcatgt ttgcctggat cttcaactac tggtatgtcg ggacggtctt gctgctgttc 5280 ctcaactttt acgtgcagac gtacatccgg aagccgagga agaaccgagg gaagaaggag 5340 cccgcccgtc cagctggact cccgccggcc acgtactacg actccctggc agtgagcggc 5400 cgcaccatgt ctcctaagcg gcaagctctg ccaatcacaa ttgatggcgc aacttatgat 5460 gtgtctgctt gggtcaatca ccaccctgga ggagctgaca ttatcgagaa ctatcgcaac 5520 cgcgatgcga ccgatgtctt catggtgatg cactctcaag aagccgtcgc caagttgaag 5580 agaatgcctg ttatggagcc ttcctctcct gacacacctg ttgcacccaa gcctaagcgt 5640 gatgagcccc aggaggattt ccgcaagttg cgggaggaat tcatctccaa gggtatgttc 5700 gagacgagtt tcctttggta tttttacaag acttcaacta ccgtcggttt gatggtcctt 5760 tccatcttga tgaccgtgta cacgaattgg tatttcaccg ctgctttggt tcttggcgtg 5820 tgctaccaac agctaggctg gttgtcccac gactattgcc atcaccaggt tttcacaaac 5880 cgcaagatta acgacgcttt cggtctcttt ttcggtaacg tgatgcaggg atactcacag 5940 acttggtgga aggataggca caatggtcac catgccgcca ccaatgtggt cggccatgac 6000 ccagatattg ataacctccc catcctggct tggtctcccg aagatgtcaa gagggctact 6060 ccttcgactc ggaatctcat caagtaccag cagtactact tcattcccac cattgcatcc 6120 cttaggttca tctggtgcct ccaatccatc ggcggcgtca tgtcctacaa gagcgaggag 6180 aggaacctgt actacaagcg ccagtacact aaggaggcga ttggtctggc cctccactgg 6240 gtgctcaagg ccactttcta ttgcagtgcc atgcctagct ttgccaccgg tttgggatgc 6300 ttcttgatct ccgagctgct cggaggattt ggcattgcca tcgttgtgtt tctgaatcac 6360 tatcctttgg acaaggttga ggagactgtc tgggatgagc acgggttcag cgccagccag 6420 atccacgaga cgttgaacat taagcccggc cttctcaccg attgggtctt tggtggtctc 6480 aactaccaga ttgagcacca cttgtggccc aacatgccca ggcacaacct cacggcagct 6540 tccctggagg tgcagaagtt gtgcgccaag cacaacctgc cctacagggc cccagccatc 6600 atccccgggg ttcagaaatt ggtcagcttc ttaggcgaga ttgcccagct ggctgctgtc 6660 cctgaatgag cggccgcaag tatgaactaa aatgcacgta ggtgtaagag ctcatggaga 6720 gcatggaata ttgtatccga ccatgtaaca gtataataac tgagctccat ctcacttctt 6780 ctatgaataa acaaaggatg ttatgatata ttaacactct atctatgcac cttattgttc 6840
tatgataaat ttcctcttat tattataaat catctgaatc gtgacggctt atggaatgct 6900
tcaaatagta caaaaacaaa tgtgtactat aagactttct aaacaattct aactttagca 6960
ttgtgaacga gacataagtg ttaagaagac ataacaatta taatggaaga agtttgtctc 7020
catttatata ttatatatta cccacttatg tattatatta ggatgttaag gagacataac 7080
aattataaag agagaagttt gtatccattt atatattata tactacccat ttatatatta 7140
tacttatcca cttatttaat gtctttataa ggtttgatcc atgatatttc taatatttta 7200
gttgatatgt atatgaaagg gtactatttg aactctctta ctctgtataa aggttggatc 7260
atccttaaag tgggtctatt taattttatt gcttcttaca gataaaaaaa aaattatgag 7320
ttggtttgat aaaatattga aggatttaaa ataataataa ataacatata atatatgtat 7380
ataaatttat tataatataa catttatcta taaaaaagta aatattgtca taaatctata 7440
caatcgttta gccttgctgg acgaatctca attatttaaa cgagagtaaa catatttgac 7500
tttttggtta tttaacaaat tattatttaa cactatatga aatttttttt tttatcagca 7560
aagaataaaa ttaaattaag gaggacaatg gtgtcccaat ccttatacaa ccaacttcca 7620
caagaaagtc aagtcagaga caacaaaaaa acaagcaaag gaaatttttt aatttgagtt 7680
gtcttgtttg ctgcataatt tatgcagtaa aacactacac ataacccttt tagcagtaaa 7740
gcaatggttg accgtgtgct tagcttcttt tattttattt ttttatcagc aaagaataaa 7800
taaaataaaa tgagacactt cagggatgtt tcaacggatc cgtcgacggc gcgcccgatc 7860
atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa 7920
ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt 7980
tgttagcagc cggatcgatc caagctgtac ctcactattc ctttgccctc ggacgagtgc 8040
tggggcgtcg gtttccacta tcggcgagta cttctacaca gccatcggtc cagacggccg 8100
cgcttctgcg ggcgatttgt gtacgcccga cagtcccggc tccggatcgg acgattgcgt 8160
cgcatcgacc ctgcgcccaa gctgcatcat cgaaattgcc gtcaaccaag ctctgataga 8220
gttggtcaag accaatgcgg agcatatacg cccggagccg cggcgatcct gcaagctccg 8280
gatgcctccg ctcgaagtag cgcgtctgct gctccataca agccaaccac ggcctccaga 834 0
agaagatgtt ggcgacctcg tattgggaat ccccgaacat cgcctcgctc cagtcaatga 8400
ccgctgttat gcggccattg tccgtcagga cattgttgga gccgaaatcc gcgtgcacga 8460
ggtgccggac ttcggggcag tcctcggccc aaagcatcag ctcatcgaga gcctgcgcga 8520
cggacgcact gacggtgtcg tccatcacag tttgccagtg atacacatgg ggatcagcaa 8580
tcgcgcatat gaaatcacgc catgtagtgt attgaccgat tccttgcggt ccgaatgggc 8640 cgaacccgct cgtctggcta agatcggccg cagcgatcgc atccatagcc tccgcgaccg 8700
gctgcagaac agcgggcagt tcggtttcag gcaggtcttg caacgtgaca ccctgtgcac 8760
ggcgggagat gcaataggtc aggctctcgc tgaattcccc aatgtcaagc acttccggaa 8820
tcgggagcgc ggccgatgca aagtgccgat aaacataacg atctttgtag aaaccatcgg 8880
cgcagctatt tacccgcagg acatatccac gccctcctac atcgaagctg aaagcacgag 8940
attcttcgcc ctccgagagc tgcatcaggt cggagacgct gtcgaacttt tcgatcagaa 9000
acttctcgac agacgtcgcg gtgagttcag gcttttccat gggtatatct ccttcttaaa 9060
gttaaacaaa attatttcta gagggaaacc gttgtggtct ccctatagtg agtcgtatta 9120
atttcgcggg atcgagatct gatcaacctg cattaatgaa tcggccaacg cgcggggaga 9180
ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 9240
gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 9300
tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 9360
aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 9420
aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 9480
ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 9540
tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc 9600
agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 9660
gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 9720
tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 9780
acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc 9840
tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 9900
caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 9960
aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 10020
aactcacgtt aagggatttt ggtcatgaca ttaacctata aaaataggcg tatcacgagg 10080
ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg 10140
gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg 10200
tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta 10260
ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta atacataacc 10320
ttatgtatca tacacatacg atttaggtga cactatagaa cggcgcgcca agcttggatc 10380
tcctgcaggt ctcaatagat taagaagttg gcgtctcatt gattgaccat gggggatcct 10440
ctagacgtac gtgaaggtta aacatggtga atatgttacc actagctggg atgcccatta 10500 gatcaaaact gtaaaattct cccgtttccc ttctattcac atgtgagccc cctccctttt 10560 CtttCtttCt caattttgat tgagttaaag tcaccagcaa tgcatcactc accctccaaa 10620 aaatttcttg tacaacttct cggactatcc caaagctcct tttcctgaga tggatggtcc 10680 tgtctcttgc ccttgatgtc ttccttgttc gattttggct tcctctaatg tctttcttgc 10740 taggaatcac cacctcactc atctatgttg tcgtagcttc tgaaagtctc atacatatcc 10800 ttagtgttgc actcatcttg tattgaagtg aaaaagaatg ttgttctcct atccaaatct 10860 ccattgaatc tctttctccc aatgttgtcc catcggttgg tcctcctctc caaccaatta 10920 attgtaaggt gtttaacata aacatggtac aattaagatt tttcatttca ttaagaaaag 10980 attgagattt gtggttctaa agtttcaatt agagtttgat gatattgaaa caaccgtaga 11040 acacattaag tattactaac ttatacatag agcattggaa tttcaccttt tatttattct 11100 gtttccgcca aaggtacatg actcaagtta ttttacacaa gtaacaaagg catctaagcc 11160 taagtattct tattcagact tttcattatt actttcattg atttggtgcg aaatgcggcc 11220 gctcagaatt cctgggcatc ctgaattgtg tgggccacgc ccaccttctg catgtagcgg 11280 agcgtggaca ttatattctc caggatggtc gggtagtatg tgtagcgcac gccatggcgc 11340 ttgcaacatt cccgcaccgt atcttggatg aaggggtagt gccaagagga catgcggggg 11400 aacaagtggt gctcgatctg caggttcaaa ccgccggtca gcatcatccc gacgctgccc 11460 ccgtaggtcg ccgcggtctc ggcctgcgcc cggtaccagt ccacccggtc ctcctctaca 11520 tcgcggacct ccttcccctc cttggccctt ttggccttgc agttcttttc gggggtccgc 11580 tcagcgcctt caaagttatg ggacagcaca aaaacaaatg tcaagaagat gcctgtgacc 11640 gttggcacca gcaggatggt gagcagcgga gaggcgcccg cgatgtacca ggggaggaag 11700 gcgttgcgga agatgaacag taggcggagg gaaacggcga aggcgcgctg ccggttgaaa 11760 aagctgcctg gtctaagaga ggtggcctct gggattgtgt cgttgtgctg catcgtgaac 11820 aagtacatcg gattgtacac catcgacaca ccatacatcc ccaaaacgat gtaçatgtac 11880 cacgcctgga accgatggta ccactttcgt gacgcatttg ttgccgggta ggagtggaac 11940 aaaatgaacg gctccgccga ggcagcgtcc aggtccaggc catgctggtt ggtgtaggtg 12000 tggtggcgca taatgtggga ctgcagccag atccagcggg aggatccaat ggcgtcgatg 12060 ccgtaggcga aaaaggcgtt cacccagggc tttttgctga tggctccgtg agaggcatcg 12120 tgttgaatcg ccaacccaat ctggctgtgg aagtacccgg tgaagatgcc ccacatcact 12180 gccccggtag tgatccaata ccattcgcct atggcagtgc acgcgatgat cagcgcggtc 12240 cgcagccaaa agccgggcgt ggcgtaccag ttgcgcccct tcatgacctc ccgaacgctc 12300 tgcttgacat cctgggcgaa gggactgtca aaggtgtaca ctgggtcctt cgagcttcgt 12360 tcgtgtttgc ctcgggcaaa ttgctgcagc agcttcgaat tctctggctt cactccaggg 12420 tgcatggagt agaacagggc ggttccgtcg gaggcaccgg cggcttcaat gaggttgcca 12480 cctggatgca ctttcgccag tccgaagaga tcgtacaaca ccccatcaat tgcaaccagt 12540 tctgggtgtt ctaaaagttg ctcagtagtc aaagagatgg tggccatggt gcggccgcgg 12600 tgatgactga tgagtgttta aggaccaatg gagagaatgt ttgagttgtg aagcggagaa 12660 cctgaggcgt ggttatttat agggaagaga ggaaggtgaa tgagggacac gtcacagaag 12720 tagggtgctg agcttgagac attcttcagt atgcatggct atggaagcct tgggtgctac 12780 acctcatgaa gttcatggtg tgaggtggct tcggcatctc aattaagtga caaagagaaa 12840 ggtgtttcag tgtttctatt gcaaatggca gaaactcgtg atgacgaggg gaccatgcat 12900 ggtttcattt cttttcttcc tggattcttt ctttcctttt atatatgcag gttcataatt 12960 taaaaattag actcgctttc aatttcttaa tttctcattt tcctcttata ttactgtact 13020 aatgttaacc acgtacactt attttttttt tagtttaatt ttgatagatt gtgttgattt 13080 aaacatatta atattttcaa ccaaataaaa atcattttag tagatacggc tttttaaata 13140 attattaaaa atattaacta tttatcctaa atggcacatt ttaattaaaa aaaatccggt 13200 gttgtaagtg ttttattaat ttgttttggc attattaaag caactttttt tttatttgtt 13260 ggcattttga gtacgtactt aggctagcct gca 13293
<210> 77
<211> 14120
<212> DNA
<213> Seqüência Artificial <220>
<223> pKR952 <220>
<221> misc_feature
<222> (11120)..(11120)
<223> n é a, c, g, ou t
<400> 77
gtaccgtacg tgggcggatc ccccgggctg caggtcgact cgacgtacgt ctagaggatc 60
cgtcgacggc gcgcccgatc atccggatat agttcctcct ttcagcaaaa aacccctcaa 120
gacccgttta gaggccccaa ggggttatgc tagttattgc tcagcggtgg cagcagccaa 180
ctcagcttcc tttcgggctt tgttagcagc cggatcgatc caagctgtac ctcactattc 240
ctttgccctc ggacgagtgc tggggcgtcg gtttccacta tcggcgagta cttctacaca 300
gccatcggtc cagacggccg cgcttctgcg ggcgatttgt gtacgcccga cagtcccggc 360
tccggatcgg acgattgcgt cgcatcgacc ctgcgcccaa gctgcatcat cgaaattgcc 420 gtcaaccaag ctctgataga gttggtcaag accaatgcgg agcatatacg cccggagccg 480 cggcgatcct gcaagctccg gatgcctccg ctcgaagtag cgcgtctgct gctccataca 540 agccaaccac ggcctccaga agaagatgtt ggcgacctcg tattgggaat ccccgaacat 600 cgcctcgctc cagtcaatga ccgctgttat gcggccattg tccgtcagga cattgttgga 660 gccgaaatcc gcgtgcacga ggtgccggac ttcggggcag tcctcggccc aaagcatcag 720 ctcatcgaga gcctgcgcga cggacgcact gacggtgtcg tccatcacag tttgccagtg 780 atacacatgg ggatcagcaa tcgcgcatat gaaatcacgc catgtagtgt attgaccgat 840 tccttgcggt ccgaatgggc cgaacccgct cgtctggcta agatcggccg cagcgatcgc 900 atccatagcc tccgcgaccg gctgcagaac agcgggcagt tcggtttcag gcaggtcttg 960 caacgtgaca ccctgtgcac ggcgggagat gcaataggtc aggctctcgc tgaattcccc 1020 aatgtcaagc acttccggaa tcgggagcgc ggccgatgca aagtgccgat aaacataacg 1080 atctttgtag aaaccatcgg cgcagctatt tacccgcagg acatatccac gccctcctac 1140 atcgaagctg aaagcacgag attcttcgcc ctccgagagc tgcatcaggt cggagacgct 1200 gtcgaacttt tcgatcagaa acttctcgac agacgtcgcg gtgagttcag gcttttccat 1260 gggtatatct ccttcttaaa gttaaacaaa attatttcta gagggaaacc gttgtggtct 1320 ccctatagtg agtcgtatta atttcgcggg atcgagatct gatcaacctg cattaatgaa 1380 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 1440 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 1500 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 1560 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 1620 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 1680 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 1740 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat 1800 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 1860 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 1920 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 1980 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 2040 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 2100 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 2160 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 2220 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaca ttaacctata 2280 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc 2340
tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca 2400
gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg 2460
cggcatcaga gcagattgta ctgagagtgc accatatgga catattgtcg ttagaacgcg 2520
gctacaatta atacataacc ttatgtatca tacacatacg atttaggtga cactatagaa 2580
cggcgcgcca agctgggtct agaactagaa acgtgatgcc acttgttatt gaagtcgatt 2640
acagcatcta ttctgtttta ctatttataa ctttgccatt tctgactttt gaaaactatc 2700
tctggatttc ggtatcgctt tgtgaagatc gagcaaaaga gacgttttgt ggacgcaatg 2760
gtccaaatcc gttctacatg aacaaattgg tcacaatttc cactaaaagt aaataaatgg 2820
caagttaaaa aaggaatatg cattttactg attgcctagg tgagctccaa gagaagttga 2880
atctacacgt ctaccaaccg ctaaaaaaag aaaaacattg atatgtaacc tgattccatt 2940
agcttttgac ttcttcaaca gattctctac ttagatttct aacagaaata ttattactag 3000
cacatcattt tcagtctcac tacagcaaaa aatccaacgg cacaatacag acaacaggag 3060
atatcagact acagagatag atagatgcta ctgcatgtag taagttaaat aaaaggaaaa 3120
taaaatgtct tgctaccaaa actactacag actatgatgc tcaccacagg ccaaatcctg 3180
caactaggac agcattatct tatatatatt gtacaaaaca agcatcaagg aacatttggt 3240
ctaggcaatc agtacctcgt tctaccatca ccctcagtta tcacatcctt gaaggatcca 3300
ttactgggaa tcatcggcaa cacatgctcc tgatggggca caatgacatc aagaaggtag 3360
gggccagggg tgtccaacat tctctgaatt gccgctctaa gctcttcctt cttcgtcact 3420
cgcgctgccg gtatcccaca agcatcagca aacttgagca tgtttgggaa tatctcgctc 3480
tcgctagacg gatctccaag ataggtgtga gctctattgg acttgtagaa cctatcctcc 3540
aactgaacca ccatacccaa atgctgattg ttcaacaaca atatcttaac tgggagattc 3600
tccactctta tagtggccaa ctcctgaaca ttcatgatga aactaccatc cccatcaatg 3660
tcaaccacaa cagccccagg gttagcaaca gcagcaccaa tagccgcagg caatccaaaa 3720
cccatggctc caagaccccc tgaggtcaac cactgcctcg gtctcttgta cttgtaaaac 3780
tgcgcagccc acatttgatg ctgcccaacc ccagtactaa caatagcatc tccattagtc 3840
aactcatcaa gaacctcgat agcatgctgc ggagaaatcg cgtcctggaa tgtcttgtaa 3900
cccaatggaa acttgtgttt ctgcacatta atctcttctc tccaacctcc aagatcaaac 3960
ttaccctcca ctcctttctc ctccaaaatc atattaattc ccttcaaggc caacttcaaa 4020
tccgcgcaaa ccgacacgtg cgcctgcttg ttcttcccaa tctcggcaga atcaatatca 4080
atgtgaacaa tcttagccct actagcaaaa gcctcaagct tcccagtaac acggtcatca 4140 aaccttaccc caaaggcaag caacaaatca ctattgtcaa cagcatagtt agcataaaca 4200 gtaccatgca tacccagcat ctgaagggaa tattcatcac caataggaaa agttccaaga 4260 cccattaaag tgctagcaac gggaatacca gtgagttcaa caaagcgcct caattcagca 4320 ctggaattca aactgccacc gccgacgtag agaacgggct tttgggcctc catgatgagt 4380 ctgacaatgt gttccaattg ggcctcggcg gggggcctgg gcagcctggc gaggtaaccg 4440 gggaggttaa cgggctcgtc ccaattaggc acggcgagtt gctgctgaac gtctttggga 4500 atgtcgatga ggaccggacc ggggcggccg gaggtggcga cgaagaaagc ctcggcgacg 4560 acgcggggga tgtcgtcgac gtcgaggatg aggtagttgt gcttcgtgat ggatctgctc 4620 acctccacga tcggggtttc ttggaaggcg tcggtgccga tcatccggcg ggcgacctgg 4680 ccggtgatgg cgacgactgg gacgctgtcc attaaagcgt cggcgaggcc gctcacgagg 4740 ttggtggcgc cggggccgga ggtggcaatg cagacgccgg ggaggccgga ggaacgcgcg 4800 tagccttcgg cggcgaagac gccgccctgc tcgtggcgcg ggagcacgtt gcggatggcg 4860 gcggagcgcg tgagcgcctg gtggatctcc atcgacgcac cgccggggta cgcgaacacc 4920 gtcgtcacgc cctgcctctc cagcgcctcc acaaggatgt ccgcgccctt gcgaggttcg 4980 ccggaggcga accgtgacac gaagggctcc gtggtcggcg cttccttggt gaagggcgcc 5040 gccgtggggg gtttggagat ggaacatttg attttgagag cgtggttggg tttggtgagg 5100 gtttgatgag agagagggag ggtggatcta gtaatgcgtt tggggaaggt ggggtgtgaa 5160 gaggaagaag agaatcgggt ggttctggaa gcggtggccg ccattgtgtt gtgtggcatg 5220 gttatacttc aaaaactgca caacaagcct agagttagta cctaaacagt aaatttacaa 5280 cagagagcaa agacacatgc aaaaatttca gccataaaaa aagttataat agaatttaaa 5340 gcaaaagttt cattttttaa acatatatac aaacaaactg gatttgaagg aagggattaa 5400 ttcccctgct caaagtttga attcctattg tgacctatac tcgaataaaa ttgaagccta 5460 aggaatgtat gagaaacaag aaaacaaaac aaaactacag acaaacaagt acaattacaa 5520 aattcgctaa aattctgtaa tcaccaaacc ccatctcagt cagcacaagg cccaaggttt 5580 attttgaaat aaaaaaaaag tgattttatt tctcataagc taaaagaaag aaaggcaatt 5640 atgaaatgat ttcgactaga tctgaaagtc caacgcgtat tccgcagata ttaaagaaag 5700 agtagagttt cacatggatc ctagatggac ccagttgagg aaaaagcaag gcaaagcaaa 5760 ccagaagtgc aagatccgaa attgaaccac ggaatctagg atttggtaga gggagaagaa 5820 aagtaccttg agaggtagaa gagaagagaa gagcagagag atatatgaac gagtgtgtct 5880 tggtctcaac tctgaagcga tacgagttta gaggggagca ttgagttcca atttataggg 5940 aaaccgggtg gcaggggtga gttaatgacg gaaaagcccc taagtaacga gattggattg 6000 tgggttagat tcaaccgttt gcatccgcgg cttagattgg ggaagtcaga gtgaatctca 6060 accgttgact gagttgaaaa ttgaatgtag caaccaattg agccaacccc agcctttgcc 6120
ctttgatttt gatttgtttg ttgcatactt tttatttgtc ttctggttct gactctcttt 6180
ctctcgtttc aatgccaggt tgcctactcc cacaccactc acaagaagat tctactgtta 6240
gtattaaata ttttttaatg tattaaatga tgaatgcttt tgtaaacaga acaagactat 6300
gtctaataag tgtcttgcaa cattttttaa gaaattaaaa aaaatatatt tattatcaaa 6360
atcaaatgta tgaaaaatca tgaataatat aattttatac atttttttaa aaaatctttt 6420
aatttcttaa ttaatatctt aaaaataatg attaatattt aacccaaaat aattagtatg 6480
attggtaagg aagatatcca tgttatgttt ggatgtgagt ttgatctaga gcaaagctta 6540
ctagagtcga cctgcaggtc gactcgacgt acgatcccac atgcaagttt ttatttcaat 6600
cccttttcct ttgaataact gaccaagaac aacaagaaaa aaaaaaaaaa agaaaaggat 6660
cattttgaaa ggatattttt cgctcctatt caaatactgt atttttacca aaaaaactgt 6720
atttttccta cactctcaag ctttgttttt cgcttcgact ctcatgattt ccttcatatg 6780
ccaatcactc tatttataaa tggcataagg tagtgtgaac aattgcaaag cttgtcatca 6840
aaagcttgca atgtacaaat taatgttttt catgcctttc aaaattatct gcacccccta 6900
gctattaatc taacatctaa gtaaggctag tgaatttttt cgaatagtca tgcagtgcat 6960
taatttcccc gtgactattt tggctttgac tccaacactg gccccgtaca tccgtccctc 7020
attacatgaa aagaaatatt gtttatattc ttaattaaaa atattgtccc ttctaaattt 7080
tcatatagtt aattattata ttactttttt ctctattcta ttagttctat tttcaaatta 7140
ttatttatgc atatgtaaag tacattatat ttttgctata tacttaaata tttctaaatt 7200
attaaaaaaa gactgatatg aaaaatttat tctttttaaa gctatatcat tttatatata 7260
ctttttcttt tcttttcttt cattttctat tcaatttaat aagaaataaa ttttgtaaat 7320
ttttatttat caatttataa aaatatttta ctttatatgt tttttcacat ttttgttaaa 7380
caaatcatat cattatgatt gaaagagagg aaattgacag tgagtaataa gtgatgagaa 7440
aaaaatgtgt tatttcctaa aaaaaaccta aacaaacatg tatctactct ctatttcatc 7500
tatctctcat ttcatttttc tctttatctc tttctttatt tttttatcat atcatttcac 7560
attaattatt tttactctct ttattttttc tctctatccc tctcttattt ccactcatat 7620
atacactcca aaattggggc atgcctttat cactactcta tctcctccac taaatcattt 7680
aaatgaaact gaaaagcatt ggcaagtctc ctcccctcct caagtgattt ccaactcagc 7740
attggcatct aattgattca gtatatctat tgcatgtgta aaagtctttc cacaatacat 7800
aactattaat taatcttaaa taaataaagg ataaaatatt tttttttctt cataaaatta 7860 aaatatgtta ttttttgttt agatgtatat tcgaataaat ctaaatatat gataatgatt 7920
ttttatattg attaaacata taatcaatat taaatatgat atttttttat ataggttgta 7980
cacataattt tataaggata aaaaatatga taaaaataaa ttttaaatat ttttatattt 8040
acgagaaaaa aaaatatttt agccataaat aaatgaccag catattttac aaccttagta 8100
attcataaat tcctatatgt atatttgaaa ttaaaaacag ataatcgtta agggaaggaa 8160
tcctacgtca tctcttgcca tttgtttttc atgcaaacag aaagggacga aaaaccacct 8220
caccatgaat cactcttcac accattttta ctagcaaaca agtctcaaca actgaagcca 8280
gctctctttc cgtttctttt tacaacactt tctttgaaat agtagtattt ttttttcaca 8340
tgatttatta acgtgccaaa agatgcttat tgaatagagt gcacatttgt aatgtactac 8400
taattagaac atgaaaaagc attgttctaa cacgataatc ctgtgaaggc gttaactcca 8460
aagatccaat ttcactatat aaattgtgac gaaagcaaaa tgaattcaca tagctgagag 8520
agaaaggaaa ggttaactaa gaagcaatac ttcagcggcc gcatgactga ggataagacg 8580
aaggtcgagt tcccgacgct cacggagctc aagcactcga tcccgaacgc gtgctttgag 8640
tcgaacctcg gcctctcgct ctactacacg gcccgcgcga tcttcaacgc gtcggcctcg 8700
gcggcgctgc tctacgcggc gcgctcgacg ccgttcattg ccgataacgt tctgctccac 8760
gcgctcgttt gcgccaccta catctacgtg cagggcgtca tcttctgggg cttcttcacg 8820
gtcggccacg actgcggcca ctcggccttc tcgcgctacc acagcgtcaa ctttatcatc 8880
ggctgcatca tgcactctgc gattttgacg ccgttcgaga gctggcgcgt gacgcaccgc 8940
caccaccaca agaacacggg caacattgat aaggacgaga tcttttaccc gcaccggtcg 9000
gtcaaggacc tccaggacgt gcgccaatgg gtctacacgc tcggcggtgc gtggtttgtc 9060
tacttgaagg tcgggtatgc cccgcgcacg atgagccact ttgacccgtg ggacccgctc 9120
ctccttcgcc gcgcgtcggc cgtcatcgtg tcgctcggcg tctgggccgc cttcttcgcc 9180
gcgtacgcgt acctcacata ctcgctcggc tttgccgtca tgggcctcta ctactatgcg 9240
ccgctctttg tctttgcttc gttcctcgtc attacgacct tcttgcacca caacgacgaa 9300
gcgacgccgt ggtacggcga ctcggagtgg acgtacgtca agggcaacct ctcgagcgtc 9360
gaccgctcgt acggcgcgtt cgtggacaac ctgagccacc acattggcac gcaccaggtc 9420
caccacttgt tcccgatcat tccgcactac aagctcaacg aagccaccaa gcactttgcg 9480
gccgcgtacc cgcacctcgt gcgcaggaac gacgagccca tcatcacggc cttcttcaag 9540
accgcgcacc tctttgtcaa ctacggcgct gtgcccgaga cggcgcagat cttcacgctc 9600
aaagagtcgg ccgcggccgc caaggccaag tcggactaag cggccgcgaa ttcaatcact 9660
agtgaattcg cggccgcatg agccgtaaag gttcaataca acgagtgctt gttttcttag 9720 ggacaagcat tgtacttatg tatgattctg tgtaaccatg agtcttccac gttgtactaa 9780 tgtgaagggc aaaaataaaa cacagaacaa gttcgttttt ctcaaataat gtgaaggtag 9840 aaaatggaac catgcctcct ctcttgcatg tgatttaaaa tattagcaga tggtaccggg 9900 ccccccctcg aggtcgcccg ggggatccgc cctaagcttc gtacgtcctc gaagagaagg 9960 gttaataaca cattttttaa catttttaac acaaatttta gttatttaaa aatttattaa 10020 aaaatttaaa ataagaagag gaactcttta aataaatcta acttacaaaa tttatgattt 10080 ttaataagtt ttcaccaata aaaaatgtca taaaaatatg ttaaaaagta tattatcaat 10140 attctcttta tgataaataa aaagaaaaaa aaaataaaag ttaagtgaaa atgagattga 10200 agtgacttta ggtgtgtata aatatatcaa ccccgccaac aatttattta atccaaatat 10260 attgaagtat attattccat agcctttatt tatttatata tttattatat aaaagcttta 10320 tttgttctag gttgttcatg aaatattttt ttggttttat ctccgttgta agaaaatcat 10380 gtgctttgtg tcgccactca ctattgcagc tttttcatgc attggtcaga ttgacggttg 10440 attgtatttt tgttttttat ggttttgtgt tatgacttaa gtcttcatct ctttatctct 10500 tcatcaggtt tgatggttac ctaatatggt ccatgggtac atgcatggtt aaattaggtg 10560 gccaactttg ttgtgaacga tagaattttt tttatattaa gtaaactatt tttatattat 10620 gaaataataa taaaaaaaat attttatcat tattaacaaa atcatattag ttaatttgtt ι η CQ η xyuuv aactctataa taaaagaaat actgtaacat tcacattaca tggtaacatc tttccaccct 10740 ttcatttgtt ttttgtttga tgactttttt tcttgtttaa atttatttcc cttcttttaa 10800 atttggaata cattatcatc atatataaac taaaatacta aaaacaggat tacacaaatg 10860 ataaataata acacaaatat ttataaatct agctgcaata tatttaaact agctatatcg 10920 atattgtaaa ataaaactag ctgcattgat actgataaaa aaatatcatg tgctttctgg 10980 actgatgatg cagtatactt ttgacattgc ctttatttta tttttcagaa aagctttctt 11040 agttctgggt tcttcattat ttgtttccca tctccattgt gaattgaatc atttgcttcg 11100 tgtcacaaat acaatttagn taggtacatg cattggtcag attcacggtt tattatgtca 11160 tgacttaagt tcatggtagt acattacctg ccacgcatgc attatattgg ttagatttga 11220 taggcaaatt tggttgtcaa caatataaat ataaataatg tttttatatt acgaaataac 11280 agtgatcaaa acaaacagtt ttatctttat taacaagatt ttgtttttgt ttgatgacgt 11340 tttttaatgt ttacgctttc ccccttcttt tgaatttaga acactttatc atcataaaat 11400 caaatactaa aaaaattaca tatttcataa ataataacac aaatattttt aaaaaatctg 11460 aaataataat gaacaatatt acatattatc acgaaaattc attaataaaa atattatata 11520 aataaaatgt aatagtagtt atatgtagga aaaaagtact gcacgcataa tatatacaaa 11580 aagattaaaa tgaactatta taaataataa cactaaatta atggtgaatc atatcaaaat 11640
aatgaaaaag taaataaaat ttgtaattaa cttctatatg tattacacac acaaataata 11700
aataatagta aaaaaaatta tgataaatat ttaccatctc ataagatatt taaaataatg 11760
ataaaaatat agattatttt ttatgcaact agctagccaa aaagagaaca cgggtatata 11820
taaaaagagt acctttaaat tctactgtac ttcctttatt cctgacgttt ttatatcaag 11880
tggacatacg tgaagatttt aattatcagt ctaaatattt cattagcact taatactttt 11940
ctgttttatt cctatcctat aagtagtccc gattctccca acattgctta ttcacacaac 12000
taactaagaa agtcttccat agccccccaa gcggccgcca attcatgacg gtgggcggcg 12060
atgaggtgta cagcatggcg caggtgcgcg accacaacac cccggacgac gcctggtgcg 12120
ccatccacgg cgaggtgtac gagctgacca agttcgcccg cacccacccc gggggggaca 12180
tcatcttgct ggccgccggc aaggaggcca ccatcctgtt cgagacgtac cacgtgcgcc 12240
ccatctccga cgcggtcctg cgcaagtacc gcatcggcaa gctcgccgcc gccggcaagg 12300
atgagccggc caacgacagc acctactaca gctgggacag cgacttttac aaggtgctcc 12360
gccagcgtgt cgtggcgcgt ctcgaggagc gcaagatcgc ccgccgcggc ggccccgaga 12420
tctggatcaa ggccgccatc ctcgtcagcg gcttctggtc catgctctac ctcatgtgca 12480
ccctggaccc gaaccgcggc gccatcctgg ccgccatcgc gctgggcatc gtcgccgcct 1254 0
tcgtcggcac gtgcattcag cacgacggca accacggcgc gttcgccttc tctccgttca 12600
tgaacaagct ctctggctgg acgctcgaca tgatcggcgc cagtgccatg acctgggaga 12660
tgcagcacgt gctgggccac cacccgtaca ccaacctgat cgagatggag aacggcaccc 12720
aaaaggtcac ccacgccgac gtcgacccca agaaggccga ccaggagagc gacccggacg 12780
tcttcagcac ctaccccatg ctccgtctgc acccgtggca ccgcaagcgc ttctaccacc 12840
gcttccagca cctgtacgcg ccgctgctct tcggtttcat gaccatcaac aaggtgatca 12900
cccaggatgt gggagttgtc ctcagcaagc gtctgtttca gatcgatgcc aactgccgtt 12960
acgccagcaa gtcgtacgtt gcgcgcttct ggatcatgaa gctgctcacc gtcctctaca 13020
tggtcgccct ccccgtgtac acccagggcc ttgtcgacgg gctcaagctc ttcttcatcg 13080
cccacttttc gtgcggcgag ctgctggcca ccatgttcat cgtcaaccac atcatcgagg 13140
gcgtctcgta cgcctccaag gactctgtca agggcaccat ggcgccgccg cgcacggtgc 13200
acggcgtgac cccgatgcat gacacccgcg acgcgctcgg caaggagaag gcagccacca 13260
agcacgtgcc gctcaacgac tgggccgcgg tccagtgcca gacctcggtc aactggtcga 13320
tcggctcgtg gttctggaac cacttctccg gcgggctcaa ccaccagatc gagcaccacc 13380
tcttccccgg cctcacccac accacctacg tgtacattca ggatgtggtg caggcgacgt 13440 gcgccgagta cggggtcccg taccagtcgg agcagagcct cttctccgcc tacttcaaga 13500
tgctctccca ccttcgggcg ctcggcaacg agccgatgcc ctcgtgggag aaggaccacc 13560
ccaagtccaa gtgaaagctt acgcgtcgac ccgggtatcc gtatgatgtg cctgactacg 13620
catgatatcg cggccgcgac acaagtgtga gagtactaaa taaatgcttt ggttgtacga 13680
aatcattaca ctaaataaaa taatcaaagc ttatatatgc cttccgctaa ggccgaatgc 13740
aaagaaattg gttctttctc gttatctttt gccactttta ctagtacgta ttaattacta 13800
cttaatcatc tttgtttacg gctcattata tccggtctag aggatccaag gccgcgaagt 13860
taaaagcaat gttgtcactt gtcgtactaa cacatgatgt gatagtttat gctagctagc 13920
tataacataa gctgtctctg agtgtgttgt atattaataa agatcatcac tggtgaatgg 13980
tgatcgtgta cgtaccctac ttagtaggca atggaagcac ttagagtgtg ctttgtgcat 14040
ggccttgcct ctgttttgag acttttgtaa tgttttcgag tttaaatctt tgcctttgcg 14100
tacgtctaga ggatccccgg 14120
<210> 78
<211> 18
<212> DNA
<213> Seqüência Artificial <220>
<223> M13F primer Universal
<400> 78
tgtaaaacga cggccagt
<210> 79
<211> 277
<212> PRT
<213> Pavlova sp.
<400> 79
Met Met Leu Ala Ala Gly Tyr Leu Leu Val Leu Ser Ala Ala Arg Gln 1 5 10 15
Ser Phe Gln Gln Asp Ile Asp Asn Pro Asn Gly Ala Tyr Ser Thr Ser 20 25 30
Trp Thr Gly Leu Pro Ile Val Met Ser Val Val Tyr Leu Ser Gly Val 35 40 45
Phe Gly Leu Thr Lys Tyr Phe Glu Asn Arg Lys Pro Met Thr Gly Leu 50 55 60
Lys Asp Tyr Met Phe Thr Tyr Asn Leu Tyr Gln Val Ile Ile Asn Val 65 70 75 80 Trp Cys Val Val Ala Phe Leu Leu Glu Val Arg Arg Ala Gly Met Ser 85 90 95
Leu Ile Gly Asn Lys Val Asp Leu Gly Pro Asn Ser Phe Arg Leu Gly 100 105 110
Phe Val Thr Trp Val His Tyr Asn Asn Lys Tyr Val Glu Leu Leu Asp 115 120 125
Thr Leu Trp Met Val Leu Arg Lys Lys Thr Gln Gln Val Ser Phe Leu 130 135 140
His Val Tyr His His Val Leu Leu Met Trp Ala Trp Phe Val Val Val 145 150 155 160
Lys Leu Gly Asn Gly Gly Asp Ala Tyr Phe Gly Gly Leu Met Asn Ser 165 170 175
Ile Ile His Val Met Met Tyr Ser Tyr Tyr Thr Met Ala Leu Leu Gly 180 185 190
Trp Ser Cys Pro Trp Lys Arg Tyr Leu Thr Gln Ala Gln Leu Val Gln 195 200 205
Phe Cys Ile Cys Leu Ala His Ser Thr Trp Ala Ala Val Thr Gly Ala 210 215 220
Tyr Pro Trp Arg Ile Cys Leu Val Glu Val Trp Val Met Val Ser Met 225 230 235 240
Leu Val Leu Phe Thr Arg Phe Tyr Arg Gln Ala Tyr Ala Lys Glu Ala 245 250 255
Lys Ala Lys Glu Ala Lys Lys Leu Ala Gln Glu Ala Ser Gln Ala Lys 260 265 270
Ala Val Lys Ala Glu 275
<210> 80
<211> 742
<212> DNA
<213> Euglena gracilís
<400> 80
attttttttc ggtgcggcgc attgtattgc ctgcacaatg gcggatagcc cagtcatcaa cctcagcacc atgtggaaac ccctttcact gatggctttg gaccttgccg ttttgggaca 120
tgtctggaag caggcacaac aggagggcag catttcggcc tatgctgatt ctgtttggac 180
tcctctcatt atgtccggtt tatacttatc aatgatcttc gtggggtgcc gctggatgaa 240
gaaccgtgaa ccctttgaga tcaaaacata catgtttgcg tataacctgt atcagacctt 300
gatgaacctt tgcatcgtgt tgggattctt gtaccaggtg catgccactg ggatgcgctt 360
ttggggaagt ggtgtcgacc gaagcccaaa aggtttgggc attggcttct tcatttatgc 420
ccactaccac aacaagtatg tggaatattt tgatacactt tttatggtgc tgcgaaagaa 480
gaacaaccag atttctttcc ttcacgtgta tcatcatgcc ctgttgacat gggcttggtt 540
tgctgttgtg tatttcgcac ctggaggtga tggctggttt ggagcttgct acaattcttc 600
catccatgtc ctgatgtact cttactactt gcttgcaact tttggcatca gttgcccatg 660
gaagaagatc ttgacacagc tccagatggt tcaattctgt ttctgtttta cacattccat 720
ttatgtgtgg atttgcgggt ca 742
<210> 81 <211> 2630 <212> DNA
<213> Euglena gracilis <400> 81
attttttttc ggtgcggcgc attgtattgc rfnrarapiri _ ---W^3 gcggatagcc 60 cctcagcacc atgtggaaac ccctttcact gatggctttg gaccttgccg ttttgggaca 120 tgtctggaag caggcacaac aggagggcag catttcggcc tatgctgatt ctgtttggac 180 tcctctcatt atgtccggtt tatacttatc aatgatcttc gtggggtgcc gctggatgaa 240 gaaccgtgaa ccctttgaga tcaaaacata catgtttgcg tataacctgt atcagacctt 300 gatgaacctt tgcatcgtgt tgggattctt gtaccaggtg catgccactg ggatgcgctt 360 ttggggaagt ggtgtcgacc gaagcccaaa aggtttgggc attggcttct tcatttatgc 420 ccactaccac aacaagtatg tggaatattt tgatacactt tttatggtgc tgcgaaagaa 480 gaacaaccag atttctttcc ttcacgtgta tcatcatgcc ctgttgacat gggcttggtt 540 tgctgttgtg tatttcgcac ctggaggtga tggctggttt ggagcttgct acaattcttc 600 catccatgtc ctgatgtact cttactactt gcttgcaact tttggcatca gttgcccatg 660 gaagaagatc ttgacacagc tccagatggt tcaattctgt ttctgtttta cacattccat 720 ttatgtgtgg atttgcgggt cagagatcta cccacggcct ctgactgctt tgcagtcgtt 780 cgtgatggtc aatatgttgg tgctgtttgg caatttctat gtcaagcaat actcccaaaa 840 gaacggcaag ccggagaacg gagccacccc tgagaacgga gcgaagccgc aaccttgcga 900 gaacggcacg gtggaaaagc gagagaatga caccgccaac gttcggcccg cccgtccagc 960 tggactcccg ccggccacgt actacgactc cctggcagtg tcggggcagg gcaaggagcg 1020 gctgttcacc accgatgagg tgaggcggca catcctcccc accgatggct ggctgacgtg 1080 ccacgaagga gtctacgatg tcactgattt ccttgccaag caccctggtg gcggtgtcat 1140 cacgctgggc cttggaaggg actgcacaat cctcgtcgag tcataccacc ctgctgggcg 1200 cccggacaag gtgatggaga agtaccgcat tggtacgctg caggacccca agacgttcta 1260 tgcttgggga gagtccgatt tctaccctga gttgaagcgc cgggcccttg caaggctgaa 1320 ggaggctggt caggcgcggc gcggcggcct tggggtgaag gccctcctgg tgctcaccct 1380 cttcttcgtg tcgtggtaca tgtgggtggc ccacaagtcc ttcctctggg ccgccgtctg 1440 gggcttcgcc ggctcccacg tcgggctgag catccagcac gacggcaacc acggcgcgtt 1500 cagccgcagc acactggtga accgcctggc ggggtggggc atggacttga tcggcgcgtc 1560 gtcaacggtg tgggagtacc agcacgtcat cggccaccac cagtacacca acctcgtgtc 1620 ggacacgcta ttcagtctgc ctgagaacga tccggacgtc ttctccagct acccgctgat 1680 gcgcatgcac ccggatacgg cgtggcagcc gcaccaccgc ttccagcacc tgttcgcgtt 1740 cccactgttc gccctgatga caatcagcaa ggtgctgacc agcgatttcg ctgtctgcct 1800 cagcatgaag aaggggtcca tcgactgctc ctccaggctc gtcccactgg aggggcagct 1860 actgttctgg ggggccaagc tggcgaactt cctgttgcag ~ V.^, ^vj ν- ^ ^ τ no rs χ yt.\J ccacgggaca gctatgggcc tggccctctt ctctgttgcc caccttgtgt cgggggagta 1980 cctcgcgatc tgcttcatca tcaaccacat cagcgagtct tgtgagttta tgaatacaag 2040 ctttcaaacc gccgcccgga ggacagagat gcttcaggca gcccatcagg cagcggaggc 2100 caagaaggtg aagcccaccc ctccaccgaa cgattgggct gtgacacagg tccaatgctg 2160 cgtgaattgg agatcaggtg gcgtgttggc caatcacctc tctggaggct tgaaccacca 2220 gatcgagcat catctgttcc ccagcatctc gcatgccaac taccccatca tcgcccgtgt 2280 tgtgaaggag gtgtgcgagg agtatgggtt gccgtacaag aactacgtca cgttctggga 2340 tgcagtctgt ggcatggttc agcacctccg gttgatgggt gctccaccgg tgccaacgaa 2400 cggggacaaa aagtcataag ccacgacatc atttggggct cactccgtgc agccttttct 2460 tgggctgccc acgaagatgc gcgatgaggc acctggtggt tgccctccgc cggcctcgga 2520 aaacggttcg acgcctgctc cttcagccca gagcactccg gcgaagagtg aaagagcact 2580 gacctgaatt ttatgatgac ccattttttt aaaaaaaaaa aaaaaaaaaa 2630
<210> 82
<211> 2382
<212> DNA
<213> Euglena gracilis <400> 82 atggcggata gcccagtcat caacctcagc accatgtgga aacccctttc actgatggct 60 ttggaccttg ccgttttggg acatgtctgg aagcaggcac aacaggaggg cagcatttcg 120 gcctatgctg attctgtttg gactcctctc attatgtccg gtttatactt atcaatgatc 180 ttcgtggggt gccgctggat gaagaaccgt gaaccctttg agatcaaaac atacatgttt 240 gcgtataacc tgtatcagac cttgatgaac ctttgcatcg tgttgggatt cttgtaccag 300 gtgcatgcca ctgggatgcg cttttgggga agtggtgtcg accgaagccc aaaaggtttg 360 ggcattggct tcttcattta tgcccactac cacaacaagt atgtggaata ttttgataca 420 ctttttatgg tgctgcgaaa gaagaacaac cagatttctt tccttcacgt gtatcatcat 480 gccctgttga catgggcttg gtttgctgtt gtgtatttcg cacctggagg tgatggctgg 540 tttggagctt gctacaattc ttccatccat gtcctgatgt actcttacta cttgcttgca 600 acttttggca tcagttgccc atggaagaag atcttgacac agctccagat ggttcaattc 660 tgtttctgtt ttacacattc catttatgtg tggatttgcg ggtcagagat ctacccacgg 720 cctctgactg ctttgcagtc gttcgtgatg gtcaatatgt tggtgctgtt tggcaatttc 780 tatgtcaagc aatactccca aaagaacggc aagccggaga acggagccac ccctgagaac 840 ggagcgaagc cgcaaccttg cgagaacggc acggtggaaa agcgagagaa tgacaccgcc 900 aacgttcggc ccgcccgtcc agctggactc ccgccggcca cgtactacga ctccctggca 960 gtgtcggggc agggcaagga gcggctgttc accaccgatg aggtgaggcg gcacatcctc 1020 cccaccgatg gctggctgac gtgccacgaa ggagtctacg atgtcactga tttccttgcc 1080 aagcaccctg gtggcggtgt catcacgctg ggccttggaa gggactgcac aatcctcgtc 1140 gagtcatacc accctgctgg gcgcccggac aaggtgatgg agaagtaccg cattggtacg 1200 ctgcaggacc ccaagacgtt ctatgcttgg ggagagtccg atttctaccc tgagttgaag 1260 cgccgggccc ttgcaaggct gaaggaggct ggtcaggcgc ggcgcggcgg ccttggggtg 1320 aaggccctcc tggtgctcac cctcttcttc gtgtcgtggt acatgtgggt ggcccacaag 1380 tccttcctct gggccgccgt ctggggcttc gccggctccc acgtcgggct gagcatccag 1440 cacgacggca accacggcgc gttcagccgc agcacactgg tgaaccgcct ggcggggtgg 1500 ggcatggact tgatcggcgc gtcgtcaacg gtgtgggagt accagcacgt catcggccac 1560 caccagtaca ccaacctcgt gtcggacacg ctattcagtc tgcctgagaa cgatccggac 1620 gtcttctcca gctacccgct gatgcgcatg cacccggata cggcgtggca gccgcaccac 1680 cgcttccagc acctgttcgc gttcccactg ttcgccctga tgacaatcag caaggtgctg 1740 accagcgatt tcgctgtctg cctcagcatg aagaaggggt ccatcgactg ctcctccagg 1800 ctcgtcccac tggaggggca gctgctgttc tggggggcca agctggcgaa cttcctgttg 1860 cagattgtgt tgccatgcta cctccacggg acagctatgg gcctggccct cttctctgtt 1920 gcccaccttg tgtcggggga gtacctcgcg atctgcttca tcatcaacca catcagcgag 1980 tcttgtgagt ttatgaatac aagctttcaa accgccgccc ggaggacaga gatgcttcag 2040 gcagcccatc aggcagcgga ggccaagaag gtgaagccca cccctccacc gaacgattgg 2100 gctgtgacac aggtccaatg ctgcgtgaat tggagatcag gtggcgtgtt ggccaatcac 2160 ctctctggag gcttgaacca ccagatcgag catcatctgt tccccagcat ctcgcatgcc 2220 aactacccca tcatcgcccg tgttgtgaag gaggtgtgcg aggagtatgg gttgccgtac 2280 aagaactacg tcacgttctg ggatgcagtc tgtggcatgg ttcagcacct ccggttgatg 2340 ggtgctccac cggtgccaac gaacggggac aaaaagtcat aa 2382
<210> 83 <211> 793 <212> PRT
<213> Euglena gracilis <400> 83
Met Ala Asp Ser Pro Val Ile Asn Leu Ser Thr Met Trp Lys Pro Leu 1 5 10 15
Ser Leu Met Ala Leu Asp Leu Ala Val Leu Gly His Val Trp Lys Gln 20 25 30
Ala Gln Gln Glu Gly Ser Ile Ser Ala Tyr Ala Asp Ser Val Trp Thr 35 40 45
Pro Leu Ile Met Ser Gly Leu Tyr Leu Ser Met Ile Phe Val Gly Cys 50 55 60
Arg Trp Met Lys Asn Arg Glu Pro Phe Glu Ile Lys Thr Tyr Met Phe 65 70 75 80
Ala Tyr Asn Leu Tyr Gln Thr Leu Met Asn Leu Cys Ile Val Leu Gly 85 90 95
Phe Leu Tyr Gln Val His Ala Thr Gly Met Arg Phe Trp Gly Ser Gly 100 105 110
Val Asp Arg Ser Pro Lys Gly Leu Gly Ile Gly Phe Phe Ile Tyr Ala 115 120 125
His Tyr His Asn Lys Tyr Val Glu Tyr Phe Asp Thr Leu Phe Met Val 130 135 140 Leu Arg Lys Lys Asn Asn Gln Ile Ser Phe Leu His Val Tyr His His 145 150 155 160
Ala Leu Leu Thr Trp Ala Trp Phe Ala Val Val Tyr Phe Ala Pro Gly 165 170 175
Gly Asp Gly Trp Phe Gly Ala Cys Tyr Asn Ser Ser Ile His Val Leu 180 185 190
Met Tyr Ser Tyr Tyr Leu Leu Ala Thr Phe Gly Ile Ser Cys Pro Trp 195 200 205
Lys Lys Ile Leu Thr Gln Leu Gln Met Val Gln Phe Cys Phe Cys Phe 210 215 220
Thr His Ser Ile Tyr Val Trp Ile Cys Gly Ser Glu Ile Tyr Pro Arg 225 230 235 240
Pro Leu Thr Ala Leu Gln Ser Phe Val Met Val Asn Met Leu Val Leu 245 250 255
Phe Gly Asn Phe Tyr Val Lys Gln Tyr Ser Gln Lys Asn Gly Lys Pro 260 265 270
Glu Asn Gly Ala Thr Pro Glu Asn Gly Ala Lys Pro Gln Pro Cys Glu 275 280 285
Asn Gly Thr Val Glu Lys Arg Glu Asn Asp Thr Ala Asn Val Arg Pro 290 295 300
Ala Arg Pro Ala Gly Leu Pro Pro Ala Thr Tyr Tyr Asp Ser Leu Ala 305 310 315 320
Val Ser Gly Gln Gly Lys Glu Arg Leu Phe Thr Thr Asp Glu Val Arg 325 330 335
Arg His Ile Leu Pro Thr Asp Gly Trp Leu Thr Cys His Glu Gly Val 340 345 350
Tyr Asp Val Thr Asp Phe Leu Ala Lys His Pro Gly Gly Gly Val Ile 355 360 365
Thr Leu Gly Leu Gly Arg Asp Cys Thr Ile Leu Val Glu Ser Tyr His 370 375 380 Ala His Leu Val Ser Gly Glu Tyr Leu Ala Ile Cys Phe Ile Ile Asn 645 650 655
His Ile Ser Glu Ser Cys Glu Phe Met Asn Thr Ser Phe Gln Thr Ala 660 665 670
Ala Arg Arg Thr Glu Met Leu Gln Ala Ala His Gln Ala Ala Glu Ala 675 680 685
Lys Lys Val Lys Pro Thr Pro Pro Pro Asn Asp Trp Ala Val Thr Gln 690 695 700
Val Gln Cys Cys Val Asn Trp Arg Ser Gly Gly Val Leu Ala Asn His 705 710 715 720
Leu Ser Gly Gly Leu Asn His Gln Ile Glu His His Leu Phe Pro Ser 725 730 735
Ile Ser His Ala Asn Tyr Pro Ile Ile Ala Arg Val Val Lys Glu Val 740 745 750
Cys Glu Glu Tyr Gly Leu Pro Tyr Lys Asn Tyr Val Thr Phe Trp Asp 755 760 765
Ala Val Cys Gly Met Val Gln His Leu Arg Leu Met Gly Ala Pro Pro 770 775 780
Val Pro Thr Asn Gly Asp Lys Lys Ser 785 790
<210> 84 <211> 541 <212> PRT
<213> Euglena gracilis <400> 84
Met Leu Val Leu Phe Gly Asn Phe Tyr Val Lys Gln Tyr Ser Gln Lys 15 10 15
Asn Gly Lys Pro Glu Asn Gly Ala Thr Pro Glu Asn Gly Ala Lys Pro 20 25 30
Gln Pro Cys Glu Asn Gly Thr Val Glu Lys Arg Glu Asn Asp Thr Ala 35 40 45
Asn Val Arg Pro Thr Arg Pro Ala Gly Pro Pro Pro Ala Thr Tyr Tyr 50 55 60 Asp Ser Leu Ala Val Ser Gly Gln Gly Lys Glu Arg Leu Phe Thr Thr 65 70 75 80
Asp Glu Val Arg Arg His Ile Leu Pro Thr Asp Gly Trp Leu Thr Cys 85 90 95
His Glu Gly Val Tyr Asp Val Thr Asp Phe Leu Ala Lys His Pro Gly 100 105 110
Gly Gly Val Ile Thr Leu Gly Leu Gly Arg Asp Cys Thr Ile Leu Ile 115 120 125
Glu Ser Tyr His Pro Ala Gly Arg Pro Asp Lys Val Met Glu Lys Tyr 130 135 140
Arg Ile Gly Thr Leu Gln Asp Pro Lys Thr Phe Tyr Ala Trp Gly Glu 145 150 155 160
Ser Asp Phe Tyr Pro Glu Leu Lys Arg Arg Ala Leu Ala Arg Leu Lys 165 170 175
Glu Ala Gly Gln Ala Arg Arg Gly Gly Leu Gly Val Lys Ala Leu Leu 180 185 190
Val Leu Thr Leu Phe Phe Val Ser Trp Tyr Met Trp Val Ala His Lys 195 200 205
Ser Phe Leu Trp Ala Ala Val Trp Gly Phe Ala Gly Ser His Val Gly 210 215 220
Leu Ser Ile Gln His Asp Gly Asn His Gly Ala Phe Ser Arg Asn Thr 225 230 235 240
Leu Val Asn Arg Leu Ala Gly Trp Gly Met Asp Leu Ile Gly Ala Ser 245 250 255
Ser Thr Val Trp Glu Tyr Gln His Val Ile Gly His His Gln Tyr Thr 260 265 270
Asn Leu Val Ser Asp Thr Leu Phe Ser Leu Pro Glu Asn Asp Pro Asp 275 280 285
Val Phe Ser Ser Tyr Pro Leu Met Arg Met His Pro Asp Thr Ala Trp 290 295 300 Gln Pro His His Arg Phe Gln His Leu Phe Ala Phe Pro Leu Phe Ala 305 310 315 320
Leu Met Thr Ile Ser Lys Val Leu Thr Ser Asp Phe Ala Val Cys Leu 325 330 335
Ser Met Lys Lys Gly Ser Ile Asp Cys Ser Ser Arg Leu Val Pro Leu 340 345 350
Glu Gly Gln Leu Leu Phe Trp Gly Ala Lys Leu Ala Asn Phe Leu Leu 355 360 365
Gln Ile Val Leu Pro Cys Tyr Leu His Gly Thr Ala Met Gly Leu Ala 370 375 380
Leu Phe Ser Val Ala His Leu Val Ser Gly Glu Tyr Leu Ala Ile Cys 385 390 395 400
Phe Ile Ile Asn His Ile Ser Glu Ser Cys Glu Phe Met Asn Thr Ser 405 410 415
Phe Gln Thr Ala Ala Arg Arg Thr Glu Met Leu Gln Ala Ala His Gln 420 425 430
Ala Ala Glu Ala Lys Lys Val Lys Pro Thr Pro Pro Pro Asn Asp Trp 435 440 445
Ala Val Thr Gln Val Gln Cys Cys Val Asn Trp Arg Ser Gly Gly Val 450 455 460
Leu Ala Asn His Leu Ser Gly Gly Leu Asn His Gln Ile Glu His His 465 470 475 480
Leu Phe Pro Ser Ile Ser His Ala Asn Tyr Pro Thr Ile Ala Pro Val 485 490 495
Val Lys Glu Val Cys Glu Glu Tyr Gly Leu Pro Tyr Lys Asn Tyr Val 500 505 510
Thr Phe Trp Asp Ala Val Cys Gly Met Val Gln His Leu Arg Leu Met 515 520 525
Gly Ala Pro Pro Val Pro Thr Asn Gly Asp Lys Lys Ser 530 535 540
<210> 85 <211> 300 <212> PRT
<213> Ostreococcus tauri <400> 85
Met Ser Ala Ser Gly Ala Leu Leu Pro Ala Ile Ala Ser Ala Ala Tyr 1 5 10 15
Ala Tyr Ala Thr Tyr Ala Tyr Ala Phe Glu Trp Ser His Ala Asn Gly 20 25 30
Ile Asp Asn Val Asp Ala Arg Glu Trp Ile Gly Ala Leu Ser Leu Arg 35 40 45
Leu Pro Ala Ile Ala Thr Thr Met Tyr Leu Leu Phe Cys Leu Val Gly 50 55 60
Pro Arg Leu Met Ala Lys Arg Glu Ala Phe Asp Pro Lys Gly Phe Met 65 70 75 80
Leu Ala Tyr Asn Ala Tyr Gln Thr Ala Phe Asn Val Val Val Leu Gly 85 90 95
Met Phe Ala Arg Glu Ile Ser Gly Leu Gly Gln Pro Val Trp Gly Ser 100 105 110
Thr Met Pro Trp Ser Asp Arg Lys Ser Phe Lys Ile Leu Leu Gly Val 115 120 125
Trp Leu His Tyr Asn Asn Lys Tyr Leu Glu Leu Leu Asp Thr Val Phe 130 135 140
Met Val Ala Arg Lys Lys Thr Lys Gln Leu Ser Phe Leu His Val Tyr 145 150 155 160
His His Ala Leu Leu Ile Trp Ala Trp Trp Leu Val Cys His Leu Met 165 170 175
Ala Thr Asn Asp Cys Ile Asp Ala Tyr Phe Gly Ala Ala Cys Asn Ser 180 185 190
Phe Ile His Ile Val Met Tyr Ser Tyr Tyr Leu Met Ser Ala Leu Gly 195 200 205
Ile Arg Cys Pro Trp Lys Arg Tyr Ile Thr Gln Ala Gln Met Leu Gln 210 215 220
Phe Val Ile Val Phe Ala His Ala Val Phe Val Leu Arg Gln Lys His 225 230 235 240 Pro Ala Gly Arg Pro Asp Lys Val Met Glu Lys Tyr Arg Ile Gly Thr 385 390 395 400
Leu Gln Asp Pro Lys Thr Phe Tyr Ala Trp Gly Glu Ser Asp Phe Tyr 405 410 415
Pro Glu Leu Lys Arg Arg Ala Leu Ala Arg Leu Lys Glu Ala Gly Gln 420 425 430
Ala Arg Arg Gly Gly Leu Gly Val Lys Ala Leu Leu Val Leu Thr Leu 435 440 445
Phe Phe Val Ser Trp Tyr Met Trp Val Ala His Lys Ser Phe Leu Trp 450 455 460
Ala Ala Val Trp Gly Phe Ala Gly Ser His Val Gly Leu Ser Ile Gln 465 470 475 480
His Asp Gly Asn His Gly Ala Phe Ser Arg Ser Thr Leu Val Asn Arg 485 490 495
Leu Ala Gly Trp Gly Met Asp Leu Ile Gly Ala Ser Ser Thr Val Trp 500 505 510
Glu Tyr Gln His Val Ile Gly His His Gln Tyr Thr Asn Leu Val Ser 515 520 525
Asp Thr Leu Phe Ser Leu Pro Glu Asn Asp Pro Asp Val Phe Ser Ser 530 535 540
Tyr Pro Leu Met Arg Met His Pro Asp Thr Ala Trp Gln Pro His His 545 550 555 560
Arg Phe Gln His Leu Phe Ala Phe Pro Leu Phe Ala Leu Met Thr Ile 565 570 575
Ser Lys Val Leu Thr Ser Asp Phe Ala Val Cys Leu Ser Met Lys Lys 580 585 590
Gly Ser Ile Asp Cys Ser Ser Arg Leu Val Pro Leu Glu Gly Gln Leu 595 600 605
Leu Phe Trp Gly Ala Lys Leu Ala Asn Phe Leu Leu Gln Ile Val Leu 610 615 620
Pro Cys Tyr Leu His Gly Thr Ala Met Gly Leu Ala Leu Phe Ser Val 625 630 635 640 Cys Pro Val Thr Leu Pro Trp Ala Gln Met Phe Val Met Thr Asn Met 245 250 255
Leu Val Leu Phe Gly Asn Phe Tyr Leu Lys Ala Tyr Ser Asn Lys Ser 260 265 270
Arg Gly Asp Gly Ala Ser Ser Val Lys Pro Ala Glu Thr Thr Arg Ala 275 280 285
Pro Ser Val Arg Arg Thr Arg Ser Arg Lys Ile Asp 290 295 300
<210> 86 <211> 358 <212> PRT
<213> Thalassiosira pseudonana <4 00> 86
Met Cys Ser Ser Pro Pro Ser Gln Ser Lys Thr Thr Ser Leu Leu Ala 15 10 15
Arg Tyr Thr Thr Ala Ala Leu Leu Leu Leu Thr Leu Thr Thr Trp Cys 20 25 30
His Phe Ala Phe Pro Ala Ala Thr Ala Thr Pro Gly Leu Thr Ala Glu 35 40 45
Met His Ser Tyr Lys Val Pro Leu Gly Leu Thr Val Phe Tyr Leu Leu 50 55 60
Ser Leu Pro Ser Leu Lys Tyr Val Thr Asp Asn Tyr Leu Ala Lys Lys 65 70 75 80
Tyr Asp Met Lys Ser Leu Leu Thr Glu Ser Met Val Leu Tyr Asn Val 85 90 95
Ala Gln Val Leu Leu Asn Gly Trp Thr Val Tyr Ala Ile Val Asp Ala 100 105 110
Val Met Asn Arg Asp His Pro Phe Ile Gly Ser Arg Ser Leu Val Gly 115 120 125
Ala Ala Leu His Ser Gly Ser Ser Tyr Ala Val Trp Val His Tyr Cys 130 135 140
Asp Lys Tyr Leu Glu Phe Phe Asp Thr Tyr Phe Met Val Leu Arg Gly 145 150 155 160 Lys Met Asp Gln Val Ser Phe Leu His Ile Tyr His His Thr Thr Ile 165 170 175
Ala Trp Ala Trp Trp Ile Ala Leu Arg Phe Ser Pro Gly Gly Asp Ile 180 185 190
Tyr Phe Gly Ala Leu Leu Asn Ser Ile Ile His Val Leu Met Tyr Ser 195 200 205
Tyr Tyr Ala Leu Ala Leu Leu Lys Val Ser Cys Pro Trp Lys Arg Tyr 210 215 220
Leu Thr Gln Ala Gln Leu Leu Gln Phe Thr Ser Val Val Val Tyr Thr 225 230 235 240
Gly Cys Thr Gly Tyr Thr His Tyr Tyr His Thr Lys His Gly Ala Asp 245 250 255
Glu Thr Gln Pro Ser Leu Gly Thr Tyr Tyr Phe Cys Cys Gly Val Gln 260 265 270
Val Phe Glu Met Val Ser Leu Phe Val Leu Phe Ser Ile Phe Tyr Lys
Arg Ser Tyr Ser Lys Lys Asn Lys Ser Gly Gly Lys Asp Ser Lys Lys 290 295 300
Asn Asp Asp Gly Asn Asn Glu Asp Gln Cys His Lys Ala Met Lys Asp 305 310 315 320
Ile Ser Glu Gly Ala Lys Glu Val Val Gly His Ala Ala Lys Asp Ala 325 330 335
Gly Lys Leu Val Ala Thr Ala Ser Lys Ala Val Lys Arg Lys Gly Thr 340 345 350
Arg Val Thr Gly Ala Met 355
<210> 87 <211> 515
<212> PRT
<213> Thraustochytrium aureum
<4 00> 87
Met Thr Val Gly Phe Asp Glu Thr Val Thr Met Asp Thr Val Arg Asn 15 10 15 His Asn Met Pro Asp Asp Ala Trp Cys Ala Ile His Gly Thr Val Tyr 20 25 30
Asp Ile Thr Lys Phe Ser Lys Val His Pro Gly Gly Asp Ile Ile Met 35 40 45
Leu Ala Ala Gly Lys Glu Ala Thr Ile Leu Phe Glu Thr Tyr His Ile 50 55 60
Lys Gly Val Pro Asp Ala Val Leu Arg Lys Tyr Lys Val Gly Lys Leu 65 70 75 80
Pro Gln Gly Lys Lys Gly Glu Thr Ser His Met Pro Thr Gly Leu Asp 85 90 95
Ser Ala Ser Tyr Tyr Ser Trp Asp Ser Glu Phe Tyr Arg Val Leu Arg 100 105 110
Glu Arg Val Ala Lys Lys Leu Ala Glu Pro Gly Leu Met Gln Arg Ala 115 120 125
Arg Met Glu Leu Trp Ala Lys Ala Ile Phe Leu Leu Ala Gly Phe Trp 130 135 140
Gly Ser Leu Tyr Ala Met Cys Val Leu Asp Pro His Gly Gly Ala Met 145 150 155 160
Val Ala Ala Val Thr Leu Gly Val Phe Ala Ala Phe Val Gly Thr Cys 165 170 175
Ile Gln His Asp Gly Ser His Gly Ala Phe Ser Lys Ser Arg Phe Met 180 185 190
Asn Lys Ala Ala Gly Trp Thr Leu Asp Met Ile Gly Ala Ser Ala Met 195 200 205
Thr Trp Glu Met Gln His Val Leu Gly His His Pro Tyr Thr Asn Leu 210 215 220
Ile Glu Met Glu Asn Gly Leu Ala Lys Val Lys Gly Ala Asp Val Asp 225 230 235 240
Pro Lys Lys Val Asp Gln Glu Ser Asp Pro Asp Val Phe Ser Thr Tyr 245 250 255 Pro Met Leu Arg Leu His Pro Trp His Arg Gln Arg Phe Tyr His Lys 260 265 270
Phe Gln His Leu Tyr Ala Pro Leu Ile Phe Gly Phe Met Thr Ile Asn 275 280 285
Lys Val Ile Ser Gln Asp Val Gly Val Val Leu Arg Lys Arg Leu Phe 290 295 300
Gln Ile Asp Ala Asn Cys Arg Tyr Gly Ser Pro Trp Asn Val Ala Arg 305 310 315 320
Phe Trp Ile Met Lys Leu Leu Thr Thr Leu Tyr Met Val Ala Leu Pro 325 330 335
Met Tyr Met Gln Gly Pro Ala Gln Gly Leu Lys Leu Phe Phe Met Ala 340 345 350
His Phe Thr Cys Gly Glu Val Leu Ala Thr Met Phe Ile Val Asn His 355 360 365
Ile Ile Glu Gly Val Ser Tyr Ala Ser Lys Asp Ala Val Lys Gly Val 370 375 380
Met Ala Pro Pro Arg Thr Val His Gly Val Thr Pro Met Gln Val Thr 385 390 395 400
Gln Lys Ala Leu Ser Ala Ala Glu Ser Thr Lys Ser Asp Ala Asp Lys 405 410 415
Thr Thr Met Ile Pro Leu Asn Asp Trp Ala Ala Val Gln Cys Gln Thr 420 425 430
Ser Val Asn Trp Ala Val Gly Ser Trp Phe Trp Asn His Phe Ser Gly 435 440 445
Gly Leu Asn His Gln Ile Glu His His Cys Phe Pro Gln Asn Pro His 450 455 460
Thr Val Asn Val Tyr Ile Ser Gly Ile Val Lys Glu Thr Cys Glu Glu 465 470 475 480
Tyr Gly Val Pro Tyr Gln Ala Glu Ile Ser Leu Phe Ser Ala Tyr Phe 485 490 495
Lys Met Leu Ser His Leu Arg Thr Leu Gly Asn Glu Asp Leu Thr Ala 500 505 510 Trp Ser Thr 515
<210> 88 <211> 509 <212> PRT
<213> Schizochytrium aggregatum <400> 88
Met Thr Val Gly Gly Asp Glu Val Tyr Ser Met Ala Gln Val Arg Asp 1 5 10 15
His Asn Thr Pro Asp Asp Ala Trp Cys Ala Ile His Gly Glu Val Tyr 20 25 30
Glu Leu Thr Lys Phe Ala Arg Thr His Pro Gly Gly Asp Ile Ile Leu 35 40 45
Leu Ala Ala Gly Lys Glu Ala Thr Ile Leu Phe Glu Thr Tyr His Val 50 55 60
Arg Pro Ile Ser Asp Ala Val Leu Arg Lys Tyr Arg Ile Gly Lys Leu 65 70 75 80
Ala Ala Ala Gly Lys Asp Glu Pro Ala Asn Asp Ser Thr Tyr Tyr Ser 85 90 95
Trp Asp Ser Asp Phe Tyr Lys Val Leu Arg Gln Arg Val Val Ala Arg 100 105 110
Leu Glu Glu Arg Lys Ile Ala Arg Arg Gly Gly Pro Glu Ile Trp Ile 115 120 125
Lys Ala Ala Ile Leu Val Ser Gly Phe Trp Ser Met Leu Tyr Leu Met 130 135 140
Cys Thr Leu Asp Pro Asn Arg Gly Ala Ile Leu Ala Ala Ile Ala Leu 145 150 155 160
Gly Ile Val Ala Ala Phe Val Gly Thr Cys Ile Gln His Asp Gly Asn 165 170 175
His Gly Ala Phe Ala Phe Ser Pro Phe Met Asn Lys Leu Ser Gly Trp 180 185 190
Thr Leu Asp Met Ile Gly Ala Ser Ala Met Thr Trp Glu Met Gln His 195 200 205 Val Leu Gly His His Pro Tyr Thr Asn Leu Ile Glu Met Glu Asn Gly 210 215 220
Thr Gln Lys Val Thr His Ala Asp Val Asp Pro Lys Lys Ala Asp Gln 225 230 235 240
Glu Ser Asp Pro Asp Val Phe Ser Thr Tyr Pro Met Leu Arg Leu His 245 250 255
Pro Trp His Arg Lys Arg Phe Tyr His Arg Phe Gln His Leu Tyr Ala 260 265 270
Pro Leu Leu Phe Gly Phe Met Thr Ile Asn Lys Val Ile Thr Gln Asp 275 280 285
Val Gly Val Val Leu Ser Lys Arg Leu Phe Gln Ile Asp Ala Asn Cys 290 295 300
Arg Tyr Ala Ser Lys Ser Tyr Val Ala Arg Phe Trp Ile Met Lys Leu 305 310 315 320
Leu Thr Val Leu Tyr Met Val Ala Leu Pro Val Tyr Thr Gln Gly Leu 325 330 335
Val Asp Gly Leu Lys Leu Phe Phe Ile Ala His Phe Ser Cys Gly Glu 340 345 350
Leu Leu Ala Thr Met Phe Ile Val Asn His Ile Ile Glu Gly Val Ser 355 360 365
Tyr Ala Ser Lys Asp Ser Val Lys Gly Thr Met Ala Pro Pro Arg Thr 370 375 380
Val His Gly Val Thr Pro Met His Asp Thr Arg Asp Ala Leu Gly Lys 385 390 395 400
Glu Lys Ala Ala Thr Lys His Val Pro Leu Asn Asp Trp Ala Ala Val 405 410 415
Gln Cys Gln Thr Ser Val Asn Trp Ser Ile Gly Ser Trp Phe Trp Asn 420 425 430
His Phe Ser Gly Gly Leu Asn His Gln Ile Glu His His Leu Phe Pro 435 440 445 Gly Leu Thr His Thr Thr Tyr Val Tyr Ile Gln Asp Val Val Gln Ala 450 455 460
Thr Cys Ala Glu Tyr Gly Val Pro Tyr Gln Ser Glu Gln Ser Leu Phe 465 470 475 480
Ser Ala Tyr Phe Lys Met Leu Ser His Leu Arg Ala Leu Gly Asn Glu 485 490 495
Pro Met Pro Ser Trp Glu Lys Asp His Pro Lys Ser Lys 500 505
<210> 89 <211> 550 <212> PRT
<213> Thalassiosira pseudonana <4 00> 89
Met Gly Asn Gly Asn Leu Pro Ala Ser Thr Ala Gln Leu Lys Ser Thr 15 10 15
Ser Lys Pro Gln Gln Gln His Glu His Arg Thr Ile Ser Lys Ser Glu 20 25 30
Leu Ala Gln His Asn Thr Pro Lys Ser Ala Trp Cys Ala Val His Ser 35 40 45
Thr Pro Ala Thr Asp Pro Ser His Ser Asn Asn Lys Gln His Ala His 50 55 60
Leu Val Leu Asp Ile Thr Asp Phe Ala Ser Arg His Pro Gly Gly Asp 65 70 75 80
Leu Ile Leu Leu Ala Ser Gly Lys Asp Ala Ser Val Leu Phe Glu Thr 85 90 95
Tyr His Pro Arg Gly Val Pro Thr Ser Leu Ile Gln Lys Leu Gln Ile 100 105 110
Gly Val Met Glu Glu Glu Ala Phe Arg Asp Ser Phe Tyr Ser Trp Thr 115 120 125
Asp Ser Asp Phe Tyr Thr Val Leu Lys Arg Arg Val Val Glu Arg Leu 130 135 140
Glu Glu Arg Gly Leu Asp Arg Arg Gly Ser Lys Glu Ile Trp Ile Lys 145 150 155 160 Ala Leu Phe Leu Leu Val Gly Phe Trp Tyr Cys Leu Tyr Lys Met Tyr 165 170 175
Thr Thr Ser Asp Ile Asp Gln Tyr Gly Ile Ala Ile Ala Tyr Ser Ile 180 185 190
Gly Met Gly Thr Phe Ala Ala Phe Ile Gly Thr Cys Ile Gln His Asp 195 200 205
Gly Asn His Gly Ala Phe Ala Gln Asn Lys Leu Leu Asn Lys Leu Ala 210 215 220
Gly Trp Thr Leu Asp Met Ile Gly Ala Ser Ala Phe Thr Trp Glu Leu 225 230 235 240
Gln His Met Leu Gly His His Pro Tyr Thr Asn Val Leu Asp Gly Val 245 250 255
Glu Glu Glu Arg Lys Glu Arg Gly Glu Asp Val Ala Leu Glu Glu Lys 260 265 270
Asp Gln Glu Ser Asp Pro Asp Val Phe Ser Ser Phe Pro Leu Met Arg 275 280 285
Met His Pro His His Thr Thr Ser Trp Tyr His Lys Tyr Gln His Leu 290 295 300
Tyr Ala Pro Pro Leu Phe Ala Leu Met Thr Leu Ala Lys Val Phe Gln 305 310 315 320
Gln Asp Phe Glu Val Ala Thr Ser Gly Arg Leu Tyr His Ile Asp Ala 325 330 335
Asn Val Arg Tyr Gly Ser Val Trp Asn Val Met Arg Phe Trp Ala Met 340 345 350
Lys Val Ile Thr Met Gly Tyr Met Met Gly Leu Pro Ile Tyr Phe His 355 360 365
Gly Val Leu Arg Gly Val Gly Leu Phe Val Ile Gly His Leu Ala Cys 370 375 380
Gly Glu Leu Leu Ala Thr Met Phe Ile Val Asn His Val Ile Glu Gly 385 390 395 400
Val Ser Tyr Gly Thr Lys Asp Leu Val Gly Gly Ala Ser His Gly Asp 405 410 415 Glu Lys Lys Ile Val Lys Pro Thr Thr Val Leu Gly Asp Thr Pro Met 420 425 430
Glu Lys Thr Arg Glu Glu Ala Leu Lys Ser Asn Ser Asn Asn Asn Lys 435 440 445
Lys Lys Gly Glu Lys Asn Ser Val Pro Ser Val Pro Phe Asn Asp Trp 450 455 460
Ala Ala Val Gln Cys Gln Thr Ser Val Asn Trp Ser Pro Gly Ser Trp 465 470 475 480
Phe Trp Asn His Phe Ser Gly Gly Leu Ser His Gln Ile Glu His His 485 490 495
Leu Phe Pro Ser Ile Cys His Thr Asn Tyr Cys His Ile Gln Asp Val 500 505 510
Val Glu Ser Thr Cys Ala Glu Tyr Gly Val Pro Tyr Gln Ser Glu Ser 515 520 525
Asn Leu Phe Val Ala Tyr Gly Lys Met Ile Ser His Leu Lys Phe Leu 530 535 540
Gly Lys Ala Lys Cys Glu 545 550 <210> 90 <211> 433 <212> PRT <213> Isochrysis galbana <400> 90
Met Cys Asn Ala Ala Gln Val
10 15
Ala Ala Lys Pro Thr Trp Thr Lys Ile His Gly Arg Thr Val Asp Val 20 25 30
Glu Thr Phe Arg His Pro Gly Gly Asn Ile Leu Asp Leu Phe Leu Gly 35 40 45
Met Asp Ala Thr Thr Ala Phe Glu Thr Phe His Gly His His LyS Gly 50 55 60
Ala Trp Lys Met Leu Lys Thr Leu Pro Glu Lys Glu Val Ala Ala Ala 65 70 75 80 Asp Ile Pro Ala Gln Lys Glu Glu His Val Ala Glu Met Thr Arg Leu 85 90 95
Met Ala Ser Trp Arg Glu Arg Gly Leu Phe Lys Pro Arg Pro Val Ala 100 105 110
Ser Ser Ile Tyr Gly Leu Cys Val Ile Phe Ala Ile Ala Ala Ser Val 115 120 125
Ala Cys Ala Pro Tyr Ala Pro Val Leu Ala Gly Ile Ala Val Gly Thr 130 135 140
Cys Trp Ala Gln Cys Gly Phe Leu Gln His Met Gly Gly His Arg Glu 145 150 155 160
Trp Gly Arg Thr Trp Ser Phe Ala Phe Gln His Leu Phe Glu Gly Leu 165 170 175
Leu Lys Gly Gly Ser Ala Ser Trp Trp Arg Asn Arg His Asn Lys His 180 185 190
His Ala Lys Thr Asn Val Leu Gly Glu Asp Gly Asp Leu Arg Thr Thr 195 200 205
Pro Phe Phe Ala Trp Asp Pro Thr Leu Ala Lys Lys Val Pro Asp Trp 210 215 220
Ser Leu Arg Thr Gln Ala Phe Thr Phe Leu Pro Ala Leu Gly Ala Tyr 225 230 235 240
Val Phe Val Phe Ala Phe Thr Val Arg Lys Tyr Ser Val Val Lys Arg 245 250 255
Leu Trp His Glu Val Ala Leu Met Val Ala His Tyr Ala Leu Phe Ser 260 265 270
Trp Ala Leu Ser Ala Ala Gly Ala Ser Leu Ser Ser Gly Leu Thr Phe 275 280 285
Tyr Cys Thr Gly Tyr Ala Trp Gln Gly Ile Tyr Leu Gly Phe Phe Phe 290 295 300
Gly Leu Ser His Phe Ala Val Glu Arg Val Pro Ser Thr Ala Thr Trp 305 310 315 320 Leu Glu Ser Thr Met Met Gly Thr Val Asp Trp Gly Gly Ser Ser Ala 325 330 335
Phe Cys Gly Tyr Leu Ser Gly Phe Leu Asn Ile Gln Ile Glu His His 340 345 350
Met Ala Pro Gln Met Pro Met Glu Asn Leu Arg Gln Ile Arg Ala Asp 355 360 365
Cys Lys Ala Ala Ala His Lys Phe Gly Leu Pro Tyr Arg Glu Leu Thr 370 375 380
Phe Val Ala Ala Thr Lys Leu Met Met Ser Gly Leu Tyr Arg Thr Gly 385 390 395 400
Lys Asp Glu Leu Lys Leu Arg Ala Asp Arg Arg Lys Phe Thr Arg Ala 405 410 415
Gln Ala Tyr Met Gly Ala Ala Ser Ala Leu Val Asp Thr Leu Lys Ala 420 425 430
Asp
<210> 91
<211> 909
<212> DNA
<213> Euglena gracilis
<400> 91
atggcggata gcccagtcat caacctcagc accatgtgga aacccctttc actgatggct 60 ttggaccttg ccgttttggg acatgtctgg aagcaggcac aacaggaggg cagcatttcg 120 gcctatgctg attctgtttg gactcctctc attatgtccg gtttatactt atcaatgatc 180 ttcgtggggt gccgctggat gaagaaccgt gaaccctttg agatcaaaac atacatgttt 240 gcgtataacc tgtatcagac cttgatgaac ctttgcatcg tgttgggatt cttgtaccag 300 gtgcatgcca ctgggatgcg cttttgggga agtggtgtcg accgaagccc aaaaggtttg 360 ggcattggct tcttcattta tgcccactac cacaacaagt atgtggaata ttttgataca 420 ctttttatgg tgctgcgaaa gaagaacaac cagatttctt tccttcacgt gtatcatcat 480 gccctgttga catgggcttg gtttgctgtt gtgtatttcg cacctggagg tgatggctgg 540 tttggagctt gctacaattc ttccatccat gtcctgatgt actcttacta cttgcttgca 600 acttttggca tcagttgccc atggaagaag atcttgacac agctccagat ggttcaattc 660 tgtttctgtt ttacacattc catttatgtg tggatttgcg ggtcagagat ctacccacgg 720 cctctgactg ctttgcagtc gttcgtgatg gtcaatatgt tggtgctgtt tggcaatttc 780
tatgtcaagc aatactccca aaagaacggc aagccggaga acggagccac ccctgagaac 840
ggagcgaagc cgcaaccttg cgagaacggc acggtggaaa agcgagagaa tgacaccgcc 900
aacgttcgg 909
<210> 92 <211> 303 <212> PRT
<213> Euglena gracilis <400> 92
Met Ala Asp Ser Pro Val Ile Asn Leu Ser Thr Met Trp Lys Pro Leu 15 10 15
Ser Leu Met Ala Leu Asp Leu Ala Val Leu Gly His Val Trp Lys Gln 20 25 30
Ala Gln Gln Glu Gly Ser Ile Ser Ala Tyr Ala Asp Ser Val Trp Thr 35 40 45
Pro Leu Ile Met Ser Gly Leu Tyr Leu Ser Met Ile Phe Val Gly Cys 50 55 60
Arg Trp Met Lys Asn Arg Glu Pro Phe Glu Ile Lys Thr Tyr Met Phe 65 70 75 80
Ala Tyr Asn Leu Tyr Gln Thr Leu Met Asn Leu Cys Ile Val Leu Gly 85 90 95
Phe Leu Tyr Gln Val His Ala Thr Gly Met Arg Phe Trp Gly Ser Gly 100 105 110
Val Asp Arg Ser Pro Lys Gly Leu Gly Ile Gly Phe Phe Ile Tyr Ala 115 120 125
His Tyr His Asn Lys Tyr Val Glu Tyr Phe Asp Thr Leu Phe Met Val 130 135 140
Leu Arg Lys Lys Asn Asn Gln Ile Ser Phe Leu His Val Tyr His His 145 150 155 160
Ala Leu Leu Thr Trp Ala Trp Phe Ala Val Val Tyr Phe Ala Pro Gly 165 170 175
Gly Asp Gly Trp Phe Gly Ala Cys Tyr Asn Ser Ser Ile His Val Leu 180 185 190 Met Tyr Ser Tyr Tyr Leu Leu Ala Thr Phe Gly Ile Ser Cys Pro Trp 195 200 205
Lys Lys Ile Leu Thr Gln Leu Gln Met Val Gln Phe Cys Phe Cys Phe 210 215 220
Thr His Ser Ile Tyr Val Trp Ile Cys Gly Ser Glu Ile Tyr Pro Arg 225 230 235 240
Pro Leu Thr Ala Leu Gln Ser Phe Val Met Val Asn Met Leu Val Leu 245 250 255
Phe Gly Asn Phe Tyr Val Lys Gln Tyr Ser Gln Lys Asn Gly Lys Pro 260 265 270
Glu Asn Gly Ala Thr Pro Glu Asn Gly Ala Lys Pro Gln Pro Cys Glu 275 280 285
Asn Gly Thr Val Glu Lys Arg Glu Asn Asp Thr Ala Asn Val Arg 290 295 300
<210> 93
<211> 4
<212> PRT
<213> Seqüência Artificial <220>
<223> ng motif
<400> 93
Lys Asn Gly Lys 1
<210> 94
<211> 5
<212> PRT
<213> Seqüência Artificial <220>
<223> unidade ng
<400> 94
Pro Glu Asn Gly Ala 1 5
<210> 95
<211> 5
<212> PRT
<213> Seqüência Artificial <220>
<223> unidade ng <400> 95
Pro Glu Asn Gly Ala 1 5
<210> 96 <211> 7 <212> PRT <213> Seqüência Artificial <220> <223> unidade ng <4 00> 96 Pro Cys 1 Glu Asn Gly Thr Val
<210> 97 <211> 54 <212> DNA <213> Seqüência Artificial <220> <223> EgDHAsynl proline-ligante <4 00> 97
cccgcccgtc cagctggact cccgccggcc acgtactacg actccctggc agtg 54
<210> 98 <211> 18 <212> PRT <213> Seqüência Artificial <220> <223> EgDHAsynl proline-ligante <400> 98
Pro Ala Arg Pro Ala Gly Leu Pro Pro Ala Thr Tyr Tyr Asp Ser Leu 15 10 15
Ala Val
<210> 99
<211> 3668
<212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo pLF121-l <220>
<221> misc_feature
<222> (3616)..(3655)
<223> η é a, c, g, ou t
<400> 99 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac 60 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta 120 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta 180 cattgcacaa gataaaaata tatcatcatg ttagaaaaac tcatcgagca tcaaatgaaa 240 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa 300 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc 360 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt 420 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg 480 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc 540 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct 600 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc 660 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc 720 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt 780 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt 840 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa 900 tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa 960 atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc gttgaatatg 1020 gctcatagat cttttctcca tcactgatag ggagtggtaa aataactcca tcaatgatag 1080 agtgtcaaca acatgaccaa aatcccttaa cgtgagttac gcgtattaat tgcgttgcgc 1140 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 1200 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 1260 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 1320 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 1380 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 1440 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 1500 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 1560 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc 1620 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 1680 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 1740 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 1800 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggttacac tagaagaaca 1860 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 1920 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 1980 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 2040 cagggaacga cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg 2100 tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc 2160 accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca 2220 ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct 2280 ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt 2340 tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggcccca aataatgatt 2400 ttattttgac tgatagtgac ctgttcgttg caacaaattg atgagcaatg cttttttata 2460 atgccaactt tgtacaaaaa agttggtttt tttcggtcta aaatggaagc agccaaagaa 2520 ttggtttcca tcgtccaaga ggagctcccc aaggtggact atgcccagct ttggcaggat 2580 gccagcagct gtgaggtcct ttacctctcg gtggcattcg tggcgatcaa gttcatgctg 2640 cgcccactgg acctgaagcg ccaggccacc ttgaagaagc tgttcacagc atacaacttc 2700 ctcatgtcga tctattcctt tggctccttc ctggccatgg cctatgccct atcagtaact 2760 ggcatcctct ccggcgactg tgagacggcg ttcaacaacg atgtgttcag gatcacaact 2820 cagctgttct acctcagcaa gttcgtagag tacatcgact ccttctacct tccccttatg 2880 gacaagccac tgtcgttcct tcagttcttc catcatttgg gggcccccat tgacatgtgg 2940 ctattctaca aataccgcaa cgaaggagtc tggatctttg tcctgttgaa tgggttcatt 3000 cactggatca tgtacggtta ctattggacg cggctcatca agctgaactt ccccatgccc 3060 aagaacctga tcacctccat gcagatcatc cagttcaatg tcgggttcta catcgtctgg 3120 aagtaccgca atgtgccatg ctaccgccag gatgggatgc gcatgtttgc ctggatcttc 3180 aactactggt atgtcgggac ggtcttgctg ctgttcctca acttttacgt gcagacgtac 3240 atccggaagc cgaggaagaa ccgagggaag aaggagtagg ccacatggcg cctgcgctgg 3300 aggaaacggt acgctcggat ggtgcactgc acttgcactc cgccgtttct agcctcccct 3360 cgctctaacc actgcggcat gcctgcttga ggcgtgacgt tgcctcgtat gatacagttt 3420 acacccttcc cacagcccac ggagctggtg actgtttcca gcgtctgcag atcattgatc 3480 tggtgcaatg tgcacagacc aagcccctct aacgtcttgc ggtgtaccgc tcgacactca 3540 ctgcaagaga cagatggctg agcatgttat agccccttac attctaccct tcgtcccaac 3600 ctgaccgtca cattcnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnaccca 3660 actttctt 3668 <210> 100 <211> 3684 <212> DNA <213> Seqüência Artificial <220> <223> pLF121-l <220> <221> misc feature <222> (3632)..(3671) <223> η é a, c, g, ou t <400> 100 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac 60 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta 120 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta 180 cattgcacaa gataaaaata tatcatcatg ttagaaaaac tcatcgagca tcaaatgaaa 240 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa 300 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc 360 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt 420 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg 480 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc 540 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct 600 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc 660 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc 720 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt 780 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt 840 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa 900 tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa 960 atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc gttgaatatg 1020 gctcatagat cttttctcca tcactgatag ggagtggtaa aataactcca tcaatgatag 1080 agtgtcaaca acatgaccaa aatcccttaa cgtgagttac gcgtattaat tgcgttgcgc 1140 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 1200 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 1260
ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 1320
ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 1380
gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 1440
gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 1500
taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 1560
accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc 1620
tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 1680
cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 1740
agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 1800
gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggttacac tagaagaaca 1860
gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 1920
tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 1980
acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 2040
cagggaacga cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg 2100
tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc 2160
accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca 2220
ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct 2280
ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt 2340
tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggcccca aataatgatt 2400
ttattttgac tgatagtgac ctgttcgttg caacaaattg atgagcaatg cttttttata 2460
atgccaactt tgtacaaaaa agttggattt tttttcggtc taaaatggaa gcagccaaag 2520
aattggtttc catcgtccaa gaggagctcc ccaaggtgga ctatgcccag ctttggcagg 2580
acgccagcag ctgtgaggtc ctttacctct cggtggcatt cgtggcgatc aagttcatgc 2640
tgcgcccact ggacctgaag cgccaggcca ccttgaagaa gctgttcaca gcatacaact 2700
tcctcatgtc gatctattcc tttggctcct tcctggccat ggcctatgcc ctatcagtaa 2760
ctggcatcct ctccggcgac tgtgagacag cgttcaacaa cgatgtgttc aggatcacaa 2820
ctcagctgtt ctacctcagc aagttcgtag agtacatcga ctccttctac cttcccctta 2880
tggacaagcc actgtcgttc cttcagttct tccatcattt gggggctccc attgacatgt 2940
ggctattcta caaataccgc aacgaaggag tctggatctt tgtcctgttg aatgggttca 3000
ttcactggat catgtacggt tactactgga cgcggctcat caagctgaac ttccccatgc 3060 ccaagaacct gatcacctcc atgcagatca tccagttcaa tgtcgggttc tacatcgtct 3120
ggaagtaccg caatgtgcca tgctaccgcc aggatgggat gcgcatgttt gcctggatct 3180
tcaactactg gtacgtcggg acggtcttgc tgctgttcct caacttttac gtgcagacgt 3240
acatccggaa gccgaggaag aaccaaggga agaaggagta ggccacatgg cgcctgcgct 3300
ggaggaaacg gtacgctcgg atggtgcact gcacttgcac tccgccgctt ctagcctccc 3360
ctcgctctaa cctctgcgac atgcctgctt gaggcgtgac gttgcctcgt gcgatacagt 3420
ttacaccctt cccatggccc acggagcagg tgactgtctc cagcgtctgc aattctgatc 3480
attggtctgg tgcaatgtgc gcagaccaag cccctctaac gtcttgcggt gtaccgctcg 3540
acactcactg cacgagacag atggctgagc atgttatagc ccctgacatt ctacccttcg 3600
tccttacctg accgtcacat tcatgcttac cnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 3660
nnnnnnnnnn nacccaactt tctt 3684
<210> 101 <211> 774 <212> DNA
<213> Euglena anabaena <4 00> 101
atggaagcag ccaaagaatt ggtttccatc gtccaagagg agctccccaa ggtggactat 60
gcccagcttt ggcaggatgc cagcagctgt gaggtccttt acctctcggt ggcattcgtg 120
gcgatcaagt tcatgctgcg cccactggac ctgaagcgcc aggccacctt gaagaagctg 180
ttcacagcat acaacttcct catgtcgatc tattcctttg gctccttcct ggccatggcc 240
tatgccctat cagtaactgg catcctctcc ggcgactgtg agacggcgtt caacaacgat 300
gtgttcagga tcacaactca gctgttctac ctcagcaagt tcgtagagta catcgactcc 360
ttctaccttc cccttatgga caagccactg tcgttccttc agttcttcca tcatttgggg 420
gcccccattg acatgtggct attctacaaa taccgcaacg aaggagtctg gatctttgtc 480
ctgttgaatg ggttcattca ctggatcatg tacggttact attggacgcg gctcatcaag 540
ctgaacttcc ccatgcccaa gaacctgatc acctccatgc agatcatcca gttcaatgtc 600
gggttctaca tcgtctggaa gtaccgcaat gtgccatgct accgccagga tgggatgcgc 660
atgtttgcct ggatcttcaa ctactggtat gtcgggacgg tcttgctgct gttcctcaac 720
ttttacgtgc agacgtacat ccggaagccg aggaagaacc gagggaagaa ggag 774
<210> 102 <211> 774 <212> DNA
<213> Euglena anabaena <400> 102
atggaagcag ccaaagaatt ggtttccatc gtccaagagg agctccccaa ggtggactat 60
gcccagcttt ggcaggacgc cagcagctgt gaggtccttt acctctcggt ggcattcgtg 120
gcgatcaagt tcatgctgcg cccactggac ctgaagcgcc aggccacctt gaagaagctg 180
ttcacagcat acaacttcct catgtcgatc tattcctttg gctccttcct ggccatggcc 240
tatgccctat cagtaactgg catcctctcc ggcgactgtg agacagcgtt caacaacgat 300
gtgttcagga tcacaactca gctgttctac ctcagcaagt tcgtagagta catcgactcc 360
ttctaccttc cccttatgga caagccactg tcgttccttc agttcttcca tcatttgggg 420
gctcccattg acatgtggct attctacaaa taccgcaacg aaggagtctg gatctttgtc 480
ctgttgaatg ggttcattca ctggatcatg tacggttact actggacgcg gctcatcaag 540
ctgaacttcc ccatgcccaa gaacctgatc acctccatgc agatcatcca gttcaatgtc 600
gggttctaca tcgtctggaa gtaccgcaat gtgccatgct accgccagga tgggatgcgc 660
atgtttgcct ggatcttcaa ctactggtac gtcgggacgg tcttgctgct gttcctcaac 720
ttttacgtgc agacgtacat ccggaagccg aggaagaacc aagggaagaa ggag 774
<210> 103 <211> 258 <212> PRT
<213> Euglena anabaena <400> 103
Met Glu Ala Ala Lys Glu Leu Val Ser Ile Val Gln Glu Glu Leu Pro 15 10 15
Lys Val Asp Tyr Ala Gln Leu Trp Gln Asp Ala Ser Ser Cys Glu Val 20 25 30
Leu Tyr Leu Ser Val Ala Phe Val Ala Ile Lys Phe Met Leu Arg Pro 35 40 45
Leu Asp Leu Lys Arg Gln Ala Thr Leu Lys Lys Leu Phe Thr Ala Tyr 50 55 60
Asn Phe Leu Met Ser Ile Tyr Ser Phe Gly Ser Phe Leu Ala Met Ala 65 70 75 80
Tyr Ala Leu Ser Val Thr Gly Ile Leu Ser Gly Asp Cys Glu Thr Ala 85 90 95
Phe Asn Asn Asp Val Phe Arg Ile Thr Thr Gln Leu Phe Tyr Leu Ser 100 105 110 Lys Phe Val Glu Tyr Ile Asp Ser Phe Tyr Leu Pro Leu Met Asp Lys 115 120 125
Pro Leu Ser Phe Leu Gln Phe Phe His His Leu Gly Ala Pro Ile Asp 130 135 140
Met Trp Leu Phe Tyr Lys Tyr Arg Asn Glu Gly Val Trp Ile Phe Val 145 150 155 160
Leu Leu Asn Gly Phe Ile His Trp Ile Met Tyr Gly Tyr Tyr Trp Thr 165 170 175
Arg Leu Ile Lys Leu Asn Phe Pro Met Pro Lys Asn Leu Ile Thr Ser 180 185 190
Met Gln Ile Ile Gln Phe Asn Val Gly Phe Tyr Ile Val Trp Lys Tyr 195 200 205
Arg Asn Val Pro Cys Tyr Arg Gln Asp Gly Met Arg Met Phe Ala Trp 210 215 220
Ile Phe Asn Tyr Trp Tyr Val Gly Thr Val Leu Leu Leu Phe Leu Asn 225 230 235 240
Phe Tyr Val Gln Thr Tyr Ile Arg Lys Pro Arg Lys Asn Arg Gly Lys 245 250 255
Lys Glu
<210> 104 <211> 258 <212> PRT
<213> Euglena anabaena <400> 104
Met Glu Ala Ala Lys Glu Leu Val Ser Ile Val Gln Glu Glu Leu Pro 1 5 10 15
Lys Val Asp Tyr Ala Gln Leu Trp Gln Asp Ala Ser Ser Cys Glu Val 20 25 30
Leu Tyr Leu Ser Val Ala Phe Val Ala Ile Lys Phe Met Leu Arg Pro 35 40 45
Leu Asp Leu Lys Arg Gln Ala Thr Leu Lys Lys Leu Phe Thr Ala Tyr 50 55 60 Asn Phe Leu Met Ser Ile Tyr Ser Phe Gly Ser Phe Leu Ala Met Ala 65 70 75 80
Tyr Ala Leu Ser Val Thr Gly Ile Leu Ser Gly Asp Cys Glu Thr Ala 85 90 95
Phe Asn Asn Asp Val Phe Arg Ile Thr Thr Gln Leu Phe Tyr Leu Ser 100 105 110
Lys Phe Val Glu Tyr Ile Asp Ser Phe Tyr Leu Pro Leu Met Asp Lys 115 120 125
Pro Leu Ser Phe Leu Gln Phe Phe His His Leu Gly Ala Pro Ile Asp 130 135 140
Met Trp Leu Phe Tyr Lys Tyr Arg Asn Glu Gly Val Trp Ile Phe Val 145 150 155 160
Leu Leu Asn Gly Phe Ile His Trp Ile Met Tyr Gly Tyr Tyr Trp Thr 165 170 175
Arg Leu Ile Lys Leu Asn Phe Pro Met Pro Lys Asn Leu Ile Thr Ser 180 185 190
Met Gln Ile Ile Gln Phe Asn Val Gly Phe Tyr Ile Val Trp Lys Tyr 195 200 205
Arg Asn Val Pro Cys Tyr Arg Gln Asp Gly Met Arg Met Phe Ala Trp 210 215 220
Ile Phe Asn Tyr Trp Tyr Val Gly Thr Val Leu Leu Leu Phe Leu Asn 225 230 235 240
Phe Tyr Val Gln Thr Tyr Ile Arg Lys Pro Arg Lys Asn Gln Gly Lys 245 250 255
Lys Glu
<210> 105
<211> 4276
<212> DNA
<213> Seqüência
<220>
<223> pLF119
Artificial <400> 105
gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac 60
tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta 120
ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta 180
cattgcacaa gataaaaata tatcatcatg ttagaaaaac tcatcgagca tcaaatgaaa 240
ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa 300
tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc 360
gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt 420
atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg 480
catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc 540
atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct 600
gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc 660
atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc 720
ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt 780
cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt 840
ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa 900
tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa 960
atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc gttgaatatg 1020
gctcatagat cttttctcca tcactgatag ggagtggtaa aataactcca tcaatgatag 1080
agtgtcaaca acatgaccaa aatcccttaa cgtgagttac gcgtattaat tgcgttgcgc 1140
tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 1200
cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 1260
ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 1320
ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 1380
gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 1440
gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 1500
taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 1560
accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc 1620
tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 1680
cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 1740
agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 1800
gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggttacac tagaagaaca 1860 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 1920
tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 1980
acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 2040
cagggaacga cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg 2100
tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc 2160
accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca 2220
ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct 2280
ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt 2340
tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggcccca aataatgatt 2400
ttattttgac tgatagtgac ctgttcgttg caacaaattg atgagcaatg cttttttata 2460
atgccaactt tgtacaaaaa agttggattt tttttcggga tggccaccat ctctttgact 2520
actgagcaac ttttagaaca cccagaactg gttgcaattg atggggtgtt gtacgatctc 2580
ttcggactgg cgaaagtgca tccaggtggc aacctcattg aagccgccgg tgcctccgac 2640
ggaaccgccc tgttctactc catgcaccct ggagtgaagc cagagaattc gaagctgctg 2700
cagcaatttg cccgaggcaa acacgaacga agctcgaagg acccagtgta cacctttgac 2760
agtcccttcg cccaggatgt caagcagagc gttcgggagg tcatgaaggg gcgcaactgg 2820
tacgccacgc ccggcttttg gctgcggacc gcgctgatca tcgcgtgcac tgccataggc 2880
gaatggtatt ggatcactac cggggcagtg atgtggggca tcttcaccgg gtacttccac 2940
agccagattg ggttggcgat tcaacacgat gcctctcacg gagccatcag caaaaagccc 3000
tgggtgaacg cctttttcgc ctacggcatc gacgccattg gatcctcccg ctggatctgg 3060
ctgcagtccc acattatgcg ccaccacacc tacaccaacc agcatggcct ggacctggac 3120
gctgcctcgg cggagccgtt cattttgttc cactcctacc cggcaacaaa tgcgtcacga 3180
aagtggtacc atcggttcca ggcgtggtac atgtacatcg ttttggggat gtatggtgtg 3240
tcgatggtgt acaatccgat gtacttgttc acgatgcagc acaacgacac aatcccagag 3300
gccacctctc ttagaccagg cagctttttc aaccggcagc gcgccttcgc cgtttccctc 3360
cgcctactgt tcatcttccg caacgccttc ctcccctggt acatcgcggg cgcctctccg 3420
ctgctcacca tcctgctggt gccaacggtc acaggcatct tcttgacatt tgtttttgtg 3480
ctgtcccata actttgaagg cgctgagcgg acccccgaaa agaactgcaa ggccaaaagg 3540
gccaaggagg ggaaggaggt ccgcgatgta gaggaggacc gggtggactg gtaccgggcg 3600
caggccgaga ccgcggcgac ctacgggggc agcgtcggga tgatgctgac cggcggtttg 3660
aacctgcaga tcgagcacca cttgttcccc cgcatgtcct cttggcacta ccccttcatc 3720 caagatacgg tgcgggaatg ttgcaagcgc catggcgtgc gctacacata ctacccgacc 3780
atcctggaga atataatgtc cacgctccgc tacatgcaga aggtgggcgt ggcccacaca 3840
attcaggatg cccaggaatt ctgagtgagt tcgatccgca tcgacgtcta ccatttttga 3900
tgctgtctat tcctgttttc agtcacctcc agcattctca tggctggtga ccactgcccc 3960
tctaacccat tgtgacacac cgccaaagac tttgcctctt ttttttccct ttcttttgtc 4020
ctcggggtgc tttggccggt gtttactcgc cttgcagtcc ccgcaaacga ccgacgttta 4080
agctccgttg ttgactgggc cgctcgtaaa cccatctgca ggttgaggct cccatggaga 4140
attgtgatgg ctgattagga ggtggcgggg catacatgcc tcgacactca aagccgggcg 4200
gcttctggat tcgaaaacgc aaatgggcgc tttggaaaaa aaaaaaaaaa aaaaaaaaaa 4260
aaaacccaac tttctt 4276
<210> 106 <211> 1362 <212> DNA
<213> Euglena anabaena <4 00> 106
atggccacca tctctttgac tactgagcaa cttttagaac acccagaact ggttgcaatt 60
gatggggtgt tgtacgatct cttcggactg gcgaaagtgc atccaggtgg caacctcatt 120
gaagccgccg gtgcctccga cggaaccgcc ctgttctact ccatgcaccc tggagtgaag 180
ccagagaatt cgaagctgct gcagcaattt gcccgaggca aacacgaacg aagctcgaag 24 0
gacccagtgt acacctttga cagtcccttc gcccaggatg tcaagcagag cgttcgggag 300
gtcatgaagg ggcgcaactg gtacgccacg cccggctttt ggctgcggac cgcgctgatc 360
atcgcgtgca ctgccatagg cgaatggtat tggatcacta ccggggcagt gatgtggggc 420
atcttcaccg ggtacttcca cagccagatt gggttggcga ttcaacacga tgcctctcac 480
ggagccatca gcaaaaagcc ctgggtgaac gcctttttcg cctacggcat cgacgccatt 54 0
ggatcctccc gctggatctg gctgcagtcc cacattatgc gccaccacac ctacaccaac 600
cagcatggcc tggacctgga cgctgcctcg gcggagccgt tcattttgtt ccactcctac 660
ccggcaacaa atgcgtcacg aaagtggtac catcggttcc aggcgtggta catgtacatc 720
gttttgggga tgtatggtgt gtcgatggtg tacaatccga tgtacttgtt cacgatgcag 780
cacaacgaca caatcccaga ggccacctct cttagaccag gcagcttttt caaccggcag 840
cgcgccttcg ccgtttccct ccgcctactg ttcatcttcc gcaacgcctt cctcccctgg 900
tacatcgcgg gcgcctctcc gctgctcacc atcctgctgg tgccaacggt cacaggcatc 960
ttcttgacat ttgtttttgt gctgtcccat aactttgaag gcgctgagcg gacccccgaa 1020 aagaactgca aggccaaaag ggccaaggag gggaaggagg tccgcgatgt agaggaggac 1080
cgggtggact ggtaccgggc gcaggccgag accgcggcga cctacggggg cagcgtcggg 1140
atgatgctga ccggcggttt gaacctgcag atcgagcacc acttgttccc ccgcatgtcc 1200
tcttggcact accccttcat ccaagatacg gtgcgggaat gttgcaagcg ccatggcgtg 1260
cgctacacat actacccgac catcctggag aatataatgt ccacgctccg ctacatgcag 1320
aaggtgggcg tggcccacac aattcaggat gcccaggaat tc 1362
<210> 107 <211> 454 <212> PRT
<213> Euglena anabaena <400> 107
Met Ala Thr Ile Ser Leu Thr Thr Glu Gln Leu Leu Glu His Pro Glu 15 10 15
Leu Val Ala Ile Asp Gly Val Leu Tyr Asp Leu Phe Gly Leu Ala Lys 20 25 30
Val His Pro Gly Gly Asn Leu Ile Glu Ala Ala Gly Ala Ser Asp Gly 35 40 45
Thr Ala Leu Phe Tyr Ser Met His Pro Gly Val Lys Pro Glu Asn Ser 50 55 60
Lys Leu Leu Gln Gln Phe Ala Arg Gly Lys His Glu Arg Ser Ser Lys 65 70 75 80
Asp Pro Val Tyr Thr Phe Asp Ser Pro Phe Ala Gln Asp Val Lys Gln 85 90 95
Ser Val Arg Glu Val Met Lys Gly Arg Asn Trp Tyr Ala Thr Pro Gly 100 105 110
Phe Trp Leu Arg Thr Ala Leu Ile Ile Ala Cys Thr Ala Ile Gly Glu 115 120 125
Trp Tyr Trp Ile Thr Thr Gly Ala Val Met Trp Gly Ile Phe Thr Gly 130 135 140
Tyr Phe His Ser Gln Ile Gly Leu Ala Ile Gln His Asp Ala Ser His 145 150 155 160
Gly Ala Ile Ser Lys Lys Pro Trp Val Asn Ala Phe Phe Ala Tyr Gly 165 170 175 Ile Asp Ala Ile Gly Ser Ser Arg Trp Ile Trp Leu Gln Ser His Ile 180 185 190
Met Arg His His Thr Tyr Thr Asn Gln His Gly Leu Asp Leu Asp Ala 195 200 205
Ala Ser Ala Glu Pro Phe Ile Leu Phe His Ser Tyr Pro Ala Thr Asn 210 215 220
Ala Ser Arg Lys Trp Tyr His Arg Phe Gln Ala Trp Tyr Met Tyr Ile 225 230 235 240
Val Leu Gly Met Tyr Gly Val Ser Met Val Tyr Asn Pro Met Tyr Leu 245 250 255
Phe Thr Met Gln His Asn Asp Thr Ile Pro Glu Ala Thr Ser Leu Arg 260 265 270
Pro Gly Ser Phe Phe Asn Arg Gln Arg Ala Phe Ala Val Ser Leu Arg 275 280 285
Leu Leu Phe Ile Phe Arg Asn Ala Phe Leu Pro Trp Tyr Ile Ala Gly 290 295 300
Ala Ser Pro Leu Leu Thr Ile Leu Leu Val Pro Thr Val Thr Gly Ile 305 310 315 320
Phe Leu Thr Phe Val Phe Val Leu Ser His Asn Phe Glu Gly Ala Glu 325 330 335
Arg Thr Pro Glu Lys Asn Cys Lys Ala Lys Arg Ala Lys Glu Gly Lys 340 345 350
Glu Val Arg Asp Val Glu Glu Asp Arg Val Asp Trp Tyr Arg Ala Gln 355 360 365
Ala Glu Thr Ala Ala Thr Tyr Gly Gly Ser Val Gly Met Met Leu Thr 370 375 380
Gly Gly Leu Asn Leu Gln Ile Glu His His Leu Phe Pro Arg Met Ser 385 390 395 400
Ser Trp His Tyr Pro Phe Ile Gln Asp Thr Val Arg Glu Cys Cys Lys 405 410 415 Arg His Gly Val Arg Tyr Thr Tyr Tyr Pro Thr Ile Leu Glu Asn Ile 420 425 430
Met Ser Thr Leu Arg Tyr Met Gln Lys Val Gly Val Ala His Thr Ile 435 440 445
Gln Asp Ala Gln Glu Phe 450
<210> 108
<211> 31
<212> DNA
<213> Seqüência Artificial <220>
<223> oligonucleotideo EaD9-5Bbs
<400> 108
gaagacacca tggaagcagc caaagaattg g 31
<210> 109
<211> 30
<212> DNA
<213> Seqüência Artificial <220>
<223> oligonucleotideo EaD9-3fusion
<4 00> 109
agctggacgg gcgggctcct tcttccctcg 30
<210> 110
<211> 30
<212> DNA
<213> Seqüência Artificial <220>
<223> oligonucleotideo de fusão EgDHAsynlLink-5
<400> 110
cgagggaaga aggagcccgc ccgtccagct 30
<210> 111
<211> 31
<212> DNA
<213> Seqüência Artificial <220>
<223> oligonucleotideo MWG511
<400> 111
gaattcgcgg ccgctcactg ccagggagtc g 31
<210> 112 <211> 852 <212> DNA
<213> Seqüência Artificial <220>
<223> EgD9eIo-EgDHAsynILink <400> 112
gaagacacca tggaagcagc caaagaattg gtttccatcg tccaagagga gctccccaag 60
gtggactatg cccagctttg gcaggatgcc agcagctgtg aggtccttta cctctcggtg 120
gcattcgtgg cgatcaagtt catgctgcgc ccactggacc tgaagcgcca ggccaccttg 180
aagaagctgt tcacagcata caacttcctc atgtcgatct attcctttgg ctccttcctg 240
gccatggcct atgccctatc agtaactggc atcctctccg gcgactgtga gacggcgttc 300
aacaacgatg tgttcaggat cacaactcag ctgttctacc tcagcaagtt cgtagagtac 360
atcgactcct tctaccttcc ccttatggac aagccactgt cgttccttca gttcttccat 420
catttggggg cccccattga catgtggcta ttctacaaat accgcaacga aggagtctgg 480
atctttgtcc tgttgaatgg gttcattcac tggatcatgt acggttacta ttggacgcgg 540
ctcatcaagc tgaacttccc catgcccaag aacctgatca cctccatgca gatcatccag 600
ttcaatgtcg ggttctacat cgtctggaag taccgcaatg tgccatgcta ccgccaggat 660
gggatgcgca tgtttgcctg gatcttcaac tactggtatg tcgggacggt cttgctgctg 720
ttcctcaact tttacgtgca gacgtacatc cggaagccga ggaagaaccg agggaagaag 780
gagcccgccc gtccagctgg actcccgccg gccacgtact acgactccct ggcagtgagc 840
ggccgcgaat tc 852
<210> 113 <211> 5559 <212> DNA
<213> Seqüência Artificial <220>
<223> pLF124 <400> 113
ggccgcaagt atgaactaaa atgcacgtag gtgtaagagc tcatggagag catggaatat 60
tgtatccgac catgtaacag tataataact gagctccatc tcacttcttc tatgaataaa 120 caaaggatgt tatgatatat taacactcta tctatgcacc ttattgttct atgataaatt 180 tcctcttatt attataaatc atctgaatcg tgacggctta tggaatgctt caaatagtac 240 aaaaacaaat gtgtactata agactttcta aacaattcta actttagcat tgtgaacgag 300 acataagtgt taagaagaca taacaattat aatggaagaa gtttgtctcc atttatatat 360 tatatattac ccacttatgt attatattag gatgttaagg agacataaca attataaaga 420 gagaagtttg tatccattta tatattatat actacccatt tatatattat acttatccac 480 ttatttaatg tctttataag gtttgatcca tgatatttct aatattttag ttgatatgta 540
tatgaaaggg tactatttga actctcttac tctgtataaa ggttggatca tccttaaagt 600
gggtctattt aattttattg cttcttacag ataaaaaaaa aattatgagt tggtttgata 660
aaatattgaa ggatttaaaa taataataaa taacatataa tatatgtata taaatttatt 720
ataatataac atttatctat aaaaaagtaa atattgtcat aaatctatac aatcgtttag 780
ccttgctgga cgaatctcaa ttatttaaac gagagtaaac atatttgact ttttggttat 840
ttaacaaatt attatttaac actatatgaa attttttttt ttatcagcaa agaataaaat 900
taaattaagg aggacaatgg tgtcccaatc cttatacaac caacttccac aagaaagtca 960
agtcagagac aacaaaaaaa caagcaaagg aaatttttta atttgagttg tcttgtttgc 1020
tgcataattt atgcagtaaa acactacaca taaccctttt agcagtaaag caatggttga 1080
ccgtgtgctt agcttctttt attttatttt tttatcagca aagaataaat aaaataaaat 1140
gagacacttc agggatgttt caacggatcc cccgggctgc aggaattcga tatcaagctt 1200
atcgataccg tcgacctcga gggggggccc ggtacccaat tcgccctata gtgagtcgta 1260
ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac 1320
ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc 1380
ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg 1440
tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc 1500
cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg 1560
ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg 1620
gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg 1680
atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt 1740
ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt 1800
gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt 1860
taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc 1920
cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc 1980
tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc 2040
gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg 2100
gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat 2160
ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc 2220
acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa 2280
ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa 2340 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt 2400
gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct 2460
tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat 2520
gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg 2580
cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg 2640
atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt 2700
attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg 2760
ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg 2820
gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg 2880
tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa 2940
aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt 3000
tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt 3060
tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt 3120
ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag 3180
ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta 3240
gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat 3300
aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg 3360
ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg 3420
agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac 3480
aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga 3540
aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt 3600
ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta 3660
cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat 3720
tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg 3780
accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct 3840
ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa 3900
gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct 3960
ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac 4020
acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa 4080
caaaagctgg agctccaccc tagaactagt ggatccttca tccatgccct tcatttgccg 4140
cttattaatt aatttggtaa cagtccgtac taatcagtta cttatccttc ccccatcata 4200 attaatcttg gtagtctcga atgccacaac actgactagt ctcttggatc ataagaaaaa 4260
gccaaggaac aaaagaagac aaaacacaat gagagtatcc tttgcatagc aatgtctaag 4320
ttcataaaat tcaaacaaaa acgcaatcac acacagtgga catcacttat ccactagctg 4380
atcaggatcg ccgcgtcaag aaaaaaaaac tggaccccaa aagccatgca caacaacacg 4440
tactcacaaa ggtgtcaatc gagcagccca aaacattcac caactcaacc catcatgagc 4500
cctcacattt gttgtttcta acccaacctc aaactcgtat tctcttccgc cacctcattt 4560
ttgtttattt caacacccgt caaactgcat gccaccccgt ggccaaatgt ccatgcatgt 4620
taacaagacc tatgactata aatagctgca atctcggccc aggttttcat catcaagaac 4680
cagttcaata tcctagtaca ccgtattaaa gaatttaaga tatactccat ggaagcagcc 4740
aaagaattgg tttccatcgt ccaagaggag ctccccaagg tggactatgc ccagctttgg 4800
caggatgcca gcagctgtga ggtcctttac ctctcggtgg cattcgtggc gatcaagttc 4860
atgctgcgcc cactggacct gaagcgccag gccaccttga agaagctgtt cacagcatac 4920
aacttcctca tgtcgatcta ttcctttggc tccttcctgg ccatggccta tgccctatca 4980
gtaactggca tcctctccgg cgactgtgag acggcgttca acaacgatgt gttcaggatc 5040
acaactcagc tgttctacct cagcaagttc gtagagtaca tcgactcctt ctaccttccc 5100
cttatggaca agccactgtc gttccttcag ttcttccatc atttgggggc ccccattgac 5160
atgtggctat tctacaaata ccgcaacgaa ggagtctgga tctttgtcct gttgaatggg 5220
ttcattcact ggatcatgta cggttactat tggacgcggc tcatcaagct gaacttcccc 5280
atgcccaaga acctgatcac ctccatgcag atcatccagt tcaatgtcgg gttctacatc 5340
gtctggaagt accgcaatgt gccatgctac cgccaggatg ggatgcgcat gtttgcctgg 5400
atcttcaact actggtatgt cgggacggtc ttgctgctgt tcctcaactt ttacgtgcag 5460
acgtacatcc ggaagccgag gaagaaccga gggaagaagg agcccgcccg tccagctgga 5520
ctcccgccgg ccacgtacta cgactccctg gcagtgagc 5559
<210> 114 <211> 5213 <212> DNA
<213> Seqüência Artificial <220>
<223> KS366 <400> 114
ctagaactag tggatccttc atccatgccc ttcatttgcc gcttattaat taatttggta 60
acagtccgta ctaatcagtt acttatcctt cccccatcat aattaatctt ggtagtctcg 120
aatgccacaa cactgactag tctcttggat cataagaaaa agccaaggaa caaaagaaga 180 caaaacacaa tgagagtatc ctttgcatag caatgtctaa gttcataaaa ttcaaacaaa 240
aacgcaatca cacacagtgg acatcactta tccactagct gatcaggatc gccgcgtcaa 300
gaaaaaaaaa ctggacccca aaagccatgc acaacaacac gtactcacaa aggtgtcaat 360
cgagcagccc aaaacattca ccaactcaac ccatcatgag ccctcacatt tgttgtttct 420
aacccaacct caaactcgta ttctcttccg ccacctcatt tttgtttatt tcaacacccg 480
tcaaactgca tgccaccccg tggccaaatg tccatgcatg ttaacaagac ctatgactat 540
aaatagctgc aatctcggcc caggttttca tcatcaagaa ccagttcaat atcctagtac 600
accgtattaa agaatttaag atatactcca tggtgagagc gcggttccca ttactgttgc 660
tgggagttgt tttcctagca tcagtttctg tctcatttgg cattgcgtat tgggaaaagc 720
agaaccccag tcacaacaag tgcctccgaa gttgcaatag cgagaaagac tcctacagga 780
accaagcatg ccacgctcgt tgcaacctcc ttaaggtgga ggaaatggtt gagaaagaga 840
aatgcaaacg cggtaagatc ccacgtccac gtcctcgtcc acagcaccca gaacgtgaac 900
cacagcagcc aggtgagaaa gaacgtgata aggatcgtca gccacgtcca atcccattcc 960
ctcgtccaca gccacgtcag aaagagaaac atgaacagcg tcgtgaacag aaatggccac 1020
gtaaagaaga gaaacgcggt gagaaaggtt ctgaacgtga aaagaaagac aaagatgagg 1080
aacaggatga acgtcagtLc coattcccgg tggcggccgc aagtatgaac taaaatgcac 1140
gtaggtgtaa gagctcatgg agagcatgga atattgtatc cgaccatgta acagtataat 1200
aactgagctc catctcactt cttctatgaa taaacaaagg atgttatgat atattaacac 1260
tctatctatg caccttattg ttctatgata aatttcctct tattattata aatcatctga 1320
atcgtgacgg cttatggaat gcttcaaata gtacaaaaac aaatgtgtac tataagactt 1380
tctaaacaat tctaacttta gcattgtgaa cgagacataa gtgttaagaa gacataacaa 1440
ttataatgga agaagtttgt ctccatttat atattatata ttacccactt atgtattata 1500
ttaggatgtt aaggagacat aacaattata aagagagaag tttgtatcca tttatatatt 1560
atatactacc catttatata ttatacttat ccacttattt aatgtcttta taaggtttga 1620
tccatgatat ttctaatatt ttagttgata tgtatatgaa agggtactat ttgaactctc 1680
ttactctgta taaaggttgg atcatcctta aagtgggtct atttaatttt attgcttctt 1740
acagataaaa aaaaaattat gagttggttt gataaaatat tgaaggattt aaaataataa 1800
taaataacat ataatatatg tatataaatt tattataata taacatttat ctataaaaaa 1860
gtaaatattg tcataaatct atacaatcgt ttagccttgc tggacgaatc tcaattattt 1920
aaacgagagt aaacatattt gactttttgg ttatttaaca aattattatt taacactata 1980
tgaaattttt ttttttatca gcaaagaata aaattaaatt aaggaggaca atggtgtccc 2040 aatccttata caaccaactt ccacaagaaa gtcaagtcag agacaacaaa aaaacaagca 2100
aaggaaattt tttaatttga gttgtcttgt ttgctgcata atttatgcag taaaacacta 2160
cacataaccc ttttagcagt aaagcaatgg ttgaccgtgt gcttagcttc ttttatttta 2220
tttttttatc agcaaagaat aaataaaata aaatgagaca cttcagggat gtttcaacgg 2280
atcccccggg ctgcaggaat tcgatatcaa gcttatcgat accgtcgacc tcgagggggg 2340
gcccggtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt 2400
tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc 2460
cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt 2520
tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg 2580
tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg 2640
ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg 2700
ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt 2760
agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt 2820
tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta 2880
tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa 2940
atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt 3000
aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca 3060
ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa 3120
aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt 3180
ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca 3240
gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag 3300
ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc 3360
ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca 3420
gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt 3480
aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct 3540
gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt 3600
aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga 3660
caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact 3720
tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc 3780
acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga 3840
gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt 3900 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga 3960 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact 4020 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga 4080 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt 4140 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca 4200 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct 4260 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta 4320 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 4380 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 4440 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca 4500 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga 4560 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 4620 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt 4680 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 4740 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 4800 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt 4860 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga 4920 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta 4980 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa 5040 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat 5100 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta 5160 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctcc acc 5213 <210> 115 <211> 5559 <212> DNA <213> Seqüência Artificial <220> <223> ρKRl177 <400> 115 ggccgcaagt atgaactaaa atgcacgtag gtgtaagagc tcatggagag catggaatat 60 tgtatccgac catgtaacag tataataact gagctccatc tcacttcttc tatgaataaa 120 caaaggatgt tatgatatat taacactcta tctatgcacc ttattgttct atgataaatt 180 tcctcttatt attataaatc atctgaatcg tgacggctta tggaatgctt caaatagtac 240 aaaaacaaat gtgtactata agactttcta aacaattcta actttagcat tgtgaacgag 300
acataagtgt taagaagaca taacaattat aatggaagaa gtttgtctcc atttatatat 360
tatatattac ccacttatgt attatattag gatgttaagg agacataaca attataaaga 420
gagaagtttg tatccattta tatattatat actacccatt tatatattat acttatccac 480
ttatttaatg tctttataag gtttgatcca tgatatttct aatattttag ttgatatgta 540
tatgaaaggg tactatttga actctcttac tctgtataaa ggttggatca tccttaaagt 600
gggtctattt aattttattg cttcttacag ataaaaaaaa aattatgagt tggtttgata 660
aaatattgaa ggatttaaaa taataataaa taacatataa tatatgtata taaatttatt 720
ataatataac atttatctat aaaaaagtaa atattgtcat aaatctatac aatcgtttag 780
ccttgctgga cgaatctcaa ttatttaaac gagagtaaac atatttgact ttttggttat 840
ttaacaaatt attatttaac actatatgaa attttttttt ttatcagcaa agaataaaat 900
taaattaagg aggacaatgg tgtcccaatc cttatacaac caacttccac aagaaagtca 960
agtcagagac aacaaaaaaa caagcaaagg aaatttttta atttgagttg tcttgtttgc 1020
tgcataattt atgcagtaaa acactacaca taaccctttt agcagtaaag caatggttga 1080
ccgtgtgctt agcttctttt attttatttt tttatcagca aagaataaat aaaataaaat 1140
gagacacttc agggatgttt caacggatcc cccgggctgc aggaattcga tatcaagctt 1200
atcgataccg tcgacctcga gggggggccc ggtacccaat tcgccctata gtgagtcgta 1260
ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac 1320
ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc 1380
ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg 1440
tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc 1500
cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg 1560
ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg 1620
gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg 1680
atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt 1740
ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt 1800
gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt 1860
taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc 1920
cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc 1980
tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc 2040
gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg 2100 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat 2160 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc 2220 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa 2280 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa 2340 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt 2400 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct 2460 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat 2520 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg 2580 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg 2640 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt 2700 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg 2760 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg 2820 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg 2880 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa 2940 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt 3000 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt 3060 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt 3120 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag 3180 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta 3240 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat 3300 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg 3360 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg 3420 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac 3480 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga 3540 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt 3600 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta 3660 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat 3720 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg 3780 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct 3840 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa 3900 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct 3960 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac 4020
acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa 4 080
caaaagctgg agctccaccc tagaactagt ggatccttca tccatgccct tcatttgccg 4140
cttattaatt aatttggtaa cagtccgtac taatcagtta cttatccttc ccccatcata 4200
attaatcttg gtagtctcga atgccacaac actgactagt ctcttggatc ataagaaaaa 4260
gccaaggaac aaaagaagac aaaacacaat gagagtatcc tttgcatagc aatgtctaag 4320
ttcataaaat tcaaacaaaa acgcaatcac acacagtgga catcacttat ccactagctg 4380
atcaggatcg ccgcgtcaag aaaaaaaaac tggaccccaa aagccatgca caacaacacg 4440
tactcacaaa ggtgtcaatc gagcagccca aaacattcac caactcaacc catcatgagc 4500
cctcacattt gttgtttcta acccaacctc aaactcgtat tctcttccgc cacctcattt 4560
ttgtttattt caacacccgt caaactgcat gccaccccgt ggccaaatgt ccatgcatgt 4620
taacaagacc tatgactata aatagctgca atctcggccc aggttttcat catcaagaac 4 680
cagttcaata tcctagtaca ccgtattaaa gaatttaaga tatactccat ggaagcagcc 4740
aaagaattgg tttccatcgt ccaagaggag ctccccaagg tggactatgc ccagctttgg 4800
caggatgcca gcagctgtga ggtcctttac ctctcggtgg cattcgtggc gatcaagttc 4860
atgctgcgcc cactggacct gaagcgccag gccaccttga agaagctgtt cacagcatac 4920
aacttcctca tgtcgatcta ttcctttggc tccttcctgg ccatggccta tgccctatca 4980
gtaactggca tcctctccgg cgactgtgag acggcgttca acaacgatgt gttcaggatc 5040
acaactcagc tgttctacct cagcaagttc gtagagtaca tcgactcctt ctaccttccc 5100
cttatggaca agccactgtc gttccttcag ttcttccatc atttgggggc ccccattgac 5160
atgtggctat tctacaaata ccgcaacgaa ggagtctgga tctttgtcct gttgaatggg 5220
ttcattcact ggatcatgta cggttactat tggacgcggc tcatcaagct gaacttcccc 5280
atgcccaaga acctgatcac ctccatgcag atcatccagt tcaatgtcgg gttctacatc 5340
gtctggaagt accgcaatgt gccatgctac cgccaggatg ggatgcgcat gtttgcctgg 5400
atcttcaact actggtatgt cgggacggtc ttgctgctgt tcctcaactt ttacgtgcag 5460
acgtacatcc ggaagccgag gaagaaccga gggaagaagg agcccgcccg tccagctgga 5520
ctcccgccgg ccacgtacta cgactccctg gcagtgagc 5559
<210> 116 <211> 7916 <212> DNA
<213> Seqüência Artificial
<220> <223> pKR1179 <400> 116
gatccgtcga cggcgcgccc gatcatccgg atatagttcc tcctttcagc aaaaaacccc 60
tcaagacccg tttagaggcc ccaaggggtt atgctagtta ttgctcagcg gtggcagcag 120
ccaactcagc ttcctttcgg gctttgttag cagccggatc gatccaagct gtacctcact 180
attcctttgc cctcggacga gtgctggggc gtcggtttcc actatcggcg agtacttcta 240
cacagccatc ggtccagacg gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc 300
cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat 360
tgccgtcaac caagctctga tagagttggt caagaccaat gcggagcata tacgcccgga 420
gccgcggcga tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca 480
tacaagccaa ccacggcctc cagaagaaga tgttggcgac ctcgtattgg gaatccccga 540
acatcgcctc gctccagtca atgaccgctg ttatgcggcc attgtccgtc aggacattgt 600
tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca 660
tcagctcatc gagagcctgc gcgacggacg cactgacggt gtcgtccatc acagtttgcc 720
agtgatacac atggggatca gcaatcgcgc atatgaaatc acgccatgta gtgtattgac 780
cgattccttg cggtccgaat gggccgaacc cgctcgtctg gctaagatcg gccgcagcga 840
tcgcatccat agcctccgcg accggctgca gaacagcggg cagttcggtt tcaggcaggt 900
cttgcaacgt gacaccctgt gcacggcggg agatgcaata ggtcaggctc tcgctgaatt 960
ccccaatgtc aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc cgataaacat 1020
aacgatcttt gtagaaacca tcggcgcagc tatttacccg caggacatat ccacgccctc 1080
ctacatcgaa gctgaaagca cgagattctt cgccctccga gagctgcatc aggtcggaga 1140
cgctgtcgaa cttttcgatc agaaacttct cgacagacgt cgcggtgagt tcaggctttt 1200
cçatgggtat atctccttct taaagttaaa caaaattatt tctagaggga aaccgttgtg 1260
gtctccctat agtgagtcgt attaatttcg cgggatcgag atcgatccaa ttccaatccc 1320
acaaaaatct gagcttaaca gcacagttgc tcctctcaga gcagaatcgg gtattcaaca 1380
ccctcatatc aactactacg ttgtgtataa cggtccacat gccggtatat acgatgactg 1440
gggttgtaca aaggcggcaa caaacggcgt tcccggagtt gcacacaaga aatttgccac 1500
tattacagag gcaagagcag cagctgacgc gtacacaaca agtcagcaaa cagacaggtt 1560
gaacttcatc cccaaaggag aagctcaact caagcccaag agctttgcta aggccctaac 1620
aagcccacca aagcaaaaag cccactggct cacgctagga accaaaaggc ccagcagtga 1680
tccagcccca aaagagatct cctttgcccc ggagattaca atggacgatt tcctctatct 1740
ttacgatcta ggaaggaagt tcgaaggtga aggtgacgac actatgttca ccactgataa 1800 tgagaaggtt agcctcttca atttcagaaa gaatgctgac ccacagatgg ttagagaggc 1860
ctacgcagca ggtctcatca agacgatcta cccgagtaac aatctccagg agatcaaata 1920
ccttcccaag aaggttaaag atgcagtcaa aagattcagg actaattgca tcaagaacac 1980
agagaaagac atatttctca agatcagaag tactattcca gtatggacga ttcaaggctt 2040
gcttcataaa ccaaggcaag taatagagat tggagtctct aaaaaggtag ttcctactga 2100
atctaaggcc atgcatggag tctaagattc aaatcgagga tctaacagaa ctcgccgtga 2160
agactggcga acagttcata cagagtcttt tacgactcaa tgacaagaag aaaatcttcg 2220
tcaacatggt ggagcacgac actctggtct actccaaaaa tgtcaaagat acagtctcag 2280
aagaccaaag ggctattgag acttttcaac aaaggataat ttcgggaaac ctcctcggat 2340
tccattgccc agctatctgt cacttcatcg aaaggacagt agaaaaggaa ggtggctcct 2400
acaaatgcca tcattgcgat aaaggaaagg ctatcattca agatgcctct gccgacagtg 2460
gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca 2520
cgtcttcaaa gcaagtggat tgatgtgaca tctccactga cgtaagggat gacgcacaat 2580
cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga 2640
cacgctcgag ctcatttctc tattacttca gccataacaa aagaactctt ttctcttctt 2700
attaaaccat gaaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa 2760
agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca 2820
gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct 2880
acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc 2940
ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg 3000
tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg 3060
ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac 3120
cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc 3180
atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc 3240
tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg 3300
atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga 3360
gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt 3420
ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag 3480
gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct 3540
tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc 3600
gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga 3660 ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga 3720
gggcaaagga atagtgaggt acctaaagaa ggagtgcgtc gaagcagatc gttcaaacat 3780
ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata 3840
atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat 3900
gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa 3960
aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatcg 4020
atgtcgaatc gatcaacctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 4080
gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 4140
ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 4200
acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 4260
cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 4320
caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 4380
gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 4440
tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt 4500
aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 4560
ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 4620
cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 4680
tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 4740
tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 4800
ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 4860
aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 4920
aagggatttt ggtcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 4980
tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 504 0
cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 5100
ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 5160
accatatgga catattgtcg ttagaacgcg gctacaatta atacataacc ttatgtatca 5220
tacacatacg atttaggtga cactatagaa cggcgcgcca agcttggatc tcctgcagga 5280
tctggccggc cggatctcgt acggatcctt catccatgcc cttcatttgc cgcttattaa 5340
ttaatttggt aacagtccgt actaatcagt tacttatcct tcccccatca taattaatct 5400
tggtagtctc gaatgccaca acactgacta gtctcttgga tcataagaaa aagccaagga 54 60
acaaaagaag acaaaacaca atgagagtat cctttgcata gcaatgtcta agttcataaa 5520 attcaaacaa aaacgcaatc acacacagtg gacatcactt atccactagc tgatcaggat 5580
cgccgcgtca agaaaaaaaa actggacccc aaaagccatg cacaacaaca cgtactcaca 5640
aaggtgtcaa tcgagcagcc caaaacattc accaactcaa cccatcatga gccctcacat 5700
ttgttgtttc taacccaacc tcaaactcgt attctcttcc gccacctcat ttttgtttat 5760
ttcaacaccc gtcaaactgc atgccacccc gtggccaaat gtccatgcat gttaacaaga 5820
cctatgacta taaatagctg caatctcggc ccaggttttc atcatcaaga accagttcaa 5880
tatcctagta caccgtatta aagaatttaa gatatactcc atggaagcag ccaaagaatt 5940
ggtttccatc gtccaagagg agctccccaa ggtggactat gcccagcttt ggcaggatgc 6000
cagcagctgt gaggtccttt acctctcggt ggcattcgtg gcgatcaagt tcatgctgcg 6060
cccactggac ctgaagcgcc aggccacctt gaagaagctg ttcacagcat acaacttcct 6120
catgtcgatc tattcctttg gctccttcct ggccatggcc tatgccctat cagtaactgg 6180
catcctctcc ggcgactgtg agacggcgtt caacaacgat gtgttcagga tcacaactca 6240
gctgttctac ctcagcaagt tcgtagagta catcgactcc ttctaccttc cccttatgga 6300
caagccactg tcgttccttc agttcttcca tcatttgggg gcccccattg acatgtggct 6360
attctacaaa taccgcaacg aaggagtctg gatctttgtc ctgttgaatg ggttcattca 6420
ctggatcatg tacggttact attggacgcg gctcatcaag ctgaacttcc ccatgcccaa 6480
gaacctgatc acctccatgc agatcatcca gttcaatgtc gggttctaca tcgtctggaa 6540
gtaccgcaat gtgccatgct accgccagga tgggatgcgc atgtttgcct ggatcttcaa 6600
ctactggtat gtcgggacgg tcttgctgct gttcctcaac ttttacgtgc agacgtacat 6660
ccggaagccg aggaagaacc gagggaagaa ggagcccgcc cgtccagctg gactcccgcc 6720
ggccacgtac tacgactccc tggcagtgag cggccgcaag tatgaactaa aatgcacgta 6780
ggtgtaagag ctcatggaga gcatggaata ttgtatccga ccatgtaaca gtataataac 6840
tgagctccat ctcacttctt ctatgaataa acaaaggatg ttatgatata ttaacactct 6900
atctatgcac cttattgttc tatgataaat ttcctcttat tattataaat catctgaatc 6960
gtgacggctt atggaatgct tcaaatagta caaaaacaaa tgtgtactat aagactttct 7020
aaacaattct aactttagca ttgtgaacga gacataagtg ttaagaagac ataacaatta 7080
taatggaaga agtttgtctc catttatata ttatatatta cccacttatg tattatatta 7140
ggatgttaag gagacataac aattataaag agagaagttt gtatccattt atatattata 7200
tactacccat ttatatatta tacttatcca cttatttaat gtctttataa ggtttgatcc 7260
atgatatttc taatatttta gttgatatgt atatgaaagg gtactatttg aactctctta 7320
ctctgtataa aggttggatc atccttaaag tgggtctatt taattttatt gcttcttaca 7380 gataaaaaaa aaattatgag ttggtttgat aaaatattga aggatttaaa ataataataa 7440
ataacatata atatatgtat ataaatttat tataatataa catttatcta taaaaaagta 7500
aatattgtca taaatctata caatcgttta gccttgctgg acgaatctca attatttaaa 7560
cgagagtaaa catatttgac tttttggtta tttaacaaat tattatttaa cactatatga 7620
aatttttttt tttatcagca aagaataaaa ttaaattaag gaggacaatg gtgtcccaat 7680
ccttatacaa ccaacttcca caagaaagtc aagtcagaga caacaaaaaa acaagcaaag 7740
gaaatttttt aatttgagtt gtcttgtttg ctgcataatt tatgcagtaa aacactacac 7800
ataacccttt tagcagtaaa gcaatggttg accgtgtgct tagcttcttt tattttattt 7860
ttttatcagc aaagaataaa taaaataaaa tgagacactt cagggatgtt tcaacg 7916
<210> 117 <211> 9190 <212> DNA
<213> Seqüência Artificial <220>
<223> pKRl183 <400> 117
ggccgcaagt atgaactaaa atgcacgtag gtgtaagagc tcatggagag catggaatat 60
tgtatccgac catgtaacag tataataact gagctccatc tcacttcttc tatgaataaa 120
caaaggatgt tatgatatat taacactcta tctatgcacc ttattgttct atgataaatt 180
tcctcttatt attataaatc atctgaatcg tgacggctta tggaatgctt caaatagtac 240
aaaaacaaat gtgtactata agactttcta aacaattcta actttagcat tgtgaacgag 300
acataagtgt taagaagaca taacaattat aatggaagaa gtttgtctcc atttatatat 360
tatatattac ccacttatgt attatattag gatgttaagg agacataaca attataaaga 420
gagaagtttg tatccattta tatattatat actacccatt tatatattat acttatccac 480
ttatttaatg tctttataag gtttgatcca tgatatttct aatattttag ttgatatgta 540
tatgaaaggg tactatttga actctcttac tctgtataaa ggttggatca tccttaaagt 600
gggtctattt aattttattg cttcttacag ataaaaaaaa aattatgagt tggtttgata 660
aaatattgaa ggatttaaaa taataataaa taacatataa tatatgtata taaatttatt 720
ataatataac atttatctat aaaaaagtaa atattgtcat aaatctatac aatcgtttag 780
ccttgctgga cgaatctcaa ttatttaaac gagagtaaac atatttgact ttttggttat 840
ttaacaaatt attatttaac actatatgaa attttttttt ttatcagcaa agaataaaat 900
taaattaagg aggacaatgg tgtcccaatc cttatacaac caacttccac aagaaagtca 960
agtcagagac aacaaaaaaa caagcaaagg aaatttttta atttgagttg tcttgtttgc 1020 tgcataattt atgcagtaaa acactacaca taaccctttt agcagtaaag caatggttga 1080
ccgtgtgctt agcttctttt attttatttt tttatcagca aagaataaat aaaataaaat 1140
gagacacttc agggatgttt caacggatcc gtcgacggcg cgcccgatca tccggatata 1200
gttcctcctt tcagcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct 1260
agttattgct cagcggtggc agcagccaac tcagcttcct ttcgggcttt gttagcagcc 1320
ggatcgatcc aagctgtacc tcactattcc tttgccctcg gacgagtgct ggggcgtcgg 1380
tttccactat cggcgagtac ttctacacag ccatcggtcc agacggccgc gcttctgcgg 1440
gcgatttgtg tacgcccgac agtcccggct ccggatcgga cgattgcgtc gcatcgaccc 1500
tgcgcccaag ctgcatcatc gaaattgccg tcaaccaagc tctgatagag ttggtcaaga 1560
ccaatgcgga gcatatacgc ccggagccgc ggcgatcctg caagctccgg atgcctccgc 1620
tcgaagtagc gcgtctgctg ctccatacaa gccaaccacg gcctccagaa gaagatgttg 1680
gcgacctcgt attgggaatc cccgaacatc gcctcgctcc agtcaatgac cgctgttatg 1740
cggccattgt ccgtcaggac attgttggag ccgaaatccg cgtgcacgag gtgccggact 1800
tcggggcagt cctcggccca aagcatcagc tcatcgagag cctgcgcgac ggacgcactg 1860
acggtgtcgt ccatcacagt ttgccagtga tacacatggg gatcagcaat cgcgcatatg 1920
aaatcacgcc atgtagtgta ttgaccgatt ccttgcggtc cgaatgggcc gaacccgctc 1980
gtctggctaa gatcggccgc agcgatcgca tccatagcct ccgcgaccgg ctgcagaaca 2040
gcgggcagtt cggtttcagg caggtcttgc aacgtgacac cctgtgcacg gcgggagatg 2100
caataggtca ggctctcgct gaattcccca atgtcaagca cttccggaat cgggagcgcg 2160
gccgatgcaa agtgccgata aacataacga tctttgtaga aaccatcggc gcagctattt 2220
acccgcagga catatccacg ccctcctaca tcgaagctga aagcacgaga ttcttcgccc 2280
tccgagagct gcatcaggtc ggagacgctg tcgaactttt cgatcagaaa cttctcgaca 2340
gacgtcgcgg tgagttcagg cttttccatg ggtatatctc cttcttaaag ttaaacaaaa 2400
ttatttctag agggaaaccg ttgtggtctc cctatagtga gtcgtattaa tttcgcggga 2460
tcgagatcga tccaattcca atcccacaaa aatctgagct taacagcaca gttgctcctc 2520
tcagagcaga atcgggtatt caacaccctc atatcaacta ctacgttgtg tataacggtc 2580
cacatgccgg tatatacgat gactggggtt gtacaaaggc ggcaacaaac ggcgttcccg 2640
gagttgcaca caagaaattt gccactatta cagaggcaag agcagcagct gacgcgtaca 2700
caacaagtca gcaaacagac aggttgaact tcatccccaa aggagaagct caactcaagc 2760
ccaagagctt tgctaaggcc ctaacaagcc caccaaagca aaaagcccac tggctcacgc 2820
taggaaccaa aaggcccagc agtgatccag ccccaaaaga gatctccttt gccccggaga 2880 ttacaatgga cgatttcctc tatctttacg atctaggaag gaagttcgaa ggtgaaggtg 2940
acgacactat gttcaccact gataatgaga aggttagcct cttcaatttc agaaagaatg 3000
ctgacccaca gatggttaga gaggcctacg cagcaggtct catcaagacg atctacccga 3060
gtaacaatct ccaggagatc aaataccttc ccaagaaggt taaagatgca gtcaaaagat 3120
tcaggactaa ttgcatcaag aacacagaga aagacatatt tctcaagatc agaagtacta 3180
ttccagtatg gacgattcaa ggcttgcttc ataaaccaag gcaagtaata gagattggag 3240
tctctaaaaa ggtagttcct actgaatcta aggccatgca tggagtctaa gattcaaatc 3300
gaggatctaa cagaactcgc cgtgaagact ggcgaacagt tcatacagag tcttttacga 3360
ctcaatgaca agaagaaaat cttcgtcaac atggtggagc acgacactct ggtctactcc 3420
aaaaatgtca aagatacagt ctcagaagac caaagggcta ttgagacttt tcaacaaagg 3480
ataatttcgg gaaacctcct cggattccat tgcccagcta tctgtcactt catcgaaagg 3540
acagtagaaa aggaaggtgg ctcctacaaa tgccatcatt gcgataaagg aaaggctatc 3600
attcaagatg cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc 3660
gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgacatctcc 3720
actgacgtaa gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa 3780
ggaagttcat ttcatttgga gaggacacgc tcgagctcat ttctctatta cttcagccat 3840
aacaaaagaa ctcttttctc ttcttattaa accatgaaaa agcctgaact caccgcgacg 3900
tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg 3960
gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg 4020
gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg 4080
gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat 414 0
tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc 4200
gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag 4260
acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat 4320
ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc 4380
gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc 4440
gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc 4500
cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc 4560
gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc 4 620
gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt 4 680
ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg 4740 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc 4 800
gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga 4860
aaccgacgcc ccagcactcg tccgagggca aaggaatagt gaggtaccta aagaaggagt 4920
gcgtcgaagc agatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg 4980
ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta 5040
acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat 5100
acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg 5160
cggtgtcatc tatgttacta gatcgatgtc gaatcgatca acctgcatta atgaatcggc 5220
caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 5280
tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 5340
cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 5400
aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 5460
gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 5520
agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 5580
cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca 5640
cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 5700
ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 5760
gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 5820
tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg 5880
acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 5940
tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 6000
attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 6060
gctcagtgga acgaaaactc acgttaaggg attttggtca tgacattaac ctataaaaat 6120
aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga 6180
cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa 6240
gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca 6300
tcagagcaga ttgtactgag agtgcaccat atggacatat tgtcgttaga acgcggctac 6360
aattaataca taaccttatg tatcatacac atacgattta ggtgacacta tagaacggcg 6420
cgccaagctt ggatctcctg caggatctgg ccggccggat ctcgtacgga tccttcatcc 6480
atgcccttca tttgccgctt attaattaat ttggtaacag tccgtactaa tcagttactt 6540
atccttcccc catcataatt aatcttggta gtctcgaatg ccacaacact gactagtctc 6600 ttggatcata agaaaaagcc aaggaacaaa agaagacaaa acacaatgag agtatccttt 6660
gcatagcaat gtctaagttc ataaaattca aacaaaaacg caatcacaca cagtggacat 6720
cacttatcca ctagctgatc aggatcgccg cgtcaagaaa aaaaaactgg accccaaaag 6780
ccatgcacaa caacacgtac tcacaaaggt gtcaatcgag cagcccaaaa cattcaccaa 6840
ctcaacccat catgagccct cacatttgtt gtttctaacc caacctcaaa ctcgtattct 6900
cttccgccac ctcatttttg tttatttcaa cacccgtcaa actgcatgcc accccgtggc 6960
caaatgtcca tgcatgttaa caagacctat gactataaat agctgcaatc tcggcccagg 7020
ttttcatcat caagaaccag ttcaatatcc tagtacaccg tattaaagaa tttaagatat 7080
actccatgga agcagccaaa gaattggttt ccatcgtcca agaggagctc cccaaggtgg 7140
actatgccca gctttggcag gatgccagca gctgtgaggt cctttacctc tcggtggcat 7200
tcgtggcgat caagttcatg ctgcgcccac tggacctgaa gcgccaggcc accttgaaga 7260
agctgttcac agcatacaac ttcctcatgt cgatctattc ctttggctcc ttcctggcca 7320
tggcctatgc cctatcagta actggcatcc tctccggcga ctgtgagacg gcgttcaaca 7380
acgatgtgtt caggatcaca actcagctgt tctacctcag caagttcgta gagtacatcg 7440
actccttcta ccttcccctt atggacaagc cactgtcgtt ccttcagttc ttccatcatt 7500
tgggggcccc cattgacatg tggctattct acaaataccg caacgaagga gtctggatct 7560
ttgtcctgtt gaatgggttc attcactgga tcatgtacgg ttactattgg acgcggctca 7620
tcaagctgaa cttccccatg cccaagaacc tgatcacctc catgcagatc atccagttca 7680
atgtcgggtt ctacatcgtc tggaagtacc gcaatgtgcc atgctaccgc caggatggga 7740
tgcgcatgtt tgcctggatc ttcaactact ggtatgtcgg gacggtcttg ctgctgttcc 7800
tcaactttta cgtgcagacg tacatccgga agccgaggaa gaaccgaggg aagaaggagc 7860
ccgcccgtcc agctggactc ccgccggcca cgtactacga ctccctggca gtgagcggcc 7920
gcaccatgtc tcctaagcgg caagctctgc caatcacaat tgatggcgca acttatgatg 7980
tgtctgcttg ggtcaatcac caccctggag gagctgacat tatcgagaac tatcgcaacc 8040
gcgatgcgac cgatgtcttc atggtgatgc actctcaaga agccgtcgcc aagttgaaga 8100
gaatgcctgt tatggagcct tcctctcctg acacacctgt tgcacccaag cctaagcgtg 8160
atgagcccca ggaggatttc cgcaagttgc gggaggaatt catctccaag ggtatgttcg 8220
agacgagttt cctttggtat ttttacaaga cttcaactac cgtcggtttg atggtccttt 8280
ccatcttgat gaccgtgtac acgaattggt atttcaccgc tgctttggtt cttggcgtgt 8340
gctaccaaca gctaggctgg ttgtcccacg actattgcca tcaccaggtt ttcacaaacc 8400
gcaagattaa cgacgctttc ggtctctttt tcggtaacgt gatgcaggga tactcacaga 8460 cttggtggaa ggataggcac aatggtcacc atgccgccac caatgtggtc ggccatgacc 8520 cagatattga taacctcccc atcctggctt ggtctcccga agatgtcaag agggctactc 8580 cttcgactcg gaatctcatc aagtaccagc agtactactt cattcccacc attgcatccc 8640 ttaggttcat ctggtgcctc caatccatcg gcggcgtcat gtcctacaag agcgaggaga 8700 ggaacctgta ctacaagcgc cagtacacta aggaggcgat tggtctggcc ctccactggg 8760 tgctcaaggc cactttctat tgcagtgcca tgcctagctt tgccaccggt ttgggatgct 8820 tcttgatctc cgagctgctc ggaggatttg gcattgccat cgttgtgttt ctgaatcact 8880 atcctttgga caaggttgag gagactgtct gggatgagca cgggttcagc gccagccaga 8940 tccacgagac gttgaacatt aagcccggcc ttctcaccga ttgggtcttt ggtggtctca 9000 actaccagat tgagcaccac ttgtggccca acatgcccag gcacaacctc acggcagctt 9060 ccctggaggt gcagaagttg tgcgccaagc acaacctgcc ctacagggcc ccagccatca 9120 tccccggggt tcagaaattg gtcagcttct taggcgagat tgcccagctg gctgctgtcc 9180 ctgaatgagc 9190 <210> 118 <211> 6615 <212> DNA <213> Seqüência Artificial <220> <223> pKR1237 <220> <221> misc feature <222> (4340)..(4340) <223> η é a, c, g, ou t <400> 118 ggccgcgaca caagtgtgag agtactaaat aaatgctttg gttgtacgaa atcattacac 60 taaataaaat aatcaaagct tatatatgcc ttccgctaag gccgaatgca aagaaattgg 120 ttctttctcg ttatcttttg ccacttttac tagtacgtat taattactac ttaatcatct 180 ttgtttacgg ctcattatat ccggtctaga ggatccaagg ccgcgaagtt aaaagcaatg 240 ttgtcacttg tcgtactaac acatgatgtg atagtttatg ctagctagct ataacataag 300 ctgtctctga gtgtgttgta tattaataaa gatcatcact ggtgaatggt gatcgtgtac 360 gtaccctact tagtaggcaa tggaagcact tagagtgtgc tttgtgcatg gccttgcctc 420 tgttttgaga cttttgtaat gttttcgagt ttaaatcttt gcctttgcgt acgtgggcgg 480 atcccccggg ctgcaggaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc 540 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata 600 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc 660 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca 720
ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac 780
ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga 840
ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac 900
gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt 960
agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct 1020
aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat 1080
attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg 1140
cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg 1200
aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc 1260
ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat 1320
gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact 1380
attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca 1440
tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact 1500
tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg 1560
atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg 1620
agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg 1680
aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg 1740
caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag 1800
ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc 1860
gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga 1920
tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat 1980
atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc 2040
tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag 2100
accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct 2160
gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac 2220
caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc 2280
tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg 2340
ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt 2400
tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt 2460 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc 2520
tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca 2580
gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata 2640
gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg 2700
ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct 2760
ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta 2820
ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag 2880
tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga 2940
ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg 3000
caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg 3060
ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc 3120
atgattacgc caagcttgca tgcctgcagg tcgactcgac gtacgtcctc gaagagaagg 3180
gttaataaca cattttttaa catttttaac acaaatttta gttatttaaa aatttattaa 3240
aaaatttaaa ataagaagag gaactcttta aataaatcta acttacaaaa tttatgattt 3300
ttaataagtt ttcaccaata aaaaatgtca taaaaatatg ttaaaaagta tattatcaat 3360
attctcttta tgataaataa aaagaaaaaa aaaataaaag ttaagtgaaa atgagattga 3420
agtgacttta ggtgtgtata aatatatcaa ccccgccaac aatttattta atccaaatat 3480
attgaagtat attattccat agcctttatt tatttatata tttattatat aaaagcttta 3540
tttgttctag gttgttcatg aaatattttt ttggttttat ctccgttgta agaaaatcat 3600
gtgctttgtg tcgccactca ctattgcagc tttttcatgc attggtcaga ttgacggttg 3660
attgtatttt tgttttttat ggttttgtgt tatgacttaa gtcttcatct ctttatctct 3720
tcatcaggtt tgatggttac ctaatatggt ccatgggtac atgcatggtt aaattaggtg 3780
gccaactttg ttgtgaacga tagaattttt tttatattaa gtaaactatt tttatattat 3840
gaaataataa taaaaaaaat attttatcat tattaacaaa atcatattag ttaatttgtt 3900
aactctataa taaaagaaat actgtaacat tcacattaca tggtaacatc tttccaccct 3960
ttcatttgtt ttttgtttga tgactttttt tcttgtttaa atttatttcc cttcttttaa 4020
atttggaata cattatcatc atatataaac taaaatacta aaaacaggat tacacaaatg 4080
ataaataata acacaaatat ttataaatct agctgcaata tatttaaact agctatatcg 4140
atattgtaaa ataaaactag ctgcattgat actgataaaa aaatatcatg tgctttctgg 4200
actgatgatg cagtatactt ttgacattgc ctttatttta tttttcagaa aagctttctt 4260
agttctgggt tcttcattat ttgtttccca tctccattgt gaattgaatc atttgcttcg 4320 tgtcacaaat acaatttagn taggtacatg cattggtcag attcacggtt tattatgtca 4380
tgacttaagt tcatggtagt acattacctg ccacgcatgc attatattgg ttagatttga 4440
taggcaaatt tggttgtcaa caatataaat ataaataatg tttttatatt acgaaataac 4500
agtgatcaaa acaaacagtt ttatctttat taacaagatt ttgtttttgt ttgatgacgt 4560
tttttaatgt ttacgctttc ccccttcttt tgaatttaga acactttatc atcataaaat 4620
caaatactaa aaaaattaca tatttcataa ataataacac aaatattttt aaaaaatctg 4680
aaataataat gaacaatatt acatattatc acgaaaattc attaataaaa atattatata 4740
aataaaatgt aatagtagtt atatgtagga aaaaagtact gcacgcataa tatatacaaa 4800
aagattaaaa tgaactatta taaataataa cactaaatta atggtgaatc atatcaaaat 4860
aatgaaaaag taaataaaat ttgtaattaa cttctatatg tattacacac acaaataata 4920
aataatagta aaaaaaatta tgataaatat ttaccatctc ataagatatt taaaataatg 4980
ataaaaatat agattatttt ttatgcaact agctagccaa aaagagaaca cgggtatata 5040
taaaaagagt acctttaaat tctactgtac ttcctttatt cctgacgttt ttatatcaag 5100
tggacatacg tgaagatttt aattatcagt ctaaatattt cattagcact taatactttt 5160
ctgttttatt cctatcctat aagtagtccc gattctccca acattgctta ttcacacaac 5220
taactaagaa agtcttccat agccccccaa gcggccgcaa accatggctc tcagtcttac 5280
cacagaacag ctgttagaac gccctgattt ggttgcgatt gatggcatcc tctacgacct 5340
tgaagggctt gccaaagttc atccaggagg agatttgatt ctcgcttctg gtgcctctga 5400
tgcctcccct ctcttttatt caatgcatcc atacgtcaaa ccggagaatt ccaaattgct 5460
tcaacagttc gtccgaggga agcatgaccg cacctcgaag gacattgtct acacgtatga 5520
ttctcccttc gcacaagacg ttaagcggac aatgcgcgag gtgatgaaag ggaggaactg 5580
gtacgcaacc cctggcttct ggctgcgcac cgttgggatc atcgccgtga cggccttttg 5640
cgagtggcac tgggctacca cggggatggt gctgtggggc ctgttgactg gattcatgca 57 00
catgcagatc ggcttatcca tccagcatga tgcgtcccac ggggccatca gcaagaagcc 5760
ttgggtcaac gccctcttcg cctacggcat tgacgtcatc ggatcgtccc ggtggatttg 5820
gctgcagtcg cacatcatgc ggcaccacac ctacaccaac cagcacggcc tcgacctgga 5880
tgcggagtcg gcagagccgt tcctggtgtt ccacaactac cccgccgcaa acaccgcccg 594 0
aaagtggttc caccgcttcc aagcttggta catgtacctt gtgctggggg catacggggt 6000
atcgctggtg tacaacccgc tctacatttt ccggatgcag cacaatgaca ccatcccaga 6060
gtctgtcacg gccatgcggg aaaatggctt tctgcggcgc taccgcacac ttgcattcgt 6120
gatgcgagct ttcttcatct tccggaccgc attcttgccc tggtacctca ctgggacctc 6180 attgctgatc accattcctc tggtgcccac cgcaactggt gccttcttga cgttcttctt 6240 cattttgtcc cacaattttg atggctccga acggatcccc gacaagaact gcaaggttaa 6300 gagatctgag aaggacgttg aggctgacca aattgactgg tatcgggcgc aggtggagac 6360 gtcctccaca tacggtggcc ccatcgccat gttcttcact ggcggtctca atttccagat 6420 cgagcaccac ctctttcccc ggatgtcgtc ttggcactac cccttcgtcc agcaggcggt 6480 ccgggagtgt tgcgaacgcc atggagtgcg atatgttttc taccctacca tcgtcggcaa 6540 catcatctcc accctgaagt acatgcataa ggtgggtgtc gtccactgcg tgaaggacgc 6600 acaggattcc taagc 6615 <210> 119 <211> 6464 <212> DNA <213> Seqüência Artificial <220> <223> pKR1252 <400> 119 cgcgcccgat catccggata tagttcctcc tttcagcaaa aaacccctca agacccgttt 60 agaggcccca aggggttatg ctagttattg ctcagcggtg gcagcagcca actcagcttc 120 ctttcgggct ttgttagcag ccggatcgat ccaagctgta cctcactatt cctttgccct 180 cggacgagtg ctggggcgtc ggtttccact atcggcgagt acttctacac agccatcggt 240 ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg ctccggatcg 300 gacgattgcg tcgcatcgac cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa 360 gctctgatag agttggtcaa gaccaatgcg gagcatatac gcccggagcc gcggcgatcc 420 tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac aagccaacca 480 cggcctccag aagaagatgt tggcgacctc gtattgggaa tccccgaaca tcgcctcgct 540 ccagtcaatg accgctgtta tgcggccatt gtccgtcagg acattgttgg agccgaaatc 600 cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc caaagcatca gctcatcgag 660 agcctgcgcg acggacgcac tgacggtgtc gtccatcaca gtttgccagt gatacacatg 720 gggatcagca atcgcgcata tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg 780 tccgaatggg ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg catccatagc 840 ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac 900 accctgtgca cggcgggaga tgcaataggt caggctctcg ctgaattccc caatgtcaag 960 cacttccgga atcgggagcg cggccgatgc aaagtgccga taaacataac gatctttgta 1020 gaaaccatcg gcgcagctat ttacccgcag gacatatcca cgccctccta catcgaagct 1080 gaaagcacga gattcttcgc cctccgagag ctgcatcagg tcggagacgc tgtcgaactt 1140
ttcgatcaga aacttctcga cagacgtcgc ggtgagttca ggcttttcca tgggtatatc 1200
tccttcttaa agttaaacaa aattatttct agagggaaac cgttgtggtc tccctatagt 1260
gagtcgtatt aatttcgcgg gatcgagatc tgatcaacct gcattaatga atcggccaac 1320
gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 1380
tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 1440
tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 1500
ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 1560
agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 1620
accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 1680
ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct 1740
gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 1800
ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 1860
gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 1920
taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag 1980
tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 2040
gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 2100
cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 2160
agtggaacga aaactcacgt taagggattt tggtcatgac attaacctat aaaaataggc 2220
gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 2280
tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 2340
gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 2400
agcagattgt actgagagtg Caccatatgg acatattgtc gttagaacgc ggctacaatt 2460
aatacataac cttatgtatc atacacatac gatttaggtg acactataga acggcgcgcc 2520
aagcttggat ctcctgcagg atctggccgg ccggatctcg tacggatcct tcatccatgc 2580
ccttcatttg ccgcttatta attaatttgg taacagtccg tactaatcag ttacttatcc 2640
ttcccccatc ataattaatc ttggtagtct cgaatgccac aacactgact agtctcttgg 2700
atcataagaa aaagccaagg aacaaaagaa gacaaaacac aatgagagta tcctttgcat 2760
agcaatgtct aagttcataa aattcaaaca aaaacgcaat cacacacagt ggacatcact 2820
tatccactag ctgatcagga tcgccgcgtc aagaaaaaaa aactggaccc caaaagccat 2880
gcacaacaac acgtactcac aaaggtgtca atcgagcagc ccaaaacatt caccaactca 2940 acccatcatg agccctcaca tttgttgttt ctaacccaac ctcaaactcg tattctcttc 3000
cgccacctca tttttgttta tttcaacacc cgtcaaactg catgccaccc cgtggccaaa 3060
tgtccatgca tgttaacaag acctatgact ataaatagct gcaatctcgg cccaggtttt 3120
catcatcaag aaccagttca atatcctagt acaccgtatt aaagaattta agatatactc 3180
catggaagca gccaaagaat tggtttccat cgtccaagag gagctcccca aggtggacta 3240
tgcccagctt tggcaggatg ccagcagctg tgaggtcctt tacctctcgg tggcattcgt 3300
ggcgatcaag ttcatgctgc gcccactgga cctgaagcgc caggccacct tgaagaagct 3360
gttcacagca tacaacttcc tcatgtcgat ctattccttt ggctccttcc tggccatggc 3420
ctatgcccta tcagtaactg gcatcctctc cggcgactgt gagacggcgt tcaacaacga 3480
tgtgttcagg atcacaactc agctgttcta cctcagcaag ttcgtagagt acatcgactc 3540
cttctacctt ccccttatgg acaagccact gtcgttcctt cagttcttcc atcatttggg 3600
ggcccccatt gacatgtggc tattctacaa ataccgcaac gaaggagtct ggatctttgt 3660
cctgttgaat gggttcattc actggatcat gtacggttac tattggacgc ggctcatcaa 3720
gctgaacttc cccatgccca agaacctgat cacctccatg cagatcatcc agttcaatgt 3780
cgggttctac atcgtctgga agtaccgcaa tgtgccatgc taccgccagg atgggatgcg 3840
catgtttgcc tggatcttca actactggta tgtcgggacg gtcttgctgc tgttcctcaa 3900
cttttacgtg cagacgtaca tccggaagcc gaggaagaac cgagggaaga aggagcccgc 3960
ccgtccagct ggactcccgc cggccacgta ctacgactcc ctggcagtga gcggccgcac 4020
catgtctcct aagcggcaag ctctgccaat cacaattgat ggcgcaactt atgatgtgtc 4080
tgcttgggtc aatcaccacc ctggaggagc tgacattatc gagaactatc gcaaccgcga 4140
tgcgaccgat gtcttcatgg tgatgcactc tcaagaagcc gtcgccaagt tgaagagaat 4200
gcctgttatg gagccttcct ctcctgacac acctgttgca cccaagccta agcgtgatga 4260
gccccaggag gatttccgca agttgcggga ggaattcatc tccaagggta tgttcgagac 4320
gagtttcctt tggtattttt acaagacttc aactaccgtc ggtttgatgg tcctttccat 4380
cttgatgacc gtgtacacga attggtattt caccgctgct ttggttcttg gcgtgtgcta 4440
ccaacagcta ggctggttgt cccacgacta ttgccatcac caggttttca caaaccgcaa 4500
gattaacgac gctttcggtc tctttttcgg taacgtgatg cagggatact cacagacttg 4560
gtggaaggat aggcacaatg gtcaccatgc cgccaccaat gtggtcggcc atgacccaga 4 620
tattgataac ctccccatcc tggcttggtc tcccgaagat gtcaagaggg ctactccttc 4680
gactcggaat ctcatcaagt accagcagta ctacttcatt cccaccattg catcccttag 4740
gttcatctgg tgcctccaat ccatcggcgg cgtcatgtcc tacaagagcg aggagaggaa 4800 cctgtactac aagcgccagt acactaagga ggcgattggt ctggccctcc actgggtgct 4860
caaggccact ttctattgca gtgccatgcc tagctttgcc accggtttgg gatgcttctt 4920
gatctccgag ctgctcggag gatttggcat tgccatcgtt gtgtttctga atcactatcc 4980
tttggacaag gttgaggaga ctgtctggga tgagcacggg ttcagcgcca gccagatcca 5040
cgagacgttg aacattaagc ccggccttct caccgattgg gtctttggtg gtctcaacta 5100
ccagattgag caccacttgt ggcccaacat gcccaggcac aacctcacgg cagcttccct 5160
ggaggtgcag aagttgtgcg ccaagcacaa cctgccctac agggccccag ccatcatccc 5220
cggggttcag aaattggtca gcttcttagg cgagattgcc cagctggctg ctgtccctga 5280
atgagcggcc gcaagtatga actaaaatgc acgtaggtgt aagagctcat ggagagcatg 5340
gaatattgta tccgaccatg taacagtata ataactgagc tccatctcac ttcttctatg 5400
aataaacaaa ggatgttatg atatattaac actctatcta tgcaccttat tgttctatga 5460
taaatttcct cttattatta taaatcatct gaatcgtgac ggcttatgga atgcttcaaa 5520
tagtacaaaa acaaatgtgt actataagac tttctaaaca attctaactt tagcattgtg 5580
aacgagacat aagtgttaag aagacataac aattataatg gaagaagttt gtctccattt 5640
atatattata tattacccac ttatgtatta tattaggatg ttaaggagac ataacaatta 5700
taaagagaga agtttgtatc catttatata ttatatacta cccatttata tattatactt 5760
atccacttat ttaatgtctt tataaggttt gatccatgat atttctaata ttttagttga 5820
tatgtatatg aaagggtact atttgaactc tcttactctg tataaaggtt ggatcatcct 5880
taaagtgggt ctatttaatt ttattgcttc ttacagataa aaaaaaaatt atgagttggt 5940
ttgataaaat attgaaggat ttaaaataat aataaataac atataatata tgtatataaa 6000
tttattataa tataacattt atctataaaa aagtaaatat tgtcataaat ctatacaatc 6060
gtttagcctt gctggacgaa tctcaattat ttaaacgaga gtaaacatat ttgacttttt 6120
ggttatttaa caaattatta tttaacacta tatgaaattt ttttttttat cagcaaagaa 6180
taaaattaaa ttaaggagga caatggtgtc ccaatcctta tacaaccaac ttccacaaga 6240
aagtcaagtc agagacaaca aaaaaacaag caaaggaaat tttttaattt gagttgtctt 6300
gtttgctgca taatttatgc agtaaaacac tacacataac ccttttagca gtaaagcaat 6360
ggttgaccgt gtgcttagct tcttttattt tattttttta tcagcaaaga ataaataaaa 6420
taaaatgaga cacttcaggg atgtttcaac ggatccgtcg acgg 6464
<210> 120
<211> 10387
<212> DNA
<213> Seqüência Artificial <220>
<223> pKR1253 <220>
<221> misc_feature <222> (9212)..(9212) <223> η é a, c, g, ou t
<400> 120
gtacggatcc ttcatccatg cccttcattt gccgcttatt aattaatttg gtaacagtcc 60
gtactaatca gttacttatc cttcccccat cataattaat cttggtagtc tcgaatgcca 120
caacactgac tagtctcttg gatcataaga aaaagccaag gaacaaaaga agacaaaaca 180
caatgagagt atcctttgca tagcaatgtc taagttcata aaattcaaac aaaaacgcaa 240
tcacacacag tggacatcac ttatccacta gctgatcagg atcgccgcgt caagaaaaaa 300
aaactggacc ccaaaagcca tgcacaacaa cacgtactca caaaggtgtc aatcgagcag 360
cccaaaacat tcaccaactc aacccatcat gagccctcac atttgttgtt tctaacccaa 420
cctcaaactc gtattctctt ccgccacctc atttttgttt atttcaacac ccgtcaaact 480
gcatgccacc ccgtggccaa atgtccatgc atgttaacaa gacctatgac tataaatagc 540
tgcaatctcg gcccaggttt tcatcatcaa gaaccagttc aatatcctag tacaccgtat 600
taaagaattt aagatatact ccatggaagc agccaaagaa ttggtttcca tcgtccaaga 660
ggagctcccc aaggtggact atgcccagct ttggcaggat gccagcagct gtgaggtcct 720
ttacctctcg gtggcattcg tggcgatcaa gttcatgctg cgcccactgg acctgaagcg 780
ccaggccacc ttgaagaagc tgttcacagc atacaacttc ctcatgtcga tctattcctt 840
tggctccttc ctggccatgg cctatgccct atcagtaact ggcatcctct ccggcgactg 900
tgagacggcg ttcaacaacg atgtgttcag gatcacaact cagctgttct acctcagcaa 960
gttcgtagag tacatcgact ccttctacct tccccttatg gacaagccac tgtcgttcct 1020
tcagttcttc catcatttgg gggcccccat tgacatgtgg ctattctaca aataccgcaa 1080
cgaaggagtc tggatctttg tcctgttgaa tgggttcatt cactggatca tgtacggtta 1140
ctattggacg cggctcatca agctgaactt ccccatgccc aagaacctga tcacctccat 1200
gcagatcatc cagttcaatg tcgggttcta catcgtctgg aagtaccgca atgtgccatg 1260
ctaccgccag gatgggatgc gcatgtttgc ctggatcttc aactactggt atgtcgggac 1320
ggtcttgctg ctgttcctca acttttacgt gcagacgtac atccggaagc cgaggaagaa 1380
ccgagggaag aaggagcccg cccgtccagc tggactcccg ccggccacgt actacgactc 1440
cctggcagtg agcggccgca ccatgtctcc taagcggcaa gctctgccaa tcacaattga 1500
tggcgcaact tatgatgtgt ctgcttgggt caatcaccac cctggaggag ctgacattat 1560 cgagaactat cgcaaccgcg atgcgaccga tgtcttcatg gtgatgcact ctcaagaagc 1620
cgtcgccaag ttgaagagaa tgcctgttat ggagccttcc tctcctgaca cacctgttgc 1680
acccaagcct aagcgtgatg agccccagga ggatttccgc aagttgcggg aggaattcat 1740
ctccaagggt atgttcgaga cgagtttcct ttggtatttt tacaagactt caactaccgt 1800
cggtttgatg gtcctttcca tcttgatgac cgtgtacacg aattggtatt tcaccgctgc 1860
tttggttctt ggcgtgtgct accaacagct aggctggttg tcccacgact attgccatca 1920
ccaggttttc acaaaccgca agattaacga cgctttcggt ctctttttcg gtaacgtgat 1980
gcagggatac tcacagactt ggtggaagga taggcacaat ggtcaccatg ccgccaccaa 2040
tgtggtcggc catgacccag atattgataa cctccccatc ctggcttggt ctcccgaaga 2100
tgtcaagagg gctactcctt cgactcggaa tctcatcaag taccagcagt actacttcat 2160
tcccaccatt gcatccctta ggttcatctg gtgcctccaa tccatcggcg gcgtcatgtc 2220
ctacaagagc gaggagagga acctgtacta caagcgccag tacactaagg aggcgattgg 2280
tctggccctc cactgggtgc tcaaggccac tttctattgc agtgccatgc ctagctttgc 2340
caccggtttg ggatgcttct tgatctccga gctgctcgga ggatttggca ttgccatcgt 2400
tgtgtttctg aatcactatc ctttggacaa ggttgaggag actgtctggg atgagcacgg 2460
gttcagcgcc agccagatcc acgagacgtt gaacattaag cccggccttc tcaccgattg 2520
ggtctttggt ggtctcaact accagattga gcaccacttg tggcccaaca tgcccaggca 2580
caacctcacg gcagcttccc tggaggtgca gaagttgtgc gccaagcaca acctgcccta 2640
cagggcccca gccatcatcc ccggggttca gaaattggtc agcttcttag gcgagattgc 2700
ccagctggct gctgtccctg aatgagcggc cgcaagtatg aactaaaatg cacgtaggtg 2760
taagagctca tggagagcat ggaatattgt atccgaccat gtaacagtat aataactgag 2820
ctccatctca cttcttctat gaataaacaa aggatgttat gatatattaa cactctatct 2880
atgcacctta ttgttctatg ataaatttcc tcttattatt ataaatcatc tgaatcgtga 2940
cggcttatgg aatgcttcaa atagtacaaa aacaaatgtg tactataaga ctttctaaac 3000
aattctaact ttagcattgt gaacgagaca taagtgttaa gaagacataa caattataat 3060
ggaagaagtt tgtctccatt tatatattat atattaccca cttatgtatt atattaggat 3120
gttaaggaga cataacaatt ataaagagag aagtttgtat ccatttatat attatatact 3180
acccatttat atattatact tatccactta tttaatgtct ttataaggtt tgatccatga 3240
tatttctaat attttagttg atatgtatat gaaagggtac tatttgaact ctcttactct 3300
gtataaaggt tggatcatcc ttaaagtggg tctatttaat tttattgctt cttacagata 3360
aaaaaaaaat tatgagttgg tttgataaaa tattgaagga tttaaaataa taataaataa 3420 catataatat atgtatataa atttattata atataacatt tatctataaa aaagtaaata 3480
ttgtcataaa tctatacaat cgtttagcct tgctggacga atctcaatta tttaaacgag 3540
agtaaacata tttgactttt tggttattta acaaattatt atttaacact atatgaaatt 3600
ttttttttta tcagcaaaga ataaaattaa attaaggagg acaatggtgt cccaatcctt 3660
atacaaccaa cttccacaag aaagtcaagt cagagacaac aaaaaaacaa gcaaaggaaa 3720
ttttttaatt tgagttgtct tgtttgctgc ataatttatg cagtaaaaca ctacacataa 3780
cccttttagc agtaaagcaa tggttgaccg tgtgcttagc ttcttttatt ttattttttt 3840
atcagcaaag aataaataaa ataaaatgag acacttcagg gatgtttcaa cggatccgtc 3900
gacggcgcgc ccgatcatcc ggatatagtt cctcctttca gcaaaaaacc cctcaagacc 3960
cgtttagagg ccccaagggg ttatgctagt tattgctcag cggtggcagc agccaactca 4020
gcttcctttc gggctttgtt agcagccgga tcgatccaag ctgtacctca ctattccttt 4080
gccctcggac gagtgctggg gcgtcggttt ccactatcgg cgagtacttc tacacagcca 4140
tcggtccaga cggccgcgct tctgcgggcg atttgtgtac gcccgacagt cccggctccg 4200
gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa attgccgtca 4260
accaagctct gatagagttg gtcaagacca atgcggagca tatacgcccg gagccgcggc 4 320
gatcctgcaa gctccggatg cctccgctcg aagtagcgcg tctgctgctc catacaagcc 4380
aaccacggcc tccagaagaa gatgttggcg acctcgtatt gggaatcccc gaacatcgcc 4440
tcgctccagt caatgaccgc tgttatgcgg ccattgtccg tcaggacatt gttggagccg 4500
aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct cggcccaaag catcagctca 4560
tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac 4 620
acatggggat cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg accgattcct 4680
tgcggtccga atgggccgaa cccgctcgtc tggctaagat cggccgcagc gatcgcatcc 4740
atagcctccg cgaccggctg cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac 4800
gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa ttccccaatg 4860
tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt gccgataaac ataacgatct 4 920
ttgtagaaac catcggcgca gctatttacc cgcaggacat atccacgccc tcctacatcg 4 980
aagctgaaag cacgagattc ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg 5040
aacttttcga tcagaaactt ctcgacagac gtcgcggtga gttcaggctt ttccatgggt 5100
atatctcctt cttaaagtta aacaaaatta tttctagagg gaaaccgttg tggtctccct 5160
atagtgagtc gtattaattt cgcgggatcg agatctgatc aacctgcatt aatgaatcgg 5220
ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 5280 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 5340
acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 5400
aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 5460
tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 5520
aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 5580
gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc 5640
acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 5700
accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 5760
ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 5820
gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 5880
gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 5940
ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 6000
gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 6060
cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgacattaa cctataaaaa 6120
taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 6180
acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 6240
agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 6300
atcagagcag attgtactga gagtgcacca tatggacata ttgtcgttag aacgcggcta 6360
caattaatac ataaccttat gtatcataca catacgattt aggtgacact atagaacggc 6420
gcgccaagct tggatctcct gcaggatctg gccggccgga tctcgtacgc aaaggcaaag 6480
atttaaactc gaaaacatta caaaagtctc aaaacagagg caaggccatg cacaaagcac 6540
actctaagtg cttccattgc ctactaagta gggtacgtac acgatcacca ttcaccagtg 6600
atgatcttta ttaatataca acacactcag agacagctta tgttatagct agctagcata 6660
aactatcaca tcatgtgtta gtacgacaag tgacaacatt gcttttaact tcgcggcctt 6720
ggatcctcta gaccggatat aatgagccgt aaacaaagat gattaagtag taattaatac 6780
gtactagtaa aagtggcaaa agataacgag aaagaaccaa tttctttgca ttcggcctta 6840
gcggaaggca tatataagct ttgattattt tatttagtgt aatgatttcg tacaaccaaa 6900
gcatttattt agtactctca cacttgtgtc gcggccgctt aggaatcctg tgcgtccttc 6960
acgcagtgga cgacacccac cttatgcatg tacttcaggg tggagatgat gttgccgacg 7020
atggtagggt agaaaacata tcgcactcca tggcgttcgc aacactcccg gaccgcctgc 7080
tggacgaagg ggtagtgcca agacgacatc cggggaaaga ggtggtgctc gatctggaaa 7140 ttgagaccgc cagtgaagaa catggcgatg gggccaccgt atgtggagga cgtctccacc 7200
tgcgcccgat accagtcaat ttggtcagcc tcaacgtcct tctcagatct cttaaccttg 7260
cagttcttgt cggggatccg ttcggagcca tcaaaattgt gggacaaaat gaagaagaac 7320
gtcaagaagg caccagttgc ggtgggcacc agaggaatgg tgatcagcaa tgaggtccca 7380
gtgaggtacc agggcaagaa tgcggtccgg aagatgaaga aagctcgcat cacgaatgca 7440
agtgtgcggt agcgccgcag aaagccattt tcccgcatgg ccgtgacaga ctctgggatg 7500
gtgtcattgt gctgcatccg gaaaatgtag agcgggttgt acaccagcga taccccgtat 7560
gcccccagca caaggtacat gtaccaagct tggaagcggt ggaaccactt tcgggcggtg 7620
tttgcggcgg ggtagttgtg gaacaccagg aacggctctg ccgactccgc atccaggtcg 7680
aggccgtgct ggttggtgta ggtgtggtgc cgcatgatgt gcgactgcag ccaaatccac 7740
cgggacgatc cgatgacgtc aatgccgtag gcgaagaggg cgttgaccca aggcttcttg 7800
ctgatggccc cgtgggacgc atcatgctgg atggataagc cgatctgcat gtgcatgaat 7860
ccagtcaaca ggccccacag caccatcccc gtggtagccc agtgccactc gcaaaaggcc 7920
gtcacggcga tgatcccaac ggtgcgcagc cagaagccag gggttgcgta ccagttcctc 7980
cctttcatca cctcgcgcat tgtccgctta acgtcttgtg cgaagggaga atcatacgtg 8040
tagacaatgt ccttcgaggt gcggtcatgc ttccctcgga cgaactgttg aagcaatttg 8100
gaattctccg gtttgacgta tggatgcatt gaataaaaga gaggggaggc atcagaggca 8160
ccagaagcga gaatcaaatc tcctcctgga tgaactttgg caagcccttc aaggtcgtag 8220
aggatgccat caatcgcaac caaatcaggg cgttctaaca gctgttctgt ggtaagactg 8280
agagccatgg tttgcggccg cttggggggc tatggaagac tttcttagtt agttgtgtga 8340
ataagcaatg ttgggagaat cgggactact tataggatag gaataaaaca gaaaagtatt 8400
aagtgctaat gaaatattta gactgataat taaaatcttc acgtatgtcc acttgatata 8460
aaaacgtcag gaataaagga agtacagtag aatttaaagg tactcttttt atatataccc 8520
gtgttctctt tttggctagc tagttgcata aaaaataatc tatattttta tcattatttt 8580
aaatatctta tgagatggta aatatttatc ataatttttt ttactattat ttattatttg 8640
tgtgtgtaat acatatagaa gttaattaca aattttattt actttttcat tattttgata 8700
tgattcacca ttaatttagt gttattattt ataatagttc attttaatct ttttgtatat 8760
attatgcgtg cagtactttt ttcctacata taactactat tacattttat ttatataata 8820
tttttattaa tgaattttcg tgataatatg taatattgtt cattattatt tcagattttt 8880
taaaaatatt tgtgttatta tttatgaaat atgtaatttt tttagtattt gattttatga 8940
tgataaagtg ttctaaattc aaaagaaggg ggaaagcgta aacattaaaa aacgtcatca 9000 aacaaaaaca aaatcttgtt aataaagata aaactgtttg ttttgatcac tgttatttcg 9060
taatataaaa acattattta tatttatatt gttgacaacc aaatttgcct atcaaatcta 9120
accaatataa tgcatgcgtg gcaggtaatg tactaccatg aacttaagtc atgacataat 9180
aaaccgtgaa tctgaccaat gcatgtacct anctaaattg tatttgtgac acgaagcaaa 9240
tgattcaatt cacaatggag atgggaaaca aataatgaag aacccagaac taagaaagct 9300
tttctgaaaa ataaaataaa ggcaatgtca aaagtatact gcatcatcag tccagaaagc 9360
acatgatatt tttttatcag tatcaatgca gctagtttta ttttacaata tcgatatagc 9420
tagtttaaat atattgcagc tagatttata aatatttgtg ttattattta tcatttgtgt 9480
aatcctgttt ttagtatttt agtttatata tgatgataat gtattccaaa tttaaaagaa 9540
gggaaataaa tttaaacaag aaaaaaagtc atcaaacaaa aaacaaatga aagggtggaa 9600
agatgttacc atgtaatgtg aatgttacag tatttctttt attatagagt taacaaatta 9660
actaatatga ttttgttaat aatgataaaa tatttttttt attattattt cataatataa 9720
aaatagttta cttaatataa aaaaaattct atcgttcaca acaaagttgg ccacctaatt 9780
taaccatgca tgtacccatg gaccatatta ggtaaccatc aaacctgatg aagagataaa 9840
gagatgaaga cttaagtcat aacacaaaac cataaaaaac aaaaatacaa tcaaccgtca 9900
atctgaccaa tgcatgaaaa agctgcaata gtgagtggcg acacaaagca catgattttc 9960
ttacaacgga gataaaacca aaaaaatatt tcatgaacaa cctagaacaa ataaagcttt 10020
tatataataa atatataaat aaataaaggc tatggaataa tatacttcaa tatatttgga 10080
ttaaataaat tgttggcggg gttgatatat ttatacacac ctaaagtcac ttcaatctca 10140
ttttcactta acttttattt tttttttctt tttatttatc ataaagagaa tattgataat 10200
atacttttta acatattttt atgacatttt ttattggtga aaacttatta aaaatcataa 10260
attttgtaag ttagatttat ttaaagagtt cctcttctta ttttaaattt tttaataaat 10320
ttttaaataa ctaaaatttg tgttaaaaat gttaaaaaat gtgttattaa cccttctctt 10380
cgaggac 10387
<210> 121 <211> 30 <212> DNA
<213> Seqüência Artificial <220>
<223> oligonucleotídeo oEAd5-l-l
<400> 121
agcggccgca ccatggccac catctctttg
30 <210> 122
<211> 30
<212> DNA
<213> Seqüência Artificial
<220>
<223> oligonucleotideo oEAd5-l-l
<400> 122
tgcggccgct cagaattcct gggcatcctg
<210> 123
<211> 4899
<212> DNA
<213> Seqüência Artificial
<220>
<223> pKR1136
<400> 123
aattccagca cactggcggc cgttactagt ggatccgagc tcggtaccaa gcttgatgca 60
tagcttgagt attctaacgc gtcacctaaa tagcttggcg taatcatggt catagctgtt 120
tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa 180
gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 240
gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 300
ggggagaggc ggtttgcgta ttgggcgctc ttccgcttce tcgctcactg actcgctgcg 360
ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 420
cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaagcccag 480
gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 54 0
tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 600
ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 660
atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 720
gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 780
tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 840
cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 900
cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt 960
tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 1020
cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 1080
cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 1140
gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 1200
gatcctttta aattaaaaat gaagttttag cacgtgtcag tcctgctcct cggccacgaa 1260 gtgcacgcag ttgccggccg ggtcgcgcag ggcgaactcc cgcccccacg gctgctcgcc 1320
gatctcggtc atggccggcc cggaggcgtc ccggaagttc gtggacacga cctccgacca 1380
ctcggcgtac agctcgtcca ggccgcgcac ccacacccag gccagggtgt tgtccggcac 1440
cacctggtcc tggaccgcgc tgatgaacag ggtcacgtcg tcccggacca caccggcgaa 1500
gtcgtcctcc acgaagtccc gggagaaccc gagccggtcg gtccagaact cgaccgctcc 1560
ggcgacgtcg cgcgcggtga gcaccggaac ggcactggtc aacttggcca tggtggccct 1620
cctcacgtgc tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 1680
tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 1740
acctgtatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa 1800
ttgtaagcgt taataattca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc 1860
gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc 1920
tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc 1980
cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag 2040
gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg 2100
aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga 2160
ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg 2220
caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc 2280
tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc 2340
cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg 2400
gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg 2460
gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag 2520
cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga 2580
gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga 2640
tcagagcttg atcccctgcg ccatcagatc cttggcggcg agaaagccat ccagtttact 2700
ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg ttcgcttgct 2760
gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc tacctgcttt 2820
ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt catccggggt 2880
cagcaccgtt tctgcggact ggctttctac gtgaaaagga tctaggtgaa gatccttttt 2940
gataatctca tgcctgacat ttatattccc cagaacatca ggttaatggc gtttttgatg 3000
tcattttcgc ggtggctgag atcagccact tcttccccga taacggagac cggcacactg 3060
gccatatcgg tggtcatcat gcgccagctt tcatccccga tatgcaccac cgggtaaagt 3120 tcacgggaga ctttatctga cagcagacgt gcactggcca gggggatcac catccgtcgc 3180 cccggcgtgt caataatatc actctgtaca tccacaaaca gacgataacg gctctctctt 3240 ttataggtgt aaaccttaaa ctgccgtacg tataggctgc gcaactgttg ggaagggcga 3300 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga 3360 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa 3420 ttgtaatacg actcactata gggcgaattg ggccctctag atgcatgctc gagcggccgc 3480 cagtgtgatg gatatctgca gaattcagga gcggccgcac catggccacc atctctttga 3540 ctactgagca acttttagaa cacccagaac tggttgcaat tgatggggtg ttgtacgatc 3600 tcttcggact ggcgaaagtg catccaggtg gcaacctcat tgaagccgcc ggtgcctccg 3660 acggaaccgc cctgttctac tccatgcacc ctggagtgaa gccagagaat tcgaagctgc 3720 tgcagcaatt tgcccgaggc aaacacgaac gaagctcgaa ggacccagtg tacacctttg 3780 acagtccctt cgcccaggat gtcaagcaga gcgttcggga ggtcatgaag gggcgcaact 3840 ggtacgccac gcccggcttt tggctgcgga ccgcgctgat catcgcgtgc actgccatag 3900 gcgaatggta ttggatcact accggggcag tgatgtgggg catcttcacc gggtacttcc 3960
acagccagat tgggttggcg attcaacacg atgcctctca cggagccatc agcaaaaagc 4020
cctgggtgaa cgcctttttc gcctacggca tcgacgccat tggatcctcc cgctggatct 4080
ggctgcagtc ccacattatg cgccaccaca cctacaccaa ccagcatggc ctggacctgg 4140
acgctgcctc ggcggagccg ttcattttgt tccactccta cccggcaaca aatgcgtcac 4200
gaaagtggta ccatcggttc caggcgtggt acatgtacat cgttttgggg atgtatggtg 4260
tgtcgatggt gtacaatccg atgtacttgt tcacgatgca gcacaacgac acaatcccag 4320
aggccacctc tcttagacca ggcagctttt tcaaccggca gcgcgccttc gccgtttccc 4380
tccgcctact gttcatcttc cgcaacgcct tcctcccctg gtacatcgcg ggcgcctctc 4440
cgctgctcac catcctgctg gtgccaacgg tcacaggcat cttcttgaca tttgtttttg 4500
tgctgtccca taactttgaa ggcgctgagc ggacccccga aaagaactgc aaggccaaaa 4560
gggccaagga ggggaaggag gtccgcgatg tagaggagga ccgggtggac tggtaccggg 4 62 0
cgcaggccga gaccgcggcg acctacgggg gcagcgtcgg gatgatgctg accggcggtt 4680
tgaacctgca gatcgagcac cacttgttcc cccgcatgtc ctcttggcac taccccttca 4740
tccaagatac ggtgcgggaa tgttgcaagc gccatggcgt gcgctacaca tactacccga 4800
ccatcctgga gaatataatg tccacgctcc gctacatgca gaaggtgggc gtggcccaca 4860
caattcagga tgcccaggaa ttctgagcgg ccgcacctg 4 899 <210> 124
<211> 5592
<212> DNA
<213> Seqüência Artificial
<220> <223> pKR1139 <400> 124 ggccgcattt cgcaccaaat caatgaaagt aataatgaaa agtctgaata agaatactta 60 ggcttagatg cctttgttac ttgtgtaaaa taacttgagt catgtacctt tggcggaaac 120 agaataaata aaaggtgaaa ttccaatgct ctatgtataa gttagtaata cttaatgtgt 180 tctacggttg tttcaatatc atcaaactct aattgaaact ttagaaccac aaatctcaat 240 cttttcttaa tgaaatgaaa aatcttaatt gtaccatgtt tatgttaaac accttacaat 300 taattggttg gagaggagga ccaaccgatg ggacaacatt gggagaaaga gattcaatgg 360 agatttggat aggagaacaa cattcttttt cacttcaata caagatgagt gcaacactaa 420 ggatatgtat gagactttca gaagctacga caacatagat gagtgaggtg gtgattccta 480 gcaagaaaga cattagagga agccaaaatc gaacaaggaa gacatcaagg gcaagagaca 540 ggaccatcca tctcaggaaa aggagctttg ggatagtccg agaagttgta caagaaattt 600 tttggagggt gagtgatgca ttgctggtga ctttaactca atcaaaattg agaaagaaag 660 aaaagggagg gggctcacat gtgaatagaa gggaaacggg agaattttac agttttgatc 720 taatgggcat cccagctagt ggtaacatat tcaccatgtt taaccttcac gtacgtctag 780 aggatccccc atggtcaatc aatgagacgc caacttctta atctattgag acctgcaggt 840 ctagaagggc ggatccccgg gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc 900 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg 960 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc 1020 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 1080 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc 1140 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt 1200 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac 1260 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga 1320 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta 1380 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat 1440 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc 1500 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga 1560 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca 1620 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt 1680
ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg 1740
gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc 1800
atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata 1860
acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt 1920
tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag 1980
ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca 2040
aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg 2100
aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg 2160
ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag 2220
atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg 2280
aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag 2340
accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga 2400
tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt 2460
tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc 2520
tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc 2580
cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac 2640
caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac 2700
cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt 2760
cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct 2820
gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat 2880
acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt 2940
atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg 3000
cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt 3060
gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt 3120
tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg 3180
tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg 3240
agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc 3300
ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg 3360
gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac 3420
actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag 3480 gaaacagcta tgaccatgat tacgccaagc ttgcatgcct gcaggctagc ctaagtacgt 3540
actcaaaatg ccaacaaata aaaaaaaagt tgctttaata atgccaaaac aaattaataa 3600
aacacttaca acaccggatt ttttttaatt aaaatgtgcc atttaggata aatagttaat 3660
atttttaata attatttaaa aagccgtatc tactaaaatg atttttattt ggttgaaaat 3720
attaatatgt ttaaatcaac acaatctatc aaaattaaac taaaaaaaaa ataagtgtac 3780
gtggttaaca ttagtacagt aatataagag gaaaatgaga aattaagaaa ttgaaagcga 3840
gtctaatttt taaattatga acctgcatat ataaaaggaa agaaagaatc caggaagaaa 3900
agaaatgaaa ccatgcatgg tcccctcgtc atcacgagtt tctgccattt gcaatagaaa 3960
cactgaaaca cctttctctt tgtcacttaa ttgagatgcc gaagccacct cacaccatga 4020
acttcatgag gtgtagcacc caaggcttcc atagccatgc atactgaaga atgtctcaag 4080
ctcagcaccc tacttctgtg acgtgtccct cattcacctt cctctcttcc ctataaataa 4140
ccacgcctca ggttctccgc ttcacaactc aaacattctc tccattggtc cttaaacact 4200
catcagtcat caccgcggcc gcaccatggc caccatctct ttgactactg agcaactttt 4260
agaacaccca gaactggttg caattgatgg ggtgttgtac gatctcttcg gactggcgaa 4320
agtgcatcca ggtggcaacc tcattgaagc cgccggtgcc tccgacggaa ccgccctgtt 4380
ctactccatg caccctggag tgaagccaga gaattcgaag ctgctgcagc aatttgcccg 4440
aggcaaacac gaacgaagct cgaaggaccc agtgtacacc tttgacagtc ccttcgccca 4500
ggatgtcaag cagagcgttc gggaggtcat gaaggggcgc aactggtacg ccacgcccgg 4560
cttttggctg cggaccgcgc tgatcatcgc gtgcactgcc ataggcgaat ggtattggat 4620
cactaccggg gcagtgatgt ggggcatctt caccgggtac ttccacagcc agattgggtt 4 680
ggcgattcaa cacgatgcct ctcacggagc catcagcaaa aagccctggg tgaacgcctt 4740
tttcgcctac ggcatcgacg ccattggatc ctcccgctgg atctggctgc agtcccacat 4800
tatgcgccac cacacctaca ccaaccagca tggcctggac ctggacgctg cctcggcgga 4860
gccgttcatt ttgttccact cctacccggc aacaaatgcg tcacgaaagt ggtaccatcg 4920
gttccaggcg tggtacatgt acatcgtttt ggggatgtat ggtgtgtcga tggtgtacaa 4 980
tccgatgtac ttgttcacga tgcagcacaa cgacacaatc ccagaggcca cctctcttag 5040
accaggcagc tttttcaacc ggcagcgcgc cttcgccgtt tccctccgcc tactgttcat 5100
cttccgcaac gccttcctcc cctggtacat cgcgggcgcc tctccgctgc tcaccatcct 5160
gctggtgcca acggtcacag gcatcttctt gacatttgtt tttgtgctgt cccataactt 5220
tgaaggcgct gagcggaccc ccgaaaagaa ctgcaaggcc aaaagggcca aggaggggaa 5280
ggaggtccgc gatgtagagg aggaccgggt ggactggtac cgggcgcagg ccgagaccgc 5340 ggcgacctac gggggcagcg tcgggatgat gctgaccggc ggtttgaacc tgcagatcga 5400 gcaccacttg ttcccccgca tgtcctcttg gcactacccc ttcatccaag atacggtgcg 5460 ggaatgttgc aagcgccatg gcgtgcgcta cacatactac ccgaccatcc tggagaatat 5520 aatgtccacg ctccgctaca tgcagaaggt gggcgtggcc cacacaattc aggatgccca 5580 ggaattctga gc 5592 <210> 125 <2ll> 7497 <212> DNA <213> Seqüência Artificial <220> <223> pKR561 <400> 125 gtacgtcgag tcgacggcgc gcccgatcat ccggatatag ttcctccttt cagcaaaaaa 60 cccctcaaga cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtggca 120 gcagccaact cagcttcctt tcgggctttg ttagcagccg gatcgatcca agctgtacct 180 cactattcct ttgccctcgg acgagtgctg gggcgtcggt ttccactatc ggcgagtact 240 tctacacagc catcggtcca gacggccgcg cttctgcggg cgatttgtgt acgcccgaca 300 gtcccggctc cggatcggac gattgcgtcg catcgaccct gcgcccaagc tgcatcatcg 360 aaattgccgt caaccaagct ctgatagagt tggtcaagac caatgcggag catatacgcc 420 cggagccgcg gcgatcctgc aagctccgga tgcctccgct cgaagtagcg cgtctgctgc 480 tccatacaag ccaaccacgg cctccagaag aagatgttgg cgacctcgta ttgggaatcc 540 ccgaacatcg cctcgctcca gtcaatgacc gctgttatgc ggccattgtc cgtcaggaca 600 ttgttggagc cgaaatccgc gtgcacgagg tgccggactt cggggcagtc ctcggcccaa 660 agcatcagct catcgagagc ctgcgcgacg gacgcactga cggtgtcgtc catcacagtt 720 tgccagtgat acacatgggg atcagcaatc gcgcatatga aatcacgcca tgtagtgtat 780 tgaccgattc cttgcggtcc gaatgggccg aacccgctcg tctggctaag atcggccgca 840 gcgatcgcat ccatagcctc cgcgaccggc tgcagaacag cgggcagttc ggtttcaggc 900 aggtcttgca acgtgacacc ctgtgcacgg cgggagatgc aataggtcag gctctcgctg 960 aattccccaa tgtcaagcac ttccggaatc gggagcgcgg ccgatgcaaa gtgccgataa 1020 acataacgat ctttgtagaa accatcggcg cagctattta cccgcaggac atatccacgc 1080 cctcctacat cgaagctgaa agcacgagat tcttcgccct ccgagagctg catcaggtcg 1140 gagacgctgt cgaacttttc gatcagaaac ttctcgacag acgtcgcggt gagttcaggc 1200 ttttccatgg gtatatctcc ttcttaaagt taaacaaaat tatttctaga gggaaaccgt 1260 tgtggtctcc ctatagtgag tcgtattaat ttcgcgggat cgagatcgat ccaattccaa 1320
tcccacaaaa atctgagctt aacagcacag ttgctcctct cagagcagaa tcgggtattc 1380
aacaccctca tatcaactac tacgttgtgt ataacggtcc acatgccggt atatacgatg 1440
actggggttg tacaaaggcg gcaacaaacg gcgttcccgg agttgcacac aagaaatttg 1500
ccactattac agaggcaaga gcagcagctg acgcgtacac aacaagtcag caaacagaca 1560
ggttgaactt catccccaaa ggagaagctc aactcaagcc caagagcttt gctaaggccc 1620
taacaagccc accaaagcaa aaagcccact ggctcacgct aggaaccaaa aggcccagca 1680
gtgatccagc cccaaaagag atctcctttg ccccggagat tacaatggac gatttcctct 1740
atctttacga tctaggaagg aagttcgaag gtgaaggtga cgacactatg ttcaccactg 1800
ataatgagaa ggttagcctc ttcaatttca gaaagaatgc tgacccacag atggttagag 1860
aggcctacgc agcaggtctc atcaagacga tctacccgag taacaatctc caggagatca 1920
aataccttcc caagaaggtt aaagatgcag tcaaaagatt caggactaat tgcatcaaga 1980
acacagagaa agacatattt ctcaagatca gaagtactat tccagtatgg acgattcaag 2040
gcttgcttca taaaccaagg caagtaatag agattggagt ctctaaaaag gtagttccta 2100
ctgaatctaa ggccatgcat ggagtctaag attcaaatcg aggatctaac agaactcgcc 2160
gtgaagaetg gcgaacagtt catacagagt cttttacgac tcaatgacaa gaagaaaatc 2220
ttcgtcaaca tggtggagca cgacactctg gtctactcca aaaatgtcaa agatacagtc 2280
tcagaagacc aaagggctat tgagactttt caacaaagga taatttcggg aaacctcctc 2340
ggattccatt gcccagctat ctgtcacttc atcgaaagga cagtagaaaa ggaaggtggc 2400
tcctacaaat gccatcattg cgataaagga aaggctatca ttcaagatgc ctctgccgac 2460
agtggtccca aagatggacc cccacccacg aggagcatcg tggaaaaaga agacgttcca 2520
accacgtctt caaagcaagt ggattgatgt gacatctcca ctgacgtaag ggatgacgca 2580
caatcccact atccttcgca agacccttcc tctatataag gaagttcatt tcatttggag 2640
aggacacgct cgagctcatt tctctattac ttcagccata acaaaagaac tcttttctct 2700
tcttattaaa ccatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc 2760
gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct 2820
ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt 2880
ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa 2940
gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag 3000
ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg 3060
gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc 3120 ggaccgcaag gaatcggtca atacactaca ccccatgtgt atcactggca aactgtgatg gctctcgatg agctgatgct ttgggccgag gcggatttcg gctccaacaa tgtcctgacg tggagcgagg cgatgttcgg ggattcccaa ccgtggttgg cttgtatgga gcagcagacg gcaggatcgc cgcggctccg ggcgtatatg agcttggttg acggcaattt cgatgatgca gtccgatccg gagccgggac tgtcgggcgt tggaccgatg gctgtgtaga agtactcgcc ccgagggcaa aggaatagtg aggtacctaa acatttggca ataaagtttc ttaagattga tataatttct gttgaattac gttaagcatg ttatgagatg ggtttttatg attagagtcc acaaaatata gcgcgcaaac taggataaat atcgatgtcg aatctgatca acctgcatta tttgcgtatt gggcgctctt ccgcttcctc gctgcggcga gcggtatcag ctcactcaaa ggataacgca ggaaagaaca tgtgagcaaa ggccgcgttg ctggcgtttt tccataggct acgctcaagt cagaggtggc gaaacccgac tggaagctcc ctcgtgcgct ctcctgttcc ctttctccct tcgggaagcg tggcgctttc ggtgtaggtc gttcgctcca agctgggctg ctgcgcctta tccggtaact atcgtcttga actggcagca gccactggta acaggattag gttcttgaag tggtggccta actacggcta tctgctgaag ccagttacct tcggaaaaag caccgctggt agcggtggtt tttttgtttg atctcaagaa gatcctttga tcttttctac acgttaaggg attttggtca tgacattaac
tggcgtgatt tcatatgcgc gattgctgat 3180
gacgacaccg tcagtgcgtc cgtcgcgcag 3240
gactgccccg aagtccggca cctcgtgcac 3300
gacaatggcc gcataacagc ggtcattgac 3360
tacgaggtcg ccaacatctt cttctggagg 3420
cgctacttcg agcggaggca tccggagctt 3480
ctccgcattg gtcttgacca actctatcag 3540
gcttgggcgc agggtcgatg cgacgcaatc 3600
acacaaatcg cccgcagaag cgcggccgtc 3660
gatagtggaa accgacgccc cagcactcgt 3720
agaaggagtg cgtcgaagca gatcgttcaa 3780
atcctgttgc cggtcttgcg atgattatca 3840
taataattaa catgtaatgc atgacgttat 3900
cgcaattata catttaatac gcgatagaaa 3960
tatcgcgcgc ggtgtcatct atgttactag 4020
atgaatcggc caacgcgcgg ggagaggcgg 4080
gctcactgac tcgctgcgct cggtcgttcg 4140
ggcggtaata cggttatcca cagaatcagg 4200
aggccagcaa aaggccagga accgtaaaaa 4260
ccgcccccct gacgagcatc acaaaaatcg 4320
aggactataa agataccagg cgtttccccc 4380
gaccctgccg cttaccggat acctgtccgc 4440
tcaatgctca cgctgtaggt atctcagttc 4500
tgtgcacgaa ccccccgttc agcccgaccg 4560
gtccaacccg gtaagacacg acttatcgcc 4620
cagagcgagg tatgtaggcg gtgctacaga 4680
cactagaagg acagtatttg gtatctgcgc 4740
agttggtagc tcttgatccg gcaaacaaac 4800
caagcagcag attacgcgca gaaaaaaagg 4860
ggggtctgac gctcagtgga acgaaaactc 4920
ctataaaaat aggcgtatca cgaggccctt 4980 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac 5040
ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc 5100
gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag 5160
agtgcaccat atggacatat tgtcgttaga acgcggctac aattaataca taaccttatg 5220
tatcatacac atacgattta ggtgacacta tagaacggcg cgccaagctt ggatcctcta 5280
gacgtacggt accatctgct aatattttaa atcacatgca agagaggagg catggttcca 5340
ttttctacct tcacattatt tgagaaaaac gaacttgttc tgtgttttat ttttgccctt 5400
cacattagta caacgtggaa gactcatggt tacacagaat catacataag tacaatgctt 5460
gtccctaaga aaacaagcac tcgttgtatt gaacctttac ggctcatgcg gccgctgaag 5520
tattgcttct tagttaacct ttcctttctc tctcagctat gtgaattcat tttgctttcg 5580
tcacaattta tatagtgaaa ttggatcttt ggagttaacg ccttcacagg attatcgtgt 5640
tagaacaatg ctttttcatg ttctaattag tagtacatta caaatgtgca ctctattcaa 5700
taagcatctt ttggcacgtt aataaatcat gtgaaaaaaa aatactacta tttcaaagaa 5760
agtgttgtaa aaagaaacgg aaagagagct ggcttcagtt gttgagactt gtttgctagt 5820
aaaaatggtg tgaagagtga ttcatggtga ggtggttttt cgtccctttc tgtttgcatg 5880
aaaaacaaat ggcaagagat gacgtaggat tccttccctt aacgattatc tgtttttaat 5940
ttcaaatata catataggaa tttatgaatt actaaggttg taaaatatgc tggtcattta 6000
tttatggcta aaatattttt ttttctcgta aatataaaaa tatttaaaat ttatttttat 6060
catatttttt atccttataa aattatgtgt acaacctata taaaaaaata tcatatttaa 6120
tattgattat atgtttaatc aatataaaaa atcattatca tatatttaga tttattcgaa 6180
tatacatcta aacaaaaaat aacatatttt aattttatga agaaaaaaaa atattttatc 6240
ctttatttat ttaagattaa ttaatagtta tgtattgtgg aaagactttt acacatgcaa 6300
tagatatact gaatcaatta gatgccaatg ctgagttgga aatcacttga ggaggggagg 6360
agacttgcca atgcttttca gtttcattta aatgatttag tggaggagat agagtagtga 6420
taaaggcatg ccccaatttt ggagtgtata tatgagtgga aataagagag ggatagagag 6480
aaaaaataaa gagagtaaaa ataattaatg tgaaatgata tgataaaaaa ataaagaaag 6540
agataaagag aaaaatgaaa tgagagatag atgaaataga gagtagatac atgtttgttt 6600
aggttttttt taggaaataa cacatttttt tctcatcact tattactcac tgtcaatttc 6660
ctctctttca atcataatga tatgatttgt ttaacaaaaa tgtgaaaaaa catataaagt 6720
aaaatatttt tataaattga taaataaaaa tttacaaaat ttatttctta ttaaattgaa 6780
tagaaaatga aagaaaagaa aagaaaaagt atatataaaa tgatatagct ttaaaaagaa 6840 taaatttttc atatcagtct ttttttaata atttagaaat atttaagtat atagcaaaaa 6900 tataatgtac tttacatatg cataaataat aatttgaaaa tagaactaat agaatagaga 6960 aaaaagtaat ataataatta actatatgaa aatttagaag ggacaatatt tttaattaag 7020 aatataaaca atatttcttt tcatgtaatg agggacggat gtacggggcc agtgttggag 7080 tcaaagccaa aatagtcacg gggaaattaa tgcactgcat gactattcga aaaaattcac 7140 tagccttact tagatgttag attaatagct agggggtgca gataattttg aaaggcatga 7200 aaaacattaa tttgtacatt gcaagctttt gatgacaagc tttgcaattg ttcacactac 7260 cttatgccat ttataaatag agtgattggc atatgaagga aatcatgaga gtcgaagcga 7320 aaaacaaagc ttgagagtgt aggaaaaata cagttttttt ggtaaaaata cagtatttga 7380 ataggagcga aaaatatcct ttcaaaatga tccttttctt tttttttttt tttcttgttg 7440 ttcttggtca gttattcaaa ggaaaaggga ttgaaataaa aacttgcatg tgggatc 7497
<210> 126 <211> 709 <212> DNA <213> Seqüência Artificial <220> <223> oligonucleotideo HPfad3A'-2 <400> 126
gattgtggcc atggaaggag atagataagc agagaaaaca tggtagccca acacagtgtt 60 gatattgcta ttccttttct ctcactgggt gggaacagat tgctgtaggg attgaagtga 120 gagccttcct ttccggggct tcttgaaaac aaataaattg gatacacaaa caatggaaat 180 ggcacagtga atctaatgag tcttgtcatg ctgtctagat tcttgtaaat cttctctgtt 240 aatggaaccc atgactcatc cttctcaatg tgtccatggt tttgatggtg agttctgtgg 300 ctaattctcc atccatggta tggcacaaga attgaggaat gcaagatgtg tcccaccagg 360 ctattcagca aagggctatc tgaaaagctt ccatggccac aatcatgtcc aagaacaaag 420 agagcccaga acattgtgcc ttgaatgggg caatagatta gccagagaag ccagttgtcg 480 aagtgaattg ctgcagccac caatgcagca attacaagca catccctgag aacataactg 540 agggatctcc atggattctt gacccagcaa tgttttggta ttgaagctct gatttctgca 600 atcttaaacg gtggaggagc gctaggatca aaatcaaaag aagaaccctt ttgttggtat 660 ccattattgg cagcataggc taaaggcttt gtgtctttaa cgcggccgc 709
<210> 127 <211> 1014 <212> DNA
<213> Seqüência Artificial <220>
<223> oligonucleotídeo HPfad3ABA'-2 <400> 127
gcggccgcgt taaagacaca aagcctttag cctatgctgc caataatgga taccaacaaa 60 agggttcttc ttttgatttt gatcctagcg ctcctccacc gtttaagatt gcagaaatca 120 gagcttcaat accaaaacat tgctgggtca agaatccatg gagatccctc agttatgttc 180 tcagggatgt gcttgtaatt gctgcattgg tggctgcagc aattcacttc gacaactggc 240 ttctctggct aatctattgc cccattcaag gcacaatgtt ctgggctctc tttgttcttg 300 gacatgattg tggccatgga aggagataga taagcagaga aaacatggta gcccaacaca 360 gtgttgatat tgctattcct tttctctcac tgggtgggaa cagattgctg tagggattga 420 agtgagagcc ttcctttccg gggcttcttg aaaacaaata aattggatac acaaacaatg 480 gaaatggcac agtgaatcta atgagtcttg tcatgctgtc tagattcttg taaatcttct 540 ctgttaatgg aacccatgac tcatccttct caatgtgtcc atggttttga tggtgagttc 600 tgtggctaat tctccatcca tggtatggca caagaattga ggaatgcaag atgtgtccca 660 ccaggctatt cagcaaaggg ctatctgaaa agcttccatg gccacaatca tgtccaagaa 720 caaagagagc ccagaacatt gtgccttgaa tggggcaata gattagccag agaagccagt 780 tgtcgaagtg aattgctgca gccaccaatg cagcaattac aagcacatcc ctgagaacat 840 aactgaggga tctccatgga ttcttgaccc agcaatgttt tggtattgaa gctctgattt 900 ctgcaatctt aaacggtgga ggagcgctag gatcaaaatc aaaagaagaa cccttttgtt 960 ggtatccatt attggcagca taggctaaag gctttgtgtc tttaacgcgg ccgc 1014 <210> 128 <211> 4526 <212> DNA <213> Seqüência Artificial <220> <223> plasmideo pLF129 <400> 128 ttgtaatacg actcactata gggcgaattg ggccctctag atgcatgctc gagcggccgc 60 cagtgtgatg gatatctgca gaattcaggg cggccgcgtt aaagacacaa agcctttagc 120 ctatgctgcc aataatggat accaacaaaa gggttcttct tttgattttg atcctagcgc 180 tcctccaccg tttaagattg cagaaatcag agcttcaata ccaaaacatt gctgggtcaa 240 gaatccatgg agatccctca gttatgttct cagggatgtg cttgtaattg ctgcattggt 300 ggctgcagca attcacttcg acaactggct tctctggcta atctattgcc ccattcaagg 360 cacaatgttc tgggctctct ttgttcttgg acatgattgt ggccatggaa ggagatagat 420 aagcagagaa aacatggtag cccaacacag tgttgatatt gctattcctt ttctctcact 480
gggtgggaac agattgctgt agggattgaa gtgagagcct tcctttccgg ggcttcttga 540
aaacaaataa attggataca caaacaatgg aaatggcaca gtgaatctaa tgagtcttgt 600
catgctgtct agattcttgt aaatcttctc tgttaatgga acccatgact catccttctc 660
aatgtgtcca tggttttgat ggtgagttct gtggctaatt ctccatccat ggtatggcac 720
aagaattgag gaatgcaaga tgtgtcccac caggctattc agcaaagggc tatctgaaaa 780
gcttccatgg ccacaatcat gtccaagaac aaagagagcc cagaacattg tgccttgaat 840
ggggcaatag attagccaga gaagccagtt gtcgaagtga attgctgcag ccaccaatgc 900
agcaattaca agcacatccc tgagaacata actgagggat ctccatggat tcttgaccca 960
gcaatgtttt ggtattgaag ctctgatttc tgcaatctta aacggtggag gagcgctagg 1020
atcaaaatca aaagaagaac ccttttgttg gtatccatta ttggcagcat aggctaaagg 1080
ctttgtgtct ttaacgcggc cgccctgaat tccagcacac tggcggccgt tactagtgga 1140
tccgagctcg gtaccaagct tgatgcatag cttgagtatt ctaacgcgtc acctaaatag 1200
cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc 1260
acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta 1320
actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca 1380
gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc 1440
cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 1500
tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 1560
gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 1620
ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 1680
aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 1740
tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 1800
ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 1860
gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 1920
tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 1980
caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 2040
ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt 2100
cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 2160
ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 2220
cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 2280 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttagcac 2340
gtgtcagtcc tgctcctcgg ccacgaagtg cacgcagttg ccggccgggt cgcgcagggc 2400
gaactcccgc ccccacggct gctcgccgat ctcggtcatg gccggcccgg aggcgtcccg 2460
gaagttcgtg gacacgacct ccgaccactc ggcgtacagc tcgtccaggc cgcgcaccca 2520
cacccaggcc agggtgttgt ccggcaccac ctggtcctgg accgcgctga tgaacagggt 2580
cacgtcgtcc cggaccacac cggcgaagtc gtcctccacg aagtcccggg agaacccgag 2640
ccggtcggtc cagaactcga ccgctccggc gacgtcgcgc gcggtgagca ccggaacggc 2700
actggtcaac ttggccatgg tggccctcct cacgtgctat tattgaagca tttatcaggg 2760
ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 2820
tccgcgcaca tttccccgaa aagtgccacc tgatgcggtg tgaaataccg cacagatgcg 2880
taaggagaaa ataccgcatc aggaaattgt aagcgttaat aattcagaag aactcgtcaa 2940
gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga 3000
agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt 3060
cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat 3120
tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt 3180
cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatyctctt 3240
cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc 3300
gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca 3360
ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct 3420
gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca 3480
cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca 3540
gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg 3600
acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga 3660
atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc 3720
gaaacgatcc tcatcctgtc tcttgatcag agcttgatcc cctgcgccat cagatccttg 3780
gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc 3840
cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg 3900
taagcccact gcaagctacc tgctttctct ttgcgcttgc gttttccctt gtccagatag 3960
cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct ttctacgtga 4020
aaaggatcta ggtgaagatc ctttttgata atctcatgcc tgacatttat attccccaga 4080
acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg gctgagatca gccacttctt 4140 ccccgataac ggagaccggc acactggcca tatcggtggt catcatgcgc cagctttcat 4200
ccccgatatg caccaccggg taaagttcac gggagacttt atctgacagc agacgtgcac 4260
tggccagggg gatcaccatc cgtcgccccg gcgtgtcaat aatatcactc tgtacatcca 4320
caaacagacg ataacggctc tctcttttat aggtgtaaac cttaaactgc cgtacgtata 4380
ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg 4440
cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac 4500
gacgttgtaa aacgacggcc agtgaa 4526
<210> 129 <211> 8503 <212> DNA
<213> Seqüência Artificial <220>
<223> pKR1189 <4 00> 129
ggccgcatga gccgtaaagg ttcaatacaa cgagtgcttg ttttcttagg gacaagcatt 60
gtacttatgt atgattctgt gtaaccatga gtcttccacg ttgtactaat gtgaagggca 120
aaaataaaac acagaacaag ttcgtttttc tcaaataatg tgaaggtaga aaatggaacc 180
atgcctcctc tcttgcatgt gatttaaaat attagcagat ggtaccgtac gtctagagga 240
tccaagcttg gcgcgccgtt ctatagtgtc acctaaatcg tatgtgtatg atacataagg 300
ttatgtatta attgtagccg cgttctaacg acaatatgtc catatggtgc actctcagta 360
caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg 420
cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg 480
ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc 54 0
tcgtgatacg cctattttta taggttaatg tcatgaccaa aatcccttaa cgtgagtttt 600
cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt 660
ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 720
tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga 780
taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag 840
caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata 900
agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 960
gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga 1020
gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 1080
ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 114 0 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 1200
tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 1260
ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt 1320
ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga 1380
ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc 1440
tccccgcgcg ttggccgatt cattaatgca ggttgatcag attcgacatc gatctagtaa 1500
catagatgac accgcgcgcg ataatttatc ctagtttgcg cgctatattt tgttttctat 1560
cgcgtattaa atgtataatt gcgggactct aatcataaaa acccatctca taaataacgt 1620
catgcattac atgttaatta ttacatgctt aacgtaattc aacagaaatt atatgataat 1680
catcgcaaga ccggcaacag gattcaatct taagaaactt tattgccaaa tgtttgaacg 1740
atctgcttcg acgcactcct tctttaggta cctcactatt cctttgccct cggacgagtg 1800
ctggggcgtc ggtttccact atcggcgagt acttctacac agccatcggt ccagacggcc 1860
gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg ctccggatcg gacgattgcg 1920
tcgcatcgac cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa gctctgatag 1980
agttggtcaa gaccaatgcg gagcatatac gcccggagcc gcggcgatcc tgcaagctcc 2040
ggatgcctcc gctcgaagta gcycytctgc tgctccatac aagccaacca cggcctccag 2100
aagaagatgt tggcgacctc gtattgggaa tccccgaaca tcgcctcgct ccagtcaatg 2160
accgctgtta tgcggccatt gtccgtcagg acattgttgg agccgaaatc cgcgtgcacg 2220
aggtgccgga cttcggggca gtcctcggcc caaagcatca gctcatcgag agcctgcgcg 2280
acggacgcac tgacggtgtc gtccatcaca gtttgccagt gatacacatg gggatcagca 2340
atcgcgcata tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg tccgaatggg 2400
ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg catccatggc ctccgcgacc 2460
ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac accctgtgca 2520
cggcgggaga tgcaataggt caggctctcg ctgaattccc caatgtcaag cacttccgga 2580
atcgggagcg cggccgatgc aaagtgccga taaacataac gatctttgta gaaaccatcg 2640
gcgcagctat ttacccgcag gacatatcca cgccctccta catcgaagct gaaagcacga 2700
gattcttcgc cctccgagag ctgcatcagg tcggagacgc tgtcgaactt ttcgatcaga 2760
aacttctcga cagacgtcgc ggtgagttca ggctttttca tggtttaata agaagagaaa 2820
agagttcttt tgttatggct gaagtaatag agaaatgagc tcgagcgtgt cctctccaaa 2880
tgaaatgaac ttccttatat agaggaaggg tcttgcgaag gatagtggga ttgtgcgtca 2940
tcccttacgt cagtggagat gtcacatcaa tccacttgct ttgaagacgt ggttggaacg 3000 tcttcttttt ccacgatgct cctcgtgggt gaggcatctt gaatgatagc ctttccttta tccttttcta ctgtcctttc gatgaagtga tttcccgaaa ttatcctttg ttgaaaagtc tctttgacat ttttggagta gaccagagtg ttcttgtcat tgagtcgtaa aagactctgt tctgttagat cctcgatttg aatcttagac taccttttta gagactccaa tctctattac cgtccatact ggaatagtac ttctgatctt gcaattagtc ctgaatcttt tgactgcatc ctggagattg ttactcgggt agatcgtctt catctgtggg tcagcattct ttctgaaatt gaacatagtg tcgtcacctt caccttcgaa atcgtccatt gtaatctccg gggcaaagga ccttttggtt cctagcgtga gccagtgggc agcaaagctc ttgggcttga gttgagcttc ttgctgactt gttgtgtacg cgtcagctgc cttgtgtgca actccgggaa cgccgtttgt tataccggca tgtggaccgt tatacacaac cgattctgct ctgagaggag caactgtgct tggatcgatc tcgatcccgc gaaattaata ccctctagaa ataattttgt ttaactttaa ctcaccgcga cgtctgtcga gaagtttctg atgcagctct cggagggcga agaatctcgt tatgtcctgc gggtaaatag ctgcgccgat cactttgcat cggccgcgct cccgattccg agcctgacct attgcatctc ccgccgtgca accgaactgc ccgctgttct gcagccggtc gatcttagcc agacgagcgg gttcggccca acatggcgtg atttcatatg cgcgattgct atggacgaca ccgtcagtgc gtccgtcgcg
gggggtccat ctttgggacc actgtcggca 3060
tcgcaatgat ggcatttgta ggagccacct 3120
cagatagctg ggcaatggaa tccgaggagg 3180
tcaatagccc tttggtcttc tgagactgta 3240
tcgtgctcca ccatgttgac gaagattttc 3300
atgaactgtt cgccagtctt cacggcgagt 3360
tccatgcatg gccttagatt cagtaggaac 3420
ttgccttggt ttatgaagca agccttgaat 3480
gagaaatatg tctttctctg tgttcttgat 3540
tttaaccttc ttgggaaggt atttgatctc 3600
gatgagacct gctgcgtagg cctctctaac 3660
gaagaggcta accttctcat tatcagtggt 3720
cttccttcct agatcgtaaa gatagaggaa 3780
gatctctttt ggggctggat cactgctggg 3840
tttttgcttt ggtgggcttg ttagggcctt 3900
tcctttgggg atgaagttca acctgtctgt 3960
tgctcttgcc tctgtaatag tggcaaattt 4020
tgccgccttt gtacaacccc agtcatcgta 4080
gtagtagttg atatgagggt gttgaatacc 4140
gttaagctca gatttttgtg ggattggaat 4200
cgactcacta tagggagacc acaacggttt 4260
gaaggagata tacccatgga aaagcctgaa 4320
atcgaaaagt tcgacagcgt ctccgacctg 4380
gctttcagct tcgatgtagg agggcgtgga 4440
ggtttctaca aagatcgtta tgtttatcgg 4500
gaagtgcttg acattgggga attcagcgag 4560
cagggtgtca cgttgcaaga cctgcctgaa 4 620
gcggaggcta tggatgcgat cgctgcggcc 4680
ttcggaccgc aaggaatcgg tcaatacact 4740
gatccccatg tgtatcactg gcaaactgtg 4800
caggctctcg atgagctgat gctttgggcc 4860 gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg 4920
acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc 4 980
caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag 5040
acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat 5100
atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat 5160
gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg 5220
cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc 5280
gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata gtgaggtaca 5340
gcttggatcg atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc 5400
gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg 5460
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ctgaaaggag gaactatatc cggatgatcg ggcgcgccgt cgactcgacg tacgatccca 5520
catgcaagtt tttatttcaa tcccttttcc tttgaataac tgaccaagaa caacaagaaa 5580
aaaaaaaaaa aagaaaagga tcattttgaa aggatatttt tcgctcctat tcaaatactg 5640
tatttttacc aaaaaaactg tatttttcct acactctcaa gctttgtttt tcgcttcgac 5700
tctcatgatt tccttcatat gccaatcact ctatttataa atggcataag gtagtgtgaa 5760
caattgcaaa gcttgtcatc aaaagcttgc aatgtacaaa ttaatgtttt tcatgccttt 5820
caaaattatc tgcaccccct agctattaat ctaacatcta agtaaggcta gtgaattttt 5880
tcgaatagtc atgcagtgca ttaatttccc cgtgactatt ttggctttga ctccaacact 5940
ggccccgtac atccgtccct cattacatga aaagaaatat tgtttatatt cttaattaaa 6000
aatattgtcc cttctaaatt ttcatatagt taattattat attacttttt tctctattct 6060
attagttcta ttttcaaatt attatttatg catatgtaaa gtacattata tttttgctat 6120
atacttaaat atttctaaat tattaaaaaa agactgatat gaaaaattta ttctttttaa 6180
agctatatca ttttatatat actttttctt ttcttttctt tcattttcta ttcaatttaa 6240
taagaaataa attttgtaaa tttttattta tcaatttata aaaatatttt actttatatg 6300
ttttttcaca tttttgttaa acaaatcata tcattatgat tgaaagagag gaaattgaca 6360
gtgagtaata agtgatgaga aaaaaatgtg ttatttccta aaaaaaacct aaacaaacat 6420
gtatctactc tctatttcat ctatctctca tttcattttt ctctttatct ctttctttat 6480
ttttttatca tatcatttca cattaattat ttttactctc tttatttttt ctctctatcc 6540
ctctcttatt tccactcata tatacactcc aaaattgggg catgccttta tcactactct 6600
atctcctcca ctaaatcatt taaatgaaac tgaaaagcat tggcaagtct cctcccctcc 6660
tcaagtgatt tccaactcag cattggcatc taattgattc agtatatcta ttgcatgtgt 6720 aaaagtcttt ccacaataca taactattaa ttaatcttaa ataaataaag gataaaatat 6780
ttttttttct tcataaaatt aaaatatgtt attttttgtt tagatgtata ttcgaataaa 6840
tctaaatata tgataatgat tttttatatt gattaaacat ataatcaata ttaaatatga 6900
tattttttta tataggttgt acacataatt ttataaggat aaaaaatatg ataaaaataa 6960
attttaaata tttttatatt tacgagaaaa aaaaatattt tagccataaa taaatgacca 7020
gcatatttta caaccttagt aattcataaa ttcctatatg tatatttgaa attaaaaaca 7080
gataatcgtt aagggaagga atcctacgtc atctcttgcc atttgttttt catgcaaaca 7140
gaaagggacg aaaaaccacc tcaccatgaa tcactcttca caccattttt actagcaaac 7200
aagtctcaac aactgaagcc agctctcttt ccgtttcttt ttacaacact ttctttgaaa 7260
tagtagtatt tttttttcac atgatttatt aacgtgccaa aagatgctta ttgaatagag 7320
tgcacatttg taatgtacta ctaattagaa catgaaaaag cattgttcta acacgataat 7380
cctgtgaagg cgttaactcc aaagatccaa tttcactata taaattgtga cgaaagcaaa 7440
atgaattcac atagctgaga gagaaaggaa aggttaacta agaagcaata cttcagcggc 7500
cgcgttaaag acacaaagcc tttagcctat gctgccaata atggatacca acaaaagggt 7560
tcttcttttg attttgatcc tagcgctcct ccaccgttta agattgcaga aatcagagct 7620
tcaataccaa aacattgctg ggtcaagaat ccatggagat ccctcagtta tgttctcagg 7680
gatgtgcttg taattgctgc attggtggct gcagcaattc acttcgacaa ctggcttctc 7740
tggctaatct attgccccat tcaaggcaca atgttctggg ctctctttgt tcttggacat 7800
gattgtggcc atggaagctt ttcagatagc cctttgctga atagcctggt gggacacatc 7860
ttgcattcct caattcttgt gccataccat ggatggagaa ttagccacag aactcaccat 7920
caaaaccatg gacacattga gaaggatgag tcatgggttc cattaacaga gaagatttac 7 980
aagaatctag acagcatgac aagactcatt agattcactg tgccatttcc attgtttgtg 8040
tatccaattt atttgttttc aagaagcccc ggaaaggaag gctctcactt caatccctac 8100
agcaatctgt tcccacccag tgagagaaaa ggaatagcaa tatcaacact gtgttgggct 8160
accatgtttt ctctgcttat ctatctcctt ccatggccac aatcatgtcc aagaacaaag 8220
agagcccaga acattgtgcc ttgaatgggg caatagatta gccagagaag ccagttgtcg 8280
aagtgaattg ctgcagccac caatgcagca attacaagca catccctgag aacataactg 8340
agggatctcc atggattctt gacccagcaa tgttttggta ttgaagctct gatttctgca 8400
atcttaaacg gtggaggagc gctaggatca aaatcaaaag aagaaccctt ttgttggtat 8460
ccattattgg cagcataggc taaaggcttt gtgtctttaa cgc 8503 <210> 130
<211> 4112
<212> DNA
<213> Seqüência Artificial <220>
<223> pKR1209 <400> 130 agcttgatgc atagcttgag tattctaacg cgtcacctaa atagcttggc gtaatcatgg 60 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 120 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 180 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 240 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 300 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 360 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 420 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 480 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 540 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 600 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 660 79η gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 780 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 840 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 900 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 960 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 1020 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 1080 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 1140 atcttcacct agatcctttt aaattaaaaa tgaagtttta gcacgtgtca gtcctgctcc 1200 tcggccacga agtgcacgca gttgccggcc gggtcgcgca gggcgaactc ccgcccccac 1260 ggctgctcgc cgatctcggt catggccggc ccggaggcgt cccggaagtt cgtggacacg 1320 acctccgacc actcggcgta cagctcgtcc aggccgcgca cccacaccca ggccagggtg 1380 ttgtccggca ccacctggtc ctggaccgcg ctgatgaaca gggtcacgtc gtcccggacc 1440 acaccggcga agtcgtcctc cacgaagtcc cgggagaacc cgagccggtc ggtccagaac 1500 tcgaccgctc cggcgacgtc gcgcgcggtg agcaccggaa cggcactggt caacttggcc 1560 atggtggccc tcctcacgtg ctattattga agcatttatc agggttattg tctcatgagc 1620 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 1680 cgaaaagtgc cacctgatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg 1740 catcaggaaa ttgtaagcgt taataattca gaagaactcg tcaagaaggc gatagaaggc 1800 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc 1860 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc 1920 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt 1980 cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt 2040 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg 2100 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg 2160 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat 2220 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc 2280 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac 2340 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc 2400 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc 2460 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca 2520 oton £.--' U tgtctcttga tcagagcttg atcccctgcg ccatcagatc cttggcggca agaaagccat 2640 ccagtttact ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg 2700 ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc 2760 tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt 2820 catccggggt cagcaccgtt tctgcggact ggctttctac gtgaaaagga tctaggtgaa 2880 gatccttttt gataatctca tgcctgacat ttatattccc cagaacatca ggttaatggc 2940 gtttttgatg tcattttcgc ggtggctgag atcagccact tcttccccga taacggagac 3000 cggcacactg gccatatcgg tggtcatcat gcgccagctt tcatccccga tatgcaccac 3060 cgggtaaagt tcacgggaga ctttatctga cagcagacgt gcactggcca gggggatcac 3120 catccgtcgc cccggcgtgt caataatatc actctgtaca tccacaaaca gacgataacg 3180 gctctctctt ttataggtgt aaaccttaaa ctgccgtacg tataggctgc gcaactgttg 3240 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc 3300 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac 3360 ggccagtgaa ttgtaatacg actcactata gggcgaattg ggccctctag atgcatgctc 3420 gagcggccgc gttaaagaca caaagccttt agcctatgct gccaataatg gataccaaca 3480 aaagggttct tcttttgatt ttgatcctag cgctcctcca ccgtttaaga ttgcagaaat 3540 cagagcttca ataccaaaac attgctgggt caagaatcca tggagatccc tcagttatgt 3600 tctcagggat gtgcttgtaa ttgctgcatt ggtggctgca gcaattcact tcgacaactg 3660 gcttctctgg ctaatctatt gccccattca aggcacaatg ttctgggctc tctttgttct 3720 tggacatgat tgtggccatg gaaggagata gataagcaga gaaaacatgg tagcccaaca 3780 cagtgttgat attgctattc cttttctctc actgggtggg aacagattgc tgtagggatt 3840 gaagtgagag ccttcctttc cggggcttct tgaaaacaaa taaattggat acacaaacaa 3900 tggaaatggc acagtgaatc taatgagtct tgtcatgctg tctagattct tgtaaatctt 3960 ctctgttaat ggaacccatg actcatcctt ctcaatgtgt ccatggtttt gatggtgagt 4020 tctgtggcta attctccatc catggtatgg cacaagaatt gaggaatgca agatgtgtcc 4080 caccaggcta ttcagcaaag ggctatctga aa 4112 <210> 131 <211> 1143 <212> DNA <213> Glycine max <4 00> 131 atggttcaag cacagcctct acaacatgtt ggtaatggtg cagggaaaga agatcaagct 60 tattttgatc caagtgctcc accacccttc aagattgcaa atatcagagc agcaattcca 120 aaacattgct gggagaagaa cacattgaga tctctgagtt atgttctgag ggatgtgttg 180 gtagtgactg cattggtagc tgcagcaatc ggcttcaata gctggttctt ctggccactc 240 tattggcctg cacaaggcac aatgttttgg gcactttttg ttcttggaca tgattgtggt 300 catggaagtt tttcaaacag tcctttgttg aacagcattg tgggccacat cttgcactct 360 tcaattcttg taccatacca tggatggaga attagccaca ggactcacca tcagaaccat 420 ggccatgttg agaaggatga atcatgggtt ccgcttacag agaaagttta caagaatcta 480 gacaacatga caagaatgat gagattcact cttcctttcc ccatctttgc ataccccttt 540 tatttgtgga gcagaagccc tggaaaagaa ggctctcatt tcaaccctta cagcaacttg 600 ttctctcctg gtgagagaag agatgtgcta acttcaactc tgtgttgggg catcatgctt 660 tctgtgcttc tctatctttc cctcacaatg ggtccacttt ttatgctcaa gctctatggg 720 gttccctatt tgatcttcgt catgtggctg gatttcgtca cgtacttgca tcatcatggt 780 tacaagcaga aactgccttg gtaccgtggc caggaatgga gttatctaag gggtggtctt 840 acaacagtag atcgcgacta tggttggatc aacaacattc accatgacat tggcacccat 900 gttatccatc accttttccc tcaaattcca cattatcatt taatcgaagc gactaaagca 960 gcaaaggcag tgctaggaaa gtattatcgt gagcctcaga aatctgggcc attgccactt 1020
catctaataa agtacttgct ccacagcata agtcaggatc acttcgttag cgactctggc 1080
gacattgtgt actaccagac tgattcccag ctccacaaag attcttggac ccagtccaac 1140
taa 1143
<210> 132
<211> 380
<212> PRT
<213> Glycine max
<400> 132
Met Val Gln Ala Gln Pro Leu Gln His Val Gly Asn Gly Ala Gly Lys 1 5 10 15
Glu Asp Gln Ala Tyr Phe Asp Pro Ser Ala Pro Pro Pro Phe Lys Ile 20 25 30
Ala Asn Ile Arg Ala Ala Ile Pro Lys His Cys Trp Glu Lys Asn Thr 35 40 45
Leu Arg Ser Leu Ser Tyr Val Leu Arg Asp Val Leu Val Val Thr Ala 50 55 60
Leu Val Ala Ala Ala Ile Gly Phe Asn Ser Trp Phe Phe Trp Pro Leu 65 70 75 80
Tyr Trp Pro Ala Gln Gly Thr Met Phe Trp Ala Leu Phe Val Leu Gly 85 90 95
His Asp Cys Gly His Gly Ser Phe Ser Asn Ser Pro Leu Leu Asn Ser 100 105 110
Ile Val Gly His Ile Leu His Ser Ser Ile Leu Val Pro Tyr His Gly 115 120 125
Trp Arg Ile Ser His Arg Thr His His Gln Asn His Gly His Val Glu 130 135 140
Lys Asp Glu Ser Trp Val Pro Leu Thr Glu Lys Val Tyr Lys Asn Leu 145 150 155 160
Asp Asn Met Thr Arg Met Met Arg Phe Thr Leu Pro Phe Pro Ile Phe 165 170 175
Ala Tyr Pro Phe Tyr Leu Trp Ser Arg Ser Pro Gly Lys Glu Gly Ser 180 185 190 His Phe Asn Pro Tyr Ser Asn Leu Phe Ser Pro Gly Glu Arg Arg Asp 195 200 205
Val Leu Thr Ser Thr Leu Cys Trp Gly Ile Met Leu Ser Val Leu Leu 210 215 220
Tyr Leu Ser Leu Thr Met Gly Pro Leu Phe Met Leu Lys Leu Tyr Gly 225 230 235 240
Val Pro Tyr Leu Ile Phe Val Met Trp Leu Asp Phe Val Thr Tyr Leu 245 250 255
His His His Gly Tyr Lys Gln Lys Leu Pro Trp Tyr Arg Gly Gln Glu 260 265 270
Trp Ser Tyr Leu Arg Gly Gly Leu Thr Thr Val Asp Arg Asp Tyr Gly 275 280 285
Trp Ile Asn Asn Ile His His Asp Ile Gly Thr His Val Ile His His 290 295 300
Leu Phe Pro Gln Ile Pro His Tyr His Leu Ile Glu Ala Thr Lys Ala 305 310 315 320
Ala Lys Ala Val Leu Gly Lys Tyr Tyr Arg Glu Pro Gln Lys Ser Gly 325 330 335
Pro Leu Pro Leu His Leu Ile Lys Tyr Leu Leu His Ser Ile Ser Gln 340 345 350
Asp His Phe Val Ser Asp Ser Gly Asp Ile Val Tyr Tyr Gln Thr Asp 355 360 365
Ser Gln Leu His Lys Asp Ser Trp Thr Gln Ser Asn 370 375 380
<210> 133 <211> 40 <212> DNA
<213> Seqüência Artificial <220>
<223> oligonucleotideo fad3c-5
<400> 133
ctcgagaagc ggccgcattg atccaagtgc tccaccaccc <210> 134
<211> 28
<212> DNA
<213> Seqüência Artificial
<220>
<223> oligonucleotideo fad3c-3
<400> 134
gatatctatg tccaagaaca aaaagtgc
<210> 135 <211> 3764 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo pKR1213 <400> 135
aattccagca cactggcggc cgttactagt ggatccgagc tcggtaccaa gcttgatgca 60
120 180 240 300 420 480 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260
tagcttgagt attctaacgc gtcacctaaa tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttyCyta ttgggcgctc ttccgcttcc tcgctcactg acucgct_gcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaagcccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttag cacgtgtcag tcctgctcct cggccacgaa gtgcacgcag ttgccggccg ggtcgcgcag ggcgaactcc cgcccccacg gctgctcgcc 1320
gatctcggtc atggccggcc cggaggcgtc ccggaagttc gtggacacga cctccgacca 1380
ctcggcgtac agctcgtcca ggccgcgcac ccacacccag gccagggtgt tgtccggcac 1440
cacctggtcc tggaccgcgc tgatgaacag ggtcacgtcg tcccggacca caccggcgaa 1500
gtcgtcctcc acgaagtccc gggagaaccc gagccggtcg gtccagaact cgaccgctcc 1560
ggcgacgtcg cgcgcggtga gcaccggaac ggcactggtc aacttggcca tggtggccct 1620
cctcacgtgc tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 1680
tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 1740
acctgtatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa 1800
ttgtaagcgt taataattca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc 1860
gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc 1920
tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc 1980
cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag 2040
gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg 2100
aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga 2160
ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg 2220
caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc 2280
tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc 2340
cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg 2400
gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg 2460
gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag 2520
cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga 2580
gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga 2640
tcagagcttg atcccctgcg ccatcagatc cttggcggcg agaaagccat ccagtttact 2700
ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg ttcgcttgct 2760
gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc tacctgcttt 2820
ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt catccggggt 2880
cagcaccgtt tctgcggact ggctttctac gtgaaaagga tctaggtgaa gatccttttt 2940
gataatctca tgcctgacat ttatattccc cagaacatca ggttaatggc gtttttgatg 3000
tcattttcgc ggtggctgag atcagccact tcttccccga taacggagac cggcacactg 3060
gccatatcgg tggtcatcat gcgccagctt tcatccccga tatgcaccac cgggtaaagt 3120 tcacgggaga ctttatctga cagcagacgt gcactggcca gggggatcac catccgtcgc 3180
cccggcgtgt caataatatc actctgtaca tccacaaaca gacgataacg gctctctctt 3240
ttataggtgt aaaccttaaa ctgccgtacg tataggctgc gcaactgttg ggaagggcga 3300
tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga 3360
ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa 3420
ttgtaatacg actcactata gggcgaattg ggccctctag atgcatgctc gagcggccgc 3480
cagtgtgatg gatatctgca gaattcaggg atatctatgt ccaagaacaa aaagtgccca 3540
aaacattgtg ccttgtgcag gccaatagag tggccagaag aaccagctat tgaagccgat 3600
tgctgcagct accaatgcag tcactaccaa cacatccctc agaacataac tcagagatct 3660
caatgtgttc ttctcccagc aatgttttgg aattgctgct ctgatatttg caatcttgaa 3720
gggtggtgga gcacttggat caatgcggcc gcttctcgag cctg 3764
<210> 136 <211> 4353 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmídeo pKR1218 <400> 136
ggccgcgtta aagacacaaa gcctttagcc tatgctgcca ataatggata ccaacaaaag 60
ggttcttctt ttgattttga tcctagcgct cctccaccgt ttaagattgc agaaatcaga 120
gcttcaatac caaaacattg ctgggtcaag aatccatgga gatccctcag ttatgttctc 180
agggatgtgc ttgtaattgc tgcattggtg gctgcagcaa ttcacttcga caactggctt 240
ctctggctaa tctattgccc cattcaaggc acaatgttct gggctctctt tgttcttgga 300
catgattgtg gccatggaag gagatagata agcagagaaa acatggtagc ccaacacagt 360
gttgatattg ctattccttt tctctcactg ggtgggaaca gattgctgta gggattgaag 420
tgagagcctt cctttccggg gcttcttgaa aacaaataaa ttggatacac aaacaatgga 480
aatggcacag tgaatctaat gagtcttgtc atgctgtcta gattcttgta aatcttctct 540
gttaatggaa cccatgactc atccttctca atgtgtccat ggttttgatg gtgagttctg 600
tggctaattc tccatccatg gtatggcaca agaattgagg aatgcaagat gtgtcccacc 660
aggctattca gcaaagggct atctgaaaag cttgatgcat agcttgagta ttctaacgcg 720
tcacctaaat agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 780
gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 840
atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 900 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 960 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 1020 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 1080 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 1140 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 1200 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 1260 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 1320 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 1380 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 1440 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 1500 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 1560 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa 1620 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 1680 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 1740 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 1800 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 1860 aagttttagc acgtgtcagt cctgctcctc ggccacgaag tgcacgcagt tgccggccgg 1920 gtcgcgcagg gcgaactccc gcccccacgg ctgctcgccg atctcggtca tggccggccc 1980 ggaggcgtcc cggaagttcg tggacacgac ctccgaccac tcggcgtaca gctcgtccag 2040 gccgcgcacc cacacccagg ccagggtgtt gtccggcacc acctggtcct ggaccgcgct 2100 gatgaacagg gtcacgtcgt cccggaccac accggcgaag tcgtcctcca cgaagtcccg 2160 ggagaacccg agccggtcgg tccagaactc gaccgctccg gcgacgtcgc gcgcggtgag 2220 caccggaacg gcactggtca acttggccat ggtggccctc ctcacgtgct attattgaag 2280 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 2340 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgatgcgg tgtgaaatac 2400 cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta ataattcaga 2460 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt 2520 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag 2580 ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag 2640 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga 2700 gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc 2760 cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg 2820 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat 2880 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg 2940 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga 3000 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg 3060 cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc 3120 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc 3180 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt 3240 gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agagcttgat cccctgcgcc 3300 atcagatcct tggcggcaag aaagccatcc agtttacttt gcagggcttc ccaaccttac 3360 cagagggcgc cccagctggc aattccggtt cgcttgctgt ccataaaacc gcccagtcta 3420 gctatcgcca tgtaagccca ctgcaagcta cctgctttct ctttgcgctt gcgttttccc 3480 ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg 3540 ctttctacgt gaaaaggatc taggtgaaga tcctttttga taatctcatg cctgacattt 3600 atattcccca gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg tggctgagat 3660 cagccacttc ttccccgata acggagaccg gcacactggc catatcggtg gtcatcatgc 3720 gccagctttc atccccgata tgcaccaccg ggtaaagttc acgggagact ttatctgaca 3780 gcagacgtgc actggccagg gggatcacca tccgtcgccc cggcgtgtca ataatatcac 3840 tctgtacatc cacaaacaga cgataacggc tctctctttt ataggtgtaa accttaaact 3900 gccgtacgta taggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 3960 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 4020 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg 4080 gcgaattggg ccctctagat gcatgctcga gaagcggccq cattgatcca agtgctccac 4140 cacccttcaa gattgcaaat atcagagcag caattccaaa acattgctgg gagaagaaca 4200 cattgagatc tctgagttat gttctgaggg atgtgttggt agtgactgca ttggtagctg 4260 cagcaatcgg cttcaatagc tggttcttct ggccactcta ttggcctgca caaggcacaa 4320 tgttttgggc actttttgtt cttggacata gat 4353
<210> 137
<211> 3742
<212> DNA
<213> Seqüência Artificial
<220> <223> plasmídeo pKR1210 <400> 137
agcttgatgc atagcttgag tattctaacg cgtcacctaa atagcttggc gtaatcatgg 60 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 120 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 180 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 240 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 300 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 360 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 420 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 480 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 540 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 600 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 660 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 720 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 780 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 840 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 900 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 960 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 1020 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 1080 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 1140 atcttcacct agatcctttt aaattaaaaa tgaagtttta gcacgtgtca gtcctgctcc 1200 tcggccacga agtgcacgca gttgccggcc gggtcgcgca gggcgaactc ccgcccccac 1260 ggctgctcgc cgatctcggt catggccggc ccggaggcgt cccggaagtt cgtggacacg 1320 acctccgacc actcggcgta cagctcgtcc aggccgcgca cccacaccca ggccagggtg 1380 ttgtccggca ccacctggtc ctggaccgcg ctgatgaaca gggtcacgtc gtcccggacc 1440 acaccggcga agtcgtcctc cacgaagtcc cgggagaacc cgagccggtc ggtccagaac 1500 tcgaccgctc cggcgacgtc gcgcgcggtg agcaccggaa cggcactggt caacttggcc 1560 atggtggccc tcctcacgtg ctattattga agcatttatc agggttattg tctcatgagc 1620 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 1680 cgaaaagtgc cacctgatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg 1740 catcaggaaa ttgtaagcgt taataattca gaagaactcg tcaagaaggc gatagaaggc 1800 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc 1860 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc 1920 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt 1980 cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt 2040 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg 2100 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg 2160 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat 2220 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc 2280 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac 2340 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc 2400 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc 2460 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca 2520 agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc 2580 tgtctcttga tcagagcttg atcccctgcg ccatcagatc cttggcggca agaaagccat 2640 ccagtttact ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg 2700 ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc 2760 tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt 2820 catccggggt cagcaccgtt tctgcggact ggctttctac gtgaaaagga tctaggtgaa 2880 gatccttttt gataatctca tgcctgacat ttatattccc cagaacatca ggttaatggc 2940 gtttttgatg tcattttcgc ggtggctgag atcagccact tcttccccga taacggagac 3000 cggcacactg gccatatcgg tggtcatcat gcgccagctt tcatccccga tatgcaccac 3060 cgggtaaagt tcacgggaga ctttatctga cagcagacgt gcactggcca gggggatcac 3120 catccgtcgc cccggcgtgt caataatatc actctgtaca tccacaaaca gacgataacg 3180 gctctctctt ttataggtgt aaaccttaaa ctgccgtacg tataggctgc gcaactgttg 3240 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc 3300 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac 3360 ggccagtgaa ttgtaatacg actcactata gggcgaattg ggccctctag atgcatgctc 3420 gagcggccgc gttaaagaca caaagccttt agcctatgct gccaataatg gataccaaca 3480 aaagggttct tcttttgatt ttgatcctag cgctcctcca ccgtttaaga ttgcagaaat 3540 cagagcttca ataccaaaac attgctgggt caagaatcca tggagatccc tcagttatgt 3600 tctcagggat gtgcttgtaa ttgctgcatt ggtggctgca gcaattcact tcgacaactg 3660 gcttctctgg ctaatctatt gccccattca aggcacaatg ttctgggctc tctttgttct 3720
tggacatgat tgtggccatg ga 3742
<210> 138 <211> 3983 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo pKR1219 <400> 138
ggccgcgtta aagacacaaa gcctttagcc tatgctgcca ataatggata ccaacaaaag 60
ggttcttctt ttgattttga tcctagcgct cctccaccgt ttaagattgc agaaatcaga 120
gcttcaatac caaaacattg ctgggtcaag aatccatgga gatccctcag ttatgttctc 180
agggatgtgc ttgtaattgc tgcattggtg gctgcagcaa ttcacttcga caactggctt 240
ctctggctaa tctattgccc cattcaaggc acaatgttct gggctctctt tgttcttgga 300
catgattgtg gccatggaag cttgatgcat agcttgagta ttctaacgcg tcacctaaat 360
agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 420
ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 480
taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 540
cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 600
tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 660
gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 720
atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 780
ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 840
cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 900
tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 960
gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 1020
aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 1080
tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 1140
aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 1200
aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc 1260
ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 1320
ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 1380
atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 1440 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttagc 1500 acgtgtcagt cctgctcctc ggccacgaag tgcacgcagt tgccggccgg gtcgcgcagg 1560 gcgaactccc gcccccacgg ctgctcgccg atctcggtca tggccggccc ggaggcgtcc 1620 cggaagttcg tggacacgac ctccgaccac tcggcgtaca gctcgtccag gccgcgcacc 1680 cacacccagg ccagggtgtt gtccggcacc acctggtcct ggaccgcgct gatgaacagg 1740 gtcacgtcgt cccggaccac accggcgaag tcgtcctcca cgaagtcccg ggagaacccg 1800 agccggtcgg tccagaactc gaccgctccg gcgacgtcgc gcgcggtgag caccggaacg 1860 gcactggtca acttggccat ggtggccctc ctcacgtgct attattgaag catttatcag 1920 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 1980 gttccgcgca catttccccg aaaagtgcca cctgatgcgg tgtgaaatac cgcacagatg 2040 cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta ataattcaga agaactcgtc 2100 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag 2160 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat 2220 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc 2280 attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc 2340 gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc 2400 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat 2460 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg 2520 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc 2580 ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag 2640 cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg 2700 cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc 2760 tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc 2820 gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat 2880 gcgaaacgat cctcatcctg tctcttgatc agagcttgat cccctgcgcc atcagatcct 2940 tggcggcaag aaagccatcc agtttacttt gcagggcttc ccaaccttac cagagggcgc 3000 cccagctggc aattccggtt cgcttgctgt ccataaaacc gcccagtcta gctatcgcca 3060 tgtaagccca ctgcaagcta cctgctttct ctttgcgctt gcgttttccc ttgtccagat 3120 agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctacgt 3180 gaaaaggatc taggtgaaga tcctttttga taatctcatg cctgacattt atattcccca 3240 gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg tggctgagat cagccacttc 3300 ttccccgata acggagaccg gcacactggc catatcggtg gtcatcatgc gccagctttc 3360 atccccgata tgcaccaccg ggtaaagttc acgggagact ttatctgaca gcagacgtgc 3420 actggccagg gggatcacca tccgtcgccc cggcgtgtca ataatatcac tctgtacatc 3480 cacaaacaga cgataacggc tctctctttt ataggtgtaa accttaaact gccgtacgta 3540 taggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct 3600 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc 3660 acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg gcgaattggg 3720 ccctctagat gcatgctcga gaagcggccg cattgatcca agtgctccac cacccttcaa 3780 gattgcaaat atcagagcag caattccaaa acattgctgg gagaagaaca cattgagatc 3840 tctgagttat gttctgaggg atgtgttggt agtgactgca ttggtagctg cagcaatcgg 3900 cttcaatagc tggttcttct ggccactcta ttggcctgca caaggcacaa tgttttgggc 3960 actttttgtt cttggacata gat 3983
<210> 139 <211> 4894 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo pKR1225 <400> 139
cgcgtcacct aaatagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 60
atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 120
cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 180
gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 240
gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 300
ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 360
acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 420
cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 480
caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 540
gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 600
tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 660
aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 720
ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 780
cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 840 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 900 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 960 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 1020 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 1080 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 1140 aatgaagttt tagcacgtgt cagtcctgct cctcggccac gaagtgcacg cagttgccgg 1200 ccgggtcgcg cagggcgaac tcccgccccc acggctgctc gccgatctcg gtcatggccg 1260 gcccggaggc gtcccggaag ttcgtggaca cgacctccga ccactcggcg tacagctcgt 1320 ccaggccgcg cacccacacc caggccaggg tgttgtccgg caccacctgg tcctggaccg 1380 cgctgatgaa cagggtcacg tcgtcccgga ccacaccggc gaagtcgtcc tccacgaagt 1440 cccgggagaa cccgagccgg tcggtccaga actcgaccgc tccggcgacg tcgcgcgcgg 1500 tgagcaccgg aacggcactg gtcaacttgg ccatggtggc cctcctcacg tgctattatt 1560 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 1620 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgat gcggtgtgaa 1680 ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaataatt 1740 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata 1800 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg 1860 gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat 1920 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg 1980 acgagatcct cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg 2040 agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta 2100 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc 2160 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga 2220 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca 2280 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc 2340 gctgcctcgt cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc 2400 gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt 2460 gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca 2520 tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcagagct tgatcccctg 2580 cgccatcaga tccttggcgg caagaaagcc atccagttta ctttgcaggg cttcccaacc 2640 ttaccagagg gcgccccagc tggcaattcc ggttcgcttg ctgtccataa aaccgcccag 2700 tctagctatc gccatgtaag cccactgcaa gctacctgct ttctctttgc gcttgcgttt 2760 tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga 2820 ctggctttct acgtgaaaag gatctaggtg aagatccttt ttgataatct catgcctgac 2880 atttatattc cccagaacat caggttaatg gcgtttttga tgtcattttc gcggtggctg 2940 agatcagcca cttcttcccc gataacggag accggcacac tggccatatc ggtggtcatc 3000 atgcgccagc tttcatcccc gatatgcacc accgggtaaa gttcacggga gactttatct 3060 gacagcagac gtgcactggc cagggggatc accatccgtc gccccggcgt gtcaataata 3120 tcactctgta catccacaaa cagacgataa cggctctctc ttttataggt gtaaacctta 3180 aactgccgta cgtataggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg 3240 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca 3300 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta 3360 tagggcgaat tgggccctct agatgcatgc tcgagaagcg gccgcattga tccaagtgct 3420 ccaccaccct tcaagattgc aaatatcaga gcagcaattc caaaacattg ctgggagaag 3480 aacacattga gatctctgag ttatgttctg agggatgtgt tggtagtgac tgcattggta 3540 gctgcagcaa tcggcttcaa tagctggttc ttctggccac tctattggcc tgcacaaggc 3600 acaatgtttt gggcactttt tgttcttgga catagatggc cgcgttaaag acacaaagcc 3660 tttagcctat gctgccaata atggatacca acaaaagggt tcttcttttg attttgatcc 3720 tagcgctcct ccaccgttta agattgcaga aatcagagct tcaataccaa aacattgctg 3780 ggtcaagaat ccatggagat ccctcagtta tgttctcagg gatgtgcttg taattgctgc 3840 attggtggct gcagcaattc acttcgacaa ctggcttctc tggctaatct attgccccat 3900 tcaaggcaca atgttctggg ctctctttgt tcttggacat gattgtggcc atggaagctt 3960 ttcagatagc cctttgctga atagcctggt gggacacatc ttgcattcct caattcttgt 4020 gccataccat ggatggagaa ttagccacag aactcaccat caaaaccatg gacacattga 4080 gaaggatgag tcatgggttc cattaacaga gaagatttac aagaatctag acagcatgac 4140 aagactcatt agattcactg tgccatttcc attgtttgtg tatccaattt atttgttttc 4200 aagaagcccc ggaaaggaag gctctcactt caatccctac agcaatctgt tcccacccag 4260 tgagagaaaa ggaatagcaa tatcaacact gtgttgggct accatgtttt ctctgcttat 4320 ctatctcctt ccatggccac aatcatgtcc aagaacaaag agagcccaga acattgtgcc 4380 ttgaatgggg caatagatta gccagagaag ccagttgtcg aagtgaattg ctgcagccac 4440 caatgcagca attacaagca catccctgag aacataactg agggatctcc atggattctt 4500 gacccagcaa tgttttggta ttgaagctct gatttctgca atcttaaacg gtggaggagc 4560 gctaggatca aaatcaaaag aagaaccctt ttgttggtat ccattattgg cagcataggc 4 620
taaaggcttt gtgtctttaa cgcggccatc tatgtccaag aacaaaaagt gcccaaaaca 4 680 ttgtgccttg tgcaggccaa tagagtggcc agaagaacca gctattgaag ccgattgctg 4740
cagctaccaa tgcagtcact accaacacat ccctcagaac ataactcaga gatctcaatg 4800
tgttcttctc ccagcaatgt tttggaattg ctgctctgat atttgcaatc ttgaagggtg 4860
gtggagcact tggatcaatg cggccgcttc tcga 4894
<210> 140 <211> 8979 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmídeo pKR1229 <400> 140
ggccgcatga gccgtaaagg ttcaatacaa cgagtgcttg ttttcttagg gacaagcatt 60
gtacttatgt atgattctgt gtaaccatga gtcttccacg ttgtactaat gtgaagggca 120
aaaataaaac acagaacaag ttcgtttttc tcaaataatg tgaaggtaga aaatggaacc 180
atgcctcctc tcttgcatgt gatttaaaat attagcagat ggtaccgtac gtctagagga 240
tccaagcttg gcgcgccgtt ctatagtgtc acctaaatcg tatgtgtatg atacataagg 300
ttatgtatta attgtagccg cgttctaacg acaatatgtc catatggtgc actctcagta 360
caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg 420
cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg 480
ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc 54 0
tcgtgatacg cctattttta taggttaatg tcatgaccaa aatcccttaa cgtgagtttt 600
cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt 660
ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 720
tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga 780
taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag 840
caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata 900
agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 960
gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga 1020
gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 1080
ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 1140
acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 1200 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 1260 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt 1320 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga 1380 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc 1440 tccccgcgcg ttggccgatt cattaatgca ggttgatcag attcgacatc gatctagtaa 1500 catagatgac accgcgcgcg ataatttatc ctagtttgcg cgctatattt tgttttctat 1560 cgcgtattaa atgtataatt gcgggactct aatcataaaa acccatctca taaataacgt 1620 catgcattac atgttaatta ttacatgctt aacgtaattc aacagaaatt atatgataat 1680 catcgcaaga ccggcaacag gattcaatct taagaaactt tattgccaaa tgtttgaacg 1740 atctgcttcg acgcactcct tctttaggta cctcactatt cctttgccct cggacgagtg 1800 ctggggcgtc ggtttccact atcggcgagt acttctacac agccatcggt ccagacggcc 1860 gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg ctccggatcg gacgattgcg 1920 tcgcatcgac cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa gctctgatag 1980 agttggtcaa gaccaatgcg gagcatatac gcccggagcc gcggcgatcc tgcaagctcc 2040 ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac aagccaacca cggcctccag 2100 aagaagatgt tggcgacctc gtattgggaa tccccgaaca tcgcctcgct ccagtcaatg 2160 accgctgtta tgcggccatt gtccgtcagg acattgttgg agccgaaatc cgcgtgcacg 2220 aggtgccgga cttcggggca gtcctcggcc caaagcatca gctcatcgag agcctgcgcg 2280 acggacgcac tgacggtgtc gtccatcaca gtttgccagt gatacacatg gggatcagca 2340 atcgcgcata tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg tccgaatggg 2400 ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg catccatggc ctccgcgacc 2460 ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac accctgtgca 2520 cggcgggaga tgcaataggt caggctctcg ctgaattccc caatgtcaag cacttccgga 2580 atcgggagcg cggccgatgc aaagtgccga taaacataac gatctttgta gaaaccatcg 2640 gcgcagctat ttacccgcag gacatatcca cgccctccta catcgaagct gaaagcacga 2700 gattcttcgc cctccgagag ctgcatcagg tcggagacgc tgtcgaactt ttcgatcaga 2760 aacttctcga cagacgtcgc ggtgagttca ggctttttca tggtttaata agaagagaaa 2820 agagttcttt tgttatggct gaagtaatag agaaatgagc tcgagcgtgt cctctccaaa 2880 tgaaatgaac ttccttatat agaggaaggg tcttgcgaag gatagtggga ttgtgcgtca 2940 tcccttacgt cagtggagat gtcacatcaa tccacttgct ttgaagacgt ggttggaacg 3000 tcttcttttt ccacgatgct cctcgtgggt gggggtccat ctttgggacc actgtcggca 3060 gaggcatctt gaatgatagc ctttccttta tcgcaatgat ggcatttgta ggagccacct 3120 tccttttcta ctgtcctttc gatgaagtga cagatagctg ggcaatggaa tccgaggagg 3180 tttcccgaaa ttatcctttg ttgaaaagtc tcaatagccc tttggtcttc tgagactgta 3240 tctttgacat ttttggagta gaccagagtg tcgtgctcca ccatgttgac gaagattttc 3300 ttcttgtcat tgagtcgtaa aagactctgt atgaactgtt cgccagtctt cacggcgagt 3360 tctgttagat cctcgatttg aatcttagac tccatgcatg gccttagatt cagtaggaac 3420 taccttttta gagactccaa tctctattac ttgccttggt ttatgaagca agccttgaat 3480 cgtccatact ggaatagtac ttctgatctt gagaaatatg tctttctctg tgttcttgat 3540 gcaattagtc ctgaatcttt tgactgcatc tttaaccttc ttgggaaggt atttgatctc 3600 ctggagattg ttactcgggt agatcgtctt gatgagacct gctgcgtagg cctctctaac 3660 catctgtggg tcagcattct ttctgaaatt gaagaggcta accttctcat tatcagtggt 3720 gaacatagtg tcgtcacctt caccttcgaa cttccttcct agatcgtaaa gatagaggaa 3780 atcgtccatt gtaatctccg gggcaaagga gatctctttt ggggctggat cactgctggg 3840 ccttttggtt cctagcgtga gccagtgggc tttttgcttt ggtgggcttg ttagggcctt 3900 agcaaagctc ttgggcttga gttgagcttc tcctttgggg atgaagttca acctgtctgt 3960 ttgctgactt gttgtgtacg cgtcagctgc tgctcttgcc tctgtaatag tggcaaattt 4020 cttgtgtgca actccgggaa cgccgtttgt tgccgccttt gtacaacccc agtcatcgta 4080 tataccggca tgtggaccgt tatacacaac gtagtagttg atatgagggt gttgaatacc 4140 cgattctgct ctgagaggag caactgtgct gttaagctca gatttttgtg ggattggaat 4200 tggatcgatc tcgatcccgc gaaattaata cgactcacta tagggagacc acaacggttt 4260 ccctctagaa ataattttgt ttaactttaa gaaggagata tacccatgga aaagcctgaa 4320 ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg 4380 atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga 4440 tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg 4500 cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga attcagcgag 4560 agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa 4620 accgaactgc ccgctgttct gcagccggtc gcggaggcta tggatgcgat cgctgcggcc 4680 gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact 4740 acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg 4800 atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc 4860 gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg 4920 acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc 4980
caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag 5040
acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat 5100
atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat 5160
gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg 5220
cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc 5280
gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata gtgaggtaca 534 0
gcttggatcg atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc 5400
gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg 5460
ctgaaaggag gaactatatc cggatgatcg ggcgcgccgt cgactcgacg tacgatccca 5520
catgcaagtt tttatttcaa tcccttttcc tttgaataac tgaccaagaa caacaagaaa 5580
aaaaaaaaaa aagaaaagga tcattttgaa aggatatttt tcgctcctat tcaaatactg 5640
tatttttacc aaaaaaactg tatttttcct acactctcaa gctttgtttt tcgcttcgac 5700
tctcatgatt tccttcatat gccaatcact ctatttataa atggcataag gtagtgtgaa 5760
caattgcaaa gcttgtcatc aaaagcttgc aatgtacaaa ttaatgtttt tcatgccttt 5820
caaaattatc tgcaccccct agctattaat ctaacatcta agtaaggcta gtgaattttt 5880
tcgaatagtc atgcagtgca ttaatttccc cgtgactatt ttggctttga ctccaacact 5940
ggccccgtac atccgtccct cattacatga aaagaaatat tgtttatatt cttaattaaa 6000
aatattgtcc cttctaaatt ttcatatagt taattattat attacttttt tctctattct 6060
attagttcta ttttcaaatt attatttatg catatgtaaa gtacattata tttttgctat 6120
atacttaaat atttctaaat tattaaaaaa agactgatat gaaaaattta ttctttttaa 6180
agctatatca ttttatatat actttttctt ttcttttctt tcattttcta ttcaatttaa 6240
taagaaataa attttgtaaa tttttattta tcaatttata aaaatatttt actttatatg 6300
ttttttcaca tttttgttaa acaaatcata tcattatgat tgaaagagag gaaattgaca 6360
gtgagtaata agtgatgaga aaaaaatgtg ttatttccta aaaaaaacct aaacaaacat 6420
gtatctactc tctatttcat ctatctctca tttcattttt ctctttatct ctttctttat 6480
ttttttatca tatcatttca cattaattat ttttactctc tttatttttt ctctctatcc 6540
ctctcttatt tccactcata tatacactcc aaaattgggg catgccttta tcactactct 6600
atctcctcca ctaaatcatt taaatgaaac tgaaaagcat tggcaagtct cctcccctcc 6660
tcaagtgatt tccaactcag cattggcatc taattgattc agtatatcta ttgcatgtgt 6720
aaaagtcttt ccacaataca taactattaa ttaatcttaa ataaataaag gataaaatat 6780 ttttttttct tcataaaatt aaaatatgtt attttttgtt tagatgtata ttcgaataaa 6840
tctaaatata tgataatgat tttttatatt gattaaacat ataatcaata ttaaatatga 6900
tattttttta tataggttgt acacataatt ttataaggat aaaaaatatg ataaaaataa 6960
attttaaata tttttatatt tacgagaaaa aaaaatattt tagccataaa taaatgacca 7020
gcatatttta caaccttagt aattcataaa ttcctatatg tatatttgaa attaaaaaca 7080
gataatcgtt aagggaagga atcctacgtc atctcttgcc atttgttttt catgcaaaca 7140
gaaagggacg aaaaaccacc tcaccatgaa tcactcttca caccattttt actagcaaac 7200
aagtctcaac aactgaagcc agctctcttt ccgtttcttt ttacaacact ttctttgaaa 7260
tagtagtatt tttttttcac atgatttatt aacgtgccaa aagatgctta ttgaatagag 7320
tgcacatttg taatgtacta ctaattagaa catgaaaaag cattgttcta acacgataat 7380
cctgtgaagg cgttaactcc aaagatccaa tttcactata taaattgtga cgaaagcaaa 7440
atgaattcac atagctgaga gagaaaggaa aggttaacta agaagcaata cttcagcggc 7500
cgcattgatc caagtgctcc accacccttc aagattgcaa atatcagagc agcaattcca 7560
aaacattgct gggagaagaa cacattgaga tctctgagtt atgttctgag ggatgtgttg 7620
gtagtgactg cattggtagc tgcagcaatc ggcttcaata gctggttctt ctggccactc 7680
tattggcctg cacaaggcac aatgttttgg gcactttttg ttcttggaca tagatggccg 7740
cgttaaagac acaaagcctt tagcctatgc tgccaataat ggataccaac aaaagggttc 7800
ttcttttgat tttgatccta gcgctcctcc accgtttaag attgcagaaa tcagagcttc 7860
aataccaaaa cattgctggg tcaagaatcc atggagatcc ctcagttatg ttctcaggga 7920
tgtgcttgta attgctgcat tggtggctgc agcaattcac ttcgacaact ggcttctctg 7980
gctaatctat tgccccattc aaggcacaat gttctgggct ctctttgttc ttggacatga 8040
ttgtggccat ggaagctttt cagatagccc tttgctgaat agcctggtgg gacacatctt 8100
gcattcctca attcttgtgc cataccatgg atggagaatt agccacagaa ctcaccatca 8160
aaaccatgga cacattgaga aggatgagtc atgggttcca ttaacagaga agatttacaa 8220
gaatctagac agcatgacaa gactcattag attcactgtg ccatttccat tgtttgtgta 8280
tccaatttat ttgttttcaa gaagccccgg aaaggaaggc tctcacttca atccctacag 8340
caatctgttc ccacccagtg agagaaaagg aatagcaata tcaacactgt gttgggctac 8400
catgttttct ctgcttatct atctccttcc atggccacaa tcatgtccaa gaacaaagag 8460
agcccagaac attgtgcctt gaatggggca atagattagc cagagaagcc agttgtcgaa 8520
gtgaattgct gcagccacca atgcagcaat tacaagcaca tccctgagaa cataactgag 8580
ggatctccat ggattcttga cccagcaatg ttttggtatt gaagctctga tttctgcaat 8640 cttaaacggt ggaggagcgc taggatcaaa atcaaaagaa gaaccctttt gttggtatcc 8700
attattggca gcataggcta aaggctttgt gtctttaacg cggccatcta tgtccaagaa 8760
caaaaagtgc ccaaaacatt gtgccttgtg caggccaata gagtggccag aagaaccagc 8820
tattgaagcc gattgctgca gctaccaatg cagtcactac caacacatcc ctcagaacat 8880
aactcagaga tctcaatgtg ttcttctccc agcaatgttt tggaattgct gctctgatat 8940
ttgcaatctt gaagggtggt ggagcacttg gatcaatgc 8979
<210> 141
<211> 10221
<212> DNA
<213> Seqüência Artificial
<220>
<223> plasmideo pKR1249
<400> 141
gtacgtctag aggatccgtc gacggcgcgc ccgatcatcc ggatatagtt cctcctttca 60 gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt tattgctcag 120 cggtggcagc agccaactca gcttcctttc gggctttgtt agcagccgga tcgatccaag 180 ctgtacctca ctattccttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg 240 cgagtacttc tacacagcca tcggtccaga cggccgcgct tctgcgggcg atttgtgtac 300 gcccgacagt cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg 360 catcatcgaa attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca 420 tatacgcccg gagccgcggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg 480 tctgctgctc catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt 540 gggaatcccc gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg 600 tcaggacatt gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct 660 cggcccaaag catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca 720 tcacagtttg ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg 780 tagtgtattg accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat 840 cggccgcagc gatcgcatcc atagcctccg cgaccggctg cagaacagcg ggcagttcgg 900 tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc 960 tctcgctgaa ttccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt 1020 gccgataaac ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat 1080 atccacgccc tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca 1140 tcaggtcgga gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga 1200 gttcaggctt ttccatgggt atatctcctt cttaaagtta aacaaaatta tttctagagg 1260 gaaaccgttg tggtctccct atagtgagtc gtattaattt cgcgggatcg agatctgatc 1320 aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 1380 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 1440 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 1500 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 1560 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 1620 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 1680 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 1740 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 1800 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 1860 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 1920 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 1980 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 2040 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 2100 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 2160 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 2220 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt 2280 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa 2340 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg 2400 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatggacata 2460 ttgtcgttag aacgcggcta caattaatac ataaccttat gtatcataca catacgattt 2520 aggtgacact atagaacggc gcgccaagct gggtctagaa ctagaaacgt gatgccactt 2580 gttattgaag tcgattacag catctattct gttttactat ttataacttt gccatttctg 2640 acttttgaaa actatctctg gatttcggta tcgctttgtg aagatcgagc aaaagagacg 2700 ttttgtggac gcaatggtcc aaatccgttc tacatgaaca aattggtcac aatttccact 2760 aaaagtaaat aaatggcaag ttaaaaaagg aatatgcatt ttactgattg cctaggtgag 2820 ctccaagaga agttgaatct acacgtctac caaccgctaa aaaaagaaaa acattgaata 2880 tgtaacctga ttccattagc ttttgacttc ttcaacagat tctctactta gatttctaac 2940 agaaatatta ttactagcac atcattttca gtctcactac agcaaaaaat ccaacggcac 3000 aatacagaca acaggagata tcagactaca gagatagata gatgctactg catgtagtaa 3060 gttaaataaa aggaaaataa aatgtcttgc taccaaaact actacagact atgatgctca 3120 ccacaggcca aatcctgcaa ctaggacagc attatcttat atatattgta caaaacaagc 3180 atcaaggaac atttggtcta ggcaatcagt acctcgttct accatcaccc tcagttatca 3240 catccttgaa ggatccatta ctgggaatca tcggcaacac atgctcctga tggggcacaa 3300 tgacatcaag aaggtagggg ccaggggtgt ccaacattct ctgaattgcc gctctaagct 3360 cttccttctt cgtcactcgc gctgccggta tcccacaagc atcagcaaac ttgagcatgt 3420 ttgggaatat ctcgctctcg ctagacggat ctccaagata ggtgtgagct ctattggact 3480 tgtagaacct atcctccaac tgaaccacca tacccaaatg ctgattgttc aacaacaata 3540 tcttaactgg gagattctcc actcttatag tggccaactc ctgaacattc atgatgaaac 3600 taccatcccc atcaatgtca accacaacag ccccagggtt agcaacagca gcaccaatag 3660 ccgcaggcaa tccaaaaccc atggctccaa gaccccctga ggtcaaccac tgcctcggtc 3720 tcttgtactt gtaaaactgc gcagcccaca tttgatgctg cccaacccca gtactaacaa 3780 tagcatctcc attagtcaac tcatcaagaa cctcgatagc atgctgcgga gaaatcgcgt 3840 cctggaatgt cttgtaaccc aatggaaact tgtgtttctg cacattaatc tcttctctcc 3900 aacctccaag atcaaactta ccctccactc ctttctcctc caaaatcata ttaattccct 3960 tcaaggccaa cttcaaatcc gcgcaaaccg acacgtgcgc ctgcttgttc ttcccaatct 4020 cggcagaatc aatatcaatg tgaacaatct tagccctact agcaaaagcc tcaagcttcc 4080 cagtaacacg gtcatcaaac cttaccccaa aggcaagcaa caaatcacta ttgtcaacag 4140 catagttagc ataaacagta ccatgcatac ccagcatctg aagggaatat tcatcaccaa 4200 taggaaaagt tccaagaccc attaaagtgc tagcaacggg aataccagtg agttcaacaa 4260 agcgcctcaa ttcagcactg gaattcaaac tgccaccgcc gacgtagaga acgggctttt 4320 gggcctccat gatgagtctg acaatgtgtt ccaattgggc ctcggcgggg ggcctgggca 4380 gcctggcgag gtaaccgggg aggttaacgg gctcgtccca attaggcacg gcgagttgct 4440 gctgaacgtc tttgggaatg tcgatgagga ccggaccggg gcggccggag gtggcgacga 4500 agaaagcctc ggcgacgacg cgggggatgt cgtcgacgtc gaggatgagg tagttgtgct 4560 tcgtgatgga tctgctcacc tccacgatcg gggtttcttg gaaggcgtcg gtgccgatca 4620 tccggcgggc gacctggccg gtgatggcga cgactgggac gctgtccatt aaagcgtcgg 4680 cgaggccgct cacgaggttg gtggcgccgg ggccggaggt ggcaatgcag acgccgggga 4740 ggccggagga acgcgcgtag ccttcggcgg cgaagacgcc gccctgctcg tggcgcggga 4800 gcacgttgcg gatggcggcg gagcgcgtga gcgcctggtg gatctccatc gacgcaccgc 4860 cggggtacgc gaacaccgtc gtcacgccct gcctctccag cgcctccaca aggatgtccg 4920 cgcccttgcg aggttcgccg gaggcgaacc gtgacacgaa gggctccgtg gtcggcgctt 4 980
ccttggtgaa gggcgccgcc gtggggggtt tggagatgga acatttgatt ttgagagcgt 5040
ggttgggttt ggtgagggtt tgatgagaga gagggagggt ggatctagta atgcgtttgg 5100
ggaaggtggg gtgtgaagag gaagaagaga atcgggtggt tctggaagcg gtggccgcca 5160
ttgtgttgtg tggcatggtt atacttcaaa aactgcacaa caagcctaga gttagtacct 5220
aaacagtaaa tttacaacag agagcaaaga cacatgcaaa aatttcagcc ataaaaaaag 5280
ttataataga atttaaagca aaagtttcat tttttaaaca tatatacaaa caaactggat 5340
ttgaaggaag ggattaattc ccctgctcaa agtttgaatt cctattgtga cctatactcg 5400
aataaaattg aagcctaagg aatgtatgag aaacaagaaa acaaaacaaa actacagaca 54 60
aacaagtaca attacaaaat tcgctaaaat tctgtaatca ccaaacccca tctcagtcag 5520
cacaaggccc aaggtttatt ttgaaataaa aaaaaagtga ttttatttct cataagctaa 5580
aagaaagaaa ggcaattatg aaatgatttc gactagatct gaaagtcaaa cgcgtattcc 5640
gcagatatta aagaaagagt agagtttcac atggatccta gatggaccca gttgaggaaa 5700
aagcaaggca aagcaaacca gaagtgcaag atccgaaatt gaaccacgga atctaggatt 57 60
tggtagaggg agaagaaaag taccttgaga ggtagaagag aagagaagag cagagagata 5820
tatgaacgag tgtgtcttgg tctcaactct gaagcgatac gagtttagag gggagcattg 5880
agttccaatt tatagggaaa ccgggtggca ggggtgagtt aatgacggaa aagcccctaa 5940
gtaacgagat tggattgtgg gttagattca accgtttgca tccgcggctt agattgggga 6000
agtcagagtg aatctcaacc gttgactgag ttgaaaattg aatgtagcaa ccaattgagc 6060
caaccccagc ctttgccctt tgattttgat ttgtttgttg catacttttt atttgtcttc 6120
tggttctgac tctctttctc tcgtttcaat gccaggttgc ctactcccac accactcaca 6180
agaagattct actgttagta ttaaatattt tttaatgtat taaatgatga atgcttttgt 6240
aaacagaaca agactatgtc taataagtgt cttgcaacat tttttaagaa attaaaaaaa 6300
atatatttat tatcaaaatc aaatgtatga aaaatcatga ataatataat tttatacatt 6360
tttttaaaaa atcttttaat ttcttaatta atatcttaaa aataatgatt aatatttaac 6420
ccaaaataat tagtatgatt ggtaaggaag atatccatgt tatgtttgga tgtgagtttg 6480
atctagagca aagcttacta gagtcgacct gcaggtcgac tcgacgtacg atcccacatg 6540
caagttttta tttcaatccc ttttcctttg aataactgac caagaacaac aagaaaaaaa 6600
aaaaaaaaga aaaggatcat tttgaaagga tatttttcgc tcctattcaa atactgtatt 6660
tttaccaaaa aaactgtatt tttcctacac tctcaagctt tgtttttcgc ttcgactctc 6720
atgatttcct tcatatgcca atcactctat ttataaatgg cataaggtag tgtgaacaat 6780 tgcaaagctt gtcatcaaaa gcttgcaatg tacaaattaa tgtttttcat gcctttcaaa 6840
attatctgca ccccctagct attaatctaa catctaagta aggctagtga attttttcga 6900
atagtcatgc agtgcattaa tttccccgtg actattttgg ctttgactcc aacactggcc 6960
ccgtacatcc gtccctcatt acatgaaaag aaatattgtt tatattctta attaaaaata 7020
ttgtcccttc taaattttca tatagttaat tattatatta cttttttctc tattctatta 7080
gttctatttt caaattatta tttatgcata tgtaaagtac attatatttt tgctatatac 7140
ttaaatattt ctaaattatt aaaaaaagac tgatatgaaa aatttattct ttttaaagct 7200
atatcatttt atatatactt tttcttttct tttctttcat tttctattca atttaataag 7260
aaataaattt tgtaaatttt tatttatcaa tttataaaaa tattttactt tatatgtttt 7320
ttcacatttt tgttaaacaa atcatatcat tatgattgaa agagaggaaa ttgacagtga 7380
gtaataagtg atgagaaaaa aatgtgttat ttcctaaaaa aaacctaaac aaacatgtat 7440
ctactctcta tttcatctat ctctcatttc atttttctct ttatctcttt ctttattttt 7500
ttatcatatc atttcacatt aattattttt actctcttta ttttttctct ctatccctct 7560
cttatttcca ctcatatata cactccaaaa ttggggcatg cctttatcac tactctatct 7620
cctccactaa atcatttaaa tgaaactgaa aagcattggc aagtctcctc ccctcctcaa 7680
gtgatttcca actcagcatt ggcatctaat tgattcagta tatctattgc atgtgtaaaa 7740
gtctttccac aatacataac tattaattaa tcttaaataa ataaaggata aaatattttt 7800
ttttcttcat aaaattaaaa tatgttattt tttgtttaga tgtatattcg aataaatcta 7860
aatatatgat aatgattttt tatattgatt aaacatataa tcaatattaa atatgatatt 7920
tttttatata ggttgtacac ataattttat aaggataaaa aatatgataa aaataaattt 7980
taaatatttt tatatttacg agaaaaaaaa atattttagc cataaataaa tgaccagcat 8040
attttacaac cttagtaatt cataaattcc tatatgtata tttgaaatta aaaacagata 8100
atcgttaagg gaaggaatcc tacgtcatct cttgccattt gtttttcatg caaacagaaa 8160
gggacgaaaa accacctcac catgaatcac tcttcacacc atttttacta gcaaacaagt 8220
ctcaacaact gaagccagct ctctttccgt ttctttttac aacactttct ttgaaatagt 8280
agtatttttt tttcacatga tttattaacg tgccaaaaga tgcttattga atagagtgca 8340
catttgtaat gtactactaa ttagaacatg aaaaagcatt gttctaacac gataatcctg 8400
tgaaggcgtt aactccaaag atccaatttc actatataaa ttgtgacgaa agcaaaatga 8460
attcacatag ctgagagaga aaggaaaggt taactaagaa gcaatacttc agcggccgca 8520
ttgatccaag tgctccacca cccttcaaga ttgcaaatat cagagcagca attccaaaac 8580
attgctggga gaagaacaca ttgagatctc tgagttatgt tctgagggat gtgttggtag 8640 tgactgcatt ggtagctgca gcaatcggct tcaatagctg gttcttctgg ccactctatt 8700
ggcctgcaca aggcacaatg ttttgggcac tttttgttct tggacataga tggccgcgtt 8760
aaagacacaa agcctttagc ctatgctgcc aataatggat accaacaaaa gggttcttct 8820
tttgattttg atcctagcgc tcctccaccg tttaagattg cagaaatcag agcttcaata 8880
ccaaaacatt gctgggtcaa gaatccatgg agatccctca gttatgttct cagggatgtg 8940
cttgtaattg ctgcattggt ggctgcagca attcacttcg acaactggct tctctggcta 9000
atctattgcc ccattcaagg cacaatgttc tgggctctct ttgttcttgg acatgattgt 9060
ggccatggaa gcttttcaga tagccctttg ctgaatagcc tggtgggaca catcttgcat 9120
tcctcaattc ttgtgccata ccatggatgg agaattagcc acagaactca ccatcaaaac 9180
catggacaca ttgagaagga tgagtcatgg gttccattaa cagagaagat ttacaagaat 9240
ctagacagca tgacaagact cattagattc actgtgccat ttccattgtt tgtgtatcca 9300
atttatttgt tttcaagaag ccccggaaag gaaggctctc acttcaatcc ctacagcaat 9360
ctgttcccac ccagtgagag aaaaggaata gcaatatcaa cactgtgttg ggctaccatg 9420
ttttctctgc ttatctatct ccttccatgg ccacaatcat gtccaagaac aaagagagcc 9480
cagaacattg tgccttgaat ggggcaatag attagccaga gaagccagtt gtcgaagtga 9540
attgctgcag ccaccaatgc agcaattaca agcacatccc tgagaacata actgagggat 9600
ctccatggat tcttgaccca gcaatgtttt ggtattgaag ctctgatttc tgcaatctta 9660
aacggtggag gagcgctagg atcaaaatca aaagaagaac ccttttgttg gtatccatta 9720
ttggcagcat aggctaaagg ctttgtgtct ttaacgcggc catctatgtc caagaacaaa 9780
aagtgcccaa aacattgtgc cttgtgcagg ccaatagagt ggccagaaga accagctatt 9840
gaagccgatt gctgcagcta ccaatgcagt cactaccaac acatccctca gaacataact 9900
cagagatctc aatgtgttct tctcccagca atgttttgga attgctgctc tgatatttgc 9960
aatcttgaag ggtggtggag cacttggatc aatgcggccg catgagccgt aaaggttcaa 10020
tacaacgagt gcttgttttc ttagggacaa gcattgtact tatgtatgat tctgtgtaac 10080
catgagtctt ccacgttgta ctaatgtgaa gggcaaaaat aaaacacaga acaagttcgt 10140
ttttctcaaa taatgtgaag gtagaaaatg gaaccatgcc tcctctcttg catgtgattt 10200
aaaatattag cagatggtac c 10221
<210> 142 <211> 1263 <212> PRT
<213> Euglena gracilis
<400> 142 Ala Thr Gly Ala Ala Gly Thr Cys Ala Ala Ala Gly Cys Gly Cys Cys 1 5 10 15
Ala Ala Gly Cys Gly Cys Thr Thr Cys Cys Cys Cys Thr Thr Ala Cys 20 25 30
Ala Ala Thr Thr Gly Ala Thr Gly Gly Ala Ala Cys Ala Ala Cys Ala 35 40 45
Thr Ala Thr Gly Ala Thr Gly Thr Gly Thr Cys Thr Gly Cys Cys Thr 50 55 60
Gly Gly Gly Thr Cys Ala Ala Thr Thr Thr Cys Cys Ala Cys Cys Cys 65 70 75 80
Thr Gly Gly Thr Gly Gly Thr Gly Cys Gly Gly Ala Ala Ala Thr Thr 85 90 95
Ala Thr Ala Gly Ala Gly Ala Ala Thr Thr Ala Cys Cys Ala Ala Gly 100 105 110
Gly Ala Ala Gly Gly Gly Ala Thr Gly Cys Cys Ala Cys Thr Gly Ala 115 120 125
Thr Gly Cys Cys Thr Thr Cys Ala Thr Gly Gly Thr Thr Ala Thr Gly 130 135 140
Cys Ala Cys Thr Cys Thr Cys Ala Ala Gly Ala Ala Gly Cys Cys Thr 145 150 155 160
Thr Cys Gly Ala Cys Ala Ala Gly Cys Thr Cys Ala Ala Gly Cys Gly 165 170 175
Cys Ala Thr Gly Cys Cys Cys Ala Ala Ala Ala Thr Cys Ala Ala Thr 180 185 190
Cys Cys Cys Ala Gly Thr Thr Cys Thr Gly Ala Gly Thr Thr Gly Cys 195 200 205
Cys Ala Cys Cys Cys Cys Ala Gly Gly Cys Thr Gly Cys Ala Gly Thr 210 215 220
Gly Ala Ala Thr Gly Ala Ala Gly Cys Thr Cys Ala Ala Gly Ala Gly 225 230 235 240
Gly Ala Thr Thr Thr Cys Cys Gly Gly Ala Ala Gly Cys Thr Cys Cys 245 250 255 Gly Ala Gly Ala Ala Gly Ala Gly Thr Thr Gly Ala Thr Cys Gly Cys 260 265 270
Ala Ala Cys Thr Gly Gly Cys Ala Thr Gly Thr Thr Thr Gly Ala Thr 275 280 285
Gly Cys Cys Thr Cys Cys Cys Cys Cys Cys Thr Cys Thr Gly Gly Thr 290 295 300
Ala Cys Thr Cys Ala Thr Ala Cys Ala Ala Ala Ala Thr Cys Ala Gly 305 310 315 320
Cys Ala Cys Cys Ala Cys Ala Cys Thr Gly Gly Gly Cys Cys Thr Thr 325 330 335
Gly Gly Ala Gly Thr Gly Cys Thr Gly Gly Gly Thr Thr Ala Thr Thr 340 345 350
Thr Cys Cys Thr Gly Ala Thr Gly Gly Thr Thr Cys Ala Gly Thr Ala 355 360 365
Thr Cys Ala Gly Ala Thr Gly Thr Ala Thr Thr Thr Cys Ala Thr Thr 370 375 380
Gly Gly Gly Gly Cys Ala Gly Thr Gly Thr Thr Gly Cys Thr Thr Gly 385 390 395 400
Gly Gly Ala Thr Gly Cys Ala Cys Thr Ala Thr Cys Ala Ala Cys Ala 405 410 415
Gly Ala Thr Gly Gly Gly Cys Thr Gly Gly Cys Thr Thr Thr Cys Thr 420 425 430
Cys Ala Thr Gly Ala Cys Ala Thr Thr Thr Gly Cys Cys Ala Cys Cys 435 440 445
Ala Cys Cys Ala Gly Ala Cys Thr Thr Thr Cys Ala Ala Gly Ala Ala 450 455 460
Cys Cys Gly Gly Ala Ala Cys Thr Gly Gly Ala Ala Cys Ala Ala Cys 465 470 475 480
Cys Thr Cys Gly Thr Gly Gly Gly Ala Cys Thr Gly Gly Thr Ala Thr 485 490 495 Thr Thr Gly Gly Cys Ala Ala Thr Gly Gly Thr Cys Thr Gly Cys Ala 500 505 510
Ala Gly Gly Thr Thr Thr Thr Thr Cys Cys Gly Thr Gly Ala Cys Ala 515 520 525
Thr Gly Gly Thr Gly Gly Ala Ala Gly Gly Ala Cys Ala Gly Ala Cys 530 535 540
Ala Cys Ala Ala Thr Gly Cys Ala Cys Ala Thr Cys Ala Thr Thr Cys 545 550 555 560
Gly Gly Cys Ala Ala Cys Cys Ala Ala Thr Gly Thr Thr Cys Ala Ala 565 570 575
Gly Gly Gly Cys Ala Cys Gly Ala Cys Cys Cys Thr Gly Ala Thr Ala 580 585 590
Thr Thr Gly Ala Cys Ala Ala Cys Cys Thr Cys Cys Cys Cys Cys Thr 595 600 605
Cys Thr Thr Ala Gly Cys Cys Thr Gly Gly Thr Cys Thr Gly Ala Gly 610 615 620
Gly Ala Thr Gly Ala Cys Gly Thr Cys Ala Cys Ala Cys Gly Gly Gly 625 630 635 640
Cys Gly Thr Cys Ala Cys Cys Gly Ala Thr Thr Thr Cys Cys Cys Gly 645 650 655
Cys Ala Ala Gly Cys Thr Cys Ala Thr Thr Cys Ala Gly Thr Thr Cys 660 665 670
Cys Ala Gly Cys Ala Gly Thr Ala Cys Thr Ala Thr Thr Thr Cys Thr 675 680 685
Thr Gly Gly Thr Cys Ala Thr Cys Thr Gly Thr Ala Thr Cys Thr Thr 690 695 700
Gly Thr Thr Gly Cys Gly Gly Thr Thr Cys Ala Thr Thr Thr Gly Gly 705 710 715 720
Thr Gly Thr Thr Thr Cys Cys Ala Gly Ala Gly Cys Gly Thr Gly Thr 725 730 735
Thr Gly Ala Cys Cys Gly Thr Gly Cys Gly Cys Ala Gly Thr Thr Thr 740 745 750 Gly Ala Ala Gly Gly Ala Cys Ala Gly Ala Gly Ala Thr Ala Ala Cys 755 760 765
Cys Ala Ala Thr Thr Cys Thr Ala Thr Cys Gly Cys Thr Cys Thr Cys 770 775 780
Ala Gly Thr Ala Thr Ala Ala Gly Ala Ala Gly Gly Ala Gly Gly Cys 785 790 795 800
Cys Ala Thr Thr Gly Gly Cys Cys Thr Cys Gly Cys Cys Cys Thr Gly 805 810 815
Cys Ala Cys Thr Gly Gly Ala Cys Cys Thr Thr Gly Ala Ala Gly Ala 820 825 830
Cys Cys Cys Thr Gly Thr Thr Cys Cys Ala Cys Thr Thr Ala Thr Thr 835 840 845
Cys Thr Thr Thr Ala Thr Gly Cys Cys Cys Ala Gly Cys Ala Thr Cys 850 855 860
Cys Thr Cys Ala Cys Ala Thr Cys Gly Cys Thr Gly Thr Thr Gly Gly 865 870 875 880
Thr Gly Thr Thr Thr Thr Thr Cys Gly Thr Thr Thr Cys Gly Gly Ala 885 890 895
Gly Cys Thr Gly Gly Thr Thr Gly Gly Cys Gly Gly Cys Thr Thr Cys 900 905 910
Gly Gly Cys Ala Thr Thr Gly Cys Gly Ala Thr Cys Gly Thr Gly Gly 915 920 925
Thr Gly Thr Thr Cys Ala Thr Gly Ala Ala Cys Cys Ala Cys Thr Ala 930 935 940
Cys Cys Cys Ala Cys Thr Gly Gly Ala Gly Ala Ala Gly Ala Thr Cys 945 950 955 960
Gly Gly Gly Gly Ala Cys Thr Cys Ala Gly Thr Cys Thr Gly Gly Gly 965 970 975
Ala Thr Gly Gly Cys Cys Ala Thr Gly Gly Ala Thr Thr Cys Thr Cys 980 985 990 Gly Gly Thr Thr Gly Gly Cys Cys Ala Gly Ala Thr Cys Cys Ala Thr 995 1000 1005
Gly Ala Gly Ala Cys Cys Ala Thr Gly Ala Ala Cys Ala Thr Thr 1010 1015 1020
Cys Gly Gly Cys Gly Ala Gly Gly Gly Ala Thr Thr Ala Thr Cys 1025 1030 1035
Ala Cys Ala Gly Ala Thr Thr Gly Gly Thr Thr Thr Thr Thr Cys 1040 1045 1050
Gly Gly Ala Gly Gly Cys Thr Thr Gly Ala Ala Thr Thr Ala Cys 1055 1060 1065
Cys Ala Gly Ala Thr Thr Gly Ala Gly Cys Ala Cys Cys Ala Thr 1070 1075 1080
Thr Thr Gly Thr Gly Gly Cys Cys Gly Ala Cys Cys Cys Thr Cys 1085 1090 1095
Cys Cys Thr Cys Gly Cys Cys Ala Cys Ala Ala Cys Cys Thr Gly 1100 1105 1110
Ala Cys Ala Gly Cys Gly Gly Thr Thr Ala Gly Cys Thr Ala Cys 1115 1120 1125
Cys Ala Gly Gly Thr Gly Gly Ala Ala Cys Ala Gly Cys Thr Gly 1130 1135 1140
Thr Gly Cys Cys Ala Gly Ala Ala Gly Cys Ala Cys Ala Ala Cys 1145 1150 1155
Cys Thr Gly Cys Cys Gly Thr Ala Thr Cys Gly Gly Ala Ala Cys 1160 1165 1170
Cys Cys Gly Cys Thr Gly Cys Cys Cys Cys Ala Thr Gly Ala Ala 1175 1180 1185
Gly Gly Gly Thr Thr Gly Gly Thr Cys Ala Thr Cys Cys Thr Gly 1190 1195 1200
Cys Thr Gly Cys Gly Cys Thr Ala Thr Cys Thr Gly Gly Cys Gly 1205 1210 1215
Gly Thr Gly Thr Thr Cys Gly Cys Cys Cys Gly Gly Ala Thr Gly 1220 1225 1230 Gly Cys Gly Gly Ala Gly Ala Ala Gly Cys Ala Ala Cys Cys Cys 1235 1240 1245
Gly Cys Gly Gly Gly Gly Ala Ala Gly Gly Cys Thr Cys Thr Ala 1250 1255 1260
<210> 143 <211> 27 <212> DNA <213> Seqüência Artificial <220> <223> oligonucleotideo pDonor222Eg5 <400> 143
gccaactttg tacaaaaaag ttggatt
<210> 144 <211> 21 <212> PRT
<213> Seqüência Artificial <220>
<223> oligonucleotideo degenerado D8DEG3-1 <400> 144
Arg Thr Thr Arg Thr Gly Asn Cys Lys Ala Thr Cys Thr Thr Thr Cys 1 5 10 15
Cys Ala Cys Cys Ala 20
<210> 145 <211> 21 <212> PRT
<213> Seqüência Artificial <220>
<223> oligonucleotideo degenerado D8DEG3-2 <400> 145
Arg Thr Thr Arg Thr Gly Asn Cys Lys Gly Thr Cys Thr Thr Thr Cys 1 5 10 15
Cys Ala Cys Cys Ala 20
<210> 146
<211> 21
<212> PRT
<213> Seqüência Artificial <220>
<223> oligonucleotídeo degenerado D8DEG3-3 <400> 146
Arg Thr Thr Arg Thr Gly Asn Cys Lys Ala Thr Cys Cys Thr Thr Cys 1 5 10 15
Cys Ala Cys Cys Ala 20
<210> 147 <211> 21 <212> PRT
<213> Seqüência Artificial <220>
<223> oligonucleotídeo degenerado D8DEG3-4 <400> 147
Arg Thr Thr Arg Thr Gly Asn Cys Lys Gly Thr Cys Cys Thr Thr Cys 1 5 10 15
Cys Ala Cys Cys Ala 20
<210> 148
<211> 4116
<212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo pHD23-l
<400> 148
cctgaattcc agcacactgg cggccgttac tagtggatcc gagctcggta ccaagcttga 60 tgcatagctt gagtattcta acgcgtcacc taaatagctt ggcgtaatca tggtcatagc 120 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 180 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 240 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 300 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 360 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 420 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 480 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 540 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 600 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 660 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 720 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 780 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 840 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 900 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 960 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 1020 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 1080 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 1140 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 1200 cctagatcct tttaaattaa aaatgaagtt ttagcacgtg tcagtcctgc tcctcggcca 1260 cgaagtgcac gcagttgccg gccgggtcgc gcagggcgaa ctcccgcccc cacggctgct 1320 cgccgatctc ggtcatggcc ggcccggagg cgtcccggaa gttcgtggac acgacctccg 1380 accactcggc gtacagctcg tccaggccgc gcacccacac ccaggccagg gtgttgtccg 1440 gcaccacctg gtcctggacc gcgctgatga acagggtcac gtcgtcccgg accacaccgg 1500 cgaagtcgtc ctccacgaag tcccgggaga acccgagccg gtcggtccag aactcgaccg 1560 ctccggcgac gtcgcgcgcg gtgagcaccg gaacggcact ggtcaacttg gccatggtgg 1620 ccctcctcac gtgctattat tgaagcattt atcagggtta ttgtctcatg agcggataca 1680 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 1740 tgccacctga tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg 1800 aaattgtaag cgttaataat tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc 1860 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca 1920 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc 1980 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag 2040 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg 2100 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca 2160 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat 2220 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact 2280 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc 2340 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc 2400 gtggccagcc acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg 2460 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca 2520 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc 2580 ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct 2640 tgatcagagc ttgatcccct gcgccatcag atccttggcg gcaagaaagc catccagttt 2700 actttgcagg gcttcccaac cttaccagag ggcgccccag ctggcaattc cggttcgctt 2760 gctgtccata aaaccgccca gtctagctat cgccatgtaa gcccactgca agctacctgc 2820 tttctctttg cgcttgcgtt ttcccttgtc cagatagccc agtagctgac attcatccgg 2880 ggtcagcacc gtttctgcgg actggctttc tacgtgaaaa ggatctaggt gaagatcctt 2940 tttgataatc tcatgcctga catttatatt ccccagaaca tcaggttaat ggcgtttttg 3000 atgtcatttt cgcggtggct gagatcagcc acttcttccc cgataacgga gaccggcaca 3060 ctggccatat cggtggtcat catgcgccag ctttcatccc cgatatgcac caccgggtaa 3120 agttcacggg agactttatc tgacagcaga cgtgcactgg ccagggggat caccatccgt 3180 cgccccggcg tgtcaataat atcactctgt acatccacaa acagacgata acggctctct 3240 cttttatagg tgtaaacctt aaactgccgt acgtataggc tgcgcaactg ttgggaaggg 3300 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg 3360 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt 3420 gaattgtaat acgactcact atagggcgaa ttgggccctc tagatgcatg ctcgagcggc 3480 cgccagtgtg atggatatct gcagaattca gggccaactt tgtacaaaaa agttggattt 3540 tttttcggcc cacgatctca catggtgaaa aggccagcac ttccgctgac cgttgatggt 3600 gtcacctatg atgtgtctgc ctggttgaac catcatccag ggggtgctga catcattgag 3660 aactaccgcg gtcgtgatgc cactgatgtc tttatggtta tgcactctga aaatgctgtg 3720 agtaaactaa gaaggatgcc tatcatggaa ccatcatctc cactgacgcc tacgccaccg 3780 aaacccaact cagacgaacc gcaggaggat ttccgcaagc tccgagatga gctcatcgca 3840 gcaggaatgt tcgacgcatc accgatgtgg tacgcatata agacgctcac tacgctgggc 3900 ctcggggtcc tcgcggtgct attgatgacc cagtggcact ggtacctcgt cggggcaatc 3960 gtgttgggca ttcacttcca acaaatgggt tggttgtcgc acgatatctg ccaccatcag 4020 ctgttcaagg accgatcgat caacaacgcc atcggcttgc ttttcgggaa cgtcttgcaa 4080 gggttctctg tgacctggtg gaaggacagt cacaac 4116
<210> 149
<211> 419
<212> PRT
<213> Euglena gracilis
<400> 149 Met Lys Ser Lys Arg Gln Ala Leu Ser Pro Leu Gln Leu Met Glu Gln 1 5 10 15
Thr Tyr Asp Val Val Asn Phe His Pro Gly Gly Ala Glu Ile Ile Glu 20 25 30
Asn Tyr Gln Gly Arg Asp Ala Thr Asp Ala Phe Met Val Met His Phe 35 40 45
Gln Glu Ala Phe Asp Lys Leu Lys Arg Met Pro Lys Ile Asn Pro Ser 50 55 60
Phe Glu Leu Pro Pro Gln Ala Ala Val Asn Glu Ala Gln Glu Asp Phe 65 70 75 80
Arg Lys Leu Arg Glu Glu Leu Ile Ala Thr Gly Met Phe Asp Ala Ser 85 90 95
Pro Leu Trp Tyr Ser Tyr Lys Ile Ser Thr Thr Leu Gly Leu Gly Val 100 105 110
Leu Gly Tyr Phe Leu Met Val Gln Tyr Gln Met Tyr Phe Ile Gly Ala 115 120 125
Val Leu Leu Gly Met His Tyr Gln Gln Met Gly Trp Leu Ser His Asp 130 135 140
Ile Cys His His Gln Thr Phe Lys Asn Arg Asn Trp Asn Asn Leu Val 145 150 155 160
Gly Leu Val Phe Gly Asn Gly Leu Gln Gly Phe Ser Val Thr Cys Trp 165 170 175
Lys Asp Arg His Asn Ala His His Ser Ala Thr Asn Val Gln Gly His 180 185 190
Asp Pro Asp Ile Asp Asn Leu Pro Pro Leu Ala Trp Ser Glu Asp Asp 195 200 205
Val Thr Arg Ala Ser Pro Ile Ser Arg Lys Leu Ile Gln Phe Gln Gln 210 215 220
Tyr Tyr Phe Leu Val Ile Cys Ile Leu Leu Arg Phe Ile Trp Cys Phe 225 230 235 240
Gln Cys Val Leu Thr Val Arg Ser Leu Lys Asp Arg Asp Asn Gln Phe 245 250 255 Tyr Arg Ser Gln Tyr Lys Lys Glu Ala Ile Gly Leu Ala Leu His Trp 260 265 270
Thr Leu Lys Ala Leu Phe His Leu Phe Phe Met Pro Ser Ile Leu Thr 275 280 285
Ser Leu Leu Val Phe Phe Val Ser Glu Leu Val Gly Gly Phe Gly Ile 290 295 300
Ala Ile Val Val Phe Met Asn His Tyr Pro Leu Glu Lys Ile Gly Asp 305 310 315 320
Pro Val Trp Asp Gly His Gly Phe Ser Val Gly Gln Ile His Glu Thr 325 330 335
Met Asn Ile Arg Arg Gly Ile Ile Thr Asp Trp Phe Phe Gly Gly Leu 340 345 350
Asn Tyr Gln Ile Glu His His Leu Trp Pro Thr Leu Pro Arg His Asn 355 360 365
Leu Thr Ala Val Ser Tyr Gln Val Glu Gln Leu Cys Gln Lys His Asn 370 375 380
Leu Pro Tyr Arg Asn Pro Leu Pro His Glu Gly Leu Val Ile Leu Leu 385 390 395 400
Arg Tyr Leu Ala Val Phe Ala Arg Met Ala Glu Lys Gln Pro Ala Gly 405 410 415
Lys Ala Leu
<210> 150 <211> 4307 <212> DNA <213> Seqüência Artificial <220> <223> plasmideo pLF118-3
<220>
<221> misc_feature <222> (4255) .. (4294) <223> η é a, c, g, ou t
<400> 150
gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta 120 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta 180 cattgcacaa gataaaaata tatcatcatg ttagaaaaac tcatcgagca tcaaatgaaa 240 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa 300 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc 360 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt 420 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg 480 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc 540 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct 600 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc 660 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc 720 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt 780 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt 840 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa 900 tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa 960 atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc gttgaatatg 1020 gctcatagat cttttctcca tcactgatag ggagtggtaa aataactcca tcaatgatag 1080 agtgtcaaca acatgaccaa aatcccttaa cgtgagttac gcgtattaat tgcgttgcgc 1140 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa 1200 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 1260 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 1320 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 1380 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 1440 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 1500 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 1560 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc 1620 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 1680 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 1740 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 1800 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggttacac tagaagaaca 1860 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 1920 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 1980
acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 2040
cagggaacga cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg 2100
tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc 2160
accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca 2220
ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct 2280
ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt 2340
tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggcccca aataatgatt 2400
ttattttgac tgatagtgac ctgttcgttg caacaaattg atgagcaatg cttttttata 2460
atgccaactt tgtacaaaaa agttggctat tttttttcgg cccacgatct cacatggtga 2520
aaaggccagc acttccgctg accgttgatg gtgtcaccta tgatgtgtct gcctggttga 2580
accatcatcc agggggtgct gacatcattg agaactaccg cggtcgtgat gccactgatg 2640
tctttatggt tatgcactct gaaaatgctg tgagtaaact aagaaggatg cctatcatgg 2700
aaccatcatc tccactgacg cctacgccac cgaaacccaa ctcagacgaa ccgcaggagg 2760
atttccgcaa gctccgagat gagctcatcg cagcaggaat gttcgacgca tcaccgatgt 2820
ggtacgcata taagacgctc agtacgctgg gcctcggggt cctcgcggtg ctattgatga 2880
cccagtggca ctggtacctc gtcggggcaa tcgtgttggg cattcacttc caacaaatgg 2940
gttggttgtc gcacgatatc tgccaccatc agctgttcaa ggaccgatcg atcaacaacg 3000
ccatcggctt gcttttcggg aacgtcttgc aagggttctc tgtgacctgg tggaaggaca 3060
ggcacaatgc acaccactcc gccaccaacg tgcaaggcca cgaccccgac attgacaacc 3120
tgccgctgct ggcatggtcc aaggaggacg tggagagggc cggcccgttc tcacggcgga 3180
tgatcaagta ccagcaatac tacttcttct tcatctgtgc cctcctgagg ttcatctggt 3240
gcttccagag catccacaca gccacgggcc tgaaggatcg cagcaaccag tactaccgca 3300
ggcagtacga gaaagagagc gtgggcctgg ccctccactg gggcctgaag gcgttgttct 3360
actactttta tatgccaagc ttcttgaccg gactcatggt gtttttcgtg tccgagttgc 3420
ttgggggctt cggcatcgcc atcgtggtgt tcatgaacca ctaccccctg gagaagatcc 3480
aggactcggt gtgggacggc cacggctttt gcgccggcca gattcacgaa acgatgaacg 3540
tccagcgggg actcgtcacg gactggttct tcggtgggct gaattaccaa atcgagcacc 3600
acctgtggcc gacgctgccc cggcacaacc tgacggcggc cagcatcaaa gtggagcagt 3660
tgtgcaagaa gcacaacttg ccgtatcgca gccccccaat gctggagggg gtgggcatcc 3720
tgatcagcta cctgggcacc tttgcccgca tggtggcaaa ggccgacaag gcgtaagtga 3780 catggcaccg ctcaggactc tgatagttgg gctgacgctt tggttgtcat cccttgcccc 3840 ttcatatcac ctctggccct actcggattc tctctagctc taacctgttc aatgtggact 3900 gctacacata tgagttcctc ggatctctgg ggaacagcct ttggaagact cggcattcct 3960 ttatgcttgg aaggcttgag acctcttctg caggactcaa ggcaaccctc ctcagtgtcg 4020 ggaaagagta tttgccttcg gcctgacctg ctatacctca cccaacatgc gtcgtggaat 4080 taatgatcat tgttaagagt ttggtgcgat ttctgattgt gcgcaaattg tgcggaggcg 4140 cggcacacac gttctcctcc ggccatcaca gtccaaggtc aaatttccaa ctctaatcac 4200 catgatgggc cacagctttg cacactattt ctggcagagc tgcaagaaac tcgcnnnnnn 4260 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnacccaa CtttCtt 4307 <210> 151 <211> 1260 <212> PRT <213> Euglena anabaena <400> 151 Ala Thr Gly 1 Gly Thr Gly Ala Ala Ala Ala Gly 5 10 Gly Cys Cys Ala Gly 15 Cys Ala Cvs Thr Thr Cys Cys Gly Cys Thr Gly 20 ~ 25 Ala Cys Cys 30 Gly Thr Thr Gly Ala 35 Thr Gly Gly Thr Gly Thr Cys Ala 40 Cys Cys Thr 45 Ala Thr Gly Ala Thr 50 Gly Thr Gly Thr Cys Thr Gly Cys 55 Cys Thr Gly 60 Gly Thr Thr Gly Ala 65 Ala Cys Cys Ala Thr Cys Ala Thr 70 75 Cys Cys Ala Gly Gly 80 Gly Gly Gly Thr Gly Cys Thr Gly Ala Cys Ala 85 90 Thr Cys Ala Thr Thr 95 Gly Ala Gly Ala Ala Cys Thr Ala Cys Cys Gly 100 105 Cys Gly Gly 110 Thr Cys Gly Thr Gly 115 Ala Thr Gly Cys Cys Ala Cys Thr 120 Gly Ala Thr 125 Gly Thr Cys Thr Thr 130 Thr Ala Thr Gly Gly Thr Thr Ala 135 Thr Gly Cys 140 Ala Cys Thr Cys Thr Gly Ala Ala Ala Ala Thr Gly Cys Thr Gly Thr Gly Ala 145 150 155 160
Gly Thr Ala Ala Ala Cys Thr Ala Ala Gly Ala Ala Gly Gly Ala Thr 165 170 175
Gly Cys Cys Thr Ala Thr Cys Ala Thr Gly Gly Ala Ala Cys Cys Ala 180 185 190
Thr Cys Ala Thr Cys Thr Cys Cys Ala Cys Thr Gly Ala Cys Gly Cys 195 200 205
Cys Thr Ala Cys Gly Cys Cys Ala Cys Cys Gly Ala Ala Ala Cys Cys 210 215 220
Cys Ala Ala Cys Thr Cys Ala Gly Ala Cys Gly Ala Ala Cys Cys Gly 225 230 235 240
Cys Ala Gly Gly Ala Gly Gly Ala Thr Thr Thr Cys Cys Gly Cys Ala 245 250 255
Ala Gly Cys Thr Cys Cys Gly Ala Gly Ala Thr Gly Ala Gly Cys Thr 260 265 270
Cys Ala Thr Cys Gly Cys Ala Gly Cys Ala Gly Gly Ala Ala Thr Gly 275 280 285
Thr Thr Cys Gly Ala Cys Gly Cys Ala Thr Cys Ala Cys Cys Gly Ala 290 295 300
Thr Gly Thr Gly Gly Thr Ala Cys Gly Cys Ala Thr Ala Thr Ala Ala 305 310 315 320
Gly Ala Cys Gly Cys Thr Cys Ala Gly Thr Ala Cys Gly Cys Thr Gly 325 330 335
Gly Gly Cys Cys Thr Cys Gly Gly Gly Gly Thr Cys Cys Thr Cys Gly 340 345 350
Cys Gly Gly Thr Gly Cys Thr Ala Thr Thr Gly Ala Thr Gly Ala Cys 355 360 365
Cys Cys Ala Gly Thr Gly Gly Cys Ala Cys Thr Gly Gly Thr Ala Cys 370 375 380
Cys Thr Cys Gly Thr Cys Gly Gly Gly Gly Cys Ala Ala Thr Cys Gly 385 390 395 400 Thr Gly Thr Thr Gly Gly Gly Cys Ala Thr Thr Cys Ala Cys Thr Thr 405 410 415
Cys Cys Ala Ala Cys Ala Ala Ala Thr Gly Gly Gly Thr Thr Gly Gly 420 425 430
Thr Thr Gly Thr Cys Gly Cys Ala Cys Gly Ala Thr Ala Thr Cys Thr 435 440 445
Gly Cys Cys Ala Cys Cys Ala Thr Cys Ala Gly Cys Thr Gly Thr Thr 450 455 460
Cys Ala Ala Gly Gly Ala Cys Cys Gly Ala Thr Cys Gly Ala Thr Cys 465 470 475 480
Ala Ala Cys Ala Ala Cys Gly Cys Cys Ala Thr Cys Gly Gly Cys Thr 485 490 495
Thr Gly Cys Thr Thr Thr Thr Cys Gly Gly Gly Ala Ala Cys Gly Thr 500 505 510
Cys Thr Thr Gly Cys Ala Ala Gly Gly Gly Thr Thr Cys Thr Cys Thr 515 520 525
Gly Thr Gly Ala Cys Cys Thr Gly Gly Thr Gly Gly Ala Ala Gly Gly 530 535 540
Ala Cys Ala Gly Gly Cys Ala Cys Ala Ala Thr Gly Cys Ala Cys Ala 545 550 555 560
Cys Cys Ala Cys Thr Cys Cys Gly Cys Cys Ala Cys Cys Ala Ala Cys 565 570 575
Gly Thr Gly Cys Ala Ala Gly Gly Cys Cys Ala Cys Gly Ala Cys Cys 580 585 590
Cys Cys Gly Ala Cys Ala Thr Thr Gly Ala Cys Ala Ala Cys Cys Thr 595 600 605
Gly Cys Cys Gly Cys Thr Gly Cys Thr Gly Gly Cys Ala Thr Gly Gly 610 615 620
Thr Cys Cys Ala Ala Gly Gly Ala Gly Gly Ala Cys Gly Thr Gly Gly 625 630 635 640 Ala Gly Ala Gly Gly Gly Cys Cys Gly Gly Cys Cys Cys Gly Thr Thr 645 650 655
Cys Thr Cys Ala Cys Gly Gly Cys Gly Gly Ala Thr Gly Ala Thr Cys 660 665 670
Ala Ala Gly Thr Ala Cys Cys Ala Gly Cys Ala Ala Thr Ala Cys Thr 675 680 685
Ala Cys Thr Thr Cys Thr Thr Cys Thr Thr Cys Ala Thr Cys Thr Gly 690 695 700
Thr Gly Cys Cys Cys Thr Cys Cys Thr Gly Ala Gly Gly Thr Thr Cys 705 710 715 720
Ala Thr Cys Thr Gly Gly Thr Gly Cys Thr Thr Cys Cys Ala Gly Ala 725 730 735
Gly Cys Ala Thr Cys Cys Ala Cys Ala Cys Ala Gly Cys Cys Ala Cys 740 745 750
Gly Gly Gly Cys Cys Thr Gly Ala Ala Gly Gly Ala Thr Cys Gly Cys 755 760 765
Ala Gly Cys Ala Ala Cys Cys Ala Gly Thr Ala Cys Thr Ala Cys Cys 770 775 780
Gly Cys Ala Gly Gly Cys Ala Gly Thr Ala Cys Gly Ala Gly Ala Ala 785 790 795 800
Ala Gly Ala Gly Ala Gly Cys Gly Thr Gly Gly Gly Cys Cys Thr Gly 805 810 815
Gly Cys Cys Cys Thr Cys Cys Ala Cys Thr Gly Gly Gly Gly Cys Cys 820 825 830
Thr Gly Ala Ala Gly Gly Cys Gly Thr Thr Gly Thr Thr Cys Thr Ala 835 840 845
Cys Thr Ala Cys Thr Thr Thr Thr Ala Thr Ala Thr Gly Cys Cys Ala 850 855 860
Ala Gly Cys Thr Thr Cys Thr Thr Gly Ala Cys Cys Gly Gly Ala Cys 865 870 875 880
Thr Cys Ala Thr Gly Gly Thr Gly Thr Thr Thr Thr Thr Cys Gly Thr 885 890 895 Gly Thr Cys Cys Gly Ala Gly Thr Thr Gly Cys Thr Thr Gly Gly Gly 900 905 910
Gly Gly Cys Thr Thr Cys Gly Gly Cys Ala Thr Cys Gly Cys Cys Ala 915 920 925
Thr Cys Gly Thr Gly Gly Thr Gly Thr Thr Cys Ala Thr Gly Ala Ala 930 935 940
Cys Cys Ala Cys Thr Ala Cys Cys Cys Cys Cys Thr Gly Gly Ala Gly 945 950 955 960
Ala Ala Gly Ala Thr Cys Cys Ala Gly Gly Ala Cys Thr Cys Gly Gly 965 970 975
Thr Gly Thr Gly Gly Gly Ala Cys Gly Gly Cys Cys Ala Cys Gly Gly 980 985 990
Cys Thr Thr Thr Thr Gly Cys Gly Cys Cys Gly Gly Cys Cys Ala Gly 995 1000 1005
Ala Thr Thr Cys Ala Cys Gly Ala Ala Ala Cys Gly Ala Thr Gly 1010 1015 1020
Ala Ala Cys Gly Thr Cys Cys Ala Gly Cys Gly Gly Gly Gly Ala 1025 1030 1035
Cys Thr Cys Gly Thr Cys Ala Cys Gly Gly Ala Cys Thr Gly Gly 1040 1045 1050
Thr Thr Cys Thr Thr Cys Gly Gly Thr Gly Gly Gly Cys Thr Gly 1055 1060 1065
Ala Ala Thr Thr Ala Cys Cys Ala Ala Ala Thr Cys Gly Ala Gly 1070 1075 1080
Cys Ala Cys Cys Ala Cys Cys Thr Gly Thr Gly Gly Cys Cys Gly 1085 1090 1095
Ala Cys Gly Cys Thr Gly Cys Cys Cys Cys Gly Gly Cys Ala Cys 1100 1105 1110
Ala Ala Cys Cys Thr Gly Ala Cys Gly Gly Cys Gly Gly Cys Cys 1115 1120 1125 Ala Gly Cys Ala Thr Cys Ala Ala Ala Gly Thr Gly Gly Ala Gly 1130 1135 1140
Cys Ala Gly Thr Thr Gly Thr Gly Cys Ala Ala Gly Ala Ala Gly 1145 1150 1155
Cys Ala Cys Ala Ala Cys Thr Thr Gly Cys Cys Gly Thr Ala Thr 1160 1165 1170
Cys Gly Cys Ala Gly Cys Cys Cys Cys Cys Cys Ala Ala Thr Gly 1175 1180 1185
Cys Thr Gly Gly Ala Gly Gly Gly Gly Gly Thr Gly Gly Gly Cys 1190 1195 1200
Ala Thr Cys Cys Thr Gly Ala Thr Cys Ala Gly Cys Thr Ala Cys 1205 1210 1215
Cys Thr Gly Gly Gly Cys Ala Cys Cys Thr Thr Thr Gly Cys Cys 1220 1225 1230
Cys Gly Cys Ala Thr Gly Gly Thr Gly Gly Cys Ala Ala Ala Gly 1235 1240 1245
Gly Cys Cys Gly Ala Cys Ala Ala Gly Gly Cys Gly 1250 1255 1260
<210> 152 <211> 420 <212> PRT
<213> Euglena anabaena <400> 152
Met Val Lys Arg Pro Ala Leu Pro Leu Thr Val Asp Gly Val Thr Tyr 1 5 10 15
Asp Val Ser Ala Trp Leu Asn His His Pro Gly Gly Ala Asp Ile Ile 20 25 30
Glu Asn Tyr Arg Gly Arg Asp Ala Thr Asp Val Phe Met Val Met His 35 40 45
Ser Glu Asn Ala Val Ser Lys Leu Arg Arg Met Pro Ile Met Glu Pro 50 55 60
Ser Ser Pro Leu Thr Pro Thr Pro Pro Lys Pro Asn Ser Asp Glu Pro 65 70 75 80 Gln Glu Asp Phe Arg Lys Leu Arg Asp Glu Leu Ile Ala Ala Gly Met 85 90 95
Phe Asp Ala Ser Pro Met Trp Tyr Ala Tyr Lys Thr Leu Ser Thr Leu 100 105 110
Gly Leu Gly Val Leu Ala Val Leu Leu Met Thr Gln Trp His Trp Tyr 115 120 125
Leu Val Gly Ala Ile Val Leu Gly Ile His Phe Gln Gln Met Gly Trp 130 135 140
Leu Ser His Asp Ile Cys His His Gln Leu Phe Lys Asp Arg Ser Ile 145 150 155 160
Asn Asn Ala Ile Gly Leu Leu Phe Gly Asn Val Leu Gln Gly Phe Ser 165 170 175
Val Thr Trp Trp Lys Asp Arg His Asn Ala His His Ser Ala Thr Asn 180 185 190
Val Gln Gly His Asp Pro Asp Ile Asp Asn Leu Pro Leu Leu Ala Trp 195 200 205
Ser Lys Glu Asp Val Glu Arg Ala Gly Pro Phe Ser Arg Arg Met Ile 210 215 220
Lys Tyr Gln Gln Tyr Tyr Phe Phe Phe Ile Cys Ala Leu Leu Arg Phe 225 230 235 240
Ile Trp Cys Phe Gln Ser Ile His Thr Ala Thr Gly Leu Lys Asp Arg 245 250 255
Ser Asn Gln Tyr Tyr Arg Arg Gln Tyr Glu Lys Glu Ser Val Gly Leu 260 265 270
Ala Leu His Trp Gly Leu Lys Ala Leu Phe Tyr Tyr Phe Tyr Met Pro 275 280 285
Ser Phe Leu Thr Gly Leu Met Val Phe Phe Val Ser Glu Leu Leu Gly 290 295 300
Gly Phe Gly Ile Ala Ile Val Val Phe Met Asn His Tyr Pro Leu Glu 305 310 315 320
Lys Ile Gln Asp Ser Val Trp Asp Gly His Gly Phe Cys Ala Gly Gln 325 330 335 Ile His Glu Thr Met Asn 340
Val Gln Arg Gly Leu Val 345
Thr Asp Trp Phe 350
Phe Gly Gly Leu Asn Tyr 355
Gln Ile Glu 360
His His Leu Trp Pro Thr Leu 365
Pro Arg 370
His Asn Leu Thr Ala Ala Ser 375
Ile Lys Val Glu Gln Leu Cys 380
Lys Lys 385
His Asn Leu Pro Tyr Arg Ser
Pro Pro Met 395
Leu Glu Gly Val 400
390
Gly Ile Leu Ile Ser Tyr Leu Gly Thr 405
Phe Ala Arg Met 410
Val Ala Lys 415
Ala Asp Lys Ala 420
<210> 153
<211> 34
<212> DNA
<213> Seqüência Artificial <220>
<223> oligonucleotideo EaD8-5
<400> 153
gcggccgcac catggtgaaa aggccagcac ttcc 34
<210> 154
<211> 30
<212> DNA
<213> Seqüência Artificial <220>
<223> oligonucleotideo EaD8-3
<400> 154
gcggccgctt acgccttgtc ggcctttgcc 30
<210> 155
<211> 4794
<212> DNA
<213> Seqüência Artificial <220>
<223> plasmídeo pLF120-3
<400> 155
ttgtaatacg actcactata gggcgaattg ggccctctag atgcatgctc gagcggccgc 60
cagtgtgatg gatatctgca gaattcaggc ctgaattcca gcacactggc ggccgttact 120 agtggatccg agctcggtac caagcttgat gcatagcttg agtattctaa cgcgtcacct 180 aaatagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 240
aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 300
gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 360
gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 420
ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 480
atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 54 0
gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 600
gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 660
gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 720
gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 780
aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 840
ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 900
taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 960
tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 1020
gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 1080
taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 1140
tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 1200
tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 1260
ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 1320
tagcacgtgt cagtcctgct cctcggccac gaagtgcacg cagttgccgg ccgggtcgcg 1380
cagggcgaac tcccgccccc acggctgctc gccgatctcg gtcatggccg gcccggaggc 1440
gtcccggaag ttcgtggaca cgacctccga ccactcggcg tacagctcgt ccaggccgcg 1500
cacccacacc caggccaggg tgttgtccgg caccacctgg tcctggaccg cgctgatgaa 1560
cagggtcacg tcgtcccgga ccacaccggc gaagtcgtcc tccacgaagt cccgggagaa 1620
cccgagccgg tcggtccaga actcgaccgc tccggcgacg tcgcgcgcgg tgagcaccgg 1680
aacggcactg gtcaacttgg ccatggtggc cctcctcacg tgctattatt gaagcattta 1740
tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 1800
aggggttccg cgcacatttc cccgaaaagt gccacctgat gcggtgtgaa ataccgcaca 1860
gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaataatt cagaagaact 1920
cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca 1980 cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg 2040 ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc 2100 ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct 2160 cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat 2220 gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct 2280 cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc 2340 gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga 2400 gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt 2460 cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt 2520 cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct 2580 gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat 2640 agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa 2700 tcatgcgaaa cgatcctcat cctgtctctt gatcagagct tgatcccctg cgccatcaga 2760 tccttggcgg caagaaagcc atccagttta ctttgcaggg cttcccaacc ttaccagagg 2820 gcgccccagc tggcaattcc ggttcgcttg ctgtccataa aaccgcccag tctagctatc 2880 gccatgtaag cccactgcaa gctacctgct ttctctttgc gcttgcgttt tcccttgtcc 2940 agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct 3000 acgtgaaaag gatctaggtg aagatccttt ttgataatct catgcctgac atttatattc 3060 cccagaacat caggttaatg gcgtttttga tgtcattttc gcggtggctg agatcagcca 3120 cttcttcccc gataacggag accggcacac tggccatatc ggtggtcatc atgcgccagc 3180 tttcatcccc gatatgcacc accgggtaaa gttcacggga gactttatct gacagcagac 3240 gtgcactggc cagggggatc accatccgtc gccccggcgt gtcaataata tcactctgta 3300 catccacaaa cagacgataa cggctctctc ttttataggt gtaaacctta aactgccgta 3360 cgtataggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc 3420 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc 3480 agtcacgacg ttgtaaaacg acggccagtg aagcggccgc accatggtga aaaggccagc 3540 acttccgctg accgttgatg gtgtcaccta tgatgtgtct gcctggttga accatcatcc 3600 agggggtgct gacatcattg agaactaccg cggtcgtgat gccactgatg tctttatggt 3660 tatgcactct gaaaatgctg tgagtaaact aagaaggatg cctatcatgg aaccatcatc 3720 tccactgacg cctacgccac cgaaacccaa ctcagacgaa ccgcaggagg atttccgcaa 3780 gctccgagat gagctcatcg cagcaggaat gttcgacgca tcaccgatgt ggtacgcata 3840 taagacgctc agtacgctgg gcctcggggt cctcgcggtg ctattgatga cccagtggca 3900
ctggtacctc gtcggggcaa tcgtgttggg cattcacttc caacaaatgg gttggttgtc 3960
gcacgatatc tgccaccatc agctgttcaa ggaccgatcg atcaacaacg ccatcggctt 4020
gcttttcggg aacgtcttgc aagggttctc tgtgacctgg tggaaggaca ggcacaatgc 4080
acaccactcc gccaccaacg tgcaaggcca cgaccccgac attgacaacc tgccgctgct 4140
ggcatggtcc aaggaggacg tggagagggc cggcccgttc tcacggcgga tgatcaagta 4200
ccagcaatac tacttcttct tcatctgtgc cctcctgagg ttcatctggt gcttccagag 4260
catccacaca gccacgggcc tgaaggatcg cagcaaccag tactaccgca ggcagtacga 4320
gaaagagagc gtgggcctgg ccctccactg gggcctgaag gcgttgttct actactttta 4380
tatgccaagc ttcttgaccg gactcatggt gtttttcgtg tccgagttgc ttgggggctt 4440
cggcatcgcc atcgtggtgt tcatgaacca ctaccccctg gagaagatcc aggactcggt 4500
gtgggacggc cacggctttt gcgccggcca gattcacgaa acgatgaacg tccagcgggg 4560
actcgtcacg gactggttct tcggtgggct gaattaccaa atcgagcacc acctgtggcc 4620
gacgctgccc cggcacaacc tgacggcggc cagcatcaaa gtggagcagt tgtgcaagaa 4 680
gcacaacttg ccgtatcgca gccccccaat gctggagggg gtgggcatcc tgatcagcta 4740
cctgggcacc tttgcccgca tggtggcaaa ggccgacaag gcgtaagcgg ccgc 4794
<210> 156
<211> 33
<212> DNA
<213> Seguência Artificial <220>
<223> oligonucleotídeo oEAd9ell-l
<400> 156
agcggccgca ccatggaagc agccaaagaa ttg
<210> 157
<211> 26
<212> DNA
<213> Seqüência Artificial <220>
<223> oligonucleotídeo oEAd9ell-2
<400> 157
tgcggccgct actccttctt ccctcg
<210> 158
<211> 4310
<212> DNA
<213> Seqüência Artificial
33
26 <220>
<223> plasmideo pKR1137 <400> 158
aattccagca cactggcggc cgttactagt ggatccgagc tcggtaccaa gcttgatgca 60 tagcttgagt attctaacgc gtcacctaaa tagcttggcg taatcatggt catagctgtt 120 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa 180 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 240 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 300 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 360 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 420 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaagcccag 480 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 540 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 600 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 660 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 720 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 780 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 840 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 900 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt 960 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 1020 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 1080 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 1140 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 1200 gatcctttta aattaaaaat gaagttttag cacgtgtcag tcctgctcct cggccacgaa 1260 gtgcacgcag ttgccggccg ggtcgcgcag ggcgaactcc cgcccccacg gctgctcgcc 1320 gatctcggtc atggccggcc cggaggcgtc ccggaagttc gtggacacga cctccgacca 1380 ctcggcgtac agctcgtcca ggccgcgcac ccacacccag gccagggtgt tgtccggcac 1440 cacctggtcc tggaccgcgc tgatgaacag ggtcacgtcg tcccggacca caccggcgaa 1500 gtcgtcctcc acgaagtccc gggagaaccc gagccggtcg gtccagaact cgaccgctcc 1560 ggcgacgtcg cgcgcggtga gcaccggaac ggcactggtc aacttggcca tggtggccct 1620 cctcacgtgc tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 1680 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 1740 acctgtatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa 1800 ttgtaagcgt taataattca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc 1860 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc 1920 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc 1980 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag 2040 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg 2100 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga 2160 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg 2220 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc 2280 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc 2340 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg 2400 gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg 2460 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag 2520 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga 2580 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga 2640 tcagagcttg atcccctgcg ccatcagatc cttggcqqcg agaaagccat ccagtttact 2700 ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg ttcgcttgct 2760 gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc tacctgcttt 2820 ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt catccggggt 2880 cagcaccgtt tctgcggact ggctttctac gtgaaaagga tctaggtgaa gatccttttt 2940 gataatctca tgcctgacat ttatattccc cagaacatca ggttaatggc gtttttgatg 3000 tcattttcgc ggtggctgag atcagccact tcttccccga taacggagac cggcacactg 3060 gccatatcgg tggtcatcat gcgccagctt tcatccccga tatgcaccac cgggtaaagt 3120 tcacgggaga ctttatctga cagcagacgt gcactggcca gggggatcac catccgtcgc 3180 cccggcgtgt caataatatc actctgtaca tccacaaaca gacgataacg gctctctctt 3240 ttataggtgt aaaccttaaa ctgccgtacg tataggctgc gcaactgttg ggaagggcga 3300 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga 3360 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa 3420 ttgtaatacg actcactata gggcgaattg ggccctctag atgcatgctc gagcggccgc 3480 cagtgtgatg gatatctgca gaattcagga gcggccgcac catggaagca gccaaagaat 3540 tggtttccat cgtccaagag gagctcccca aggtggacta tgcccagctt tggcaggatg 3600 ccagcagctg tgaggtcctt tacctctcgg tggcattcgt ggcgatcaag ttcatgctgc 3660
gcccactgga cctgaagcgc caggccacct tgaagaagct gttcacagca tacaacttcc 3720
tcatgtcgat ctattccttt ggctccttcc tggccatggc ctatgcccta tcagtaactg 3780
gcatcctctc cggcgactgt gagacggcgt tcaacaacga tgtgttcagg atcacaactc 3840
agctgttcta cctcagcaag ttcgtagagt acatcgactc cttctacctt ccccttatgg 3900
acaagccact gtcgttcctt cagttcttcc atcatttggg ggcccccatt gacatgtggc 3960
tattctacaa ataccgcaac gaaggagtct ggatctttgt cctgttgaat gggttcattc 4020
actggatcat gtacggttac tattggacgc ggctcatcaa gctgaacttc cccatgccca 4080
agaacctgat cacctccatg cagatcatcc agttcaatgt cgggttctac atcgtctgga 4140
agtaccgcaa tgtgccatgc taccgccagg atgggatgcg catgtttgcc tggatcttca 4200
actactggta tgtcgggacg gtcttgctgc tgttcctcaa cttttacgtg cagacgtaca 4260
tccggaagcc gaggaagaac cgagggaaga aggagtagcg gccgcacctg 4310
<210> 159 <211> 7872 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmídeo pKR1140 <400> 159
ggccgcaagt atgaactaaa atgcatgtag gtgtaagagc tcatggagag catggaatat 60
tgtatccgac catgtaacag tataataact gagctccatc tcacttcttc tatgaataaa 120
caaaggatgt tatgatatat taacactcta tctatgcacc ttattgttct atgataaatt 180
tcctcttatt attataaatc atctgaatcg tgacggctta tggaatgctt caaatagtac 240
aaaaacaaat gtgtactata agactttcta aacaattcta accttagcat tgtgaacgag 300
acataagtgt taagaagaca taacaattat aatggaagaa gtttgtctcc atttatatat 360
tatatattac ccacttatgt attatattag gatgttaagg agacataaca attataaaga 420
gagaagtttg tatccattta tatattatat actacccatt tatatattat acttatccac 480
ttatttaatg tctttataag gtttgatcca tgatatttct aatattttag ttgatatgta 540
tatgaaaggg tactatttga actctcttac tctgtataaa ggttggatca tccttaaagt 600
gggtctattt aattttattg cttcttacag ataaaaaaaa aattatgagt tggtttgata 660
aaatattgaa ggatttaaaa taataataaa taacatataa tatatgtata taaatttatt 720
ataatataac atttatctat aaaaaagtaa atattgtcat aaatctatac aatcgtttag 780
ccttgctgga cgaatctcaa ttatttaaac gagagtaaac atatttgact ttttggttat 840 ttaacaaatt attatttaac actatatgaa attttttttt ttatcagcaa agaataaaat 900 taaattaaga aggacaatgg tgtcccaatc cttatacaac caacttccac aagaaagtca 960 agtcagagac aacaaaaaaa caagcaaagg aaatttttta atttgagttg tcttgtttgc 1020 tgcataattt atgcagtaaa acactacaca taaccctttt agcagtagag caatggttga 1080 ccgtgtgctt agcttctttt attttatttt tttatcagca aagaataaat aaaataaaat 1140 gagacacttc agggatgttt caacaagctt ggcgcgccgt tctatagtgt cacctaaatc 1200 gtatgtgtat gatacataag gttatgtatt aattgtagcc gcgttctaac gacaatatgt 1260 ccatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac 1320 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca 1380 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga 1440 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgacca 1500 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 1560 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 1620 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 1680 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc 1740 1800 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 1860 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 1920 gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc 1980 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 2040 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 2100 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 2160 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct 2220 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata 2280 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc 2340 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc aggttgatcg 2400 attcgacatc gatctagtaa catagatgac accgcgcgcg ataatttatc ctagtttgcg 2460 cgctatattt tgttttctat cgcgtattaa atgtataatt gcgggactct aatcataaaa 2520 acccatctca taaataacgt catgcattac atgttaatta ttacatgctt aacgtaattc 2580 aacagaaatt atatgataat catcgcaaga ccggcaacag gattcaatct taagaaactt 2640 tattgccaaa tgtttgaacg atctgcttcg acgcactcct tctttaggta cctcactatt 2700 cctttgccct cggacgagtg ctggggcgtc ggtttccact atcggcgagt acttctacac 2760 agccatcggt ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg 2820 ctccggatcg gacgattgcg tcgcatcgac cctgcgccca agctgcatca tcgaaattgc 2880 cgtcaaccaa gctctgatag agttggtcaa gaccaatgcg gagcatatac gcccggagcc 2940 gcggcgatcc tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac 3000 aagccaacca cggcctccag aagaagatgt tggcgacctc gtattgggaa tccccgaaca 3060 tcgcctcgct ccagtcaatg accgctgtta tgcggccatt gtccgtcagg acattgttgg 3120 agccgaaatc cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc caaagcatca 3180 gctcatcgag agcctgcgcg acggacgcac tgacggtgtc gtccatcaca gtttgccagt 3240 gatacacatg gggatcagca atcgcgcata tgaaatcacg ccatgtagtg tattgaccga 3300 ttccttgcgg tccgaatggg ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg 3360 catccatggc ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt 3420 gcaacgtgac accctgtgca cggcgggaga tgcaataggt caggctctcg ctgaattccc 3480 caatgtcaag cacttccgga atcgggagcg cggccgatgc aaagtgccga taaacataac 3540 gatctttgta gaaaccatcg gcgcagctat ttacccgcag gacatatcca cgccctccta 3600 catcgaagct gaaagcacga gattcttcgc cctccgagag ctgcatcagg tcggagacgc 3660 tgtcgaactt ttcgatcaga aacttctcga cagacgtcgc ggtgagttca ggctttttca 3720 tggtttaata agaagagaaa agagttcttt tgttatggct gaagtaatag agaaatgagc 3780 tcgagcgtgt cctctccaaa tgaaatgaac ttccttatat agaggaaggg tcttgcgaag 3840 gatagtggga ttgtgcgtca tcccttacgt cagtggagat gtcacatcaa tccacttgct 3900 ttgaagacgt ggttggaacg tcttcttttt ccacgatgct cctcgtgggt gggggtccat 3960 ctttgggacc actgtcggca gaggcatctt gaatgatagc ctttccttta tcgcaatgat 4020 ggcatttgta ggagccacct tccttttcta ctgtcctttc gatgaagtga cagatagctg 4080 ggcaatggaa tccgaggagg tttcccgaaa ttatcctttg ttgaaaagtc tcaatagccc 4140 tttggtcttc tgagactgta tctttgacat ttttggagta gaccagagtg tcgtgctcca 4200 ccatgttgac gaagattttc ttcttgtcat tgagtcgtaa aagactctgt atgaactgtt 4260 cgccagtctt cacggcgagt tctgttagat cctcgatttg aatcttagac tccatgcatg 4320 gccttagatt cagtaggaac taccttttta gagactccaa tctctattac ttgccttggt 4380 ttatgaagca agccttgaat cgtccatact ggaatagtac ttctgatctt gagaaatatg 4440 tctttctctg tgttcttgat gcaattagtc ctgaatcttt tgactgcatc tttaaccttc 4500 ttgggaaggt atttgatctc ctggagattg ttactcgggt agatcgtctt gatgagacct 4560 gctgcgtagg cctctctaac catctgtggg tcagcattct ttctgaaatt gaagaggcta 4 620
accttctcat tatcagtggt gaacatagtg tcgtcacctt caccttcgaa cttccttcct 4680
agatcgtaaa gatagaggaa atcgtccatt gtaatctccg gggcaaagga gatctctttt 4740
ggggctggat cactgctggg ccttttggtt cctagcgtga gccagtgggc tttttgcttt 4800
ggtgggcttg ttagggcctt agcaaagctc ttgggcttga gttgagcttc tcctttgggg 4860
atgaagttca acctgtctgt ttgctgactt gttgtgtacg cgtcagctgc tgctcttgcc 4920
tctgtaatag tggcaaattt cttgtgtgca actccgggaa cgccgtttgt tgccgccttt 4980
gtacaacccc agtcatcgta tataccggca tgtggaccgt tatacacaac gtagtagttg 5040
atatgagggt gttgaatacc cgattctgct ctgagaggag caactgtgct gttaagctca 5100
gatttttgtg ggattggaat tggatcgatc tcgatcccgc gaaattaata cgactcacta 5160
tagggagacc acaacggttt ccctctagaa ataattttgt ttaactttaa gaaggagata 5220
tacccatgga aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt 5280
tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct 5340
tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca 5400
aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg 5460
acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca 5520
cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcta 5580
tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc 5640
aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg 5700
tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg 5760
atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt 5820
tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg 5880
aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt 5940
tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat 6000
cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg 6060
ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat 6120
ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg 6180
atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg 6240
caaaggaata gtgaggtaca gcttggatcg atccggctgc taacaaagcc cgaaaggaag 6300
ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac 6360
gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatgatcg ggcgcgccgt 6420 cgacggatcc gtacgagatc cggccggcca gatcctgcag gagatccaag cttttgatcc 6480
atgcccttca tttgccgctt attaattaat ttggtaacag tccgtactaa tcagttactt 6540
atccttcccc catcataatt aatcttggta gtctcgaatg ccacaacact gactagtctc 6600
ttggatcata agaaaaagcc aaggaacaaa agaagacaaa acacaatgag agtatccttt 6660
gcatagcaat gtctaagttc ataaaattca aacaaaaacg caatcacaca cagtggacat 6720
cacttatcca ctagctgatc aggatcgccg cgtcaagaaa aaaaaactgg accccaaaag 6780
ccatgcacaa caacacgtac tcacaaaggt gtcaatcgag cagcccaaaa cattcaccaa 6840
ctcaacccat catgagccct cacatttgtt gtttctaacc caacctcaaa ctcgtattct 6900
cttccgccac ctcatttttg tttatttcaa cacccgtcaa actgcatgcc accccgtggc 6960
caaatgtcca tgcatgttaa caagacctat gactataaat agctgcaatc tcggcccagg 7020
ttttcatcat caagaaccag ttcaatatcc tagtacaccg tattaaagaa tttaagatat 7080
actgcggccg caccatggaa gcagccaaag aattggtttc catcgtccaa gaggagctcc 7140
ccaaggtgga ctatgcccag ctttggcagg atgccagcag ctgtgaggtc ctttacctct 7200
cggtggcatt cgtggcgatc aagttcatgc tgcgcccact ggacctgaag cgccaggcca 7260
ccttgaagaa gctgttcaca gcatacaact tcctcatgtc gatctattcc tttggctcct 7320
tcctggccat ggcctatgcc ctatcagtaa ctggcatcct ctccggcgac tgtgagacgg 7380
cgttcaacaa cgatgtgttc aggatcacaa ctcagctgtt ctacctcagc aagttcgtag 7440
agtacatcga ctccttctac cttcccctta tggacaagcc actgtcgttc cttcagttct 7500
tccatcattt gggggccccc attgacatgt ggctattcta caaataccgc aacgaaggag 7560
tctggatctt tgtcctgttg aatgggttca ttcactggat catgtacggt tactattgga 7620
cgcggctcat caagctgaac ttccccatgc ccaagaacct gatcacctcc atgcagatca 7680
tccagttcaa tgtcgggttc tacatcgtct ggaagtaccg caatgtgcca tgctaccgcc 7740
aggatgggat gcgcatgttt gcctggatct tcaactactg gtatgtcggg acggtcttgc 7800
tgctgttcct caacttttac gtgcagacgt acatccggaa gccgaggaag aaccgaggga 7860
agaaggagta gc 7872
<210> 160 <211> 5146 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmídeo pKRH73
<400> 160
cgcgcccgat catccggata tagttcctcc tttcagcaaa aaacccctca agacccgttt
60 agaggcccca aggggttatg ctagttattg ctcagcggtg gcagcagcca actcagcttc 120 ctttcgggct ttgttagcag ccggatcgat ccaagctgta cctcactatt cctttgccct 180 cggacgagtg ctggggcgtc ggtttccact atcggcgagt acttctacac agccatcggt 240 ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg ctccggatcg 300 gacgattgcg tcgcatcgac cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa 360 gctctgatag agttggtcaa gaccaatgcg gagcatatac gcccggagcc gcggcgatcc 420 tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac aagccaacca 480 cggcctccag aagaagatgt tggcgacctc gtattgggaa tccccgaaca tcgcctcgct 540 ccagtcaatg accgctgtta tgcggccatt gtccgtcagg acattgttgg agccgaaatc 600 cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc caaagcatca gctcatcgag 660 agcctgcgcg acggacgcac tgacggtgtc gtccatcaca gtttgccagt gatacacatg 720 gggatcagca atcgcgcata tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg 780 tccgaatggg ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg catccatagc 840 ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac 900 accctgtgca cggcgggaga tgcaataggt caggctctcg ctgaattccc caatgtcaag 960 cacttccgga atcgggagcg cggccgatgc aaagtgccga taaacataac gatctttgta 1020 gaaaccatcg gcgcagctat ttacccgcag gacatatcca cgccctccta catcgaagct 1080 gaaagcacga gattcttcgc cctccgagag ctgcatcagg tcggagacgc tgtcgaactt 1140 ttcgatcaga aacttctcga cagacgtcgc ggtgagttca ggcttttcca tgggtatatc 1200 tccttcttaa agttaaacaa aattatttct agagggaaac cgttgtggtc tccctatagt 1260 gagtcgtatt aatttcgcgg gatcgagatc tgatcaacct gcattaatga atcggccaac 1320 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 1380 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 1440 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 1500 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 1560 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 1620 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 1680 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct 1740 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 1800 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 1860 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 1920 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag 1980
tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 2040
gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 2100
cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 2160
agtggaacga aaactcacgt taagggattt tggtcatgac attaacctat aaaaataggc 2220
gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 2280
tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 2340
gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 2400
agcagattgt actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt 2460
aatacataac cttatgtatc atacacatac gatttaggtg acactataga acggcgcgcc 2520
gtcgacggat ccgtacgaga tccggccggc cagatcctgc aggagatcca agcttttgat 2580
ccatgccctt catttgccgc ttattaatta atttggtaac agtccgtact aatcagttac 2640
ttatccttcc cccatcataa ttaatcttgg tagtctcgaa tgccacaaca ctgactagtc 2700
tcttggatca taagaaaaag ccaaggaaca aaagaagaca aaacacaatg agagtatcct 2760
ttgcatagca atgtctaagt tcataaaatt caaacaaaaa cgcaatcaca cacagtggac 2820
atcacttatc cactagctga tcaggatcgc cgcgtcaaga aaaaaaaact ggaccccaaa 2880
agccatgcac aacaacacgt actcacaaag gtgtcaatcg agcagcccaa aacattcacc 2940
aactcaaccc atcatgagcc ctcacatttg ttgtttctaa cccaacctca aactcgtatt 3000
ctcttccgcc acctcatttt tgtttatttc aacacccgtc aaactgcatg ccaccccgtg 3060
gccaaatgtc catgcatgtt aacaagacct atgactataa atagctgcaa tctcggccca 3120
ggttttcatc atcaagaacc agttcaatat cctagtacac cgtattaaag aatttaagat 3180
atactgcggc cgcaccatgg aagcagccaa agaattggtt tccatcgtcc aagaggagct 3240
ccccaaggtg gactatgccc agctttggc.a ggatgccagc agctgtgagg tcctttacct 3300
ctcggtggca ttcgtggcga tcaagttcat gctgcgccca ctggacctga agcgccaggc 3360
caccttgaag aagctgttca cagcatacaa cttcctcatg tcgatctatt cctttggctc 3420
cttcctggcc atggcctatg ccctatcagt aactggcatc ctctccggcg actgtgagac 3480
ggcgttcaac aacgatgtgt tcaggatcac aactcagctg ttctacctca gcaagttcgt 3540
agagtacatc gactccttct accttcccct tatggacaag ccactgtcgt tccttcagtt 3600
cttccatcat ttgggggccc ccattgacat gtggctattc tacaaatacc gcaacgaagg 3660
agtctggatc tttgtcctgt tgaatgggtt cattcactgg atcatgtacg gttactattg 3720
gacgcggctc atcaagctga acttccccat gcccaagaac ctgatcacct ccatgcagat 3780 catccagttc aatgtcgggt tctacatcgt ctggaagtac cgcaatgtgc catgctaccg 3840
ccaggatggg atgcgcatgt ttgcctggat cttcaactac tggtatgtcg ggacggtctt 3900
gctgctgttc ctcaactttt acgtgcagac gtacatccgg aagccgagga agaaccgagg 3960
gaagaaggag tagcggccgc aagtatgaac taaaatgcat gtaggtgtaa gagctcatgg 4020
agagcatgga atattgtatc cgaccatgta acagtataat aactgagctc catctcactt 4080
cttctatgaa taaacaaagg atgttatgat atattaacac tctatctatg caccttattg 4140
ttctatgata aatttcctct tattattata aatcatctga atcgtgacgg cttatggaat 4200
gcttcaaata gtacaaaaac aaatgtgtac tataagactt tctaaacaat tctaacctta 4260
gcattgtgaa cgagacataa gtgttaagaa gacataacaa ttataatgga agaagtttgt 4 320
ctccatttat atattatata ttacccactt atgtattata ttaggatgtt aaggagacat 4380
aacaattata aagagagaag tttgtatcca tttatatatt atatactacc catttatata 4440
ttatacttat ccacttattt aatgtcttta taaggtttga tccatgatat ttctaatatt 4500
ttagttgata tgtatatgaa agggtactat ttgaactctc ttactctgta taaaggttgg 4560
atcatcctta aagtgggtct atttaatttt attgcttctt acagataaaa aaaaaattat 4620
gagttggttt gataaaatat tgaaggattt aaaataataa taaataacat ataatatatg 4680
tatataaatt tattataata taacatttat ctataaaaaa gtaaatattg tcataaatct 4740
atacaatcgt ttagccttgc tggacgaatc tcaattattt aaacgagagt aaacatattt 4800
gactttttgg ttatttaaca aattattatt taacactata tgaaattttt ttttttatca 4860
gcaaagaata aaattaaatt aagaaggaca atggtgtccc aatccttata caaccaactt 4920
ccacaagaaa gtcaagtcag agacaacaaa aaaacaagca aaggaaattt tttaatttga 4980
gttgtcttgt ttgctgcata atttatgcag taaaacacta cacataaccc ttttagcagt 5040
agagcaatgg ttgaccgtgt gcttagcttc ttttatttta tttttttatc agcaaagaat 5100
aaataaaata aaatgagaca cttcagggat gtttcaacaa gcttgg 514 6
<210> 161 <211> 5250 <212> DNA
<213> Seguência Artificial <220>
<223> plasmídeo pKR393 <400> 161
gatcccccgg gctgcaggaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac 60
cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat 120
agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg 180 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc 240 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca 300 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg 360 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga 420 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct 480 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc 540 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa 600 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt 660 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct 720 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc 780 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta 840 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac 900 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc 960 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac 1020 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg 1080 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac 1140 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc 1200 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt 1260 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga 1320 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc 1380 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag 1440 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca 1500 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc 1560 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca 1620 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc 1680 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta 1740 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt 1800 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc 1860 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg 1920 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg 1980 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag 2040 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc 2100
agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat 2160
agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg 2220
gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc 2280
tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt 2340
accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca 2400
gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg 2460
attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac 2520
gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg 2580
gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac 2640
catgattacg ccaagcttgc atgcctgcag gctagcctaa gtacgtactc aaaatgccaa 2700
caaataaaaa aaaagttgct ttaataatgc caaaacaaat taataaaaca cttacaacac 2760
cggatttttt ttaattaaaa tgtgccattt aggataaata gttaatattt ttaataatta 2820
tttaaaaagc cgtatctact aaaatgattt ttatttggtt gaaaatatta atatgtttaa 2880
atcaacacaa tctatcaaaa ttaaactaaa aaaaaaataa gtgtacgtgg ttaacattag 2940
tacagtaata taagaggaaa atgagaaatt aagaaattga aagcgagtct aatttttaaa 3000
ttatgaacct gcatatataa aaggaaagaa agaatccagg aagaaaagaa atgaaaccat 3060
gcatggtccc ctcgtcatca cgagtttctg ccatttgcaa tagaaacact gaaacacctt 3120
tctctttgtc acttaattga gatgccgaag ccacctcaca ccatgaactt catgaggtgt 3180
agcacccaag gcttccatag ccatgcatac tgaagaatgt ctcaagctca gcaccctact 3240
tctgtgacgt gtccctcatt caccttcctc tcttccctat aaataaccac gcctcaggtt 3300
ctccgcttca caactcaaac attctctcca ttggtcctta aacactcatc agtcatcacc 3360
gcggccgcca attcaggtgc ccatgatgtt ggccgcaggc tatcttctag tgctctcggc 3420
cgctcgccag agcttccagc aggacattga caaccccaac ggggcctact cgacctcgtg 3480
gactggcctg cccattgtga tgtctgtggt ctatctcagc ggtgtgtttg ggctcacaaa 3540
gtacttcgag aaccggaagc ccatgacggg gctgaaggac tacatgttca cttacaatct 3600
ctaccaggtg atcatcaacg tgtggtgcgt ggtggccttt ctcctggagg tgcggcgtgc 3660
gggcatgtca ctcatcggca ataaggtgga ccttgggccc aactccttca ggctcggctt 3720
cgtcacgtgg gtgcactaca acaacaagta cgtggagctc ctcgacaccc tatggatggt 3780
gctgcgcaag aagacgcagc aggtctcctt cctccacgtc tatcatcacg tgcttctgat 3840
gtgggcctgg ttcgttgtcg tcaagctcgg caatggtggt gacgcatatt ttggcggtct 3900 catgaactcg atcatccacg tgatgatgta ttcctactac accatggcgc tcctgggctg 3960
gtcatgcccc tggaagcgct acctcacgca ggcacagctc gtgcagtttt gcatctgcct 4020
cgcccactcc acatgggcgg cagtaacggg tgcctacccg tggcgaattt gcttggtgga 4080
ggtgtgggtg atggtgtcca tgctggtgct cttcacacgc ttctaccgcc aggcctatgc 4140
caaggaggcg aaggccaagg aggcgaaaaa gctcgcacag gaggcatcac aggccaaggc 4200
ggtcaaggcg gagtaagtca ctggaggtgg accgcacatg caccacgggc ccggcgagca 4260
gcatggttcg gcgagtcagg cccggtcatg cgtcatggtt ggagtttgca gggcggcagg 4320
tgatcgcctc cgccatgcac ggccacaggc acagccggtc ctctggacgt cccaactttc 4380
aaccgtggtg caaagcacgc tggcgaccgc gagcagcagt cagcgcagcg tgttatcaca 4440
gtgtcgctgg ctgcacgtgc tctctccatc gcggccgcat ttcgcaccaa atcaatgaaa 4500
gtaataatga aaagtctgaa taagaatact taggcttaga tgcctttgtt acttgtgtaa 4560
aataacttga gtcatgtacc tttggcggaa acagaataaa taaaaggtga aattccaatg 4620
ctctatgtat aagttagtaa tacttaatgt gttctacggt tgtttcaata tcatcaaact 4680
ctaattgaaa ctttagaacc acaaatctca atcttttctt aatgaaatga aaaatcttaa 4740
ttgtaccatg tttatgttaa acaccttaca attggttgga gaggaggacc aaccgatggg 4800
acaacattgg gagaaagaga ttcaatggag atttggatag gagaacaaca ttctttttca 4860
cttcaataca agatgagtgc aacactaagg atatgtatga gactttcaga agctacgaca 4920
acatagatga gtgaggtggt gattcctagc aagaaagaca ttagaggaag ccaaaatcga 4 980
acaaggaaga catcaagggc aagagacagg accatccatc tcaggaaaag gagctttggg 5040
atagtccgag aagttgtaca agaaattttt tggagggtga gtgatgcatt gctggtgact 5100
ttaactcaat caaaattgag aaagaaagaa aagggagggg gctcacatgt gaatagaagg 5160
gaaacgggag aattttacag ttttgatcta atgggcat;cc cagctagtgg taacatattc 5220
accatgttta accttcacgt acgtctagag 5250
<210> 162
<211> 4140
<212> DNA
<213> Seguência Artificial
<220>
<223> plasmídeo pKR407
<400> 162
ggccgcattt cgcaccaaat caatgaaagt aataatgaaa agtctgaata agaatactta 60
ggcttagatg cctttgttac ttgtgtaaaa taacttgagt catgtacctt tggcggaaac 120
agaataaata aaaggtgaaa ttccaatgct ctatgtataa gttagtaata cttaatgtgt 180 tctacggttg tttcaatatc atcaaactct aattgaaact ttagaaccac aaatctcaat 240 cttttcttaa tgaaatgaaa aatcttaatt gtaccatgtt tatgttaaac accttacaat 300 tggttggaga ggaggaccaa ccgatgggac aacattggga gaaagagatt caatggagat 360 ttggatagga gaacaacatt ctttttcact tcaatacaag atgagtgcaa cactaaggat 420 atgtatgaga ctttcagaag ctacgacaac atagatgagt gaggtggtga ttcctagcaa 480 gaaagacatt agaggaagcc aaaatcgaac aaggaagaca tcaagggcaa gagacaggac 540 catccatctc aggaaaagga gctttgggat agtccgagaa gttgtacaag aaattttttg 600 gagggtgagt gatgcattgc tggtgacttt aactcaatca aaattgagaa agaaagaaaa 660 gggagggggc tcacatgtga atagaaggga aacgggagaa ttttacagtt ttgatctaat 720 gggcatccca gctagtggta acatattcac catgtttaac cttcacgtac gtctagagga 780 tcccccgggc tgcaggaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc 840 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag 900 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg 960 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac 1020 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc 1080 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac 1140 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg 1200 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta 1260 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 1320 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 1380 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc 1440 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga 1500 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct 1560 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg 1620 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta 1680 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat 1740 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt 1800 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga 1860 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga 1920 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga 1980 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc 2040 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc 2100
cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg 2160
tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat 2220
cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata 2280
tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct 2340
ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 2400
ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 2460
cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 2520
aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct 2580
agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 2640
tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 2700
ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 2760
cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 2820
atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 2880
ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 2940
tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 3000
gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 3060
gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac 3120
cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt 3180
gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat 3240
tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc 3300
aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc 3360
tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca 3420
tgattacgcc aagcttgcat gcctgcaggc tagcctaagt acgtactcaa aatgccaaca 3480
aataaaaaaa aagttgcttt aataatgcca aaacaaatta ataaaacact tacaacaccg 3540
gatttttttt aattaaaatg tgccatttag gataaatagt taatattttt aataattatt 3600
taaaaagccg tatctactaa aatgattttt atttggttga aaatattaat atgtttaaat 3660
caacacaatc tatcaaaatt aaactaaaaa aaaaataagt gtacgtggtt aacattagta 3720
cagtaatata agaggaaaat gagaaattaa gaaattgaaa gcgagtctaa tttttaaatt 3780
atgaacctgc atatataaaa ggaaagaaag aatccaggaa gaaaàgaaat gaaaccatgc 3840
atggtcccct cgtcatcacg agtttctgcc atttgcaata gaaacactga aacacctttc 3900 tctttgtcac ttaattgaga tgccgaagcc acctcacacc atgaacttca tgaggtgtag 3960 cacccaaggc ttccatagcc atgcatactg aagaatgtct caagctcagc accctacttc 4020 tgtgacgtgt ccctcattca ccttcctctc ttccctataa ataaccacgc ctcaggttct 4080 ccgcttcaca actcaaacat tctctccatt ggtccttaaa cactcatcag tcatcaccgc 4140 <210> 163 <211> 5414 <212> DNA <213> Seqüência Artificial <220> <223> plasmideo pKR1176 <4 00> 163 ggccgcattt cgcaccaaat caatgaaagt aataatgaaa agtctgaata agaatactta 60 ggcttagatg cctttgttac ttgtgtaaaa taacttgagt catgtacctt tggcggaaac 120 agaataaata aaaggtgaaa ttccaatgct ctatgtataa gttagtaata cttaatgtgt 180 tctacggttg tttcaatatc atcaaactct aattgaaact ttagaaccac aaatctcaat 240 cttttcttaa tgaaatgaaa aatcttaatt gtaccatgtt tatgttaaac accttacaat 300 tggttggaga ggaggaccaa ccgatgggac aacattggga gaaagagatt caatggagat 360 ttggatagga gaacaacatt ctttttcact tcaatacaag atgagtgcaa cactaaggat 420 atgtatgaga ctttcagaag ctacgacaac atagatgagt gaggtggtga ttcctagcaa 480 gaaagacatt agaggaagcc aaaatcgaac aaggaagaca tcaagggcaa gagacaggac 540 catccatctc aggaaaagga gctttgggat agtccgagaa gttgtacaag aaattttttg 600 gagggtgagt gatgcattgc tggtgacttt aactcaatca aaattgagaa agaaagaaaa 660 gggagggggc tcacatgtga atagaaggga aacgggagaa ttttacagtt ttgatctaat 720 gggcatccca gctagtggta acatattcac catgtttaac cttcacgtac gtctagagga 780 tcccccgggc tgcaggaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc 840 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag 900 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg 960 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac 1020 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc 1080 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac 1140 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg 1200 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta 1260 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 1320 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 1380 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc 1440 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga 1500 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct 1560 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg 1620 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta 1680 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat 1740 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt 1800 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga 1860 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga 1920 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga 1980 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc 2040 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc 2100 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg 2160 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat 2220 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata 2280 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct 2340 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 2400 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 2460 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 2520 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct 2580 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 2640 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 2700 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 2760 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 2820 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 2880 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 2940 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 3000 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 3060 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac 3120 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt 3180 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat 3240 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc 3300 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc 3360 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca 3420 tgattacgcc aagcttgcat gcctgcaggc tagcctaagt acgtactcaa aatgccaaca 3480 aataaaaaaa aagttgcttt aataatgcca aaacaaatta ataaaacact tacaacaccg 3540 gatttttttt aattaaaatg tgccatttag gataaatagt taatattttt aataattatt 3600 taaaaagccg tatctactaa aatgattttt atttggttga aaatattaat atgtttaaat 3660 caacacaatc tatcaaaatt aaactaaaaa aaaaataagt gtacgtggtt aacattagta 3720 cagtaatata agaggaaaat gagaaattaa gaaattgaaa gcgagtctaa tttttaaatt 3780 atgaacctgc atatataaaa ggaaagaaag aatccaggaa gaaaagaaat gaaaccatgc 3840 atggtcccct cgtcatcacg agtttctgcc atttgcaata gaaacactga aacacctttc 3900 tctttgtcac ttaattgaga tgccgaagcc acctcacacc atgaacttca tgaggtgtag 3960 cacccaaggc ttccatagcc atgcatactg aagaatgtct caagctcagc accctacttc 4020 tgtgacgtgt ccctcattca ccttcctctc ttccctataa ataaccacgc ctcaggttct 4080 ccgcttcaca actcaaacat tctctccatt ggtccttaaa cactcatcag tcatcaccgc 4140 ggccgcacca tggtgaaaag gccagcactt ccgctgaccg ttgatggtgt cacctatgat 4200 gtgtctgcct ggttgaacca tcatccaggg ggtgctgaca tcattgagaa ctaccgcggt 4260 cgtgatgcca ctgatgtctt tatggttatg cactctgaaa atgctgtgag taaactaaga 4320 aggatgccta tcatggaacc atcatctcca ctgacgccta cgccaccgaa acccaactca 4380 gacgaaccgc aggaggattt ccgcaagctc cgagatgagc tcatcgcagc aggaatgttc 4440 gacgcatcac cgatgtggta cgcatataag acgctcagta cgctgggcct cggggtcctc 4500 gcggtgctat tgatgaccca gtggcactgg tacctcgtcg gggcaatcgt gttgggcatt 4560 cacttccaac aaatgggttg gttgtcgcac gatatctgcc accatcagct gttcaaggac 4620 cgatcgatca acaacgccat cggcttgctt ttcgggaacg tcttgcaagg gttctctgtg 4680 acctggtgga aggacaggca caatgcacac cactccgcca ccaacgtgca aggccacgac 4740 cccgacattg acaacctgcc gctgctggca tggtccaagg aggacgtgga gagggccggc 4800 ccgttctcac ggcggatgat caagtaccag caatactact tcttcttcat ctgtgccctc 4860 ctgaggttca tctggtgctt ccagagcatc cacacagcca cgggcctgaa ggatcgcagc 4920 aaccagtact accgcaggca gtacgagaaa gagagcgtgg gcctggccct ccactggggc 4980 ctgaaggcgt tgttctacta cttttatatg ccaagcttct tgaccggact catggtgttt 5040 ttcgtgtccg agttgcttgg gggcttcggc atcgccatcg tggtgttcat gaaccactac 5100 cccctggaga agatccagga ctcggtgtgg gacggccacg gcttttgcgc cggccagatt 5160 cacgaaacga tgaacgtcca gcggggactc gtcacggact ggttcttcgg tgggctgaat 5220 taccaaatcg agcaccacct gtggccgacg ctgccccggc acaacctgac ggcggccagc 5280 atcaaagtgg agcagttgtg caagaagcac aacttgccgt atcgcagccc cccaatgctg 5340 gagggggtgg gcatcctgat cagctacctg ggcacctttg cccgcatggt ggcaaaggcc 5400 gacaaggcgt aagc 5414
<210> 164 <211> 7907 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo pKR1178 <400> 164
ggagatccaa gcttttgatc catgcccttc atttgccgct tattaattaa tttggtaaca 60
gtccgtacta atcagttact tatccttccc ccatcataat taatcttggt agtctcgaat 120
gccacaacac tgactagtct cttggatcat aagaaaaagc caaggaacaa aagaagacaa 180
aacacaatga gagtatcctt tgcatagcaa tgtctaagtt cataaaattc aaacaaaaac 240
gcaatcacac acagtggaca tcacttatcc actagctgat caggatcgcc gcgtcaagaa 300
aaaaaaactg gaccccaaaa gccatgcaca acaacacgta ctcacaaagg tgtcaatcga 360
gcagcccaaa acattcacca actcaaccca tcatgagccc tcacatttgt tgtttctaac 420
ccaacctcaa actcgtattc tcttccgcca cctcattttt gtttatttca acacccgtca 480
aactgcatgc caccccgtgg ccaaatgtcc atgcatgtta acaagaccta tgactataaa 540
tagctgcaat ctcggcccag gttttcatca tcaagaacca gttcaatatc ctagtacacc 600
gtattaaaga atttaagata tactgcggcc gcaccatgga agcagccaaa gaattggttt 660
ccatcgtcca agaggagctc cccaaggtgg actatgccca gctttggcag gatgccagca 720
gctgtgaggt cctttacctc tcggtggcat tcgtggcgat caagttcatg ctgcgcccac 780
tggacctgaa gcgccaggcc accttgaaga agctgttcac agcatacaac ttcctcatgt 840
cgatctattc ctttggctcc ttcctggcca tggcctatgc cctatcagta actggcatcc 900
tctccggcga ctgtgagacg gcgttcaaca acgatgtgtt caggatcaca actcagctgt 960
tctacctcag caagttcgta gagtacatcg actccttcta ccttcccctt atggacaagc 1020
cactgtcgtt ccttcagttc ttccatcatt tgggggcccc cattgacatg tggctattct 1080
acaaataccg caacgaagga gtctggatct ttgtcctgtt gaatgggttc attcactgga 1140 tcatgtacgg ttactattgg acgcggctca tcaagctgaa cttccccatg cccaagaacc 1200 tgatcacctc catgcagatc atccagttca atgtcgggtt ctacatcgtc tggaagtacc 1260 gcaatgtgcc atgctaccgc caggatggga tgcgcatgtt tgcctggatc ttcaactact 1320 ggtatgtcgg gacggtcttg ctgctgttcc tcaactttta cgtgcagacg tacatccgga 1380 agccgaggaa gaaccgaggg aagaaggagt agcggccgca agtatgaact aaaatgcatg 1440 taggtgtaag agctcatgga gagcatggaa tattgtatcc gaccatgtaa cagtataata 1500 actgagctcc atctcacttc ttctatgaat aaacaaagga tgttatgata tattaacact 1560 ctatctatgc accttattgt tctatgataa atttcctctt attattataa atcatctgaa 1620 tcgtgacggc ttatggaatg cttcaaatag tacaaaaaca aatgtgtact ataagacttt 1680 ctaaacaatt ctaaccttag cattgtgaac gagacataag tgttaagaag acataacaat 1740 tataatggaa gaagtttgtc tccatttata tattatatat tacccactta tgtattatat 1800 taggatgtta aggagacata acaattataa agagagaagt ttgtatccat ttatatatta 1860 tatactaccc atttatatat tatacttatc cacttattta atgtctttat aaggtttgat 1920 ccatgatatt tctaatattt tagttgatat gtatatgaaa gggtactatt tgaactctct 1980 tactctgtat aaaggttgga tcatccttaa agtgggtcta tttaatttta ttgcttctta 2040 cagataaaaa aaaaattatg agttggtttg ataaaatatt gaaggattta aaataataat 2100 aaataacata taatatatgt atataaattt attataatat aacatttatc tataaaaaag 2160 taaatattgt cataaatcta tacaatcgtt tagccttgct ggacgaatct caattattta 2220 aacgagagta aacatatttg actttttggt tatttaacaa attattattt aacactatat 2280 gaaatttttt tttttatcag caaagaataa aattaaatta agaaggacaa tggtgtccca 2340 atccttatac aaccaacttc cacaagaaag tcaagtcaga gacaacaaaa aaacaagcaa 2400 aggaaatttt ttaatttgag ttgtcttgtt tgctgcataa tttatgcagt aaaacactac 2460 acataaccct tttagcagta gagcaatggt tgaccgtgtg cttagcttct tttattttat 2520 ttttttatca gcaaagaata aataaaataa aatgagacac ttcagggatg tttcaacaag 2580 cttggcgcgc ccgatcatcc ggatatagtt cctcctttca gcaaaaaacc cctcaagacc 2640 cgtttagagg ccccaagggg ttatgctagt tattgctcag cggtggcagc agccaactca 2700 gcttcctttc gggctttgtt agcagccgga tcgatccaag ctgtacctca ctattccttt 2760 gccctcggac gagtgctggg gcgtcggttt ccactatcgg cgagtacttc tacacagcca 2820 tcggtccaga cggccgcgct tctgcgggcg atttgtgtac gcccgacagt cccggctccg 2880 gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa attgccgtca 2940 accaagctct gatagagttg gtcaagacca atgcggagca tatacgcccg gagccgcggc 3000 gatcctgcaa gctccggatg cctccgctcg aagtagcgcg tctgctgctc catacaagcc 3060 aaccacggcc tccagaagaa gatgttggcg acctcgtatt gggaatcccc gaacatcgcc 3120 tcgctccagt caatgaccgc tgttatgcgg ccattgtccg tcaggacatt gttggagccg 3180 aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct cggcccaaag catcagctca 3240 tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac 3300 acatggggat cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg accgattcct 3360 tgcggtccga atgggccgaa cccgctcgtc tggctaagat cggccgcagc gatcgcatcc 3420 atagcctccg cgaccggctg cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac 3480 gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa ttccccaatg 3540 tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt gccgataaac ataacgatct 3600 ttgtagaaac catcggcgca gctatttacc cgcaggacat atccacgccc tcctacatcg 3660 aagctgaaag cacgagattc ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg 3720 aacttttcga tcagaaactt ctcgacagac gtcgcggtga gttcaggctt ttccatgggt 3780 atatctcctt cttaaagtta aacaaaatta tttctagagg gaaaccgttg tggtctccct 3840 atagtgagtc gtattaattt cgcgggatcg agatctgatc aacctgcatt aatgaatcgg 3900 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 3960 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 4020 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 4080 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 4140 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 4200 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 4260 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc 4320 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 4380 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 4440 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 4500 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 4560 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 4620 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 4680 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 4740 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgacattaa cctataaaaa 4800 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 4860 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 4920 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 4980 atcagagcag attgtactga gagtgcacca tatggacata ttgtcgttag aacgcggcta 5040 caattaatac ataaccttat gtatcataca catacgattt aggtgacact atagaacggc 5100 gcgccgtcga cggatccgta cgagatccgg ccggccagat cctgcagccc gggggatcct 5160 ctagacgtac gtgaaggtta aacatggtga atatgttacc actagctggg atgcccatta 5220 gatcaaaact gtaaaattct cccgtttccc ttctattcac atgtgagccc cctccctttt 5280 CtttCtttCt caattttgat tgagttaaag tcaccagcaa tgcatcactc accctccaaa 5340 aaatttcttg tacaacttct cggactatcc caaagctcct tttcctgaga tggatggtcc 5400 tgtctcttgc ccttgatgtc ttccttgttc gattttggct tcctctaatg tctttcttgc 5460 taggaatcac cacctcactc atctatgttg tcgtagcttc tgaaagtctc atacatatcc 5520 ttagtgttgc actcatcttg tattgaagtg aaaaagaatg ttgttctcct atccaaatct 5580 ccattgaatc tctttctccc aatgttgtcc catcggttgg tcctcctctc caaccaattg 5640 taaggtgttt aacataaaca tggtacaatt aagatttttc atttcattaa gaaaagattg 5700 agatttgtgg ttctaaagtt tcaattagag tttgatgata ttgaaacaac cgtagaacac 5760 attaagtatt actaacttat acatagagca ttggaatttc accttttatt tattctgttt 5820 ccgccaaagg tacatgactc aagttatttt acacaagtaa caaaggcatc taagcctaag 5880 tattcttatt cagacttttc attattactt tcattgattt ggtgcgaaat gcggccgctt 5940 acgccttgtc ggcctttgcc accatgcggg caaaggtgcc caggtagctg atcaggatgc 6000 ccaccccctc cagcattggg gggctgcgat acggcaagtt gtgcttcttg cacaactgct 6060 ccactttgat gctggccgcc gtcaggttgt gccggggcag cgtcggccac aggtggtgct 6120 cgatttggta attcagccca ccgaagaacc agtccgtgac gagtccccgc tggacgttca 6180 tcgtttcgtg aatctggccg gcgcaaaagc cgtggccgtc ccacaçcgag tcctggatct 6240 tctccagggg gtagtggttc atgaacacca cgatggcgat gccgaagccc ccaagcaact 6300 cggacacgaa aaacaccatg agtccggtca agaagcttgg catataaaag tagtagaaca 6360 acgccttcag gccccagtgg agggccaggc ccacgctctc tttctcgtac tgcctgcggt 6420 agtactggtt gctgcgatcc ttcaggcccg tggctgtgtg gatgctctgg aagcaccaga 6480 tgaacctcag gagggcacag atgaagaaga agtagtattg ctggtacttg atcatccgcc 6540 gtgagaacgg gccggccctc tccacgtcct ccttggacca tgccagcagc ggcaggttgt 6600 caatgtcggg gtcgtggcct tgcacgttgg tggcggagtg gtgtgcattg tgcctgtcct 6660 tccaccaggt cacagagaac ccttgcaaga cgttcccgaa aagcaagccg atggcgttgt 6720 tgatcgatcg gtccttgaac agctgatggt ggcagatatc gtgcgacaac caacccattt 6780 gttggaagtg aatgcccaac acgattgccc cgacgaggta ccagtgccac tgggtcatca 6840 atagcaccgc gaggaccccg aggcccagcg tactgagcgt cttatatgcg taccacatcg 6900 gtgatgcgtc gaacattcct gctgcgatga gctcatctcg gagcttgcgg aaatcctcct 6960 gcggttcgtc tgagttgggt ttcggtggcg taggcgtcag tggagatgat ggttccatga 7020 taggcatcct tcttagttta ctcacagcat tttcagagtg cataaccata aagacatcag 7080 tggcatcacg accgcggtag ttctcaatga tgtcagcacc ccctggatga tggttcaacc 7140 aggcagacac atcataggtg acaccatcaa cggtcagcgg aagtgctggc cttttcacca 7200 tggtgcggcc gcggtgatga ctgatgagtg tttaaggacc aatggagaga atgtttgagt 7260 tgtgaagcgg agaacctgag gcgtggttat ttatagggaa gagaggaagg tgaatgaggg 7320 acacgtcaca gaagtagggt gctgagcttg agacattctt cagtatgcat ggctatggaa 7380 gccttgggtg ctacacctca tgaagttcat ggtgtgaggt ggcttcggca tctcaattaa 7440 gtgacaaaga gaaaggtgtt tcagtgtttc tattgcaaat ggcagaaact cgtgatgacg 7500 aggggaccat gcatggtttc atttcttttc ttcctggatt ctttctttcc ttttatatat 7560 gcaggttcat aatttaaaaa ttagactcgc tttcaatttc ttaatttctc attttcctct 7620 tatattactg tactaatgtt aaccacgtac acttattttt tttttagttt aattttgata 7680 gattgtgttg atttaaacat attaatattt tcaaccaaat aaaaatcatt ttagtagata 7740 cggcttttta aataattatt aaaaatatta actatttatc ctaaatggca cattttaatt 7800 aaaaaaaatc cggtgttgta agtgttttat taatttgttt tggcattatt aaagcaactt 7860 tttttttatt tgttggcatt ttgagtacgt acttaggcta gcctgca 7907
<210> 165 <211> 29 <212> DNA <213> Seqüência Artificial
<220> <223> oligonucleotideo oKti5 <400> 165 atctagacgt acgtcctcga agagaaggg 29
<210> 166 <211> 27 <212> DNA <213> Seqüência Artificial
<220> <223> oligonucleotideo oKti6 <400> 166
ttctagacgt acggatataa tgagccg 27
<210> 167
<211> 4474
<212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo pKR193
<400> 167 gtacgtctag aggatccccg ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt 60 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc 120 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc 180 ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca 240 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc 300 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct 360 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca 420 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg 480 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct 540 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga 600 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc 660 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg 720 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc 780 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact 840 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc 900 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag 960 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat 1020 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt 1080 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa 1140 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc 1200 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg 1260 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt 1320 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca 1380 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat 1440 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca 1500 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg 1560 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg 1620 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt 1680 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg 1740 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata 1800 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca 1860 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag 1920 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc 1980 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga 2040 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg 2100 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac 2160 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg 2220 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg 2280 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct 2340 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc 2400 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc 2460 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg 2520 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta 2580 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca 2640 ggaaacagct atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctcgacgtac 2700 gatccatgcc cttcatttgc cgcttattaa ttaatttggt aacagtccgt actaatcagt 2760 tacttatcct tcccccatca taattaatct tggtagtctc gaatgccaca acactgacta 2820 gtctcttgga tcataagaaa aagccaagga acaaaagaag acaaaacaca atgagagtat 2880 cctttgcata gcaatgtcta agttcataaa attcaaacaa aaacgcaatc acacacagtg 2940 gacatcactt atccactagc tgatcaggat cgccgcgtca agaaaaaaaa actggacccc 3000 aaaagccatg cacaacaaca cgtactcaca aaggtgtcaa tcgagcagcc caaaacattc 3060 accaactcaa cccatcatga gccctcacat ttgttgtttc taacccaacc tcaaactcgt 3120 attctcttcc gccacctcat ttttgtttat ttcaacaccc gtcaaactgc atgccacccc 3180 gtggccaaat gtccatgcat gttaacaaga cctatgacta taaatagctg caatctcggc 3240 ccaggttttc atcatcaaga accagttcaa tatcctagta caccgtatta aagaatttaa 3300 gatatactgc ggccgcaagt atgaactaaa atgcatgtag gtgtaagagc tcatggagag 3360 catggaatat tgtatccgac catgtaacag tataataact gagctccatc tcacttcttc 3420
tatgaataaa caaaggatgt tatgatatat taacactcta tctatgcacc ttattgttct 3480
atgataaatt tcctcttatt attataaatc atctgaatcg tgacggctta tggaatgctt 3540
caaatagtac aaaaacaaat gtgtactata agactttcta aacaattcta accttagcat 3600
tgtgaacgag acataagtgt taagaagaca taacaattat aatggaagaa gtttgtctcc 3660
atttatatat tatatattac ccacttatgt attatattag gatgttaagg agacataaca 3720
attataaaga gagaagtttg tatccattta tatattatat actacccatt tatatattat 3780
acttatccac ttatttaatg tctttataag gtttgatcca tgatatttct aatattttag 3840
ttgatatgta tatgaaaggg tactatttga actctcttac tctgtataaa ggttggatca 3900
tccttaaagt gggtctattt aattttattg cttcttacag ataaaaaaaa aattatgagt 3960
tggtttgata aaatattgaa ggatttaaaa taataataaa taacatataa tatatgtata 4020
taaatttatt ataatataac atttatctat aaaaaagtaa atattgtcat aaatctatac 4080
aatcgtttag ccttgctgga cgaatctcaa ttatttaaac gagagtaaac atatttgact 4140
ttttggttat ttaacaaatt attatttaac actatatgaa attttttttt ttatcagcaa 4200
agaataaaat taaattaaga aggacaatgg tgtcccaatc cttatacaac caacttccac 4260
aagaaagtca agtcagagac aacaaaaaaa caagcaaagg aaatttttta atttgagttg 4320
tcttgtttgc tgcataattt atgcagtaaa acactacaca taaccctttt agcagtagag 4380
caatggttga ccgtgtgctt agcttctttt attttatttt tttatcagca aagaataaat 4440
aaaataaaat gagacacttc agggatgttt caac 4474
<210> 168 <211> 5851 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmídeo pKR1174 <400> 168
ggccgcaagt atgaactaaa atgcatgtag gtgtaagagc tcatggagag catggaatat 60
tgtatccgac catgtaacag tataataact gagctccatc tcacttcttc tatgaataaa 120
caaaggatgt tatgatatat taacactcta tctatgcacc ttattgttct atgataaatt 180
tcctcttatt attataaatc atctgaatcg tgacggctta tggaatgctt caaatagtac 240
aaaaacaaat gtgtactata agactttcta aacaattcta accttagcat tgtgaacgag 300
acataagtgt taagaagaca taacaattat aatggaagaa gtttgtctcc atttatatat 360
tatatattac ccacttatgt attatattag gatgttaagg agacataaca attataaaga 420 gagaagtttg tatccattta tatattatat actacccatt tatatattat acttatccac 480
ttatttaatg tctttataag gtttgatcca tgatatttct aatattttag ttgatatgta 540
tatgaaaagg tactatttga actctcttac tctgtataaa ggttggatca tccttaaagt 600
gggtctattt aattttattg cttcttacag ataaaaaaaa aattatgagt tggtttgata 660
aaatattgaa ggatttaaaa taataataaa taacatataa tatatgtata taaatttatt 720
ataatataac atttatctat aaaaaagtaa atattgtcat aaatctatac aatcgtttag 780
ccttgctgga acgaatctca attatttaaa cgagagtaaa catatttgac tttttggtta 840
tttaacaaat tattatttaa cactatatga aatttttttt tttatcagca aagaataaaa 900
ttaaattaag aaggacaatg gtgtcccaat ccttatacaa ccaacttcca caagaaagtc 960
aagtcagaga caacaaaaaa acaagcaaag gaaatttttt aatttgagtt gtcttgtttg 1020
ctgcataatt tatgcagtaa aacactacac ataacccttt tagcagtaga gcaatggttg 1080
accgtgtgct tagcttcttt tattttattt ttttatcagc aaagaataaa taaaataaaa 1140
tgagacactt cagggatgtt tcaacgtacg tctagaggat ccccgggtac cgagctcgaa 1200
ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa 1260
tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga 1320
tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct 1380
ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc 1440
tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg 1500
ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat 1560
gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg 1620
cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt 1680
tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 1740
tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat 1800
gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt 1860
ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg 1920
agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga 1980
agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg 2040
tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt 2100
tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg 2160
cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg 2220
aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga 2280 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc 2340
tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc 2400
ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc 2460
ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg 2520
cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac 2580
gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc 2640
actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt 2700
aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac 2760
caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa 2820
aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc 2880
accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt 2940
aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg 3000
ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 3060
agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 3120
accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 3180
gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 3240
tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 3300
cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca 3360
cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 3420
cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt 3480
ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 3540
taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 3600
gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 3660
cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct 3720
cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat 3780
tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc 3840
atgcctgcag gtcgactcga cgtacgatcc atgcccttca tttgccgctt attaattaat 3900
ttggtaacag tccgtactaa tcagttactt atccttcccc catcataatt aatcttggta 3960
gtctcgaatg ccacaacact gactagtctc ttggatcata agaaaaagcc aaggaacaaa 4020
agaagacaaa acacaatgag agtatccttt gcatagcaat gtctaagttc ataaaattca 4080
aacaaaaacg caatcacaca cagtggacat cacttatcca ctagctgatc aggatcgccg 4140 cgtcaagaaa aaaaaactgg accccaaaag ccatgcacaa caacacgtac tcacaaaggt 4200
gtcaatcgag cagcccaaaa cattcaccaa ctcaacccat catgagccct cacatttgtt 4260
gtttctaacc caacctcaaa ctcgtattct cttccgccac ctcatttttg tttatttcaa 4320
cacccgtcaa actgcatgcc accccgtggc caaatgtcca tgcatgttaa caagacctat 4380
gactataaat agctgcaatc tcggcccagg ttttcatcat caagaaccag ttcaatatcc 4440
tagtacaccg tattaaagaa tttaagatat actgcggccg caccatggcc accatctctt 4500
tgactactga gcaactttta gaacacccag aactggttgc aattgatggg gtgttgtacg 4560
atctcttcgg actggcgaaa gtgcatccag gtggcaacct cattgaagcc gccggtgcct 4620
ccgacggaac cgccctgttc tactccatgc accctggagt gaagccagag aattcgaagc 4680
tgctgcagca atttgcccga ggcaaacacg aacgaagctc gaaggaccca gtgtacacct 4740
ttgacagtcc cttcgcccag gatgtcaagc agagcgttcg ggaggtcatg aaggggcgca 4800
actggtacgc cacgcccggc ttttggctgc ggaccgcgct gatcatcgcg tgcactgcca 4860
taggcgaatg gtattggatc actaccgggg cagtgatgtg gggcatcttc accgggtact 4 920
tccacagcca gattgggttg gcgattcaac acgatgcctc tcacggagcc atcagcaaaa 4980
agccctgggt gaacgccttt ttcgcctacg gcatcgacgc cattggatcc tcccgctgga 5040
tctggctgca gtcccacatt atgcgccacc acacctacac caaccagcat ggcctggacc 5100
tggacgctgc ctcggcggag ccgttcattt tgttccactc ctacccggca acaaatgcgt 5160
cacgaaagtg gtaccatcgg ttccaggcgt ggtacatgta catcgttttg gggatgtatg 5220
gtgtgtcgat ggtgtacaat ccgatgtact tgttcacgat gcagcacaac gacacaatcc 5280
cagaggccac ctctcttaga ccaggcagct ttttcaaccg gcagcgcgcc ttcgccgttt 5340
ccctccgcct actgttcatc ttccgcaacg ccttcctccc ctggtacatc gcgggcgcct 5400
ctccgctgct caccatcctg ctggtgccaa cggtcacagg catcttcttg acatttgttt 5460
ttgtgctgtc ccataacttt gaaggcgctg agcggacccc cgaaaagaac tgcaaggcca 5520
aaagggccaa ggaggggaag gaggtccgcg atgtagagga ggaccgggtg gactggtacc 5580
gggcgcaggc cgagaccgcg gcgacctacg ggggcagcgt cgggatgatg ctgaccggcg 5640
gtttgaacct gcagatcgag caccacttgt tcccccgcat gtcctcttgg cactacccct 5700
tcatccaaga tacggtgcgg gaatgttgca agcgccatgg cgtgcgctac acatactacc 57 60
cgaccatcct ggagaatata atgtccacgc tccgctacat gcagaaggtg ggcgtggccc 5820
acacaattca ggatgcccag gaattctgag c 5851
<210> <211>
169 11013 <212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo pKR1186 <400> 169
gtacgtgaag gttaaacatg gtgaatatgt taccactagc tgggatgccc attagatcaa 60
aactgtaaaa ttctcccgtt tcccttctat tcacatgtga gccccctccc ttttctttct 120
ttctcaattt tgattgagtt aaagtcacca gcaatgcatc actcaccctc caaaaaattt 180
cttgtacaac ttctcggact atcccaaagc tccttttcct gagatggatg gtcctgtctc 240
ttgcccttga tgtcttcctt gttcgatttt ggcttcctct aatgtctttc ttgctaggaa 300
tcaccacctc actcatctat gttgtcgtag cttctgaaag tctcatacat atccttagtg 360
ttgcactcat cttgtattga agtgaaaaag aatgttgttc tcctatccaa atctccattg 420
aatctctttc tcccaatgtt gtcccatcgg ttggtcctcc tctccaacca attgtaaggt 480
gtttaacata aacatggtac aattaagatt tttcatttca ttaagaaaag attgagattt 540
gtggttctaa agtttcaatt agagtttgat gatattgaaa caaccgtaga acacattaag 600
tattactaac ttatacatag agcattggaa tttcaccttt tatttattct gtttccgcca 660
aaggtacatg actcaagtta ttttacacaa gtaacaaagg catctaagcc taagtattct 720
tattcagact tttcattatt actttcattg atttggtgcg aaatgcggcc gcttacgcct 780
tgtcggcctt tgccaccatg cgggcaaagg tgcccaggta gctgatcagg atgcccaccc 840
cctccagcat tggggggctg cgatacggca agttgtgctt cttgcacaac tgctccactt 900
tgatgctggc cgccgtcagg ttgtgccggg gcagcgtcgg ccacaggtgg tgctcgattt 960
ggtaattcag cccaccgaag aaccagtccg tgacgagtcc ccgctggacg ttcatcgttt 1020
cgtgaatctg gccggcgcaa aagccgtggc cgtcccacac cgagtcctgg atcttctcca 1080
gggggtagtg gttcatgaac accacgatgg cgatgccgaa gcccccaagc aactcggaca 1140
cgaaaaacac catgagtccg gtcaagaagc ttggcatata aaagtagtag aacaacgcct 1200
tcaggcccca gtggagggcc aggcccacgc tctctttctc gtactgcctg cggtagtact 1260
ggttgctgcg atccttcagg cccgtggctg tgtggatgct ctggaagcac cagatgaacc 1320
tcaggagggc acagatgaag aagaagtagt attgctggta cttgatcatc cgccgtgaga 1380
acgggccggc cctctccacg tcctccttgg accatgccag cagcggcagg ttgtcaatgt 1440
cggggtcgtg gccttgcacg ttggtggcgg agtggtgtgc attgtgcctg tccttccacc 1500
aggtcacaga gaacccttgc aagacgttcc cgaaaagcaa gccgatggcg ttgttgatcg 1560
atcggtcctt gaacagctga tggtggcaga tatcgtgcga caaccaaccc atttgttgga 1620
agtgaatgcc caacacgatt gccccgacga ggtaccagtg ccactgggtc atcaatagca 1680 ccgcgaggac cccgaggccc agcgtactga gcgtcttata tgcgtaccac atcggtgatg 1740
cgtcgaacat tcctgctgcg atgagctcat ctcggagctt gcggaaatcc tcctgcggtt 1800
cgtctgagtt gggtttcggt ggcgtaggcg tcagtggaga tgatggttcc atgataggca 1860
tccttcttag tttactcaca gcattttcag agtgcataac cataaagaca tcagtggcat 1920
cacgaccgcg gtagttctca atgatgtcag caccccctgg atgatggttc aaccaggcag 1980
acacatcata ggtgacacca tcaacggtca gcggaagtgc tggccttttc accatggtgc 2040
ggccgcggtg atgactgatg agtgtttaag gaccaatgga gagaatgttt gagttgtgaa 2100
gcggagaacc tgaggcgtgg ttatttatag ggaagagagg aaggtgaatg agggacacgt 2160
cacagaagta gggtgctgag cttgagacat tcttcagtat gcatggctat ggaagccttg 2220
ggtgctacac ctcatgaagt tcatggtgtg aggtggcttc ggcatctcaa ttaagtgaca 2280
aagagaaagg tgtttcagtg tttctattgc aaatggcaga aactcgtgat gacgagggga 2340
ccatgcatgg tttcatttct tttcttcctg gattctttct ttccttttat atatgcaggt 2400
tcataattta aaaattagac tcgctttcaa tttcttaatt tctcattttc ctcttatatt 2460
actgtactaa tgttaaccac gtacacttat ttttttttta gtttaatttt gatagattgt 2520
gttgatttaa acatattaat attttcaacc aaataaaaat cattttagta gatacggctt 2580
tttaaataat tattaaaaat attaactatt tatcctaaat ggcacatttt aattaaaaaa 2640
aatccggtgt tgtaagtgtt ttattaattt gttttggcat tattaaagca actttttttt 2700
tatttgttgg cattttgagt acgtacttag gctagcctgc aggagatcca agcttttgat 2760
ccatgccctt catttgccgc ttattaatta atttggtaac agtccgtact aatcagttac 2820
ttatccttcc cccatcataa ttaatcttgg tagtctcgaa tgccacaaca ctgactagtc 2880
tcttggatca taagaaaaag ccaaggaaca aaagaagaca aaacacaatg agagtatcct 2940
ttgcatagca atgtctaagt tcataaaatt caaacaaaaa cgcaatcaca cacagtggac 3000
atcacttatc cactagctga tcaggatcgc cgcgtcaaga aaaaaaaact ggaccccaaa 3060
agccatgcac aacaacacgt actcacaaag gtgtcaatcg agcagcccaa aacattcacc 3120
aactcaaccc atcatgagcc ctcacatttg ttgtttctaa cccaacctca aactcgtatt 3180
ctcttccgcc acctcatttt tgtttatttc aacacccgtc aaactgcatg ccaccccgtg 3240
gccaaatgtc catgcatgtt aacaagacct atgactataa atagctgcaa tctcggccca 3300
ggttttcatc atcaagaacc agttcaatat cctagtacac cgtattaaag aatttaagat 3360
atactgcggc cgcaccatgg aagcagccaa agaattggtt tccatcgtcc aagaggagct 3420
ccccaaggtg gactatgccc agctttggca ggatgccagc agctgtgagg tcctttacct 3480
ctcggtggca ttcgtggcga tcaagttcat gctgcgccca ctggacctga agcgccaggc 3540 caccttgaag aagctgttca cagcatacaa cttcctcatg tcgatctatt cctttggctc 3600
cttcctggcc atggcctatg ccctatcagt aactggcatc ctctccggcg actgtgagac 3660
ggcgttcaac aacgatgtgt tcaggatcac aactcagctg ttctacctca gcaagttcgt 3720
agagtacatc gactccttct accttcccct tatggacaag ccactgtcgt tccttcagtt 3780
cttccatcat ttgggggccc ccattgacat gtggctattc tacaaatacc gcaacgaagg 3840
agtctggatc tttgtcctgt tgaatgggtt cattcactgg atcatgtacg gttactattg 3900
gacgcggctc atcaagctga acttccccat gcccaagaac ctgatcacct ccatgcagat 3960
catccagttc aatgtcgggt tctacatcgt ctggaagtac cgcaatgtgc catgctaccg 4020
ccaggatggg atgcgcatgt ttgcctggat cttcaactac tggtatgtcg ggacggtctt 4080
gctgctgttc ctcaactttt acgtgcagac gtacatccgg aagccgagga agaaccgagg 4140
gaagaaggag tagcggccgc aagtatgaac taaaatgcat gtaggtgtaa gagctcatgg 4200
agagcatgga atattgtatc cgaccatgta acagtataat aactgagctc catctcactt 4260
cttctatgaa taaacaaagg atgttatgat atattaacac tctatctatg caccttattg 4320
ttctatgata aatttcctct tattattata aatcatctga atcgtgacgg cttatggaat 4380
gcttcaaata gtacaaaaac aaatgtgtac tataagactt tctaaacaat tctaacctta 4440
gcattgtgaa cgagacataa gtgttaagaa gacataacaa ttataatgga agaagtttgt 4500
ctccatttat atattatata ttacccactt atgtattata ttaggatgtt aaggagacat 4560
aacaattata aagagagaag tttgtatcca tttatatatt atatactacc catttatata 4 620
ttatacttat ccacttattt aatgtcttta taaggtttga tccatgatat ttctaatatt 4680
ttagttgata tgtatatgaa agggtactat ttgaactctc ttactctgta taaaggttgg 4740
atcatcctta aagtgggtct atttaatttt attgcttctt acagataaaa aaaaaattat 4800
gagttggttt gataaaatat tgaaggattt aaaataataa taaataacat ataatatatg 4860
tatataaatt tattataata taacatttat ctataaaaaa gtaaatattg tcataaatct 4920
atacaatcgt ttagccttgc tggacgaatc tcaattattt aaacgagagt aaacatattt 4980
gactttttgg ttatttaaca aattattatt taacactata tgaaattttt ttttttatca 5040
gcaaagaata aaattaaatt aagaaggaca atggtgtccc aatccttata caaccaactt 5100
ccacaagaaa gtcaagtcag agacaacaaa aaaacaagca aaggaaattt tttaatttga 5160
gttgtcttgt ttgctgcata atttatgcag taaaacacta cacataaccc ttttagcagt 5220
agagcaatgg ttgaccgtgt gcttagcttc ttttatttta tttttttatc agcaaagaat 5280
aaataaaata aaatgagaca cttcagggat gtttcaacaa gcttggcgcg cccgatcatc 5340
cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag gccccaaggg 5400 gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt cgggctttgt 5460 tagcagccgg atcgatccaa gctgtacctc actattcctt tgccctcgga cgagtgctgg 5520 ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag acggccgcgc 5580 ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg attgcgtcgc 5640 atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc tgatagagtt 5700 ggtcaagacc aatgcggagc atatacgccc ggagccgcgg cgatcctgca agctccggat 5760 gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc ctccagaaga 5820 agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag tcaatgaccg 5880 ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg tgcacgaggt 5940 gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc tgcgcgacgg 6000 acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga tcagcaatcg 6060 cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg aatgggccga 6120 acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catagcctcc gcgaccggct 6180 gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc tgtgcacggc 6240 gggagatgca ataggtcagg ctctcgctga attccccaat gtcaagcact tccggaatcg 6300 ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa ccatcggcgc 6360 agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa gcacgagatt 6420 cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg atcagaaact 6480 tctcgacaga cgtcgcggtg agttcaggct tttccatggg tatatctcct tcttaaagtt 6540 aaacaaaatt atttctagag ggaaaccgtt gtggtctccc tatagtgagt cgtattaatt 6600 tcgcgggatc gagatctgat caacctgcat taatgaatcg gccaacgcgc ggggagaggc 6660 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 6720 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 6780 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 6840 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 6900 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 6960 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 7020 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt 7080 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 7140 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 7200 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 7260 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc 7320
gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 7380
accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 7440
ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 7500
tcacgttaag ggattttggt catgacatta acctataaaa ataggcgtat cacgaggccc 7560
tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag 7620
acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca 7680
gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg 7740
agagtgcacc atatggacat attgtcgtta gaacgcggct acaattaata cataacctta 7800
tgtatcatac acatacgatt taggtgacac tatagaacgg cgcgccgtcg acggatccgt 7860
acgttgaaac atccctgaag tgtctcattt tattttattt attctttgct gataaaaaaa 7920
taaaataaaa gaagctaagc acacggtcaa ccattgctct actgctaaaa gggttatgtg 7980
tagtgtttta ctgcataaat tatgcagcaa acaagacaac tcaaattaaa aaatttcctt 8040
tgcttgtttt tttgttgtct ctgacttgac tttcttgtgg aagttggttg tataaggatt 8100
gggacaccat tgtccttctt aatttaattt tattctttgc tgataaaaaa aaaaatttca 8160
tatagtgtta aataataatt tgttaaataa ccaaaaagtc aaatatgttt actctcgttt 8220
aaataattga gattcgttcc agcaaggcta aacgattgta tagatttatg acaatattta 8280
cttttttata gataaatgtt atattataat aaatttatat acatatatta tatgttattt 8340
attattattt taaatccttc aatattttat caaaccaact cataattttt tttttatctg 8400
taagaagcaa taaaattaaa tagacccact ttaaggatga tccaaccttt atacagagta 8460
agagagttca aatagtacct tttcatatac atatcaacta aaatattaga aatatcatgg 8520
atcaaacctt ataaagacat taaataagtg gataagtata atatataaat gggtagtata 8580
taatatataa atggatacaa acttctctct ttataattgt tatgtctcct taacatccta 8640
atataataca taagtgggta atatataata tataaatgga gacaaacttc ttccattata 8700
attgttatgt cttcttaaca cttatgtctc gttcacaatg ctaaggttag aattgtttag 8760
aaagtcttat agtacacatt tgtttttgta ctatttgaag cattccataa gccgtcacga 8820
ttcagatgat ttataataat aagaggaaat ttatcataga acaataaggt gcatagatag 8880
agtgttaata tatcataaca tcctttgttt attcatagaa gaagtgagat ggagctcagt 8940
tattatactg ttacatggtc ggatacaata ttccatgctc tccatgagct cttacaccta 9000
catgcatttt agttcatact tgcggccgct cagaattcct gggcatcctg aattgtgtgg 9060
gccacgccca ccttctgcat gtagcggagc gtggacatta tattctccag gatggtcggg 9120 tagtatgtgt agcgcacgcc atggcgcttg caacattccc gcaccgtatc ttggatgaag 9180 gggtagtgcc aagaggacat gcgggggaac aagtggtgct cgatctgcag gttcaaaccg 9240 ccggtcagca tcatcccgac gctgcccccg taggtcgccg cggtctcggc ctgcgcccgg 9300 taccagtcca cccggtcctc ctctacatcg cggacctcct tcccctcctt ggcccttttg 9360 gccttgcagt tcttttcggg ggtccgctca gcgccttcaa agttatggga cagcacaaaa 9420 acaaatgtca agaagatgcc tgtgaccgtt ggcaccagca ggatggtgag cagcggagag 9480 gcgcccgcga tgtaccaggg gaggaaggcg ttgcggaaga tgaacagtag gcggagggaa 9540 acggcgaagg cgcgctgccg gttgaaaaag ctgcctggtc taagagaggt ggcctctggg 9600 attgtgtcgt tgtgctgcat cgtgaacaag tacatcggat tgtacaccat cgacacacca 9660 tacatcccca aaacgatgta catgtaccac gcctggaacc gatggtacca ctttcgtgac 9720 gcatttgttg ccgggtagga gtggaacaaa atgaacggct ccgccgaggc agcgtccagg 9780 tccaggccat gctggttggt gtaggtgtgg tggcgcataa tgtgggactg cagccagatc 9840 cagcgggagg atccaatggc gtcgatgccg taggcgaaaa aggcgttcac ccagggcttt 9900 ttgctgatgg ctccgtgaga ggcatcgtgt tgaatcgcca acccaatctg gctgtggaag 9960 tacccggtga agatgcccca catcactgcc ccggtagtga tccaatacca ttcgcctatg 10020 gcagtgcacg cgatgatcag cgcggtccgc agccaaaagc cgggcgtggc gtaccagttg 10080 cgccccttca tgacctcccg aacgctctgc ttgacatcct gggcgaaggg actgtcaaag 10140 gtgtacactg ggtccttcga gcttcgttcg tgtttgcctc gggcaaattg ctgcagcagc 10200 ttcgaattct ctggcttcac tccagggtgc atggagtaga acagggcggt tccgtcggag 10260 gcaccggcgg cttcaatgag gttgccacct ggatgcactt tcgccagtcc gaagagatcg 10320 tacaacaccc catcaattgc aaccagttct gggtgttcta aaagttgctc agtagtcaaa 10380 gagatggtgg ccatggtgcg gccgcagtat atcttaaatt ctttaatacg gtgtactagg 10440 atattgaact ggttcttgat gatgaaaacc tgggccgaga ttgcagctat ttatagtcat 10500 aggtcttgtt aacatgcatg gacatttggc cacggggtgg catgcagttt gacgggtgtt 10560 gaaataaaca aaaatgaggt ggcggaagag aatacgagtt tgaggttggg ttagaaacaa 10620 caaatgtgag ggctcatgat gggttgagtt ggtgaatgtt ttgggctgct cgattgacac 10680 ctttgtgagt acgtgttgtt gtgcatggct tttggggtcc agtttttttt tcttgacgcg 10740 gcgatcctga tcagctagtg gataagtgat gtccactgtg tgtgattgcg tttttgtttg 10800 aattttatga acttagacat tgctatgcaa aggatactct cattgtgttt tgtcttcttt 10860 tgttccttgg ctttttctta tgatccaaga gactagtcag tgttgtggca ttcgagacta 10920 ccaagattaa ttatgatggg ggaaggataa gtaactgatt agtacggact gttaccaaat 10980 taattaataa gcggcaaatg aagggcatgg ate 11013
<210> 170
<211> 21767
<212> DNA
<213> Seqüência Artificial <220>
<223> plasmideo pKR1193
<400> 170
cgcgccagat cctctagagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata 60 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 120 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 180 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 240 acgcgcgggg agaggcggtt tgcgtattgg atcgatccct gaaagcgacg ttggatgtta 300 acatctacaa attgcctttt cttatcgacc atgtacgtaa gcgcttacgt ttttggtgga 360 cccttgagga aactggtagc tgttgtgggc ctgtggtctc aagatggatc attaatttcc 420 accttcacct acgatggggg gcatcgcacc ggtgagtaat attgtacggc taagagcgaa 480 tttggcctgt agacctcaat tgcgagcttt ctaatttcaa actattcggg cctaactttt 540 ggtgtgatga tgctgactgg caggatatat accgttgtaa tttgagctcg tgtgaataag 600 tcgctgtgta tgtttgtttg attgtttctg ttggagtgca gcccatttca ccggacaagt 660 cggctagatt gatttagccc tgatgaactg ccgaggggaa gccatcttga gcgcggaatg 720 ggaatggatt tcgttgtaca acgagacgac agaacaccca cgggaccgag cttcgcgagc 780 ttttgtatcc gtggcatcct tggtccgggc gatttgttca cgtccatgag gcgctctcca 840 aaggaacgca tattttccgg tgcaaccttt ccggttcttc ctctactcga cctcttgaag 900 tcccagcatg aatgttcgac cgctccgcaa gcggatcttt ggcgcaacca gccggtttcg 960 cacgtcgatt ctcgcgagcc tgcatacttt ggcaagattg ctgaatgacg ctgatgcttc 1020 atcgcaatct gcgataatgg ggtaagtatc cggtgaaggc cgcaggtcag gccgcctgag 1080 cactcagtgt cttggatgtc cagttccacg gcagctgttg ctcaagcctg ctgatcggag 1140 cgtccgcaag gtcggcgcgg acgtcggcaa gccaggcctg cggatcgatg ttattgagct 1200 tggcgctcat gatcagtgtc gccatgaacg ccgcacgttc agcacaacga tccgatccgg 1260 caaacagcca tgacttcctg ccgagtacat agcctctgag cgttcgttcg gcagcattgt 1320 tcgtcaggca aatcgggccg tcatcgagga atgacgtaat gccatcccat cgcttgagca 1380 tgtaatttat cgcctcggcg acgggagaac tgcgcgacaa tttcccccgc tcggtttcga 1440 gccaatcatg cagctcttcg gcgagtgacc ttgatcaggc caccgccacg accgcggaag 1500 acgaacagat gcctgcgcat cggatcgcgc ttcagcgtct cttgcaccat cagcgacaaa 1560
ccgggaaagc ctttgcgcat gtccgtactt atgtcgccac ttgggagggc ttcgtctacg 1620
tggccttcgt gatcgacgtc ttcgcccgtc gcattgtcgg atggcgggcg agccggacag 1680
cacatgcagg ctttgtcctc gatgccctcg aggaggctca tcatgatcgg cgtcccgctc 1740
atggcggcct agtgcatcac tcggatcgcg gtgttcaata cgtgtccttt cgctattccg 1800
agcggttggc agaagcaggt atcgagccat ctatcggaag cgtcggcgac agcacgacaa 1860
cgccctcgca gaagcgatca acggtcttta caaggccgag gtcattcatc ggcgtggacc 1920
atggaggagc ttcgaagcgg tcgagttcgc taccttggaa tggatagact ggttcaacca 1980
cggcggcttt tgaagcccat cggcaatata ccgccagccg aagacgagga tcagtattac 2040
gccatgctgg acgaagcagc catggctgcg cattttaacg aaatggcctc cggcaaaccc 2100
ggtgcggttc acttgttgcg tgggaaagtt cacgggactc cgcgcacgag ccttcttcgt 2160
aatagccata tcgaccgaat tgacctgcag gggggggggg gaaagccacg ttgtgtctca 2220
aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc 2280
tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg 2340
ctcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg 2400
cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc 2460
agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt 2520
cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac 2580
tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt 2640
agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg 2700
gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc 2760
tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg 2820
taatggctgg cctgttgaac aagtctggaa agaaatgcat aagcttttgc cattctcacc 2880
ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa 2940
attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc 3000
catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa 3060
atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt 3120
tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg 3180
ggacggcggc tttgttgaat aaatcgaact tttgctgagt tgaaggatca gatcacgcat 3240
cttcccgaca acgcagaccg ttccgtggca aagcaaaagt tcaaaatcac caactggtcc 3300
acctacaaca aagctctcat caaccgtggc tccctcactt tctggctgga tgatggggcg 3360 attcaggcct ggtatgagtc agcaacacct tcttcacgag gcagacctca gcgccccccc 3420
ccccctgcag gtcttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat 3480
tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg 3540
acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag 3600
aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa 3660
cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc 3720
gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca 3780
cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc 3840
tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc 3900
tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg 3960
ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta 4020
tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag 4080
gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga 4140
ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc 4200
tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa 4260
agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 4320
aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 4380
cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt 4440
agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 4500
tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 4560
gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 4 620
gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 4 680
ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 4740
gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 4800
ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 4860
ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc 4920
acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt 4980
gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag 5040
cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca 5100
tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc 5160
gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc 5220 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg 5280
gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agggtgcctt 5340
gatgtgggcg ccggcggtcg agtggcgacg gcgcggcttg tccgcgccct ggtagattgc 5400
ctggccgtag gccagccatt tttgagcggc cagcggccgc gataggccga cgcgaagcgg 5460
cggggcgtag ggagcgcagc gaccgaaggg taggcgcttt ttgcagctct tcggctgtgc 5520
gctggccaga cagttatgca caggccaggc gggttttaag agttttaata agttttaaag 5580
agttttaggc ggaaaaatcg ccttttttct cttttatatc agtcacttac atgtgtgacc 5640
ggttcccaat gtacggcttt gggttcccaa tgtacgggtt ccggttccca atgtacggct 5700
ttgggttccc aatgtacgtg ctatccacag gaaagagacc ttttcgacct ttttcccctg 5760
ctagggcaat ttgccctagc atctgctccg tacattagga accggcggat gcttcgccct 5820
cgatcaggtt gcggtagcgc atgactagga tcgggccagc ctgccccgcc tcctccttca 5880
aatcgtactc cggcaggtca tttgacccga tcagcttgcg cacggtgaaa cagaacttct 5940
tgaactctcc ggcgctgcca ctgcgttcgt agatcgtctt gaacaaccat ctggcttctg 6000
ccttgcctgc ggcgcggcgt gccaggcggt agagaaaacg gccgatgccg ggatcgatca 6060
aaaagtaatc ggggtgaacc gtcagcacgt ccgggttctt gccttctgtg atctcgcggt 6120
acatccaatc agctagctcg atctcgatgt actccggccg cccggtttcg ctctttacga 6180
tcttgtagcg gctaatcaag gcttcaccct cggataccgt caccaggcgg ccgttcttgg 6240
ccttcttcgt acgctgcatg gcaacgtgcg tggtgtttaa ccgaatgcag gtttctacca 6300
ggtcgtcttt ctgctttccg ccatcggctc gccggcagaa cttgagtacg tccgcaacgt 6360
gtggacggaa cacgcggccg ggcttgtctc ccttcccttc ccggtatcgg ttcatggatt 6420
cggttagatg ggaaaccgcc atcagtacca ggtcgtaatc ccacacactg gccatgccgg 6480
ccggccctgc ggaaacctct acgtgcccgt ctggaagctc gtagcggatc acctcgccag 6540
ctcgtcggtc acgcttcgac agacggaaaa cggccacgtc catgatgctg cgactatcgc 6600
gggtgcccac gtcatagagc atcggaacga aaaaatctgg ttgctcgtcg cccttgggcg 6660
gcttcctaat cgacggcgca ccggctgccg gcggttgccg ggattctttg cggattcgat 6720
cagcggccgc ttgccacgat tcaccggggc gtgcttctgc ctcgatgcgt tgccgctggg 6780
cggcctgcgc ggccttcaac ttctccacca ggtcatcacc cagcgccgcg ccgatttgta 6840
ccgggccgga tggtttgcga ccgctcacgc cgattcctcg ggcttggggg ttccagtgcc 6900
attgcagggc cggcagacaa cccagccgct tacgcctggc caaccgcccg ttcctccaca 6960
catggggcat tccacggcgt cggtgcctgg ttgttcttga ttttccatgc cgcctccttt 7020
agccgctaaa attcatctac tcatttattc atttgctcat ttactctggt agctgcgcga 7080 tgtattcaga tagcagctcg gtaatggtct tgccttggcg taccgcgtac atcttcagct 7140 tggtgtgatc ctccgccggc aactgaaagt tgacccgctt catggctggc gtgtctgcca 7200 ggctggccaa cgttgcagcc ttgctgctgc gtgcgctcgg acggccggca cttagcgtgt 7260 ttgtgctttt gctcattttc tctttacctc attaactcaa atgagttttg atttaatttc 7320 agcggccagc gcctggacct cgcgggcagc gtcgccctcg ggttctgatt caagaacggt 7380 tgtgccggcg gcggcagtgc ctgggtagct cacgcgctgc gtgatacggg actcaagaat 7 4 4 0 gggcagctcg tacbcggcca gcgcctcggc aacctcaccg ccgatgcgcg tgcctttgat 7500 cgcccgcgac acgacaaagg ccgcttgtag ccttccatcc gtgacctcaa tgcgctgctt 7560 aaccagctcc accaggtcgg cggtggccca tatgtcgtaa gggcttggct gcaccggaat 7620 cagcacgaag tcggctgcct tgatcgcgga cacagccaag tccgccgcct ggggcgctcc 7680 gtcgatcact acgaagtcgc gccggccgat ggccttcacg tcgcggtcaa tcgtcgggcg 7740 gtcgatgccg acaacggtta gcggttgatc ttcccgcacg gccgcccaat cgcgggcact 7800 gccctgggga tcggaatcga ctaacagaac atcggccccg gcgagttgca gggcgcgggc 7860 tagatgggtt gcgatggtcg tcttgcctga cccgcctttc tggttaagta cagcgataac 7920 ttcatgcgtt cccttgcgta tttgtttatt tactcatcgc atcatatacg cagcgaccgc 7980 atgacgcaag ctgttttact caaatacaca tcaccttttt agacggcggc gctcggtttc 8040 ttcagcggcc aagctggccg gccaggccgc cagcttggca tcagacaaac cggccaggat 8100 ttcatgcagc cgcacggttg agacgtgcgc gggcggctcg aacacgtacc cggccgcgat 8160 catctccgcc tcgatctctt cggtaatgaa aaacggttcg tcctggccgt cctggtgcgg 8220 tttcatgctt gttcctcttg gcgttcattc tcggcggccg ccagggcgtc ggcctcggtc 8280 aatgcgtcct cacggaaggc accgcgccgc ctggcctcgg tgggcgtcac ttcctcgctg 8340 cgctcaagtg cgcggtacag ggtcgagcga tgcacgccaa gcagtgcagc cgcctctttc 8400 acggtgcggc cttcctggtc gatcagctcg cgggcgtgcg cgatctgtgc cggggtgagg 8460 gtagggcggg ggccaaactt cacgcctcgg gccttggcgg cctcgcgccc gctccgggtg 8520 cggtcgatga ttagggaacg ctcgaactcg gcaatgccgg cgaacacggt caacaccatg 8580 cggccggccg gcgtggtggt gtcggcccac ggctctgcca ggctacgcag gcccgcgccg 8640 gcctcctgga tgcgctcggc aatgtccagt aggtcgcggg tgctgcgggc caggcggtct 8700 agcctggtca ctgtcacaac gtcgccaggg cgtaggtggt caagcatcct ggccagctcc 8760 gggcggtcgc gcctggtgcc ggtgatcttc tcggaaaaca gcttggtgca gccggccgcg 8820 tgcagttcgg cccgttggtt ggtcaagtcc tggtcgtcgg tgctgacgcg ggcatagccc 8880 agcaggccag cggcggcgct cttgttcatg gcgtaatgtc tccggttcta gtcgcaagta 8940 ttctacttta tgcgactaaa acacgcgaca agaaaacgcc aggaaaaggg cagggcggca 9000 gcctgtcgcg taacttagga cttgtgcgac atgtcgtttt cagaagacgg ctgcactgaa 9060 cgtcagaagc cgactgcact atagcagcgg aggggttgga ccacaggacg ggtgtggtcg 9120 ccatgatcgc gtagtcgata gtggctccaa gtagcgaagc gagcaggact gggcggcggc 9180 caaagcggtc ggacagtgct ccgagaacgg gtgcgcatag aaattgcatc aacgcatata 9240 gcgctagcag cacgccatag tgactggcga tgctgtcgga atggacgata tcccgcaaga 9300 ggcccggcag taccggcata accaagccta tgcctacagc atccagggtg acggtgccga 9360 ggatgacgat gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta 9420 actgtgataa actaccgcat taaagctagc ttgcttggtc gttccgcgtg aacgtcggct 9480 cgattgtacc tgcgttcaaa tactttgcga tcgtgttgcg cgcctgcccg gtgcgtcggc 9540 tgatctcacg gatcgactgc ttctctcgca acgccatccg acggatgatg tttaaaagtc 9600 ccatgtggat cactccgttg ccccgtcgct caccgtgttg gggggaaggt gcacatggct 9660 cagttctcaa tggaaattat ctgcctaacc ggctcagttc tgcgtagaaa ccaacatgca 9720 agctccaccg ggtgcaaagc ggcagcggcg gcaggatata ttcaattgta aatggcttca 9780 tgtccgggaa atctacatgg atcagcaatg agtatgatgg tcaatatgga gaaaaagaaa 9840 gagtaattac caattttttt tcaattcaaa aatgtagatg tccgcagcgt tattataaaa 9900 tgaaagtaca ttttgataaa acgacaaatt acgatccgtc gtatttatag gcgaaagcaa 9960 taaacaaatt attctaattc ggaaatcttt atttcgacgt gtctacattc acgtccaaat 10020 gggggcttag atgagaaact tcacgatcga tgccttgatt tcgccattcc cagataccca 10080 tttcatcttc agattggtct gagattatgc gaaaatatac actcatatac ataaatactg 10140 acagtttgag ctaccaattc agtgtagccc attacctcac ataattcact caaatgctag 10200 gcagtctgtc aactcggcgt caatttgtcg gccactatac gatagttgcg caaattttca 10260 aagtcctggc ctaacatcac acctctgtcg gcggcgggtc ccatttgtga taaatccacc 10320 atatcgaatt aattcagact cctttgcccc agagatcaca atggacgact tcctctatct 10380 ctacgatcta gtcaggaagt tcgacggaga aggtgacgat accatgttca ccactgataa 10440 tgagaagatt agccttttca atttcagaaa gaatgctaac ccacagatgg ttagagaggc 10500 ttacgcagca ggtctcatca agacgatcta cccgagcaat aatctccagg agatcaaata 10560 ccttcccaag aaggttaaag atgcagtcaa aagattcagg actaactgca tcaagaacac 10620 agagaaagat atatttctca agatcagaag tactattcca gtatggacga ttcaaggctt 10680 gcttcacaaa ccaaggcaag taatagagat tggagtctct aaaaaggtag ttcccactga 10740 atcaaaggcc atggagtcaa agattcaaat agaggaccta acagaactcg ccgtaaagac 10800 tggcgaacag ttcatacaga gtctcttacg actcaatgac aagaagaaaa tcttcgtcaa 10860
catggtggag cacgacacgc ttgtctactc caaaaatatc aaagatacag tctcagaaga 10920
ccaaagggca attgagactt ttcaacaaag ggtaatatcc ggaaacctcc tcggattcca 10980
ttgcccagct atctgtcact ttattgtgaa gatagtggaa aaggaaggtg gctcctacaa 11040
atgccatcat tgcgataaag gaaaggccat cgttgaagat gcctctgccg acagtggtcc 11100
caaagatgga cccccaccca cgaggagcat cgtggaaaaa gaagacgttc caaccacgtc 11160
ttcaaagcaa gtggattgat gtgatatctc cactgacgta agggatgacg cacaatccca 11220
ctatccttcg caagaccctt cctctatata aggaagttca tttcatttgg agaggacacg 11280
ctgaaatcac cagtctccaa gcttgcgggg atcgtttcgc atgattgaac aagatggatt 11340
gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca 11400
gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct 11460
ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct 11520
atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc 11580
gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct 11640
tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga 11700
tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg 11760
gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc 11820
agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac 11880
ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat 11940
cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga 12000
tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc 12060
cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg 12120
actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat 12180
tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg 12240
atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccgg atcgatccaa 12300
cacttacgtt tgcaacgtcc aagagcaaat agaccacgaa cgccggaagg ttgccgcagc 12360
gtgtggattg cgtctcaatt ctctcttgca ggaatgcaat gatgaatatg atactgacta 12420
tgaaactttg agggaatact gcctagcacc gtcacctcat aacgtgcatc atgcatgccc 12480
tgacaacatg gaacatcgct atttttctga agaattatgc tcgttggagg atgtcgcggc 12540
aattgcagct attgccaaca tcgaactacc cctcacgcat gcattcatca atattattca 12600
tgcggggaaa ggcaagatta atccaactgg caaatcatcc agcgtgattg gtaacttcag 12660 ttccagcgac ttgattcgtt ttggtgctac ccacgttttc aataaggacg agatggtgga 12720 gtaaagaagg agtgcgtcga agcagatcgt tcaaacattt ggcaataaag tttcttaaga 12780 ttgaatcctg ttgccggtct tgcgatgatt atcatataat ttctgttgaa ttacgttaag 12840 catgtaataa ttaacatgta atgcatgacg ttatttatga gatgggtttt tatgattaga 12900 gtcccgcaat tatacattta atacgcgata gaaaacaaaa tatagcgcgc aaactaggat 12960 aaattatcgc gcgcggtgtc atctatgtta ctagatcgat caaacttcgg tactgtgtaa 13020 tgacgatgag caatcgagag gctgactaac aaaaggtaca tcgcgatgga tcgatccatt 13080 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac 13140 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt 13200 cccagtcacg acgttgtaaa acgacggcca gtgaattcct gcagcccggg ggatccgccc 13260 actcgaggcg cgccgtcgac ggatccgtac gttgaaacat ccctgaagtg tctcatttta 13320 ttttatttat tctttgctga taaaaaaata aaataaaaga agctaagcac acggtcaacc 13380 attgctctac tgctaaaagg gttatgtgta gtgttttact gcataaatta tgcagcaaac 13440 aagacaactc aaattaaaaa atttcctttg cttgtttttt tgttgtctct gacttgactt 13500 tcttgtggaa gttggttgta taaggattgg gacaccattg tccttcttaa tttaatttta 13560 ttctttgctg ataaaaaaaa aaatttcata tagtgttaaa taataatttg ttaaataacc 13620 aaaaagtcaa atatgtttac tctcgtttaa ataattgaga ttcgttccag caaggctaaa 13680 cgattgtata gatttatgac aatatttact tttttataga taaatgttat attataataa 13740 atttatatac atatattata tgttatttat tattatttta aatccttcaa tattttatca 13800 aaccaactca taattttttt tttatctgta agaagcaata aaattaaata gacccacttt 13860 aaggatgatc caacctttat acagagtaag agagttcaaa tagtaccttt tcatatacat 13920 atcaactaaa atattagaaa tatcatggat caaaccttat aaagacatta aataagtgga 13980 taagtataat atataaatgg gtagtatata atatataaat ggatacaaac ttctctcttt 14040 ataattgtta tgtctcctta acatcctaat ataatacata agtgggtaat atataatata 14100 taaatggaga caaacttctt ccattataat tgttatgtct tcttaacact tatgtctcgt 14160 tcacaatgct aaggttagaa ttgtttagaa agtcttatag tacacatttg tttttgtact 14220 atttgaagca ttccataagc cgtcacgatt cagatgattt ataataataa gaggaaattt 14280 atcatagaac aataaggtgc atagatagag tgttaatata tcataacatc ctttgtttat 14340 tcatagaaga agtgagatgg agctcagtta ttatactgtt acatggtcgg atacaatatt 14400 ccatgctctc catgagctct tacacctaca tgcattttag ttcatacttg cggccgctca 14460 gaattcctgg gcatcctgaa ttgtgtgggc cacgcccacc ttctgcatgt agcggagcgt 14520 ggacattata ttctccagga tggtcgggta gtatgtgtag cgcacgccat ggcgcttgca 14580
acattcccgc accgtatctt ggatgaaggg gtagtgccaa gaggacatgc gggggaacaa 14640
gtggtgctcg atctgcaggt tcaaaccgcc ggtcagcatc atcccgacgc tgcccccgta 14700
ggtcgccgcg gtctcggcct gcgcccggta ccagtccacc cggtcctcct ctacatcgcg 14760
gacctccttc ccctccttgg cccttttggc cttgcagttc ttttcggggg tccgctcagc 14820
gccttcaaag ttatgggaca gcacaaaaac aaatgtcaag aagatgcctg tgaccgttgg 14880
caccagcagg atggtgagca gcggagaggc gcccgcgatg taccagggga ggaaggcgtt 14940
gcggaagatg aacagtaggc ggagggaaac ggcgaaggcg cgctgccggt tgaaaaagct 15000
gcctggtcta agagaggtgg cctctgggat tgtgtcgttg tgctgcatcg tgaacaagta 15060
catcggattg tacaccatcg acacaccata catccccaaa acgatgtaca tgtaccacgc 15120
ctggaaccga tggtaccact ttcgtgacgc atttgttgcc gggtaggagt ggaacaaaat 15180
gaacggctcc gccgaggcag cgtccaggtc caggccatgc tggttggtgt aggtgtggtg 15240
gcgcataatg tgggactgca gccagatcca gcgggaggat ccaatggcgt cgatgccgta 15300
ggcgaaaaag gcgttcaccc agggcttttt gctgatggct ccgtgagagg catcgtgttg 15360
aatcgccaac ccaatctggc tgtggaagta cccggtgaag atgccccaca tcactgcccc 15420
ggtagtgatc caataccatt cgcctatggc agtgcacgcg atgatcagcg cggtccgcag 15480
ccaaaagccg ggcgtggcgt accagttgcg ccccttcatg acctcccgaa cgctctgctt 15540
gacatcctgg gcgaagggac tgtcaaaggt gtacactggg tccttcgagc ttcgttcgtg 15600
tttgcctcgg gcaaattgct gcagcagctt cgaattctct ggcttcactc cagggtgcat 15660
ggagtagaac agggcggttc cgtcggaggc accggcggct tcaatgaggt tgccacctgg 15720
atgcactttc gccagtccga agagatcgta caacacccca tcaattgcaa ccagttctgg 15780
gtgttctaaa agttgctcag tagtcaaaga gatggtggcc atggtgcggc cgcagtatat 15840
cttaaattct ttaatacggt gtactaggat attgaactgg ttcttgatga tgaaaacctg 15900
ggccgagatt gcagctattt atagtcatag gtcttgttaa catgcatgga catttggcca 15960
cggggtggca tgcagtttga cgggtgttga aataaacaaa aatgaggtgg cggaagagaa 16020
tacgagtttg aggttgggtt agaaacaaca aatgtgaggg ctcatgatgg gttgagttgg 16080
tgaatgtttt gggctgctcg attgacacct ttgtgagtac gtgttgttgt gcatggcttt 16140
tggggtccag tttttttttc ttgacgcggc gatcctgatc agctagtgga taagtgatgt 16200
ccactgtgtg tgattgcgtt tttgtttgaa ttttatgaac ttagacattg ctatgcaaag 16260
gatactctca ttgtgttttg tcttcttttg ttccttggct ttttcttatg atccaagaga 16320
ctagtcagtg ttgtggcatt cgagactacc aagattaatt atgatggggg aaggataagt 16380 aactgattag tacggactgt taccaaatta attaataagc ggcaaatgaa gggcatggat 16440 cgtacgtgaa ggttaaacat ggtgaatatg ttaccactag ctgggatgcc cattagatca 16500 aaactgtaaa attctcccgt ttcccttcta ttcacatgtg agccccctcc cttttctttc 16560 tttctcaatt ttgattgagt taaagtcacc agcaatgcat cactcaccct ccaaaaaatt 16620 tcttgtacaa cttctcggac tatcccaaag ctccttttcc tgagatggat ggtcctgtct 16680 cttgcccttg atgtcttcct tgttcgattt tggcttcctc taatgtcttt cttgctagga 16740 atcaccacct cactcatcta tgttgtcgta gcttctgaaa gtctcataca tatccttagt 16800 gttgcactca tcttgtattg aagtgaaaaa gaatgttgtt ctcctatcca aatctccatt 16860 gaatctcttt ctcccaatgt tgtcccatcg gttggtcctc ctctccaacc aattgtaagg 16920 tgtttaacat aaacatggta caattaagat ttttcatttc attaagaaaa gattgagatt 16980 tgtggttcta aagtttcaat tagagtttga tgatattgaa acaaccgtag aacacattaa 17040 gtattactaa cttatacata gagcattgga atttcacctt ttatttattc tgtttccgcc 17100 aaaggtacat gactcaagtt attttacaca agtaacaaag gcatctaagc ctaagtattc 17160 ttattcagac ttttcattat tactttcatt gatttggtgc gaaatgcggc cgcttacgcc 17220 ttgtcggcct ttgccaccat gcgggcaaag gtgcccaggt agctgatcag gatgcccacc 17280 ccctccagca ttggggggct gcgatacggc aagttgtgct tcttgcacaa ctgctccact 17340 ttgatgctgg ccgccgtcag gttgtgccgg ggcagcgtcg gccacaggtg gtgctcgatt 17400 tggtaattca gcccaccgaa gaaccagtcc gtgacgagtc cccgctggac gttcatcgtt 17460 tcgtgaatct ggccggcgca aaagccgtgg ccgtcccaca ccgagtcctg gatcttctcc 17520 agggggtagt ggttcatgaa caccacgatg gcgatgccga agcccccaag caactcggac 17580 acgaaaaaca ccatgagtcc ggtcaagaag cttggcatat aaaagtagta gaacaacgcc 17640 ttcaggcccc agtggagggc caggcccacg ctctctttct cgtactgcct gcggtagtac 17700 tggttgctgc gatccttcag gcccgtggct gtgtggatgc tctggaagca ccagatgaac 17760 ctcaggaggg cacagatgaa gaagaagtag tattgctggt acttgatcat ccgccgtgag 17820 aacgggccgg ccctctccac gtcctccttg gaccatgcca gcagcggcag gttgtcaatg 17880 tcggggtcgt ggccttgcac gttggtggcg gagtggtgtg cattgtgcct gtccttccac 17940 caggtcacag agaacccttg caagacgttc ccgaaaagca agccgatggc gttgttgatc 18000 gatcggtcct tgaacagctg atggtggcag atatcgtgcg acaaccaacc catttgttgg 18060 aagtgaatgc ccaacacgat tgccccgacg aggtaccagt gccactgggt catcaatagc 18120 accgcgagga ccccgaggcc cagcgtactg agcgtcttat atgcgtacca catcggtgat 18180 gcgtcgaaca ttcctgctgc gatgagctca tctcggagct tgcggaaatc ctcctgcggt 18240 tcgtctgagt tgggtttcgg tggcgtaggc gtcagtggag atgatggttc catgataggc 18300 atccttctta gtttactcac agcattttca gagtgcataa ccataaagac atcagtggca 18360 tcacgaccgc ggtagttctc aatgatgtca gcaccccctg gatgatggtt caaccaggca 18420
gacacatcat aggtgacacc atcaacggtc agcggaagtg ctggcctttt caccatggtg 18480
cggccgcggt gatgactgat gagtgtttaa ggaccaatgg agagaatgtt tgagttgtga 18540
agcggagaac ctgaggcgtg gttatttata gggaagagag gaaggtgaat gagggacacg 18 600
tcacagaagt agggtgctga gcttgagaca ttcttcagta tgcatggcta tggaagcctt 18660
gggtgctaca cctcatgaag ttcatggtgt gaggtggctt cggcatctca attaagtgac 18720
aaagagaaag gtgtttcagt gtttctattg caaatggcag aaactcgtga tgacgagggg 18780
accatgcatg gtttcatttc ttttcttcct ggattctttc tttcctttta tatatgcagg 18840
ttcataattt aaaaattaga ctcgctttca atttcttaat ttctcatttt cctcttatat 18900
tactgtacta atgttaacca cgtacactta tttttttttt agtttaattt tgatagattg 18960
tgttgattta aacatattaa tattttcaac caaataaaaa tcattttagt agatacggct 19020
ttttaaataa ttattaaaaa tattaactat ttatcctaaa tggcacattt taattaaaaa 19080
aaatccggtg ttgtaagtgt tttattaatt tgttttggca ttattaaagc aacttttttt 19140
ttatttgttg gcattttgag tacgtactta ggctagcctg caggagatcc aagcttttga 19200
tccatgccct tcatttgccg cttattaatt aatttggtaa cagtccgtac taatcagtta 19260
cttatccttc ccccatcata attaatcttg gtagtctcga atgccacaac actgactagt 19320
ctcttggatc ataagaaaaa gccaaggaac aaaagaagac aaaacacaat gagagtatcc 19380
tttgcatagc aatgtctaag ttcataaaat tcaaacaaaa acgcaatcac acacagtgga 19440
catcacttat ccactagctg atcaggatcg ccgcgtcaag aaaaaaaaac tggaccccaa 19500
aagccatgca caacaacacg tactcacaaa ggtgtcaatc gagcagccca aaacattcac 19560
caactcaacc catcatgagc cctcacattt gttgtttcta acccaacctc aaactcgtat 19620
tctcttccgc cacctcattt ttgtttattt caacacccgt caaactgcat gccaccccgt 19680
ggccaaatgt ccatgcatgt taacaagacc tatgactata aatagctgca atctcggccc 19740
aggttttcat catcaagaac cagttcaata tcctagtaca ccgtattaaa gaatttaaga 19800
tatactgcgg ccgcaccatg gaagcagcca aagaattggt ttccatcgtc caagaggagc 19860
tccccaaggt ggactatgcc cagctttggc aggatgccag cagctgtgag gtcctttacc 19920
tctcggtggc attcgtggcg atcaagttca tgctgcgccc actggacctg aagcgccagg 19980
ccaccttgaa gaagctgttc acagcataca acttcctcat gtcgatctat tcctttggct 20040
ccttcctggc catggcctat gccctatcag taactggcat cctctccggc gactgtgaga 20100 cggcgttcaa caacgatgtg ttcaggatca caactcagct gttctacctc agcaagttcg 20160 tagagtacat cgactccttc taccttcccc ttatggacaa gccactgtcg ttccttcagt 20220 tcttccatca tttgggggcc cccattgaca tgtggctatt ctacaaatac cgcaacgaag 20280 gagtctggat ctttgtcctg ttgaatgggt tcattcactg gatcatgtac ggttactatt 20340 ggacgcggct catcaagctg aacttcccca tgcccaagaa cctgatcacc tccatgcaga 20400 tcatccagtt caatgtcggg ttctacatcg tctggaagta ccgcaatgtg ccatgctacc 20460 gccaggatgg gatgcgcatg tttgcctgga tcttcaacta ctggtatgtc gggacggtct 20520 tgctgctgtt cctcaacttt tacgtgcaga cgtacatccg gaagccgagg aagaaccgag 20580 ggaagaagga gtagcggccg caagtatgaa ctaaaatgca tgtaggtgta agagctcatg 20640 gagagcatgg aatattgtat ccgaccatgt aacagtataa taactgagct ccatctcact 20700 tcttctatga ataaacaaag gatgttatga tatattaaca ctctatctat gcaccttatt 20760 gttctatgat aaatttcctc ttattattat aaatcatctg aatcgtgacg gcttatggaa 20820 tgcttcaaat agtacaaaaa caaatgtgta ctataagact ttctaaacaa ttctaacctt 20880 agcattgtga acgagacata agtgttaaga agacataaca attataatgg aagaagtttg 20940 tctccattta tatattatat attacccact tatgtattat attaggatgt taaggagaca 21000 taacaattat aaagagagaa gtttgtatcc atttatatat tatatactac ccatttatat 21060 attatactta tccacttatt taatgtcttt ataaggtttg atccatgata tttctaatat 21120 tttagttgat atgtatatga aagggtacta tttgaactct cttactctgt ataaaggttg 21180 gatcatcctt aaagtgggtc tatttaattt tattgcttct tacagataaa aaaaaaatta 21240 tgagttggtt tgataaaata ttgaaggatt taaaataata ataaataaca tataatatat 21300 gtatataaat ttattataat ataacattta tctataaaaa agtaaatatt gtcataaatc 21360 tatacaatcg tttagccttg ctggacgaat ctcaattatt taaacgagag taaacatatt 21420 tgactttttg gttatttaac aaattattat ttaacactat atgaaatttt tttttttatc 21480 agcaaagaat aaaattaaat taagaaggac aatggtgtcc caatccttat acaaccaact 21540 tccacaagaa agtcaagtca gagacaacaa aaaaacaagc aaaggaaatt ttttaatttg 21600 agttgtcttg tttgctgcat aatttatgca gtaaaacact acacataacc cttttagcag 21660 tagagcaatg gttgaccgtg tgcttagctt cttttatttt atttttttat cagcaaagaa 21720 taaataaaat aaaatgagac acttcaggga tgtttcaaca agcttgg 21767
<210> 171
<211> 20
<212> DNA
<213> Seqüência Artificial <220>
<223> primer SeqE
<400> 171
cgacacactc caatctttcc
<210> 172
<211> 20
<212> DNA
<213> Seqüência Artificial
<220>
<223> primer SeqW
<400> 172
ggtggctgga gttagacatc

Claims (46)

1. PLANTA OLEAGINOSA TRANSGÊNICA, que produz sementes maduras em que o perfil total de ácidos graxos da semente compreende pelo menos 10,0% de ácido araquidônico e menos de 5% de ácidos graxos ômega 3 totais que contêm pelo menos dezoito átomos de carbono e pelo menos quatro uniões duplas.
2. PLANTA OLEAGINOSA TRANSGÊNICA, que produz sementes maduras em que o perfil total de ácidos graxos da semente compreende pelo menos 10,0% de ácido araquidônico e menos de 1% de ácidos graxos ômega 3 totais que contêm pelo menos dezoito átomos de carbono e pelo menos quatro uniões duplas.
3. PLANTA TRANSGÊNICA, de acordo com qualquer das reivindicações 1 ou 2, em que a planta oleaginosa é selecionada a partir do grupo que consiste de soja, espécies de Brassica, girassol, milho, algodão, linho e açafrão.
4. SEMENTE, obtida a partir da planta transgênica de acordo com qualquer das reivindicações 1 ou 2.
5. ÓLEO, obtido a partir da semente de acordo com a reivindicação 4.
6. PRODUTO ALIMENTÍCIO OU ANÁLOGO A ALIMENTO, que possui nele incorporado o óleo de acordo com a reivindicação 5.
7. PRODUTO ALIMENTÍCIO MEDICINAL OU ANÁLOGO ALIMENTÍCIO MEDICINAL, que possui nele incorporado o óleo de acordo com a reivindicação 5.
8. PRODUTO FARMACÊUTICO, que possui nele incorporado o óleo de acordo com a reivindicação 5.
9. PRODUTO ALIMENTÍCIO, de acordo com a reivindicação -6, em que o mencionado produto é selecionado a partir do grupo que consiste de uma partícula alimentícia seca por pulverização, partícula alimentícia seca por congelamento, produto de carne, alimento de cereal, alimento de lanche, produto assado, alimento extrudado, alimento frito, alimento saudável, alimento lácteo, análogos de carne, análogos de queijo, análogos de leite, ração para animais domésticos, ração animal ou alimento para aquacultura.
10. BEBIDA, que possui nela incorporado o óleo de acordo com a reivindicação 5.
11. FÓRMULA PARA BEBÊS, que possui nela incorporado o óleo de acordo com a reivindicação 5.
12. SUPLEMENTO NUTRICIONAL, que possui nele incorporado o óleo de acordo com a reivindicação 5.
13. RAÇÃO PARA ANIMAIS DOMÉSTICOS, que possui nela incorporada a semente de acordo com a reivindicação 4.
14. RAÇÃO PARA ANIMAIS DOMÉSTICOS, que possui nela incorporado o óleo de acordo com a reivindicação 5.
15. ALIMENTO PARA AQUACULTURA, que possui nele incorporada a semente de acordo com a reivindicação 4.
16. ALIMENTO PARA AQUACULTURA, que possui nele incorporado o óleo de acordo com a reivindicação 5.
17. RAÇÃO ANIMAL, que possui nela incorporada a semente de acordo com a reivindicação 4.
18. RAÇÃO ANIMAL, que possui nela incorporado o óleo de acordo com a reivindicação 5.
19. PRODUTO DE SEMENTE INTEIRA, fabricado a partir da semente de acordo com a reivindicação 4.
20. PRODUTO DE ÓLEO MISTURADO, que compreende o óleo de acordo com a reivindicação 5.
21. PRODUTO, elaborado por meio da hidrogenação, fracionamento, interesterificação ou hidrólise do óleo de acordo com a reivindicação 5.
22. SUBPRODUTO, elaborado durante a produção do óleo de acordo com a reivindicação 5.
23. SUBPRODUTO PARCIALMENTE PROCESSADO, fabricado durante a produção do óleo de acordo com a reivindicação 5.
24. PLANTA OLEAGINOSA TRANSGÊNICA, que produz sementes maduras nas quais o perfil total de ácidos graxos de semente compreende pelo menos 10,0% de ácido araquidônico e menos de 5% de ácido gamalinolênico.
25. PLANTA OLEAGINOSA TRANSGÊNICA, que produz sementes maduras nas quais o perfil total de ácidos graxos de semente compreende pelo menos 10,0% de ácido araquidônico e até 1% de ácido gamalinolênico, inclusive.
26. PLANTA OLEAGINOSA TRANSGÊNICA, que produz sementes maduras nas quais o perfil total de ácidos graxos de semente compreende pelo menos 10,0% de ácido araquidônico, até 1% de ácido gamalinolênico, inclusive, e menos de 1% de ácidos graxos ômega 3 totais que contêm pelo menos dezoito átomos de carbono e pelo menos quatro uniões duplas.
27. SEMENTE, obtida a partir da planta transgênica de acordo com qualquer das reivindicações 24, 25 ou 26.
28. ÓLEO, obtido a partir da semente de acordo com a reivindicação 27.
29. PRODUTO ALIMENTÍCIO OU ANÁLOGO A ALIMENTO, que possui nele incorporado o óleo de acordo com a reivindicação 28.
30. PRODUTO ALIMENTÍCIO MEDICINAL OU ANÁLOGO A ALIMENTO MEDICINAL, que possui nele incorporado o óleo de acordo com a reivindicação 28.
31. PRODUTO FARMACÊUTICO, que possui nele incorporado o óleo de acordo com a reivindicação 28.
32. PRODUTO ALIMENTÍCIO, de acordo com a reivindicação -29, em que o mencionado produto é selecionado a partir do grupo que consiste de uma partícula alimentícia seca por pulverização, partícula alimentícia seca por congelamento, produto de carne, alimento de cereal, alimento de lanche, alimento assado, alimento extrudado, alimento frito, alimento saudável, alimento lácteo, análogos de carne, análogos de queijo, análogos de leite, ração para animais domésticos, ração animal ou alimento para aquacultura.
33. BEBIDA, que possui nela incorporado o óleo de acordo com a reivindicação 28.
34. FÓRMULA PARA BEBÊS, que possui nela incorporado o óleo de acordo com a reivindicação 28.
35. SUPLEMENTO NUTRICIONAL, que possui nele incorporado o óleo de acordo com a reivindicação 28.
36. RAÇÃO PARA ANIMAIS DOMÉSTICOS, que possui nela incorporada a semente de acordo com a reivindicação 27.
37. RAÇÃO PARA ANIMAIS DOMÉSTICOS, que possui nela incorporado o óleo de acordo com a reivindicação 28.
38. ALIMENTO PARA AQUACULTURA, que possui nele incorporada a semente de acordo com a reivindicação 27.
39. ALIMENTO PARA AQUACULTURA, que possui nele incorporado o óleo de acordo com a reivindicação 28.
40. RAÇÃO ANIMAL, que possui nela incorporada a semente de acordo com a reivindicação 27.
41. RAÇÃO ANIMAL, que possui nela incorporado o óleo de acordo com a reivindicação 28.
42. PRODUTO DE SEMENTE INTEIRA, fabricado a partir da semente de acordo com a reivindicação 27.
43. PRODUTO DE ÓLEO MISTURADO, que compreende o óleo de acordo com a reivindicação 28.
44. PRODUTO, elaborado por meio da hidrogenação, fracionamento, interesterificação ou hidrólise do óleo de acordo com a reivindicação 28.
45. SUBPRODUTO, elaborado durante a produção do óleo de acordo com a reivindicação 28.
46. SUBPRODUTO PARCIALMENTE PROCESSADO, fabricado durante a produção do óleo de acordo com a reivindicação 28.
BRPI0806354-0A 2007-02-12 2008-02-12 plantas oleaginosas transgências, sementes, óleos, produtos alimentìcios ou análogos a alimento, produtos alimentìcios medicinais ou análogos alimentìcios medicinais, produtos farmacêuticos, bebidas fórmulas para bebês, suplementos nutricionais, rações para animais domésticos, alimentos para aquacultura, rações animais, produtos de sementes inteiras, produtos de óleos misturados, produtos, subprodutos e subprodutos parcialmente processados BRPI0806354A2 (pt)

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Families Citing this family (66)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8084074B2 (en) * 2003-02-12 2011-12-27 E. I. Du Pont De Nemours And Company Production of very long chain polyunsaturated fatty acids in oil seed plants
GB0718706D0 (en) 2007-09-25 2007-11-07 Creative Physics Ltd Method and apparatus for reducing laser speckle
US7790156B2 (en) * 2007-04-10 2010-09-07 E. I. Du Pont De Nemours And Company Δ-8 desaturases and their use in making polyunsaturated fatty acids
US7794701B2 (en) * 2007-04-16 2010-09-14 E.I. Du Pont De Nemours And Company Δ-9 elongases and their use in making polyunsaturated fatty acids
US8957280B2 (en) 2007-05-03 2015-02-17 E. I. Du Pont De Nemours And Company Delta-5 desaturases and their use in making polyunsaturated fatty acids
WO2009130291A2 (de) * 2008-04-25 2009-10-29 Basf Plant Science Gmbh Pflanzensamenöl
US11726332B2 (en) 2009-04-27 2023-08-15 Digilens Inc. Diffractive projection apparatus
US9335604B2 (en) 2013-12-11 2016-05-10 Milan Momcilo Popovich Holographic waveguide display
US8188335B2 (en) 2009-07-17 2012-05-29 Abbott Laboratories Δ9-elongase for production of polyunsaturated fatty acid-enriched oils
CN104982920A (zh) * 2009-11-03 2015-10-21 帝斯曼知识产权资产管理有限公司 包含细胞和具有至少20个碳原子的多不饱和脂肪酸(lc-pufa)的组合物
PL2496091T3 (pl) * 2009-11-03 2020-02-28 Dsm Ip Assets B.V. Olej roślinny zawierający wielonienasycony kwas tłuszczowy mający co najmniej 20 atomów węgla
DK2496092T3 (en) * 2009-11-03 2018-07-16 Dsm Ip Assets Bv COMPOSITION WITH CELLS AND A POLYUM Saturated FAT ACID WITH AT LEAST 20 CARBON ATOMS (LC-PUFA)
ES2750241T3 (es) 2009-11-03 2020-03-25 Dsm Ip Assets Bv Aceite vegetal que comprende un ácido graso poliinsaturado que tiene al menos 20 átomos de carbono
EP2516638A1 (en) 2009-12-24 2012-10-31 E.I. Du Pont De Nemours And Company Plant membrane bound o-acyl transferase (mboat) family protein sequences and their uses for altering fatty acid compositions
CA2808002A1 (en) 2010-08-26 2012-03-01 E. I. Du Pont De Nemours And Company Mutant hpgg motif and hdash motif delta-5 desaturases and their use in making polyunsaturated fatty acids
US9274349B2 (en) 2011-04-07 2016-03-01 Digilens Inc. Laser despeckler based on angular diversity
EP2748670B1 (en) 2011-08-24 2015-11-18 Rockwell Collins, Inc. Wearable data display
WO2016020630A2 (en) 2014-08-08 2016-02-11 Milan Momcilo Popovich Waveguide laser illuminator incorporating a despeckler
US10670876B2 (en) 2011-08-24 2020-06-02 Digilens Inc. Waveguide laser illuminator incorporating a despeckler
US20150010265A1 (en) 2012-01-06 2015-01-08 Milan, Momcilo POPOVICH Contact image sensor using switchable bragg gratings
BR112014018009A2 (pt) 2012-01-23 2018-06-26 Du Pont sequência de ácido nucleico isolada, construção recombinante, célula vegetal, método para reduzir a expressão de pelo menos um gene biossintético de ácido graxo vegetal, método para reduzir a expressão de dois ou mais genes biossintéticos de ácido graxo vegetal e planta ou semente transgênica
EP2842003B1 (en) 2012-04-25 2019-02-27 Rockwell Collins, Inc. Holographic wide angle display
US9456744B2 (en) 2012-05-11 2016-10-04 Digilens, Inc. Apparatus for eye tracking
US9933684B2 (en) 2012-11-16 2018-04-03 Rockwell Collins, Inc. Transparent waveguide display providing upper and lower fields of view having a specific light output aperture configuration
US10209517B2 (en) 2013-05-20 2019-02-19 Digilens, Inc. Holographic waveguide eye tracker
CA2918911A1 (en) 2013-07-31 2015-02-05 E.I. Du Pont De Nemours And Company Modification of soybean seed composition to enhance feed, food and other industrial applications of soybean products
US9727772B2 (en) 2013-07-31 2017-08-08 Digilens, Inc. Method and apparatus for contact image sensing
WO2016020632A1 (en) 2014-08-08 2016-02-11 Milan Momcilo Popovich Method for holographic mastering and replication
WO2016042283A1 (en) 2014-09-19 2016-03-24 Milan Momcilo Popovich Method and apparatus for generating input images for holographic waveguide displays
EP3198192A1 (en) 2014-09-26 2017-08-02 Milan Momcilo Popovich Holographic waveguide opticaltracker
EP3245551B1 (en) 2015-01-12 2019-09-18 DigiLens Inc. Waveguide light field displays
US10437064B2 (en) 2015-01-12 2019-10-08 Digilens Inc. Environmentally isolated waveguide display
CN107533137A (zh) 2015-01-20 2018-01-02 迪吉伦斯公司 全息波导激光雷达
US9632226B2 (en) 2015-02-12 2017-04-25 Digilens Inc. Waveguide grating device
WO2016146963A1 (en) 2015-03-16 2016-09-22 Popovich, Milan, Momcilo Waveguide device incorporating a light pipe
WO2017060665A1 (en) 2015-10-05 2017-04-13 Milan Momcilo Popovich Waveguide display
EP3398007B1 (en) 2016-02-04 2024-09-11 DigiLens, Inc. Waveguide optical tracker
US9950328B2 (en) * 2016-03-23 2018-04-24 Alfa Laval Corporate Ab Apparatus for dispersing particles in a fluid
JP6895451B2 (ja) 2016-03-24 2021-06-30 ディジレンズ インコーポレイテッド 偏光選択ホログラフィー導波管デバイスを提供するための方法および装置
EP3433658B1 (en) 2016-04-11 2023-08-09 DigiLens, Inc. Holographic waveguide apparatus for structured light projection
WO2017208173A1 (en) 2016-06-01 2017-12-07 National Research Council Of Canada Winter aconite fatty acid elongase and uses thereof in the production of fatty acids
EP3548939A4 (en) 2016-12-02 2020-11-25 DigiLens Inc. UNIFORM OUTPUT LIGHTING WAVEGUIDE DEVICE
EP3554265A1 (en) 2016-12-15 2019-10-23 DSM IP Assets B.V. Blend formulation comprising silicate and microbial and / or plant cells comprising a polyunsaturated fatty acid having at least 20 carbon atoms (lc-pufa)
US10545346B2 (en) 2017-01-05 2020-01-28 Digilens Inc. Wearable heads up displays
CN111386495B (zh) 2017-10-16 2022-12-09 迪吉伦斯公司 用于倍增像素化显示器的图像分辨率的系统和方法
WO2019136476A1 (en) 2018-01-08 2019-07-11 Digilens, Inc. Waveguide architectures and related methods of manufacturing
CN114721242B (zh) 2018-01-08 2025-08-15 迪吉伦斯公司 用于制造光学波导的方法
JP7456929B2 (ja) 2018-01-08 2024-03-27 ディジレンズ インコーポレイテッド 導波管セルを製造するためのシステムおよび方法
JP7404243B2 (ja) 2018-01-08 2023-12-25 ディジレンズ インコーポレイテッド 導波管セル内のホログラフィック格子の高スループット記録のためのシステムおよび方法
US10690851B2 (en) 2018-03-16 2020-06-23 Digilens Inc. Holographic waveguides incorporating birefringence control and methods for their fabrication
US11402801B2 (en) 2018-07-25 2022-08-02 Digilens Inc. Systems and methods for fabricating a multilayer optical structure
CN111248268A (zh) * 2018-11-30 2020-06-09 内蒙古伊利实业集团股份有限公司 一种发酵乳及其制备方法
WO2020149956A1 (en) 2019-01-14 2020-07-23 Digilens Inc. Holographic waveguide display with light control layer
US20200247017A1 (en) 2019-02-05 2020-08-06 Digilens Inc. Methods for Compensating for Optical Surface Nonuniformity
US20220283377A1 (en) 2019-02-15 2022-09-08 Digilens Inc. Wide Angle Waveguide Display
EP3924759B1 (en) 2019-02-15 2025-07-30 Digilens Inc. Methods and apparatuses for providing a holographic waveguide display using integrated gratings
JP2022525165A (ja) 2019-03-12 2022-05-11 ディジレンズ インコーポレイテッド ホログラフィック導波管バックライトおよび関連する製造方法
CN110106019A (zh) * 2019-05-20 2019-08-09 无限极（中国）有限公司 一种利用裂壶藻生产复合型多不饱和脂肪酸油脂的方法
EP3980825A4 (en) 2019-06-07 2023-05-03 Digilens Inc. WAVEGUIDES WITH TRANSMITTING AND REFLECTING GRIDS AND RELATED MANUFACTURING PROCESSES
JP2022543571A (ja) 2019-07-29 2022-10-13 ディジレンズ インコーポレイテッド 画素化されたディスプレイの画像解像度および視野を乗算するための方法および装置
JP2022546413A (ja) 2019-08-29 2022-11-04 ディジレンズ インコーポレイテッド 真空回折格子および製造方法
EP4252048A4 (en) 2020-12-21 2024-10-16 Digilens Inc. EYE LUMINESCENCE SUPPRESSION IN WAVEGUIDE-BASED DISPLAYS
WO2022150841A1 (en) 2021-01-07 2022-07-14 Digilens Inc. Grating structures for color waveguides
KR20230153459A (ko) 2021-03-05 2023-11-06 디지렌즈 인코포레이티드. 진공 주기적 구조체 및 제조 방법
EP4658259A1 (en) 2023-01-30 2025-12-10 Cold Spring Harbor Laboratory Use of arachidonic acid for amelioration of cytotoxic effects from chemotherapy and radiation therapy
WO2025231097A1 (en) 2024-05-02 2025-11-06 Cold Spring Harbor Laboratory Use of arachidonic acid for promoting endometrial epithelium regeneration and stem cell plasticity

Family Cites Families (46)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4670285A (en) 1982-08-06 1987-06-02 The University Of Toronto Innovations Foundation Infant formula
US5107065A (en) 1986-03-28 1992-04-21 Calgene, Inc. Anti-sense regulation of gene expression in plant cells
US5188958A (en) 1986-05-29 1993-02-23 Calgene, Inc. Transformation and foreign gene expression in brassica species
US5004863B2 (en) 1986-12-03 2000-10-17 Agracetus Genetic engineering of cotton plants and lines
JP2527340B2 (ja) 1986-12-15 1996-08-21 アプライド バイオシステムズ インコーポレイテッド ロ―ダミン染料の５−及び６−スクシニミジルカルボキシレ―ト異性体
US5416011A (en) 1988-07-22 1995-05-16 Monsanto Company Method for soybean transformation and regeneration
US5057419A (en) 1988-09-22 1991-10-15 Rutgers University Genetically engineered plasmid and organisms for the production of specialized oils
US5231020A (en) 1989-03-30 1993-07-27 Dna Plant Technology Corporation Genetic engineering of novel plant phenotypes
US5366860A (en) 1989-09-29 1994-11-22 Applied Biosystems, Inc. Spectrally resolvable rhodamine dyes for nucleic acid sequence determination
ATE241007T1 (de) 1990-03-16 2003-06-15 Calgene Llc Dnas, die für pflanzliche desaturasen kodieren und deren anwendungen
WO1991018985A1 (en) 1990-05-25 1991-12-12 E.I. Du Pont De Nemours And Company Nucleotide sequence of soybean stearoyl-acp desaturase gene
US5658767A (en) 1991-01-24 1997-08-19 Martek Corporation Arachidonic acid and methods for the production and use thereof
WO1992017598A1 (en) 1991-03-29 1992-10-15 The Board Of Trustees Of The University Of Illinois Production fo transgenic soybean plants
CA2124673C (en) 1991-12-04 2008-08-05 John Browse Fatty acid desaturase genes from plants
DK0668919T3 (da) 1992-11-17 2003-09-15 Du Pont Gener for mikorsomale delta-12-fedtsyredesaturaser og beslægtede enzymer fra planter
GB9703146D0 (en) 1997-02-14 1997-04-02 Innes John Centre Innov Ltd Methods and means for gene silencing in transgenic plants
US5972664A (en) 1997-04-11 1999-10-26 Abbott Laboratories Methods and compositions for synthesis of long chain poly-unsaturated fatty acids
US5968809A (en) 1997-04-11 1999-10-19 Abbot Laboratories Methods and compositions for synthesis of long chain poly-unsaturated fatty acids
US6075183A (en) 1997-04-11 2000-06-13 Abbott Laboratories Methods and compositions for synthesis of long chain poly-unsaturated fatty acids in plants
CN1202246C (zh) 1998-04-08 2005-05-18 联邦科学和工业研究组织 获得修饰表型的方法和措施
US6403349B1 (en) 1998-09-02 2002-06-11 Abbott Laboratories Elongase gene and uses thereof
US6677145B2 (en) 1998-09-02 2004-01-13 Abbott Laboratories Elongase genes and uses thereof
CA2791961A1 (en) 1998-12-07 2000-06-15 Washington State University Research Foundation Desaturases and methods of using them for synthesis of polyunsaturated fatty acids
EE200200443A (et) 2000-02-09 2003-12-15 Basf Aktiengesellschaft Uus elongaasigeen ning polüküllastumata rasvhapete saamise meetod
JP2004512020A (ja) 2000-06-23 2004-04-22 イー・アイ・デュポン・ドウ・ヌムール・アンド・カンパニー 組換え構築物ならびにこれを遺伝子発現を減少させる際に用いる方法
DE60127967T2 (de) 2000-07-21 2008-01-17 E.I. Du Pont De Nemours And Co., Wilmington Ein mit der synthese von delta-12 epoxy-gruppen in fettsäuren von pflanzen assoziiertes cytochrom p450 enzym
GB0107510D0 (en) 2001-03-26 2001-05-16 Univ Bristol New elongase gene and a process for the production of -9-polyunsaturated fatty acids
JP2002296554A (ja) 2001-03-30 2002-10-09 Minebea Co Ltd ファラデー回転子
US7045683B2 (en) 2001-05-04 2006-05-16 Abbott Laboratories Δ4-desaturase genes and uses thereof
US7211656B2 (en) * 2002-01-30 2007-05-01 Abbott Laboratories Desaturase genes, enzymes encoded thereby, and uses thereof
WO2003099216A2 (en) 2002-05-22 2003-12-04 Monsanto Technology Llc Fatty acid desaturases from fungi
CA2510462A1 (en) 2002-12-19 2004-07-08 University Of Bristol Method for producing polyunsaturated fatty acids in a transgenic plant or microorganism comprising an introduced delta-9-elongase and a delta-8-desaturase
US7129089B2 (en) 2003-02-12 2006-10-31 E. I. Du Pont De Nemours And Company Annexin and P34 promoters and use in expression of transgenic genes in plants
US20040172682A1 (en) 2003-02-12 2004-09-02 Kinney Anthony J. Production of very long chain polyunsaturated fatty acids in oilseed plants
US7238482B2 (en) 2003-05-07 2007-07-03 E. I. Du Pont De Nemours And Company Production of polyunsaturated fatty acids in oleaginous yeasts
ES2713548T3 (es) * 2003-08-01 2019-05-22 Basf Plant Science Gmbh Procedimiento para la producción de ácidos grasos poliinsaturados en organismos transgénicos
US7504259B2 (en) 2003-11-12 2009-03-17 E. I. Du Pont De Nemours And Company Δ12 desaturases suitable for altering levels of polyunsaturated fatty acids in oleaginous yeast
AU2004290051A1 (en) 2003-11-12 2005-05-26 E.I. Dupont De Nemours And Company Delta-15 desaturases suitable for altering levels of polyunsaturated fatty acids in oilseed plants and oleaginous yeast
US7943816B2 (en) * 2003-12-17 2011-05-17 Suntory Holdings Limited Arachidonic acid-containing plants and use of the plants
EP2623584B1 (de) * 2004-02-27 2019-04-10 BASF Plant Science GmbH Verfahren zur Herstellung mehrfach ungesättigter Fettsäuren in transgenen Pflanzen
CA2559360A1 (en) * 2004-02-27 2005-09-09 Basf Plant Science Gmbh Method for producing c18, c20 and c22 polyunsaturated fatty acids in transgenic plants
US7550651B2 (en) 2004-06-25 2009-06-23 E.I. Du Pont De Nemours And Company Delta-8 desaturase and its use in making polyunsaturated fatty acids
US7550286B2 (en) 2004-11-04 2009-06-23 E. I. Du Pont De Nemours And Company Docosahexaenoic acid producing strains of Yarrowia lipolytica
DK1951866T3 (da) 2005-11-23 2014-10-27 Du Pont Delta-9-elongaser og anvendelse heraf til fremstilling af flerumættede fedtsyrer
US7678560B2 (en) 2006-05-17 2010-03-16 E.I. Du Pont De Nemours And Company Δ 5 desaturase and its use in making polyunsaturated fatty acids
PL2219476T3 (pl) * 2007-11-14 2017-01-31 Loders Croklaan B.V. Kompozycja preparatu dla niemowląt

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US8013215B2 (en) 2011-09-06
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CA2675207A1 (en) 2008-07-21
US8389808B2 (en) 2013-03-05
US20110268861A1 (en) 2011-11-03
EP2441842A1 (en) 2012-04-18
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