CZ288536B6 - Bifunkční varianty urokinázy se zlepšenými fibrinolytickými vlastnostmi a s trombininhibičním účinkem, plazmidy pro jejich získávání, způsob jejich výroby a trombolytika tyto látky obsahující - Google Patents
Bifunkční varianty urokinázy se zlepšenými fibrinolytickými vlastnostmi a s trombininhibičním účinkem, plazmidy pro jejich získávání, způsob jejich výroby a trombolytika tyto látky obsahující Download PDFInfo
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Abstract
eÜen se t²k bifunk n ch variant urokin zy se zlepÜen²mi fibrinolytick²mi vlastnostmi a s trombininhibi n m · inkem obecn ho vzorce I M4-X.sub.1.n.-Y.sub.1.n., ve kter m M4 zna aminokyselinovou sekvenci .sup.47.n.Ser a .sup.411.n.Leu glykosidu zbaven prourokin zy podle obr. 1, plazmid pou it²ch p°i z sk v n t chto polypeptid , zp sobu jejich v²roby, jako i trombolytik, kter obsahuj jako · innou l tku bifunk n varianty urokin zy obecn ho vzorce I.\
Description
Oblast techniky
Vynález se týká bifunkčních variant urokinázy se zlepšenými fibrinolytickými vlastnostmi a s trombininhibičním účinkem, plazmidů použitých při získávání těchto polypeptidů, způsobu jejich výroby, jakož i trombolytik, která obsahují jako účinnou látku bifunkční varianty urokinázy.
Dosavadní stav techniky
Důležitou vlastností lidské krve je schopnost uzavřít poškozené krevní řečiště vytvořením trombů. Srážení krve je vyvoláváno řadou enzymů, obsažených v krvi, které v takzvané srážecí kaskádě vedou k tomu, že enzym trombin proteolyticky přemění předchozí fibrinogen na fibrin. Fibrin je polymerován v místě poškození vlivem trombocytů, erytrocytů a dalších krevních součástí za vytvoření trombů.
Kromě toho obsahuje krev také řadu enzymů, které působí proti srážení a zajišťují po regeneraci cévní stěny průtok krve. Nejdůležitějším enzymem pro trombolýzu je plazmin, který proteolyticky napadá fibrinová vlákna a tím ovlivňuje rozpouštění trombů. Plazmin je tvořen proteolytickým štěpením plazminogenu, svého neaktivního bílkovinného předchůdce (prekurzoru). Aktivace je způsobena proteolytickým štěpením plazminogenu plazminogenaktivátorem. Jsou známé dva lidské plazminogenaktivátory, totiž urokináza, plazminogenaktivátor, nacházející se v moči, a tkáňový plazminogenaktivátor.
Srdeční infarkt a mozkový infarkt jsou úzce spojeny s patologickým obrazem trombů. Při obou formách infarktu vznikají - často z důvodu arteriosklerotických změn arterií - za určitých podmínek tromby na stěnách cév. Tyto tromby porušují krevní proud v cévách, takže tkáně nemohou být dostatečně zásobovány kyslíkem. Toto vede při srdečním infarktu k částečnému nebo úplnému odumření srdečního svalu. Obdobně vede blokáda mozkových cév k těžkému poškození mozkové tkáně.
Při terapii nemocných s infarktem se nasazují aktivátory plazminogenu jako trombolytika, která startují odbourávání trombů plazminem. V současné době slouží k léčbě streptokináza, APSAC (Anisolated Plazminogen Streptokinase Activator Complex), dvouřetězcová urokináza (UK), rekombinantní jednořetězcová urokináza (rekombinante Prourokinase) a tkáňový plazminogenaktivátor (tPA) (Collen a Lijnen, Blood 78,3114-3124 /1991/).
U streptokinázy se jedná o bílkovinu hemolytických streptokoků. Streptokináza aktivuje plazminogen tím, že s ním vytváří komplex, čímž převede plazminogen na aktivní konformaci. Tento komplex sám přemění volný plazminogen na plazmin, který potom opět štěpí streptokinázou vázaný plazminogen. Dalším vývojem streptokinázy je APSAC, což je in vitro připravená sloučenina ze streptokinázy a lidského plazminogenu. APSAC vykazuje na základě chemické modifikace aktivního centra plazminogenu oproti streptokináze zvýšené biologické poločasové hodnoty.
Urokináza je lidská bílkovina, kterou je možno získat z moče ve dvou formách jako biologicky účinný protein, totiž vysokomolekulámí urokinázu (HUK) a nízkomolekulámí urokinázu (LUK) (Stump a kol., J. Biol. Chem. 261, 1267-1273 /1986/). HUK a LUK jsou dvouřetězcové molekuly. Urokináza se jako jednořetězcová urokináza (prourokináza) tvoří v různých tkáních a může být jako proenzym zjištěna v nepatrných množstvích v lidské krvi (Wun a kol., J. Biol.
-1 CZ 288536 B6
Chem. 257, 3276-3283 /1982/). Aktivovaná forma prourokinázy má jako HUK molekulovou hmotnost 54 kilodaltonů a sestává ze tri domén: amino-terminální Growth-Factor-Dománe, z kruhu a ze Serin-Protease-Dománe (Giinzler a kol., Hoppe-Seyler's Z. Physiol. Chem. 363, 1155-1165 /1982/); Steffens a kol., Hoppe-Seyler's Z. Physiol. Chem. 363,1043-1058 /1982/).
Ačkoliv prourokináza a plazminogen se vyskytují jako proenzymy, je prourokináza schopna na základě vnitřní aktivity přeměňovat plazminogen na plazmin. Plnou aktivitu získá ale tento plazminogenaktivátor teprve tehdy, když vytvořený plazmin štěpí prourokinázu mezi 158lysinem a 59isoleucinem (Lijnen a kol., J. Biol. Chem. 261, 1253-1258 /1986/). Genetickou technikou získaná urokináza v Escherichia coli byla poprvé popsaná Heynekerem (Procedings of the IVth Intemational Symposium on Genetics of Industrial Microorganismu 1982). Neglykosylovaná prourokináza (Saruplase) byla připravena za použití syntetického genu (Brigerius-Flohé a kol. Appl. Micobiol. Biotech. 36, 640-649 /1992/).
Tkáňový plazminogenaktivátor je bílkovina, vyskytující se v tkáních a v krvi o molekulové hmotnosti 72 kilodaltonů. Tento plazminogenaktivátor sestává z pěti domén: z aminoterminální prstové domény, z Growth-Factor domény, kruhu 1, kruhu 2 a ze Serin-Protease domény. Na rozdíl od prourokinázy je tPA teprve navázáním na fibrin schopná štěpit plazminogen. Stejně jako prourokináza je tPA schopná vlivem štěpení mezi kruhem 2 a Serin-Protease doménou, katalyzovaného plazminem, přejít do aktivní formy. Při tom se váže tkáňový plazminogenaktivátor na fibrin, ne však na fíbrinogen, čímž je plazminogen trombusspecifícky aktivován. Na rozdíl od dvouřetězcové urokinázy je všeobecná plazminogenaktivace značně snížena (Collen a Lijnen, Blood 78,3114-3124 /1991/).
Od počátku osmdesátých let se ukázalo aktivní léčení infarktu myokardu trombolytiky jako účinné a užitečné. V řadě studií bylo ukázáno, že léčení nemocných se srdečním infarktem pomocí streptokinázy, APSAC, UK, rekombinantní prourokinázy nebo tPA vedlo ve srovnání s neléčenými pacienty ke značnému snížení úmrtnosti.
Aby se zlepšila účinnost těchto preparátů, byla připravena za pomoci metod genové techniky řada derivátů tkáňového plazminogenaktivátoru a prourokinázy. Vedle zvýšené fibrinolytické aktivity a omezení vedlejších účinků, byl v centru pozornosti vývoj forem, které by byly způsobilé pro bolus-apliaci. Přehled o vkladech ke zlepšení plazminogenaktivátorů je uveden v Thrombosis and Haemostasis 66, 88-110 (1991), jakož i v Trends in Biotech. 9, 86-90 (1991).
Aby se zlepšila účinnost plazminogenaktivátorů při lýzové terapii a zvláště pak aby se zvýšily jejich hodnoty biologického poločasu, byly připraveny deleční a substituční varianty tkáňového plazminogenaktivátoru, u kterých byly například odstraněny prstové domény a Growth-Factor domény, nebo byla vyměněna Serin-Protease doména za Serin-Protease doménu urokinázy (Collen a kol., Thromb. Haemostasis 65, 174-180 /1991/; Fromage a kol., Fibrinolysis 5, 187— 190 /1991/; Lu a kol., Blood 78,125-131 /1991/).
Skutečně se ukázalo, že odstraněním prstové domény a Growth-Factor domény se zvýšily hodnoty biologického poločasu tPA-variant (Lijnen a Collen, Thromb. Haemostasis 66, 94-95 /1991/). Deleční a substituční varianty, které sestávají zobou kruhových domén tPA a SerinProtease domény UK, byly výhodnější při trombolýze na základě podstatného prodloužení hodnot původního poločasu, to znamená hodnot dosahovaných s nepozměněným plazminogenaktivátorem. Tyto varianty plazminogenaktivátoru mají však pouze velmi malou fibrinovou specifitu (Lu a kol., Blood 78, 125-131 /1991/).
Byly provedeny různé pokusy na výrobu plazminogenaktivátorů se zvýšenou fibrinovou specifitou. Pro snížení nebezpečí krvácení by takové účinné látky měly aktivovat pokud možno výhradně plazminogen v blízkosti trombu, neměly by však vyvolávat systemickou aktivaci plazminogenu. Je například známá varianta tPAS, při které byl vyměněn u původní molekuly kruh 1 za kruh 2. Tato varianta vykazuje zvýšenou afinitu k N-terminálním lysinovým zbytkům,
-2CZ 288536 B6 ale ne k fibrinu. V pokusech na zvířatech nebyla tato varianta, se zřetelem na tromblýzu, účinnější než původní tkáňový plazminogenaktivátor (Collen a kol., Thromb. Haemostasis 65, 174-180/1991/).
Ostatní známé varianty, které vznikají fúzí mezi trombus specifickými protilátkami a plazminogenaktivátorem, jsou na zvířatech účinnější, než původní plazminogenaktivátory (Lijnen a Collen, Thromb. Haemostasis 66, 88-110 /1991/). Velmi vysokou fibrinovou specifítu vykazuje plazminogenaktivátor, izolovaný z netopýra Desmodus retundus (Gardell a kol., J. Biol. Chem. 264,17947-17952 /1989/). Tento plazminogenaktivátor vykazuje v pokusech na zvířatech zlepšenou trombolýzu oproti tPA při zvýšených hodnotách poločasu a snížené systemické aktivaci plazminogenu (Gardell a kol., Circulation 84, 244-253 /1991/); Mellot a kol., Arterioscl. Thromb. 12, 212-221 /1992/).
Úspěch léčení pacientů s infarktem plazminogenaktivátory nezávisí však pouze na trombolýze, ale také na tom, jak dalece se může zabránit opětovnému ucpávání průchodných cév. Různé nálezy ukazují na to, že v trombech vázaný trombin se při trombolýze opět uvolňuje jako aktivní enzym a může způsobovat opětné ucpání cév (Szczeklik a kol., Arterioscl. Thromb. 12, 548-553 /1992/; Eisenberg, Circulation 84, 2601-2603 /1991/). Skutečně je působení trombolytika zlepšeno současnou nebo předřazenou dávkou trombinového inhibitoru heparinu. Také dávkou argatrobanu, hirugenu nebo proteinu C se může snížit výskyt opětovného ucpání cév při lýzové terapii (Schneider, Thromb. Res. 64, 677-689 /1990/; Yao a kol., Am. Physiol. 262; Heart Circ. Physiol. 31, H 374 - H 379 /1992/; Gruber a kol., Circulation 84, 2454-2462 /1991/). Kromě toho je známé, že úmrtnost pacientů po srdečním infarktu je oproti kontrolní skupině signifikantně redukována po předcházející dávce heparinu a následné aplikaci prourokinázy oproti kontrolní skupině (aplikace prourokinázy bez předchozí dávky heparinu) (Tebbe a kol., Z. Kardiol. 80. Suppl. 3, 32 /1991/).
Jedním z nejúčinnějších inhibitorů trombinu je hirudin, izolovaný z krve pijavice Hirudo medicinales, který se váže svojí karboxy-terminální polovinou specificky na takzvané aniontové vazebné místo trombinu. Určité aminokyseliny amino-terminální poloviny molekuly hirudinu uzavírají přístup k substrátovému vazebnému místu trombinu (Rydel a kol., Science 249, 277280 /1990/). Kromě toho je známo, že i menší deriváty hirudinu mohou brzdit účinek trombinu, obzvláště pak ty, které popsal Maraganore a kol. v Biochemistiy 29, 7095-7101 (1990), zdůrazněné jsou Hirulog-molekuly (Krstenansky a kol., m J. med. Chem. 30, 1688-1691 /1987/; Yue a kol., Protein Engineering 5, 77-85 /1992/).
Použití hirudinu v kombinaci s plazminogenaktivátorem k léčení trombolyticky podmíněných onemocnění cév byla popsána v evropských patentových přihláškách EP 328 057 a EP 365 468. Terapeutická použití hirudinových derivátů v kombinaci s trombolytiky jsou známa z PCT přihlášky WO 91/01142.
Trombin může být také blokován peptidem, který se odvozuje z amino-terminální sekvence lidského trombinového receptorů (Vu a kol., Nátuře 253, 674-678 /1991/). Trombinový receptor obsahuje v amino-terminální oblasti trombin vázající sekvenci se sousedním místem pro štěpení trombinu. Oblast receptorů, která váže trombin, je velmi podobná strukturou karboxy-terminální oblasti hirudinu. Receptor je trombinem aktivován tak, že je receptorová sekvence štěpena. Na základě afinity mezi receptorem a trombinem působí fragment receptorů s vazebnou oblastí a modifikovaným místem štěpení jako trombininhibitor.
Stejně tak může být trombin blokován peptidem, který je odvozen od aminokyselin 41 až 57 hemadinu (Strube a kol., J. Biol. Chem. 268, 8590-8595 /1993/).
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Podstata vynálezu
Úkolem předloženého vynálezu je objevení účinné látky pro léčení trombolyticky podmíněného ucpání cév, která během krátké doby způsobí úplnou trombolýzu a zároveň zabrání opětnému ucpání cévy po zprvu úspěšné trombolýze. Kromě toho by se mělo pomocí těchto účinných látek zabránit systemické aktivaci plazminogenu.
Nyní bylo zjištěno, že vysoké nároky, které jsou kladeny na tyto účinné látky, jsou splněny určitými bifunkčními variantami urokinázy.
Předmětem předloženého vynálezu tedy jsou bifúnkční varianty urokinázy obecného vzorce I
M4-Xl-Y1 (I), ve kterém
M4 značí aminokyselinovou sekvenci 47Ser až 411Leu glykosidu zbavené prourokinázy podle obr. 1,
Xi značí přímou vazbu mezi M4 a Yi nebo peptid o sekvenci
Ser-Pro-Pro-Ser-Pro-Pro-Gly-Gly-Phe nebo
Ser-Pro-Pro-Ser-Pro-Pro-Ser-Pro-Pro-Gly-Gly-Phe nebo
Ser-Pro-Pro-Ser-Pro-Pro-Ser-Pro-Pro-Gly-Gly-Phe-Gly nebo peptidovou sekvenci obecného vzorce II
Ser-Xr-Xj-Xr-Xj-Xff-X? (II), přičemž
X2 značí Pro nebo Leu,
X3 značí Val nebo Pro,
X» značí Lys, Val, Arg, Gly nebo Glu,
X5 značí Ala, Val, Gly, Leu nebo Ile,
X6 značí Phe, Trp, Tyr nebo Val a
X7 značí Gly nebo přímou vazbu mezi X6 a Yi a
Yi značí peptid sekvence
Y 2-Arg-Pro-Yj-Gly-Gly-Gly-Gly-Asn-Gly-Asp-Phe-Glu-Glu-Ile-Pro-Glu-GluTyr-Leu-Y4 nebo
Y2-Arg-Pro-Phe-Leu-Leu-Arg-Asn-Pro-Asn-Asp-Lys-Tyr-Glu-Pro-Phe-T rpGlu-Asp-Glu-Glu-Lys-Asn-Glu nebo
Yr-Arg-Pro-Ser-Ser-Glu-Phe-Glu-Glu-Phe-Glu-Ile-Asp-Glu-Glu-Glu-Lys, přičemž
Y2 značí Pro nebo Val,
Y3 značí Leu nebo přímou vazbu mezi Pro a Gly a
Y4 značí Gin nebo hydroxylovou skupinu.
V bifunkčních variantách urokinázy obecného vzorce I, kde Y] značí peptid sekvence
Y2-Arg-Pro-Y3-Gly-Gly-Gly-Gly-Asn-Gly-Asp-Phe-Glu-Glu-Ile-Pro-Glu-Glu-Tyr-Leuγ4, přičemž
Y2 značí Pro nebo Val,
Y3 značí Leu nebo přímou vazbu mezi Pro a Gly a
Y4 značí Gin nebo hydroxylovou skupinu, značí Xi výhodně peptidovou sekvenci obecného vzorce II
Ser-X2-X3-X4-X5-X6-X7 (II), ve kterém
X2 značí Pro nebo Leu,
X3 značí Val,
X4 značí Lys, Val nebo Arg,
X5 značí Ala, Val nebo Gly,
X6 značí Phe, Trp, Tyr nebo Val a
X7 značí Gly nebo přímou vazbu mezi Xe a Yj.
Obzvláště výhodné jsou v těchto bifunkčních variantách urokinázy peptidové sekvence obecného vzorce II, ve kterém
X2 značí Pro nebo Leu,
X3 značí Val,
X4 značí Lys, Val,
X5 značí Ala, Val,
X6 značí Phe, Trp, nebo Tyr a a obzvláště takové, ve kterých X7 značí přímou vazbu mezi X6 a Y^
V bifunkčních variantách urokinázy obecného vzorce I, ve kterém Yi značí peptid sekvence
Yr-Arg-Pro-Phe-Leu-Leu-Arg-Asn-Pro-Asn-Asp-Lys-Tyr-Glu-Pro-Phe-Trp-Glu-AspGlu-Glu-Lys-Asn-Glu ve kterém značí Y2 Pro nebo Val, představuje Xi výhodně peptidovou sekvenci obecného vzorce II
Ser-X2-X3-X4-X5-X6-X7 (II), ve kterém
X2 značí Pro nebo Leu,
X3 značí Val,
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X4 značí Lys, nebo Val,
X5 značí Ala nebo Val,
Xé značí Phe nebo Trp a
X7 značí přímou vazbu mezi X6 a Y,.
Ve srovnání se známými plazminogenaktivátory a známými směsemi plazminogenaktivátorů a inhibitoru trombinu, se vyznačují bifunkční varianty urokinázy podle předloženého vynálezu silnějším fibrinolytickým účinkem, spojeným s nepředvídaně dobrým trombin-inhibičním účinkem. Kromě toho se polypeptidů plazmatického fibrinogenu podle předloženého vynálezu překvapivě spotřebuje podstatně menší množství. Z toho vyplývající signifikantně vyšší fibrinová specifíta, obzvláště také ve srovnání se známými směsmi plazminogenaktivátorů a trombinového inhibioru, způsobuje, že je srážlivost krve ovlivněna jen mírně a nebezpečí nekontrolovatelného krvácení jako možné komplikace systemického odbourávání fiblinogenu je minimalizované.
Vysoká fibrinová specifíta variant urokinázy podle předloženého vynálezu umožňuje tedy bolusaplikaci s jasné sníženým rizikem krvácení ve srovnání s bolusaplikací známých trombolytik.
Bifunkční varianty urokinázy obecného vzorce I jsou toxikologicky neškodné, takže jsou jako takové ve vhodných farmaceutických přípravcích způsobilé k aplikaci pacientům s tromboticky podmíněným ucpáním cév.
Dalším předmětem předloženého vynálezu jsou odpovídající trombolytika, která obsahují jako účinnou látku bifunkční varianty urokinázy obecného vzorce I.
Pro léčení tromboticky podmíněných ucpání cév, například srdečního infarktu, mozkového infarktu, periferního akutního ucpání arterií, plicní embolie a hluboké trombózy vén nohy a pánve, je zapotřebí 0,1 až 1 mg/kg polypeptidu podle předloženého vynálezu. Bifunkční varianty urokinázy mohou být aplikovány intravenózně a obzvláště pomocí bolus-injekce.
Trombolytika podle předloženého vynálezu obsahují vedle alespoň jedné bifunkční varianty urokinázy pomocné látky, jako jsou například nosiče, rozpouštědla, ředidla, barviva a pojivá. Volba pomocných látek, jakož i jejich používané množství, závisí na tom, jak má být léčivo aplikováno a jak nepřipravit odborníkovi při aplikaci problémy.
Výroba bifunkčních variant urokinázy probíhá pomocí postupů genové techniky. Předmětem předloženého vynálezu jsou tedy také plazmidy pro použití při získávání bifunkčních variant urokinázy obecného vzorce I, jejichž operon má regulovatelný promotor, účinnou ShineDalgamo sekvenci jako vazebné místo pro ribozomy, startovací kodon, syntetický strukturní gen se sekvencemi ID č. 1 až 24 pro bifunkční varianty urokinázy obecného vzorce I a dále od strukturního genu jeden nebo dva terminátoiy, přičemž tyto plazmidy jsou vhodné pro expresi bifunkčních variant urokinázy ve kmenech Escherichia coli.
Jako regulovatelný promotor je vhodný obzvláště trp-promotor nebo tac-promotor. Jako terminátor se výhodně používá trp A terminátor a/nebo tet A /orf L terminátor z Tn 10.
V kontrolním regionu plazmidů podle předloženého vynálezu činí odstup mezi Shine-Dalgamosekvencí a startovacím kodonem 6 až 12, výhodně 8 až 10 nukleotidů.
Výhodné jsou plazmidy, které jsou vybrané ze skupiny, zahrnující pSJ 69 podle obr. 2e, pSJ 76 podle obr. 2h, pSJ 77 podle obr. 2h, pSJ 78 podle obr. 2h, pSJ 79 podle obr. 2h, pSJ 81 podle obr. 2j, pSJ 83 podle obr. 21, pSJ 90 podle obr. 21, pSJ 91 podle obr. 21, pSJ 92 podle obr. 21, pSJ 93 podle obr. 21, pSJ 94 podle obr. 21, pSJ 95 podle obr. 21, pSJ 101 podle obr. 21, pSJ 102 podle obr. 21, pSJ 103 podle obr. 21, pSJ 104 podle obr. 21, pSJ 105 podle obr. 21, pSJ 106 podle obr. 2o, pSJ 109 podle obr. 21, pSJ 111 podle obr. 21, pSJ 114 podle obr. 2n a pSJ 113 podle ob. 2p.
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Obzvláště výhodné jsou plazmidy, které jsou vybrané ze skupiny, zahrnující pSJ 76 podle obr. 2h, pSJ 81 podle obr. 2j, pSJ 83 podle obr. 21, pSJ 90 podle obr. 21, pSJ 91 podle obr. 21, pSJ 92 podle obr. 21, pSJ 93 podle obr. 21, pSJ 94 podle obr. 21, pSJ 95 podle obr. 21, pSJ 101 podle obr. 21, pSJ 102 podle obr. 21, pSJ 103 podle obr. 21, pSJ 105 podle obr. 21, pSJ 106 podle obr. 2o, pSJ 109 podle obr. 21, pSJ 111 podle obr. 21 a pSJ 114 podle obr. 2n.
Dále jsou obzvláště výhodné plazmidy, které jsou vybrané ze skupiny, zahrnující pSJ 76 podle obr. 2h, pSJ 81 podle obr. 2j, pSJ 83 podle obr. 21, pSJ 91 podle obr. 21, pSJ 92 podle obr. 21, pSJ 94 podle obr. 21, pSJ 95 podle obr. 21, pSJ 101 podle obr. 21, pSJ 102 podle obr. 21, pSJ 103 podle obr. 21, pSJ 105 podle obr. 21, pSJ 106 podle obr. 2o, pSJ 109 podle obr. 21, pSJ 111 podle obr. 21 a pSJ 114 podle obr. 2n.
Obzvláště je třeba uvést plazmidy, které jsou vybrané ze skupiny, zahrnující pSJ 76 podle obr. 2h, pSJ 94 podle obr. 21, pSJ 95 podle obr. 21, pSJ 101 podle obr. 21, pSJ 102 podle obr. 21, pSJ 103 podle obr. 21, pSJ 106 podle obr. 2o, pSJ 109 podle obr. 21, pSJ 111 podle obr. 21 a pSJ 114 podle obr. 2n.
Uváděné plazmidy jsou vyrobitelné z plazmidů pBlueskript KS II+, pUC 8 a pGR 201 podle obr. 2 a 2a až 2p.
Předmětem předloženého vynálezu je dále způsob výroby bifunkčních variant urokinázy obecného vzorce I, při kterém se uvedeným plazmidem transformuje kmen Escherichia coli, indukuje se exprese strukturního genu, vytvořený předstupeň proteinu bifunkční varianty urokinázy obecného vzorce I se z média a lýzovaných bakteriálních buněk oddělí, tento předstupeň proteinu se solubilizuje a potom se působením redox systému převede na bifunkční variantu urokinázy obecného vzorce I.
Exprese plazmidů podle předloženého vynálezu byla prováděna v kmenech Escherichia coli, zvláště pak ve kmenech Escherichia coli skupiny K 12, jako je například Escherichia coli K 13 JM 101 (ATC 33876), Escherichia coli K 12 JM 103 (ATCC 39403), Escherichia coli K 12 JM 105 (DSM 4162) a Escherichia coli K 12 DH 1 (ATCC 33849). V bakteriálních buňkách vznikají bifunkční varianty urokinázy podle předloženého vynálezu obecného vzorce I s vysokou výtěžností ve shlucích, ve kterých se vyskytuje protein v denaturované formě. Po izolaci shluků se denaturovaný protein pomocí postupů z chemie proteinů za působení redox systému zpětně převede na požadovanou terciární strukturu.
Příklady povedení vynálezu
Příklad 1
Příprava, izolace a čištění bifunkčních variant urokinázy podle předloženého vynálezu
a) Postup klonování
Plazmidy pro expresi pro výrobu polypeptidů podle předloženého vynálezu pomocí genové techniky v Escherichia coli byly připraveny pomocí o sobě známých metod. Sled jednotlivých kroků přípravy je znázorněn v tabulce 2 a 2a až 2p. Výchozí produkty pro přípravu plazmidů byly plazmidy pBlueskript KS 11+ (firma Stratagene, Heidelberg), pUC8 (firma Pharmacia, Freiburg) a pGR201. Plazmid pGR210 je identický s plazmidem pBF 160, který je popsaný v EP 408 945 a vAppl. Micobiol. Biotechn. 36, 640-649 (1992). Restrikční endonukleázy Bánil, BamHI, Clal, HindlII, Ncol, Ndel, Nhel a Notl, jakož i DNA-modifikující enzymy, jako je
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Změny plazmidů během jejich přípravy byly zkoušeny pomocí restrikční analýzy a DNA5 sekvencování. DNA-sekvencování bylo prováděno podle návodu výrobce se sadou reagencií firmy Pharmacia. Při přípravě plazmidů byly použity různé oligodesoxyribonukleotidy (Oligos), jejichž sekvence jsou uvedeny spolu s příslušným označením v následující tabulce 1.
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Tabulka 1
| Označení | oligosekvence od 5’ po 3’ |
| 0105 | TATGAGCAAAACTTGCTACGAAGGTAACGGTCACTTCTA CCGTGGTAAGGCTTCTACCGACAC |
| 0106 | CATGGTGTCGGTAGAAGCCTTACCACGGTAGAAGTGACC GTTACCTTCGTAGCAAGTTTTGCTCA |
| 0220 | CGGITAAGGCTTTCCCGAGGCCTGGTGGTGGTGGTAACG GTGACTTCGAAGAAATCCCGGAAGAGTACCTGTGATAGG ATCAA |
| 0221 | CTAGTTGATCCTATCACAGGTACTCTTCCGGGATTTCTT CGAAGTCACCGTTACCACCACCACCAGGCCTCGGGAAAG CCTTAACCGGGCT |
| 0222 | CGCCGAGCCCGCCGAGCCCGCCGGGTGGTTTCCCGAGGC CTGGTGGTGGTGGTAACGGTGACTTCGAAGAAATCCCGG AAGAGTACCTGTGATAGGATCAA |
| 0223 | CTAGTTGATCCTATCACAGGTACTCTTCCGGGATTTCTT CGAAGTCACCGTTACCACCACCACCAGGCCTCGGGAAAC CACCCGGCGGGCTCGGCGGGCTCGGCGGGCT |
| 0224 | CGCCGGGTGGTTTCCCGAGGCCTGGTGGTGGTGGTAACG GTGACTTCGAAGAAATCCCGGAAGAGTACCTGTGATAGG ATCAA |
| 0225 | CTAGTTGATCCTATCACAGGTACTCTTCCGGGATTTCTT CGAAGTCACCGTTACCACCACCACCAGGCCTCGGGAAAC CACCCGGCGGGCT |
| 0226 | CGCCGAGCCCGCCGAGCCCGCCGGGTGGTTTCGGTCCGA GGCCTGGTGGTGGTGGTAACGGTGACTTCGAAGAAATCC CGGAAGAGTACCTGTGATAGGATCAA |
| 0227 | CTAGTTGATCCTATCACAGGTACTCTTCCGGGATTTCTT CGAAGTCACCGTTACCACCACCACCAGGCCTCGGACCGA AACCACCCGGCGGGCTCGGCGGGCTCGGCGGGCT |
| 0265 | CACCCGGCGGAGACGGCGGGCTCAGAGCCAGACCGTTTT CTTCTTTGGTGTGAGAACG |
| 0281 | CGTCCGGGTGGTGGTGGTAACGGTGACTTCGAAGAAATC CCGGAAGAATACCTGTAAG |
| 0282 | GATCCGTTCTCACACCAAAGAAGAAAACGGTCTGGCTCT GAGCCCGCCGTCTCCGCCGGGTGGTTTCCCG |
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Tabulka 1 - pokračování
| 0283 | CTAGCTTACAGGTATTCTTCCGGGATTTCTTCGAAGTCA CCGTTACCACCACCACCCGGACGCGGGAAAC |
| 0329 | AAGAAATCCCGGAAGAATACCTGCAATAAG |
| 0330 | CGGTTAAGGCTTGGGGACCGCGGCCGCTGGGTGGTGGTG GTAACGGTGACTTCG |
| 0331 | ACCACCACCCAGCGGCCGCGGTCCCCAAGCCTTAACCGG GCT |
| 0332 | CTAGCTTATTGCAGGTATTCTTCCGGGATTTCTTCGAAG TCACCGTTACC |
| 0333 | CGGTTAAGGCTTTCGGACCGC |
| 0334 | GGCCGCGGTCCGAAAGCCTTAACCGGGCT |
| 0335 | CGGTTCGGGCTTTCGGTCCGC |
| 0336 | GGCCGCGGACCGAAAGCCCGAACCGGGCT |
| 0337 | CGGTTAAGGCTTACGGACCGC |
| 0338 | GGCCGCGGTCCGTAAGCCTTAACCGGGCT |
| 0339 | CGGTTGTTGCTTTCGGTCCGC |
| 0340 | GGCCGCGGACCGAAAGCAACAACCGGGCT |
| 0341 | CGGTTCGGGCTTTCCCGC |
| 0342 | GGCCGCGGGAAAGCCCGAACCGGGCT |
| 0343 | CGGTTAAGGCTTACCCGC |
| 0344 | GGCCGCGGGTAAGCCTTAACCGGGCT |
| 0347 | CGGTTGTTGCTTTCCCGC |
| 0348 | GGCCGCGGGAAAGCAACAACCGGGCT |
| 0381 | CGGTTAAGGCTTGGCCGC |
| 0383 | GGCCGCGGCCAAGCCTTAACCGGGCT |
| 0384 | CGGTTAAGGCTTTCCCGC |
| 0385 | GGCCGCGGGAAAGCCTTAACCGGGCT |
| 0386 | CGGTTGTAGTTTTCCCGC |
| 0387 | CGGTTGAAGTTTTCCCGC |
| 0388 | GGCCGCACTACAACTACAACCGGGCT |
| 0389 | CGGTTGTAGTTGTAGTGC |
| 0390 | GGCCGCGGGAAAACTTCAACCGGGCT |
| 0391 | GGCCGCGGGAAAACTACAACCGGGCT |
| 0392 | CrAGOTATTCGTTTTTTTCTTCGTCTTCCCAGAACGGT TCGTATTTGTCGTTCGGGTTCCGCAGCAGGAAC |
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Tabulka 1 - pokračování
| 0393 | GGCCGTTCCTGCTGCGGAACCCGAACGACAAATACGAAC CGTTCTGGGAAGACGAAGAAAAAAACGAATAAG |
| 0453 | TGGTTAAAGCTTTCCCGC |
| 0454 | GGCCGCGGGAAAGCTTTAACCAGGCT |
| 0455 | TGGTTGTTGCTTTCCCGC |
| 0456 | GGCCGCGGGAAAGCAACAACCAGGCT |
| 0465 | GGCCGCGGGAACAGAGCCAGACCGTTTTCTTCTTTGGTG TGAGAACG |
| 0466 | GATCCGTTCTCACACCAAAGAAGAAAACGGTCTGGCTCT GTTCCCGC |
| 0467 | CGGTTAAGGCTTTCCCGCGGCCGTTCCTGCTGCGGAAC |
| 0468 | TTTGTCGTTCGGGTTCCGCAGCAGGAACGGCCGCGGGAA AGCCTTAACCGGGCT |
| 0469 | CTAGCTTATTCGTITTTITCTTCGTCTTCCCAGAACGGT TCGTA |
| 0470 | CCGAACGACAAATACGAACCGTTCTGGGAAGACGAAGAA AAAAACGAATAAG |
Oligodesoxyribonukleotidy byly v detritylované formě v měřítku 0,1 pmol připraveny pomocí syntetizéru firmy Appield Biosystems, Weiterstadt (model 391) podle údajů výrobce za použití b-kyanoethylovými skupinami chráněného diisopropylaminofosfoamiditu. Vždy 100 pmol oligodesoxyribonukleotidu bylo fosforylováno v 50 mM chloridu hořečnatého a 5 mM dithiothreitolu při pH 7,5 s jednou enzymovou jednotkou T4-kinázy za přítomnosti 10 mM adenosintrifosfátu a konečně ve stejném pufru konvertováno na dvouvláknové molekuly DNA. Získané syntetické dvouvláknové molekuly DNA byly vyčištěny pomocí gelové elektroforézy na polyakrylamidovém gelu (5 % polyakrylamidu) a následně převedeny do ligace s odpovídajícím způsobem připravenými plazmidy. Příprava plazmidů štěpením restrikčními enzymy, izolace odpovídajících restrikčních fragmentů a defosforylace 5'-konců, následující ligace a transformace v Escherichia coli K 12 JM 103, jakož i všechny další genově technické práce, byly provedeny o sobě známým způsobem a jsou uvedeny v práci Sambrooka a kol., „Molecular Cloning: A Laboratory Manual“, 2. vydání, Cold Spring Harbor Laboratory Press, Cold Spring Harbour, USA, 1989.
b) Příprava trvalých kultur a fermantace
Rekombinantní expresní plazmidy pSJ69, pSJ76, pSJ77, pSJ78, pSJ79, pSJ81, pSJ83, pSJ90, pSJ91, pSJ92, pSJ93, pSJ94, pSJ95, pSJÍOl, pSJl02, pSJ103, pSJ104, pSJ105, pSJ106, pSJ109, pSJlll, pSJl 14 a pSJl 13 byly přeneseny do Escherichia coli K12 JMI03 (ATCC 39403) a natřeny na Standard-I-agar (150mg/l ampicilinu) (Sambrook a kol., „Molecular Cloning: A Laboratory Manual“). Nyní byla každá jednotlivá kolonie každé transformace kultivována ve Standard-I-médiu (pH 7,0; 150 ml/1 ampicilinu) při teplotě 20 °C až do optické hustoty (OD) 1 při 578 nm, v pěti dávkách po 2 ml jako trvalá kultura za přídavku dimethylsulfoxidu (DMSO) (7,5 °/o konečná koncentrace) v tekutém dusíku hluboce zmrazená a ponechána při teplotě -70 °C.
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K získání bifúnkčních variant urokinázy byl nyní vždy 1 ml každé trvalé kultury suspendován ve 20 ml Standard-I-media (pH 7,0; 150 mg/1 ampicilinu) a kultivován při teplotě 37 °C až do OD 1 při 578 nm.
Potom bylo celé množství získané kultury suspendováno v 1 1 Standard-I-média (pH 7,0; 150 mg/1 ampicilinu) a fermentováno v baňkách na třepačce při teplotě 37 °C. Indukce následovala po přídavku 2 ml roztoku kyseliny indolakiylové (60 mg ve 2 ml ethylalkoholu) při OD 0,5 až 1 při 578 nm.
c) Testování exprese
Pro testování řádu exprese (Ploug-jednotky pro OD pro ml) byly bezprostředně před indukcí a každou hodinu po indukci (celkem 6 hodin) buňky, odpovídající 1 ml buněčné suspense s OD 1 při 578 nm odcentrifugovány. Odstředěné buňky byly rozloženy pomocí lysozymu (1 mg 15 lysozymu pro 1 ml v 50 mM Tris/HCl pufru,pH 8,0, 50 mM kyseliny ethylendiamintetraoctové (EDTA) a 15% objemových sacharózy). Lýzované buňky byly solubilizovány ve 4 až 5 M roztoku guanidiniumhydrochloridu a po naředění na 1,2 M guanidiniumhydrochloridu a za přídavku redukčního činidla (glutathion nebo cystein) podrobeny po dobu 2 až 5 hodin reakci (Winkler a kol., Biochemistry 25, 4041-4045 (1986)). Získané jednořetězcové bifunkční deriváty 20 urokinázy byly přídavkem plazminu přeměněny na odpovídající dvouřetězcové deriváty urokinázy a jejich aktivita zjištěna za pomoci chromogenního substrátu pyro-Glu-Gly-Arg-pnitroanilidu, který je štěpen pouze dvouřetězcovou aktivní urokinázou.
Aktivace bifúnkčních variant urokinázy podle předloženého vynálezu plazminem se provádí v 25 50 mM Tris/HCl pufru, 12 mM chloridu sodného, 0,02 % tweenu 80 při pH 7,4 a teplotě 37 °C.
Poměr bifúnkčních variant urokinázy k plazminu činí asi 100 až 1 500 ku 1, vztaženo na molaritu, nebo asi 8 000 až 36 000 ku 1, vztaženo na enzymové jednotky. Inkubační test se provádí v 50 mM Tris/HCl pufru a 38 mM chloridu sodného při pH 8,8 za přítomnosti 0,36 μΜ aprotininu (pro blokování plazminu) a 0,27 mM substrátu pyro-GIu-Gly-Arg-p-nitroanilidu při 30 teplotě 37 °C. V závislosti na koncentraci bifúnkčních variant urokinázy se reakce přeruší po 5 až minutové inkubaci přídavkem 50% kyseliny octové a měří se extinkce při 405 nm. Podle údajů výrobce substrátu (Kabi Vitrum, Švédsko) odpovídá podle tohoto návodu změna extinkce 0,05 pro minutu při 405 nm aktivitě urokinázy 25 Ploug-jednotek pro 1 ml testovaného roztoku. Bifunkční deriváty urokinázy podle předloženého vynálezu vykazují specifickou aktivitu 35 v rozmezí 120 000 až 155 000 Ploug-jednotek pro jeden mg vyčištěného proteinu. Bílkovinný obsah roztoků byl zjištěn pomocí BCA-Assay firmy Pierce.
d) Izolace a čištění
5 až 6 hodin po indukci se ukončí fermentace, prováděná za podmínek popsaných v odstavci lb) (hustota 5 až 6 OD při 578 nm), načež se buňky odstředí. Buněčný sediment se resuspenduje ve 200 ml vody a rozloží se ve vysokotlakém homogenizátoru. Po nové centrifugaci se sediment, který obsahuje celé množství jednořetězcové bifunkční varianty urokinázy, rozpustí v 500 ml 5 M guanidiniumhydrochloridu, 40 mM cysteinu, 1 mM EDTA při hodnotě pH 8,0 a zředí se
2 000 ml 25 mM Tris/HCl pufru s hodnotou pH 9,0. Reake se ukončí po asi 12 hodinách.
Získané bifunkční varianty urokinázy se po přídavku 8 g křemeliny a dvouhodinovém míchání úplně navážou na křemelinu. Navázaná křemelina se oddělí a promyje se acetátovým pufrem (pH4). Varianty urokinázy se eluují 0,5 M tetramethylamoniumchloridem (TMAC) v 0,1 M 50 acetátového pufru (pH 4). Po dvojím chromatografickém dělení (sloupec měď-chelát a kationtoměnič) se varianty urokinázy získají v čisté formě. Po N-terminální sekvenční analýze byla jednak dokázána jednořetězcová povaha a jednak požadovaná amino-terminální sekvence. Charakterizace změněné karboxy-terminální oblasti jednotlivých variant bílkovinářskou chemií bylo po modifikovaném CNBr-štěpení proteinu (rozpuštěného v 1 ml 90% kyseliny mravenčí a 55 1 ml kyseliny heptafluormáselné), dosaženo štěpením peptidového řetězce za tryptofanovými
-12CZ 288536 B6 zbytky. Karboxy-terminální peptid byl před sekvenční analýzou oddělen a vyčištěn pomocí HPLC (High Pressure Liquid Chromatography).
Všechny izolované bifunkční varianty urokinázy, uvedené v tabulce 2, nevykazují v přímém testu 5 aktivity na chromogenním substrátu pro urokinázu žádnou aktivitu nebo vykazují jen velmi slabou aktivitu (pod 1 200 Ploug-jednotek projeden mg vyčištěného proteinu). Teprve po štěpení plazminem (podmínky jsou uvedeny v odstavci lc)) bylo dosaženo enzymové aktivity v rozmezí 12 000 až 155 000 Ploug-jednotek pro jeden mg vyčištěného proteinu. Všechny varianty urokinázy byly pro v E. coli K12 JM103 exprimovány jako jednořetězcové proteiny.
Tabulka 2
Bifunkční varianty urokinázy (bU) podle vynálezu obecného vzorce I M4-X^-Y-^
| bU | Xi | Y1 |
| Mil | Ser-Pro-Pro-Ser-ProPro-Gly-Gly-Phe | A3-3 Y2=Pro, Y3= vazba mezi Pro a Gly, Y4=OH |
| M12 | Ser-Pro-Val-Lys-AlaPhe | A3·) Y2=Pro, ^3=· vazba mezi Pro a Gly, Y4=OH |
| Ml 3 | Ser-Pro-Pro-Ser-ProPro-Ser-Pro-Pro-GlyGly-Phe | A1) Y2=Pro, Y3=. vazba mezi .. Pro a Gly, Y4=OH |
| Ml 4 | Ser-Pro-Pro-Gly-GlyPhe | Al) Y2=Pro, Y3- vazba mezi Pro a Gly, Y4=OH |
| Ml 5 | Ser-Pro-Pro-Ser-Pro- Pro-Ser-Pro-Pro-Gly- Gly-Phe-Gly | Al) Y2=Pro, Y3= vazba mezi Pro a Gly, Y4=OH |
| Ml 6 | Ser-Pro-Val-Lys-AlaTrp-Gly | Al) Y2-Pro, Y3=Leu, Y4=Gln |
| M17 | Ser-Pro-Val-Lys-AlaPhe-Gly | A1) Y2=Pro, Y3=Leu, Y4=Gln |
| Ml 8 | Ser-Pro-Val-Arg-AlaPhe-Gly | A3·) Y2=Pro, Y3=Leu, Y4=GIn |
| Ml 9 | Ser-Pro-Val-Lys-Ala- Tyr-Gly | A3·) . Y2=Pro, Y3=Leu, Y4=Gln |
| M20 | Ser-Pro-Val-Val-AlaPhe-Gly | A3·) Y2=Pro, Y3=Leu, Y4=Gln |
| M21 | Ser-Pro-Val-Arg-Ala- Phe | Al) Y2=Pro, Y3=Leu, Y4»Gln |
-13CZ 288536 B6
Tabulka 2 - pokračování
| bU | xi | *1 |
| M22 | Ser-Pro-Val-Lys-AlaTyr | A1) Υ2βΡΓο, Y3=Leu, Y4=Gln |
| M23 | Ser-Pro-Val-Val-Ala- Phe | A1) Y2=Proz Y3=Leuz Y4=Gln |
| M24 | Ser-Pro-Val-Lys-AlaTrp | AD Y2=Pro, Y3=Leu, Y4=Gln |
| M25 | Ser-Pro-Val-Lys-AlaPhe | Al) Y2=Pro, Y3=Leu, Y4=Gln |
| M26 | Ser-Pro-Val-Val-ValPhe | A1) Y2=Pro, Y3=Leu, Y4=Gln |
| M27 | Ser-Pro-Val-Glu-Val- Phe | A1) Y2=Pro, Y3=Leu, Y4=Gln |
| M28 | Ser-Pro-Val-Val-ValVal | aD Y2=Valz Y3=Leuz Y4=Gln |
| M29 | Ser-Pro-Val-Val-Ala- Phe | β2) Y2=Pro |
| M30 | Ser-Leu-Val-Val-AlaPhe | AÍ) Y2=sProz Y3=Leuz Y4=Gln |
| M31 | Ser-Leu-Val-Lys-AlaPhe | aD Y2=Proz Y3=Leuz Y4=Gln |
| M32 | Ser-Pro-Val-Lys-AlaPhe | B2) Y2=Pro |
| M33 | přímá vazba mezi M4 a Y1 | A1) Y2=Proz Y3=Leu, Y4=Gln |
A = Y2”Arg-Pro-Y3-Gly-Gly-Gly-Gly-Asn-Gly-Asp-PheGlu-Glu-Ile-Pro-Glu-Glu-Tyr-Leu-Y4
2) b = Y2_Ary-Pro-Phe-Leu-Leu-Arg-Asn-Pro-Asn-Asp-LysTyr-Glu-Pro-Phe-Trp-Glu-Asp-Glu-Glu-Lys-Asn-Glu
-14CZ 288536 B6
Příklad 2
Farmakologické zkoušky
Zjišťování trombininhibičního účinku
Inhibiční aktivita bifunkčních variant urokinázy byla zjišťována měřením trombinového času ve směsi 200 μΐ lidské citrátové plazmy, zředěné 1:10 veronalovým pufrem, s 50 μΐ trombinového ίο roztoku (0,2 jednotky) a 50 μΐ vodného roztoku, obsahujícího 0,5 až 50 μg bifunkční varianty urokinázy. Potom byla měřena doba až do vytvoření fibrinových vloček. V následující tabulce 3 jsou uvedeny naměřené inhibiční faktory, které udávají prodloužení trombinového času za přítomnosti vždy 10 pg bifunkční varianty urokinázy podle předloženého vynálezu.
Bylo rovněž zjišťováno prodloužení trombinového času v závislosti na koncentraci a je pro bifunkční varianty urokinázy M12, M23, M29, M32, M33 a jako srovnání pro M4 znázorněno graficky na obr. 3. Na rozdíl od bifunkčních variant urokinázy podle předloženého vynálezu nebyla prodloužena doba do sražení ani vlivem M4, to znamená pro aminokyselinovou sekvenci 47Ser až 41'Leu prourokinázy bez glykosidu podle obr. 1, ani vlivem prourokinázy bez glykosidu 20 (saruplase), ani vlivem LUK samotné v dávkování 1 mg.
Tabulka 3
Prodloužení trombinového času vlivem bifunkčních variant urokinázy podle vynálezu obecného 25 vzorce I M4-X]Yi
| Bifunkční varianta urokinázy | inhibiční faktor1) |
| Mil | 1,8 |
| M12 | 4,6 |
| M13 | 1,7 |
| M14 | 1,8 |
| M15 | 2,5 |
| M16 | 3,2 |
| M17 | 3,1 |
| M18 | 2,9 |
| M19 | 2,0 |
| M20 | 2,2 |
| M21 | 2,3 |
| M22 | 3,7 |
| M23 | 5,3 |
| M24 | 6,2 |
| M25 | 2,9 |
| M26 | 3,2 |
| M27 | 2,0 |
| M28 | 2,1 |
| M29 | 2,6 |
| M30 | 3,4 |
| M31 | 2,0 |
| M32 | 3,0 |
| M33 | 2,0 |
') vztaženo na účinek 10 pg proteinu
Inhibiční faktor = kvocient z trombinového času za přítomnosti inhibitoru a trombinového času 30 za nepřítomnosti inhibitoru.
-15CZ 288536 B6
Farmakologické vlastnosti bifunkčních variant urokinázy M12 a M23 v pokusech na zvířatech
Na farmakologickém in vivo modelu byl testován účinek bifunkčních variant urokinázy Ml2 a M23 na trombolýzu arterielní krevní ucpávky ve srovnání se saruplasou (prourokináza bez 5 glykosidu). K tomu byl narkotizovaným králíkům temporálně izolován cca 1 cm dlouhý segment arterie a vedlejší větší injikován trombin a lidský fibrinogen, značený jodem125. Tím se vytvořil trombus, který vedl ke kompletnímu ucpání cévy. Velikost vytvořeného trombu byla zjišťována pomocí inkorporované radioaktivity lidského fíbrinu extrakorporálním gama-detektorem. Elektromagnetické měření průtoku krve a evidence radioaktivity trombu se provádělo 10 kontinuálně po celou dobu trvání pokusu.
Fibrinolytický účinek tedy byl nejen kvantifikován jako zprůchodnění trombolyticky uzavřených cév, tak také jako odbourávání radioaktivně značeného inkorporovaného fíbrinu trombu.
Před aplikací bifunkčních variant urokinázy podle předloženého vynálezu, jakož i 30, 60 a 90 minut po aplikaci variant urokinázy, byly odebrány vzorky krve, ve kterých byly zjišťovány koncentrace plazmatického fíbrinogenu. Bylo vždy aplikováno 6 mg/kg Ml2, M23 a saruplasy intravenózně jako bolusinjekce. Protože M12 a M13 mají na rozdíl od saruplasy doplňkový antikoagulační účinek, byla saruplasa kombinovaná ve 4. pokusné skupině s antikoagulantem 20 heparinem (150 U/kg i.v.-bolus). Pokusné skupiny měly vždy 6 zvířat.
Po devadesátiminutovém trvání pokusu vykazovala trombolýza značeného fibrinového trombu ± 11 % pro Ml2, 43 ± 12 % pro M23, 22 ± 5 % pro saruplasu a 39 ±15 % pro kombinaci saruplasy a heparinu. Bolusaplikace M12 a M23 vedla u všech šesti zvířat ke zprůchodnění 25 trombolyticky ucpaných cév; při aplikaci saruplasy mohla být uvolněna céva u pěti ze šesti zvířat a při aplikaci saruplasy a heparinu u čtyřech ze šesti zvířat. Maximální množství reperfusiovaného toku (v % výchozí hodnoty) činilo 95 ± 10 % pro M12 a 82 ± 9 % pro M23 a lišilo se signifikantně od maximálního množství reperfusiované tekutiny 43 ± 12 % pro saruplasu. Maximální množství reperfusiované tekutiny 58 ± 8 % při použití saruplasy a heparinu 30 leželo mezi výsledky s M12 a M23 z jedné strany a se saruplasou ze strany druhé.
Celkový fibrinolytický účinek byl vyjádřen jako plocha reperfusiovaného toku (jako % výchozího toku) po devadesátiminutovém trvání pokusu. Tento celkový efekt činil 4.502 ± 1.127 %. min pro Ml2 a 4.270 ± 855 % . min pro M23 a byl pro obě varianty urokinázy 35 podle předloženého vynálezu signifikantně větší než je hodnota pro saruplasu 1.519 ±643 % .
min. Pro kombinované podání saruplasa-heparin byl naměřen celkový efekt 2.217 ± 761 % . min, který nebyl signifikantně lepší než při podání samotné saruplasy a byl podstatně vzdálený od výsledků s M12 a M23. Výsledky jsou uvedené v následující tabulce 4.
Tabulka 4
Trombolytický účinek po i.v.-bolusaplikaci; trombóza femorální arterie, narkotizovaní králíci
| Polypeptid | dávka | % 125J-fibrinolýza | max. reperfusní tok (% pův. hodn.) | kumulativní reperf. tok (%.min) |
| M12 | 6 mg/kg | 46 ± 11 | 95 ± 10* | 4502 ±1127 |
| M23 | 6 mg/kg | 43 ±12 | 82 ±9* | 4270 ± 885* |
| saruplasa | 6 mg/kg | 22 ±5 | 43 ± 12 | 119±643 |
| saruplasa + heparin | 6 mg/kg 150 U/kg | 39± 15 | 58 ±8 | 2217 ±761 |
* p < 0,05 vs saruplasa
-16CZ 288536 B6
Překvapivě bylo zjištěno, že jak po bolusaplikaci Ml2, tak také po bolusaplikaci M23 klesaly koncentrace fibrinogenu v plazmě signifikantně méně, než po bolusaplikaci saruplasy. Výsledky jsou shrnuty v následující tabulce 5.
Tabulka 5
Účinek bolusaplikace M12 a M23 ve srovnání se saruplasou bez heparinu a s heparinem na pokles koncentrace fibrinogenu v plazmě; narkotizovaní králíci
| Polypeptid | dávka | pokles plazmového fibrinogenu (% změny oproti výchozí hodnotě) | ||
| doba po aplikaci | ||||
| 30 min | 60 min | 90 min | ||
| M12 | 6 mg/kg | -19 ±9 | -20 ±9* | -19 ±9* |
| M23 | 6 mg/kg | -20±11* | -21 ±11* | -20±11* |
| saruplasa | 6 mg/kg | -64 ±7 | —66 ± 6 | -67 ±6 |
| saruplasa + heparin | 6 mg/kg 150U/kg | n.s.1) | -46±8 | -45 ±9 |
* p < 0,05 vs saruplasa ') n.s. = nebylo stanoveno
Výsledky ukazují, že bifunkční deriváty urokinázy M12 a M23 uvolní a upraví opět plný průchod krve trombotickými cévami, které byly ucpány tromby. Tohoto účinku bylo dosaženo jednorázovou bolusaplikaci M12, popřípadě M23 na zvířatech bez podání heparinu. Překvapující bylo, že tyto látky měly ve srovnání se saruplasou silnější fíbrinolytický účinek, spojený s menší spotřebou plazmatického fibrinogenu. To znamená, že M12 a M23 vykazují signifikantně vyšší fibrinovou specifítu ve srovnání se saruplasou.
Lepší ochrana plazmatického fibrinogenu pomocí M12 a M23 ve srovnání se saruplasou znamená, že schopnost srážení krve zůstane lépe zachován a tím je sníženo nebezpečí nekontrolovatelného krvácení jako možné komplikace systemického odbourávání fibrinogenu. M12 a M23 jsou ve vztahu k riziku hemostazeologických vedlejších účinků tedy jistější než saruplasa.
-17CZ 288536 B6
Sekv. ID č. 1
Délka: 1182 BP, 393 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein M24 (pSJÍOl)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| '1 | ATG AGC AAA ACT TGC TAC GAA GGT AAC | GGT CAC | TTC TAC CGT GGT AAG | |||||||||||||
| Met Ser Lys Thr Cys | Tyr | Glu | Gly | Asn | Gly 10 | His | Phe Tyr | Arg Gly Lys 15 | ||||||||
| 1 | 5 | |||||||||||||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly | Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | |
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly | Lys | His | Asn | Tyr Cys | Arg | Asn | Pro | Asp | Asn | Arg Arg | |||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | Val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA | AAA | CCG | TCT | TCT | CCG | CCG |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp | Gly | Lys | Lys | Pro | Ser | Ser | Pro | Pro | |
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT |
| Glu | Glu | Leu | Lys | Phe | Gin | Cys | Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | |
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA | AAC | CAG | CCG | TGG | TTC |
| Lys | Ile | Ile | Gly Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | ||
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | ile | Tyr Arg | Arg | His | Arg | Gly Gly | Ser | Val | Thr | Tyr | Val | Cys | |||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly Gly | Ser | Leu | Ile | Ser | Pro | Cys | Trp | Val | Ile | Ser | Ala | Thr | His Cys | |||
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC | ATC | GAC | TAC | CCG | AAA | AAA | GAA | GAC | TAC | ATC | GTT TAC | CTC | GGC | CGT | |
| Phe | Ile | Asp Tyr | Pro | Lys | Lys | Glu | Asp | Tyr | Ile | Val Tyr | Leu | Gly Arg | ||||
| 165 | 170 | 175 |
-18CZ 288536 B6
| 528 | TCT CGT Ser Arg | TTA Leu | AAC TCT Asn Ser 180 | AAC ACC CAG GGT GAA ATG | AAA TTC GAA GTT GAA | |||||||||||
| Asn | Thr Gin | Gly 185 | Glu Met | Lys Phe Glu 190 | Val Glu | |||||||||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC |
| Asn | Leu | Ile | Leu | His | Lys | Asp | Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | |
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA | AAA | ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT |
| Asn | Asp | Ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | Glu | Gly | Arg | Cys | Ala | |
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | Ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | Glu | Ile | Thr | Gly | Phe | Gly | Lys | Glu | |
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | Tyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Met | Thr | Val | Val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr | Tyr | Gly | Ser | |
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| Glu | Val | Thr | Thr | Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | |
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA | GTT | TGC | TCT | CTC | CAG |
| Asp | Ser | Cys | Gin | Gly | Asp | Ser | Gly Gly | Pro | Leu | Val | Cys | Ser | Leu | Gin | ||
| 305 | 310 | 315 | 320 | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly Arg | Met | Thr | Leu | Thr | Gly | Ile | Val | Ser | Trp | Gly Arg | Gly | Cys | Ala | |||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | val | Ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT | CAC | ACC | AAA | GAA | GAA | AAC | GGT | CTG | GCT | CTG | AGC | CCG |
| Trp | Ile | Arg | Ser | His | Thr | Lys | Glu | Glu | Asn | Gly | Leu | Ala | Leu | Ser | Pro | |
| 355 | 360 | 365 | ||||||||||||||
| 1104 | GTT | AAG | GCT | TGG | CCG | CGG | CCG | CTG | GGT | GGT | GGT | GGT | AAC | GGT | GAC | TTC |
| Val | Lys | Ala | Trp | Pro | Arg | Pro | Leu | Gly Gly | Gly | Gly | Asn | Gly Asp | Phe | |||
| 370 | 375 | 380 | ||||||||||||||
| 1152 | GAA | GAA | ATC | CCG | GAA | GAA | TAC | CTG | CAA TAA | |||||||
| G1U | Glu | Ile | Pro | Glu | Glu | Tyr | Leu | Gin | ||||||||
| 385 | 390 |
-19CZ 288536 B6
Sekv. ID č. 2
Délka: 1182 BP,393 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein M25 (pSJ102)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG AGC AAA ACT TGC TAC GAA GGT AAC | GGT CAC | TTC TAC CGT GGT AAG | |||||||||||||
| Met Ser Lys Thr Cys | Tyr | Glu | Gly | Asn | Gly 10 | His | Phe Tyr | Arg Gly Lys 15 | ||||||||
| 1 | 5 | |||||||||||||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly Lys | His | Asn | Tyr Cys | Arg | Asn | Pro | Asp | Asn | Arg Arg | ||||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | Val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA AAA | CCG | TCT | TCT | CCG | CCG | |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp | Gly | Lys | Lys | Pro | Ser | Ser | Pro | Pro | |
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT |
| Glu | Glu | Leu | Lys | Phe | Gin | Cys | Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | |
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA AAC | CAG | CCG | TGG | TTC | |
| Lys | Ile | Ile | Gly | Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | |
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | Ile | Tyr Arg Arg | His | Arg | Gly Gly | Ser | Val | Thr | Tyr | Val | Cys | ||||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly Gly | Ser | Leu | Ile | Ser | Pro Cys Trp | Val | Ile | Ser | Ala | Thr | His | Cys | ||||
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC ATC | GAC | TAC | CCG | AAA | AAA GAA | GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT | ||
| Phe | Ile | Asp | Tyr | Pro | Lys | Lys Glu | Asp | Tyr | Ile | Val | Tyr | Leu | Gly Arg | |||
| 165 | 170 | 175 |
-20CZ 288536 B6
| 528 | TCT CGT | TTA AAC | TCT Ser | AAC ACC CAG GGT GAA ATG AAA TTC GAA GTT GAA | ||||||||||||
| Ser | Arg | Leu | Asn 180 | Asn Thr | Gin | Gly 185 | Glu Met | Lys Phe | Glu Val 190 | Glu | ||||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC |
| Asn | Leu | Ile | Leu | His | Lys | Asp | Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | |
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA | AAA | ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT |
| Asn | Asp | Ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | Glu | Gly Arg | Cys | Ala | ||
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | Ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | G1U | Ile | Thr | Gly | Phe | Gly | Lys | Glu | |
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | Tyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Met | Thr | Val | Val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr | Tyr | Gly | Ser | |
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| Glu | Val | Thr | Thr | Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | |
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA | GTT | TGC | TCT | CTC | CAG |
| Asp | Ser | Cys | Gin | Gly | Asp | Ser | Gly Gly | Pro | Leu | Val | Cys | Ser | Leu | Gin | ||
| 305 | 310 | 315 | 320 | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ΑΤΓ | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly Arg | Met | Thr | Leu | Thr | Gly | Ile | Val | Ser | Trp | Gly Arg | Gly | Cys | Ala | |||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | Val | Ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT CAC | ACC | AAA | GAA | GAA | AAC | GGT | CTG | GCT | CTG | AGC | CCG | |
| Trp | Ile | Arg | Ser | His | Thr | Lys | Glu | Glu | Asn | Gly | Leu | Ala | Leu | Ser | Pro | |
| 355 | 360 | 365 | ||||||||||||||
| 1104 | GTT | AAG | GCT | TTC | CCG | CGG | CCG | CTG | GGT | GGT | GGT | GGT | AAC | GGT | GAC | TTC |
| Val | Lys | Ala | Phe | Pro | Arg | Pro | Leu | Gly Gly Gly Gly | Asn | Gly Asp | Phe | |||||
| 370 | 375 | 380 | ||||||||||||||
| 1152 | GAA | GAA | ATC | CCG | GAA | GAA | TAC | CTG | CAA TAA | |||||||
| Glu | Glu | Ile | Pro | Glu | Glu | Tyr | Leu | Gin |
385 390
-21CZ 288536 B6
Sekv. ID č. 3
Délka: 1182 BP, 393 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein M26 (pSJl 03)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG AGC AAA ACT TGC TAC GAA GGT AAC GGT CAC TTC TAC CGT GGT AAG | |||||||||||||||
| Met 1 | Ser Lys | Thr | Cys Tyr 5 | Glu | Gly | Asn | Gly 10 | His | Phe | Tyr | Arg Gly Lys 15 | |||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly | Lys | His | Asn | Tyr | Cys | Arg | Asn | Pro | Asp | Asn | Arg Arg | ||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | Val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA | AAA | CCG | TCT | TCT | CCG | CCG |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp | Gly | Lys | Lys | Pro | Ser | Ser | Pro | Pro | |
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT |
| Glu | Glu | Leu | Lys | Phe | Gin | Cys Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | ||
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA | AAC | CAG | CCG | TGG | TTC | |
| Lys | Ile | Ile | Gly Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | ||
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | Ile | Tyr Arg Arg | His | Arg | Gly | Gly | Ser | Val | Thr | Tyr | Val | Cys | |||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly Gly | Ser | Leu | Ile | Ser | Pro | Cys | Trp | Val | Ile | Ser | Ala | Thr | His | Cys | ||
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC | ATC | GAC | TAC | CCG | AAA | AAA | GAA | GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT |
| Phe | Ile | Asp | Tyr | Pro | Lys | Lys | Glu | Asp | Tyr | Ile | Val | Tyr | Leu | Gly Arg | ||
| 165 | 170 | 175 |
-22CZ 288536 B6
| 528 | TCT Ser | CGT TTA AAC TCT AAC ACC CAG GGT GAA ATG AAA TTC GAA GTT GAA | ||||||||||||||
| Arg | Leu | Asn 180 | Ser | Asn | Thr Gin | Gly 185 | Glu | Met Lys | Phe | Glu Val Glu 190 | ||||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC |
| Asn | Leu | Ile | Leu | His | Lys | Asp | Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | |
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA | AAA | ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT |
| Asn | Asp | ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | Glu | Gly Arg | Cys | Ala | ||
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | Ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | Glu | Ile | Thr | Gly | Phe | Gly | Lys | Glu | |
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | Tyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Met | Thr | Val | val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr Tyr | Gly | Ser | ||
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| Glu | Val | Thr | Thr | Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | |
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA | GTT | TGC | TCT | CTC | CAG |
| Asp | Ser | Cys | Gin | Gly Asp | Ser | Gly Gly | Pro | Leu | Val | Cys | Ser | Leu | Gin | |||
| 305 | 310 | 315 | 320 | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly Arg | Met | Thr | Leu | Thr | Gly | Ile | Val | Ser | Trp | Gly Arg | Gly | Cys | Ala | |||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GIT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | Val | Ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT | CAC | ACC | AAA | GAA | GAA AAC | GGT | CTG | GCT | CTG | AGC | CCG | |
| Trp | Ile | Arg | Ser | His | Thr | Lys | Glu | Glu | Asn | Gly | Leu | Ala | Leu | Ser | Pro | |
| 355 | 360 | 365 | ||||||||||||||
| 1104 | GTT | GTA | GTT | TTC | CCG | CGG | CCG | CTG | GGT | GGT | GGT | GGT | AAC | GGT | GAC | TTC |
| Val | Val | Val | Phe | Pro | Arg | Pro | Leu | Gly Gly | Gly Gly | Asn | Gly Asp | Phe | ||||
| 370 | 375 | 380 | ||||||||||||||
| 1152 | GAA | . GAA | ATC | CCG | GAA | GAA | TAC | CTG | CAA TAA | |||||||
| Glu | G1U | ile | Pro | G1U | Glu | Tyr | Leu | Gin |
385 393
-23CZ 288536 B6
Sekv. ID č. 4
Délka: 1182 BP, 393 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein M27 (pSJ104)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG AGC AAA ACT | TGC TAC GAA | GGT AAC GGT CAC TTC TAC CGT GGT AAG | |||||||||||||
| Met 1 | Ser | Lys | Thr | Cys Tyr 5 | Glu | Gly Asn | Gly 10 | His | Phe Tyr | Arg | Gly 15 | Lys | ||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | .CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly Lys | His | Asn | Tyr | Cys | Arg | Asn | Pro | Asp | Asn | Arg Arg | |||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA | AAA | CCG | TCT | TCT | CCG | CCG |
| Cys | Met | val | His | Asp | Cys | Ala | Asp | Gly | Lys | Lys | Pro | Ser | Ser | Pro | Pro | |
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT |
| Glu | Glu | Leu | Lys | Phe | Gin | Cys | Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | |
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA | AAC | CAG | CCG | TGG | TTC |
| Lys | Ile | Ile | Gly Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | ||
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | Ile | Tyr Arg Arg | His | Arg | Gly | Gly | Ser | Val | Thr | Tyr | Val | Cys | |||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly Gly | Ser | Leu | Ile | Ser | Pro | Cys | Trp | val | Ile | Ser | Ala | Thr | His | Cys | ||
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC ATC | GAC | TAC | CCG | AAA | AAA | GAA | GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT | |
| Phe | Ile | Asp | Tyr | Pro | Lys | Lys | Glu | Asp | Tyr | Ile | Val | Tyr | Leu | Gly Arg |
165 170 175
-24CZ 288536 B6
| 528 | TCT CGT Ser Arg | TTA Leu | AAC TCT Asn Ser 180 | AAC ACC CAG GGT GAA ATG | AAA TTC GAA GTT GAA | |||||||||||
| Asn | Thr Gin | Gly 185 | Glu Met | Lys Phe Glu 190 | Val Glu | |||||||||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC |
| Asn | Leu | Ile | Leu | His | Lys | Asp | Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | |
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA | AAA | ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT |
| Asn | Asp | Ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | Glu | Gly Arg | Cys | Ala | ||
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | Ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | Glu | Ile | Thr | Gly | Phe | Gly | Lys | Glu | |
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | Tyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Met | Thr | Val | Val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr Tyr | Gly | Ser | ||
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| Glu | Val | Thr | Thr | Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | |
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA | GTT | TGC | TCT | CTC | CAG |
| Asp | Ser | Cys | Gin | Gly | Asp | Ser | Gly Gly | Pro | Leu | Val | Cys | Ser | Leu | Gin | ||
| 305 | 310 | 315 | 320 | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly Arg | Met | Thr | Leu | Thr | Gly | Ile | Val | Ser | Trp | Gly Arg | Gly | Cys | Ala | |||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | Val | Ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT | CAC | ACC | AAA | GAA | GAA | AAC | GGT | CTG | GCT | CTG | AGC | CCG |
| Trp | Ile | Arg | Ser | His | Thr | Lys | Glu | Glu | Asn | Gly | Leu | Ala | Leu | Ser | Pro | |
| 355 | 360 | 365 | ||||||||||||||
| 1104 | GTT | GAA | GTT | TTC | CCG | CGG | CCG | CTG | GGT | GGT | GGT | GGT | AAC | GGT | GAC | TTC |
| Val | Glu | Val | Phe | Pro | Arg | Pro | Leu | Gly Gly Gly Gly | Asn | Gly Asp | Phe | |||||
| 370 | 375 | 380 | ||||||||||||||
| 1152 | GAA | GAA | ATC | CCG | GAA | GAA | TAC | CTG | CAA | TAA | ||||||
| Glu | Glu | Ile | Pro | Glu | Glu | Tyr | Leu | Gin | ||||||||
| 385 | 390 |
-25CZ 288536 B6
Sekv. ID č. 5
Délka: 1182 BP, 393 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein M28 (pSJ 105)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG AGC AAA ACT | TGC TAC GAA | GGT AAC GGT CAC TTC TAC CGT GGT AAG | |||||||||||||
| Met 1 | Ser | Lys | Thr | Cys Tyr 5 | Glu | Gly Asn | Gly 10 | His Phe | Tyr Arg Gly Lys 15 | |||||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly | Lys | His | Asn | Tyr | Cys | Arg | Asn | Pro | Asp | Asn | Arg Arg | ||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | Val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA | AAA | CCG | TCT | TCT | CCG | CCG |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp | Gly | Lys | Lys | Pro | Ser | Ser | Pro | Pro | |
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT |
| Glu | Glu | Leu | Lys | Phe | Gin | Cys | Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | |
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA | AAC | CAG | CCG | TGG | TTC |
| Lys | Ile | Ile | Gly | Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | |
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | Ile | Tyr Arg Arg | His | Arg | Gly | Gly | Ser | Val | Thr | Tyr | Val | Cys | |||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly Gly | Ser | Leu | Ile | Ser | Pro | Cys | Trp | Val | Ile | Ser | Ala | Thr | His | Cys | ||
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC ATC | GAC | TAC | CCG | AAA | AAA | GAA | GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT | |
| Phe | Ile | Asp | Tyr | Pro | Lys | Lys | Glu | Asp | Tyr | Ile | Val | Tyr | Leu | Gly Arg |
165 , 170 175
-26CZ 288536 B6
| 528 | TCT CGT Ser Arg | TTA Leu | AAC TCT Asn Ser 180 | AAC ACC CAG GGT GAA ATG AAA TTC GAA GTT GAA | ||||||||||||
| Asn Thr | Gin Gly 185 | Glu | Met | Lys | Phe | Glu Val Glu 190 | ||||||||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC |
| Asn | Leu | Ile | Leu | His | Lys | Asp | Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | |
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA | AAA | ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT |
| Asn | Asp | Ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | Glu | Gly Arg | Cys | Ala | ||
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | Ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | Glu | Ile | Thr | Gly | Phe | Gly | Lys | Glu | |
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | Tyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Meť | Thr | Val | Val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr Tyr | Gly | Ser | ||
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| Glu | Val | Thr | Thr | Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | |
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA | GTT | TGC | TCT | CTC | CAG |
| Asp | Ser | Cys | Gin | Gly | Asp | Ser | Gly | Gly | Pro | Leu | Val | Cys | Ser | Leu | Gin | |
| 305 | 310 | 315 | 320 | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly | Arg | Met | Thr | Leu | Thr | Gly | Ile | Val | Ser | Trp | Gly Arg | Gly | Cys | Ala | ||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | Val | Ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT | CAC | ACC | AAA | GAA | GAA AAC | GGT | CTG | GCT | CTG | AGC | CCG | |
| Trp | Ile | Arg | Ser | His | Thr | Lys | Glu | Glu Asn | Gly | Leu | Ala | Leu | Ser | Pro | ||
| 355 | 360 | 365 | ||||||||||||||
| 1104 | GTT | GTA GTT | GTA | GTG | CGG | CCG | CTG | GGT | GGT | GGT | GGT | AAC | GGT | GAC | TTC | |
| Val | Val | Val | Val | Val | Arg | Pro | Leu | Gly Gly | Gly Gly | Asn | Gly Asp | Phe | ||||
| 370 | 375 | 380 | ||||||||||||||
| 1152 | GAA | GAA ATC | CCG | GAA | GAA | TAC | CTG | CAA TAA | ||||||||
| Glu | Glu | Ile | Pro | Glu | Glu | Tyr | Leu | Gin | ||||||||
| 385 | 390 |
-27CZ 288536 B6
Sekv. ID č. 6
Délka: 1191 BP,396AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein M29 (pSJ106)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG AGC ΆΆΑ ACT TGC TAC GAA GGT AAC | GGT CAC | TTC TAC CGT GGT AAG | |||||||||||||
| Met Ser Lys Thr Cys | Tyr | Glu | Gly | Asn | Gly 10 | His | Phe Tyr | Arg Gly Lys 15 | ||||||||
| 1 | 5 | |||||||||||||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly | Lys | His | Asn | Tyr | Cys | Arg | Asn | Pro | Asp | Asn | Arg | Arg | |
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | Val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA | AAA | CCG | TCT | TCT | CCG | CCG |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp Gly | Lys | Lys | Přo | Ser | Ser | Pro | Pro | ||
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT |
| Glu | Glu | Leu | Lys | Phe | Gin | Cys | Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | |
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA | AAC | CAG | CCG | TGG | TTC |
| Lys | Ile | Ile | Gly Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | ||
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | Ile | Tyr Arg Arg | His | Arg Gly Gly | Ser | Val | Thr | Tyr | Val | Cys | |||||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly Gly | Ser | Leu | Ile | Ser | Pro | Cys | Trp | Val | Ile | Ser | Ala | Thr | His | Cys | ||
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC | ATC | GAC | TAC | CCG | AAA AAA | GAA | GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT | |
| Phe | Ile | Asp | Tyr | Pro | Lys Lys | Glu | Asp | Tyr | Ile | Val | Tyr | Leu | Gly | Arg | ||
| 165 | 170 | 175 |
-28CZ 288536 B6
| 528 | TCT CGT | TTA AAC | TCT Ser | AAC ACC CAG GGT GAA ATG AAA TTC GAA GTT GAA | ||||||||||||
| Ser | Arg | Leu | Asn 180 | Asn Thr | Gin | Gly 185 | Glu Met | Lys Phe | Glu Val 190 | Glu | ||||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC |
| Asn | Leu | Ile | Leu | His | Lys | Asp | Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | |
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA | AAA | ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT |
| Asn | Asp | Ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | Glu | Gly Arg | Cys | Ala | ||
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | Ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | Glu | Ile | Thr | Gly | Phe | Gly | Lys | Glu | |
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | iyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Met | Thr | Val | Val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr | Tyr Gly | Ser | ||
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| Glu | Val | Thr | Thr | Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | |
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA | GTT | TGC | TCT | CTC | CAG |
| Asp | Ser | Cys | Gin | Gly Asp | Ser | Gly | Gly | Pro | Leu | Val | Cys | Ser | Leu | Gin | ||
| 305 | 310 | 315 | 32Ó | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly Arg | Met | Thr | Leu | Thr | Gly | ile | Val | Ser | Trp | Gly Arg Gly Cys | Ala | |||||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | Val | Ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT | CAC | ACC | AAA | GAA | GAA | AAC | GGT | CTG | GCT | CTG | AGC | CCG |
| Trp | Ile | Arg | Ser | His | Thr | Lys | Glu | Glu | Asn | Gly | Leu | Ala | Leu | Ser | Pro | |
| 355 | 360 | 365 | ||||||||||||||
| 1104 | GTT | GTT | GCT | TTC | CCG | CGG | CCG | TTC | CTG | CTG | CGG | AAC | CCG | AAC | GAC | AAA |
| Val | Val | Ala | Phe | Pro | Arg | Pro | Phe | Leu | Leu | Arg | Asn | Pro | Asn | Asp | Lys | |
| 370 | 375 | 380 | ||||||||||||||
| 1152 | TAC | GAA | CCG | TTC | TGG | GAA | GAC | GAA | GAA | AAA | AAC | GAA | TAA | |||
| Tyr | Glu | Pro | Phe | Trp | Glu | Asp | Glu | Glu | Lys | Asn | Glu |
385 390 395
-29CZ 288536 B6
Sekv. ID č. 7
Délka: 1182 BP, 393 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineami
Druh molekuly: syntetický gen pro protein M30 (pSJ109)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG AGC AAA ACT TGC TAC GAA GGT AAC GGT CAC TTC TAC CGT GGT AAG | |||||||||||||||
| Met 1 | Ser Lya | Thr | Cys Tyr 5 | Glu | Gly | Asn | Gly 10 | His | Phe | Tyr | Arg | Gly 15 | Lys | |||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly | Lys | His | Asn | Tyr Cys Arg | Asn | Pro | Asp | Asn | Arg Arg | ||||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | Val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA | AAA | CCG | TCT | TCT | CCG | CCG |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp | Gly | Lys | Lys | Pro | Ser | Ser | Pro | Pro | |
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT |
| Glu | Glu | Leu | Lys | Phe | Gin | Cys Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | ||
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA | AAC | CAG | CCG | TGG | TTC |
| Lys | Ile | Ile | Gly Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | ||
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | Ile | Tyr Arg | Arg | His | Arg | Gly Gly | Ser | Val | Thr | Tyr | Val | Cys | |||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly Gly | Ser | Leu | Ile | Ser | Pro | Cys | Trp | Val | Ile | Ser | Ala | Thr | His | Cys | ||
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC ATC | GAC | TAC | CCG | AAA | AAA | GAA GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT | ||
| Phe | Ile | Aap Tyr | Pro | Lys | Lys | Glu Asp | Tyr | ile | Val | Tyr | Leu | Gly Arg | ||||
| 165 | 170 | 175 |
-30CZ 288536 B6
| 528 | TCT CGT Ser Arg | TTA Leu | AAC TCT Asn Ser 180 | AAC ACC CAG GGT GAA ATG AAA TTC GAA GTT GAA | ||||||||||||
| Asn Thr | Gin Gly 185 | Glu | Met | Lys | Phe | Glu Val Glu 190 | ||||||||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC |
| Asn | Leu | Ile | Leu | His | Lys | Asp | Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | |
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA | AAA | ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT |
| Asn | Asp | Ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | Glu | Gly Arg | Cys | Ala | ||
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | Ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | Glu | Ile | Thr | Gly | Phe | Gly | Lys | Glu | |
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | Tyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Met | Thr | Val | Val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr | Tyr | Gly | Ser | |
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| Glu | Val | Thr | Thr | Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | |
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA | GTT | TGC | TCT | CTC | CAG |
| Asp | Ser | Cys | Gin | Gly Asp | Ser | Gly Gly | Pro | Leu | Val | Cys | Ser | Leu | Gin | |||
| 305 | 310 | 315 | 320 | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly Arg | Met | Thr | Leu | Thr | Qly | Ile | Val | Ser | Trp | Gly Arg | Gly | Cys | Ala | |||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | Val | Ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT | CAC | ACC | AAA | GAA | GAA | AAC | GGT | CTG | GCT | CTG | AGC | GTG |
| Trp | Ile | Arg | Seř | His | Thr | Lys | Glu | Glu | Asn | Gly | Leu | Ala | Leu | Ser | Leu | |
| 355 | 360 | 365 | ||||||||||||||
| 1104 | GIT | GTT | GCT | TTC | CCG | CGG | CCG | CTG | GGT | GGT | GGT | GGT | AAC | GGT | GAC | TTC |
| Val | Val | Ala | Phe | Pro | Arg | Pro | Leu | Gly Gly Gly | Gly | Asn | Gly Asp | Phe | ||||
| 370 | 375 | 380 | ||||||||||||||
| 1152 | GAA | GAA | ATC | CCG | GAA | GAA | TAC | CTG | CAA | TAA | ||||||
| Glu | Glu | Ile | Pro | Glu | Glu | Tyr | Leu | Gin | ||||||||
| 385 | 390 |
-31CZ 288536 B6
Sekv. ID č. 8
Délka: 1182 BP, 393 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein M31 (pSJl 11)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG AGC AAA ACT | TGC TAC GAA | GGT AAC GGT CAC TTC TAC CGT GGT AAG | |||||||||||||
| Met 1 | Ser | Lys | Thr | Cys Tyr 5 | Glu | Gly Asn | Gly 10 | His Phe | Tyr Arg | Gly Lys 15 | ||||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly Lys | His | Asn | Tyr Cys | Arg | Asn | Pro | Asp | Asn | Arg | Arg | |||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | Val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA | AAA | CCG | TCT | TCT | CCG | CCG |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp | Gly | Lys | Lys | Pro | Ser | Ser | Pro | Pro | |
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT |
| Glu | Glu | Leu | Lys | Phe | Gin | Cys | Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | |
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA | AAC | CAG | CCG | TGG | TTC |
| Lys | Ile | Ile | Gly | Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | |
| 115 | 120 | .125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | Ile | Tyr Arg Arg | His | Arg Gly Gly | Ser | Val | Thr | Tyr | Val | Cys | |||||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly | Gly | Ser | Leu | Ile | Ser | Pro | Cys | Trp | Val | Ile | Ser | Ala | Thr | His | Cys | |
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC | ATC | GAC | TAC | CCG | AAA | AAA | GAA | GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT |
| Phe | Ile | Asp | Tyr | Pro | Lys | Lys | Glu | Asp | Tyr | Ile | Val | Tyr | Leu | Gly Arg |
165 170 175
-32CZ 288536 B6
| 528 | TCT CGT | TTA AAC | TCT Ser | AAC ACC CAG GGT GAA ATG AAA TTC GAA GTT GAA | ||||||||||||
| Ser | Arg | Leu | Asn 180 | Asn Thr | Gin | Gly 185 | Glu Met | Lys Phe | Glu Val 190 | Glu | ||||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC |
| Asn | Leu | Ile | Leu | His | Lys | Asp | Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | |
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA AAA | ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT | |
| Asn | Asp | Ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | Glu | Gly | Arg | Cys | Ala | |
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | Glu | Ile | Thr | Gly | Phe | Gly Lys | Glu | ||
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | Tyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Met | Thr | Val | Val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr | Tyr | Gly | Ser | |
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| Glu | Val | Thr | Thr Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | ||
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA | GTT | TGC | TCT | CTC | CAG |
| Asp | Ser | Cys | Gin | Gly Asp | Ser | Gly | Gly | Pro | Leu | Val | Cys | Ser | Leu | Gin | ||
| 305 | 310 | 315 | 320 | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly Arg | Met | Thr | Leu | Thr | Gly | Ile | Val | Ser | Trp | Gly Arg | Gly | Cys | Ala | |||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | Val | Ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT | CAC | ACC | AAA | GAA | GAA | AAC | GGT | CTG | GCT | CTG | AGC | GTG |
| Trp | ile | Arg | Ser | His | Thr | Lys Glu | Glu | Asn | Gly | Leu | Ala | Leu | Ser | Leu | ||
| 355 | 360 | 365 | ||||||||||||||
| 1104 | GTT | AAA | GCT | TTC | CCG | CGG | CCG | CTG | GGT | GGT | GGT | GGT | AAC | GGT | GAC | TTC |
| Val | Lys | Ala | Phe | Pro | Arg | Pro | Leu | Gly Gly | Gly | Gly | Asn | Gly Asp | Phe | |||
| 370 | 375 | 380 | ||||||||||||||
| 1152 | GAA | GAA | ATC | CCG | GAA | GAA | TAC | CTG | CAA TAA | |||||||
| Glu | Glu | ile | Pro | Glu | Glu | Tyr | Leu | Gin |
385 390
-33CZ 288536 B6
Sekv. ID č. 9
Délka: 1167 BP, 388 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein M33 (pSJl 13)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG Met 1 | AGC AAA ACT TGC | TAC GAA GGT AAC GGT CAC TTC TAC CGT | |||||||||||
| Ser | Lys Thr | Cys 5 | Tyr | Glu | Gly | Asn | Gly His Phe 10 | Tyr Arg | ||||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | ||
| 20 | 25 | 30 | ||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | |
| 35 | 40 | 45 | ||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC |
| Leu | Gly | Leu | Gly | Lys | His | Asn | Tyr | Cys Arg | Asn | Pro | Asp | Asn | ||
| 50 | 55 | 60 | ||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | val | |
| 65 | 70 | 75 | ||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA AAA | CCG | TCT | TCT | |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp Gly | Lys Lys | Pro | Ser | Ser | |||
| 85 | 90 | |||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA AAA ACC | CTA | CGT | CCG | ||
| Glu | Glu | Leu | Lys | Phe | Gin | Cys | Gly | Gin | Lys | Thr | Leu | Arg | Pro | |
| 100 | 105 | 110 | ||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA AAC | CAG | CCG | |
| Lys | Ile | Ile | Gly | Gly Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | ||
| 115 | 120 | 125 | ||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC |
| Ala | Ala | Ile | Tyr Arg Arg | His | Arg | Gly Gly | Ser | Val | Thr | Tyr | ||||
| 130 | 135 | 140 | ||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC |
| Gly Gly | Ser | Leu | Ile | Ser | Pro | Cys | Trp | Val | Ile | Ser | Ala | Thr | ||
| 145 | 150 | 155 | ||||||||||||
| 480 | TTC ATC | GAC | TAC | CCG | AAA | AAA | GAA GAC | TAC | ATC | GTT | TAC | CTC | ||
| Phe | Ile | Asp Tyr | Pro | Lys | Lys | Glu Asp | Tyr | Ile | Val | Tyr | Leu | |||
| 165 | 170 |
GGT AAG Gly Lys 15
TCT GCT Ser Ala
TTG CAG Leu Gin
CGT CGT Arg Arg
CAG GAA
Gin Glu 80
CCG CCG Pro Pro 95
CGT TTT Arg Phe
TGG -TTC Trp Phe
GTT TGC Val Cys
CAC TGC His Cys
160
GGC CGT
Gly Arg
175
-34CZ 288536 B6
| 528 | TCT CGT Ser Arg | TTA Leu | AAC TCT Asn Ser 180 | AAC ACC CAG GGT GAA ATG AAA TTC GAA GTT GAA | ||||||||||||
| Asn Thr | Gin Gly 185 | Glu | Met | Lys | Phe | Glu Val 190 | Glu | |||||||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC |
| Asn | Leu | Ile | Leu | His | Lys | Asp | Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | |
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA | AAA | ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT |
| Asn | Asp | ile | Ala | Leu | Leu | Lys | ile | Arg | Ser | Lys | Glu | Gly Arg | Cys | Ala | ||
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | Ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | Glu | Ile | Thr | Gly | Phe | Gly | Lys | Glu | |
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | Tyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Met | Thr | Val | Val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr | Tyr Gly | Ser | ||
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| G1U | Val | Thr | Thr | Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | |
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA | GTT | TGC | TCT | CTC | CAG |
| Asp | Ser | Cys | Gin | Gly | Asp | Ser | Gly | Gly | Pro | Leu | Val | Cys | Ser | Leu | Gin | |
| 305 | 310 | 315 | 320 | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly Arg | Met | Thr | Leu | Thr | Gly | Ile | Val | Ser | Trp | Gly Arg | Gly | Cys | Ala | |||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | Val | Ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT | CAC | ACC | AAA | GAA | GAA | AAC | GGT | CTG | GCT | CTG | TTC | CCG |
| Trp | ile | Arg | Ser | His | Thr | Lys | Glu | Glu | Asn | Gly | Leu | Ala | Leu | Phe | Pro | |
| 355 | 360 | 365 | ||||||||||||||
| 1104 | CGG | CCG | CTG | GGT | GGT | GGT | GGT | AAC | GGT | GAC | TTC | GAA | GAA | ATC | CCG | GAA |
| Arg | Pro | Leu | Gly | Gly | Gly | Gly | Asn | Gly Asp | Phe | G1U | Glu | Ile | Pro | Glu | ||
| 370 | 375 | 380 | ||||||||||||||
| 1152 | GAA | TAC | CTG | CAA | TAA | |||||||||||
| Glu | Tyr | Leu | Gin | |||||||||||||
| 385 |
-35CZ 288536 B6
Sekv. ID č. 10
Délka: 1191 BP,396AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein M32 (pSJl 14)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG AGC AAA ACT | TGC TAC GAA | GGT AAC GGT CAC TTC TAC CGT GGT AAG | |||||||||||||
| Met 1 | Ser | Lys | Thr | Cys Tyr 5 | Glu | Gly Asn | Gly 10 | His Phe | Tyr Arg Gly Lys 15 | |||||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly | Lys | His | Asn | Tyr | Cys | Arg | Asn | Pro | Asp | Asn | Arg | Arg | |
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | Val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA AAA | CCG | TCT | TCT | CCG | CCG | |
| cys | Met | Val | His | Asp | Cys | Ala | Asp Gly | Lys | Lys | Pro | Ser | Ser | Pro | Pro | ||
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT |
| Glu | Glu | Leu | Lys | Phe | Gin | Cys | Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | |
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA | AAC | CAG | CCG | TGG | TTC |
| Lys | Ile | Ile | Gly Gly | Glu Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | |||
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | Ile | Tyr Arg | Arg | His | Arg | Gly | Gly | Ser | Val | Thr | Tyr | Val | Cys | ||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly | Gly | Ser | Leu | Ile | Ser | Pro | Cys | Trp | Val | Ile | Ser | Ala | Thr | His | Cys | |
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC | ATC | GAC | TAC | CCG | AAA | AAA | GAA | GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT |
| Phe | Ile | Asp | Tyr | Pro | Lys | Lys | Glu | Asp | Tyr | Ile | Val | Tyr | Leu | Gly Arg |
165 170 175
-36CZ 288536 B6
| 528 | TCT CGT Ser Arg | TTA Leu | AAC TCT AAC ACC CAG GGT GAA ATG AAA TTC GAA GTT GAA | |||||||||||||
| Asn Ser Asn 180 | Thr Gin | Gly 185 | Glu Met | Lys | Phe Glu 190 | Val Glu | ||||||||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC |
| Asn | Leu | Ile | Leu | His | Lys | Asp | Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | |
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA | AAA | ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT |
| Asn | Asp | Ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | Glu | Gly Arg | Cys | Ala | ||
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | Ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | Glu | Ile | Thr | Gly | Phe | Gly | Lys | Glu | |
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | Tyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Met | Thr | Val | Val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr | Tyr | Gly | Ser | |
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| Glu | Val | Thr | Thr | Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | |
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA GTT | TGC | TCT | CTC | CAG | |
| Asp | Ser | Cys | Gin | Gly | Asp | Ser | Gly | Gly | Pro | Leu | Val | Cys | Ser | Leu | Gin | |
| 305 | 310 | 315 | 320 | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly Arg | Met | Thr | Leu | Thr | Gly | Ile | Val | Ser | Trp | Gly Arg | Gly | Cys | Ala | |||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | Val | Ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT | CAC | ACC | AAA | GAA | GAA | AAC | GGT | CTG | GCT | CTG | AGC | CCG |
| Trp | Ile | Arg | Ser | His | Thr | Lys | Glu | Glu | Asn | Gly | Leu | Ala | Leu | Ser | Pro | |
| 355 | 360 | 365 | ||||||||||||||
| 1104 | GTT | AAG | GCT | TTC | CCG | CGG | CCG | TTC | CTG | CTG | CGG | AAC | CCG AAC | GAC | AAA | |
| Val | Lys | Ala | Phe | Pro | Arg | Pro | Phe | Leu | Leu | Arg | Asn | Pro | Asn | Asp | Lys | |
| 370 | 375 | 380 | ||||||||||||||
| 1152 | TAC | GAA | CCG | TTC | TGG | GAA | GAC | GAA | GAA AAA | AAC | GAA | TAA | ||||
| Tyr | Glu | Pro | Phe | Trp | Glu | Asp | Glu | Glu Lys | Asn | Glu | ||||||
| 385 | 390 | 395 |
-37CZ 288536 B6
Sekv.IDč. 11
Délka: 1101 BP, 366 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin Topologie: lineární
Druh molekuly: syntetický gen pro protein M4 (pSJ41)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG AGC AAA ACT | TGC TAC GAA | GGT AAC GGT CAC TTC TAC CGT GGT AAG | |||||||||||||
| Met 1 | Ser | Lys | Thr | Cys Tyr 5 | Glu | Gly Asn | Gly 10 | His Phe | Tyr Arg Gly Lys 15 | |||||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly | Lys | His | Asn | Tyr | Cys | Arg | Asn | Pro | Asp | Asn | Arg Arg | ||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | Val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA | AAA | CCG | TCT | TCT | CCG | CCG |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp | Gly | Lys | Lys | Pro | Ser | Ser | Pro | Pro | |
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT |
| Glu | Glu | Leu | Lys | Phe | Gin | Cys | Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | |
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA | AAC | CAG | CCG | TGG | TTC |
| Lys | Xle | Ile | Gly | Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | |
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | Ile | Tyr Arg | Arg | His | Arg | Gly Gly | Sér | Val | Thr | Tyr | Val | Cys | |||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly Gly | Ser | Leu | Ile | Ser | Pro | Cys | Trp | Val | Ile | Ser | Ala | Thr | His | Cys | ||
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC | ATC | GAC | TAC | CCG | AAA | AAA | GAA | GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT |
| Phe | Ile | Asp Tyr | Pro | Lys | Lys | Glu | Asp Tyr | Ile | Val | Tyr | Leu | Gly Arg |
165 170 175
-38CZ 288536 B6
| TCT | CGT | TTA | AAC | TCT | AAC | ACC | CAG | GGT | GAA | ATG | AAA | TTC GAA | GTT | GAA |
| Ser | Arg | Leu | Asn | Ser | Asn | Thr | Gin | Gly | Glu | Met | Lys | Phe Glu | Val | Glu |
| 180 | 185 | 190 | ||||||||||||
| AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG GCT | CAC | CAC |
| Asn | Leu | ile | Leu | His | Lys | Asp | Tyr | Ser | Ala | Asp | Thr | Leu Ala | His | His |
| 195 | 200 | 205 | ||||||||||||
| AAC | GAC | ATC | GCT | CTG | CTA | AAA | ATC | CGT | TCT | AAA | GAA | GGT CGT | TGC | GCT |
| Asn | Asp | Ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | Glu | Gly Arg | Cys | Ala |
| 210 | 215 | 220 | ||||||||||||
| CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | Ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser Met | Tyr | Asn |
| 225 | 230 | 235 | 240 | |||||||||||
| GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | G1U | Ile | Thr | Gly | Phe Gly | Lys | Glu |
| 245 | 250 | 255 | ||||||||||||
| AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | Tyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Met Thr | Val | Val |
| 260 | 265 | 270 | ||||||||||||
| AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC TAC | GGT | TCT |
| Lys | Leu | Ile | Seř | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr Tyr Gly | Ser | |
| 275 | 280 | 285 | ||||||||||||
| GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG TGG | AAA | ACC |
| Glu | Val | Thr | Thr | Lys | Met | Leu | (tys | Ala | Ala | Asp | Pro | Gin Trp | Lys | Thr |
| 290 | 295 | 300 | ||||||||||||
| GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA GTT | TGC TCT | CTC | CAG | |
| Asp | Ser | Cys | Gin | Gly Asp | Ser | Gly | Gly | Pro | Leu Val | Cys Ser | Leu | Gin | ||
| 305 | 310 | 315 | 320 | |||||||||||
| GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT GGT | TGC | GCT |
| Gly Arg | Met | Thr | Leu | Thr | Gly | ile | Val | Ser | Trp Gly Arg Gly Cys | Ala | ||||
| 325 | 330 | 335 | ||||||||||||
| CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | Val | Ser | His Phe | Leu | Pro |
| 340 | 345 | 350 | ||||||||||||
| TGG | ATC | CGT | TCT | CAC | ACC | AAA | GAA | GAA | AAC | GGT | CTG | GCT CTG | ||
| Trp | Ile | Arg | Ser | His | Thr | Lys | Glu | Glu | Asn | Gly | Leu | Ala Leu | ||
| 355 | 360 | 365 |
Sekv. IDč. 12
Délka: 1185 BP, 394 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein Mil (pSJ69)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG AGC AAA ACT | TGC TAC GAA | GGT AAC GGT CAC TTC TAC CGT GGT AAG | |||||||||||||
| Met 1 | Ser | Lys | Thr | Cys Tyr 5 | Glu | Gly Asn | Gly 10 | His | Phe Tyr | Arg | Gly Lys 15 | |||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly | Lys | His | Asn | Tyr | Cys Arg | Asn | Pro | Asp | Asn | Arg Arg | |||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | Val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA | AAA | CCG | TCT | TCT | CCG | CCG |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp | Gly Lys | Lys | Pro | Ser | Ser | Pro | Pro | ||
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT |
| Glu | Glu | Leu | Lys | Phe | Gin | Cys | Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | |
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA | AAC | CAG | CCG | TGG | TTC |
| Lys | ile | Ile | Gly | Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | |
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | ile | Tyr Arg | Arg | His | Arg | Gly | Gly | Ser | Val | Thr | Tyr | Val | Cys | ||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly Gly | Ser | Leu | Ile | Ser | Pro | Cys | Trp | Val | Ile | Ser | Ala | Thr | His | Cys | ||
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC ATC | GAC | TAC | CCG | AAA | AAA GAA | GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT | ||
| Phe | Ile | Asp | Tyr | Pro | Lys | Lys Glu | Asp Tyr | Ile | Val | Tyr | Leu | Gly Arg |
165 170 175
-40CZ 288536 B6
| 528 | TCT CGT TTA AAC TCT AAC ACC CAG GGT GAA ATG AAA TTC GAA GTT GAA | |||||||||||||||
| Ser | Arg | Leu Asn 180 | Ser | Asn | Thr Gin Gly 185 | Glu | Met | Lys | Phe | Glu Val 190 | Glu | |||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC |
| Asn | Leu | Ile | Leu | His | Lys | Asp | Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | |
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA | AAA | ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT |
| Asn | Asp | Ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | Glu | Gly Arg | Cys | Ala | ||
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | Ile | Gin | Thr | Ile | Cys. | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | Glu | Ile | Thr | Gly | Phe | Gly | Lys | Glu | |
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | ASp | Tyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Met | Thr | Val | Val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr | Tyr Gly | Ser | ||
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| Glu | Val | Thr | Thr | Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | |
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA | GTT | TGC | TCT | CTC | CAG |
| Asp | Ser | Cys | Gin | Gly | Asp | Ser | Gly | Gly | Pro | Leu | Val | Cys | Ser | Leu | Gin | |
| 305 | 310 | 315 | 320 | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly Arg | Met | Thr | Leu | Thr | Gly | Ile | Val | Ser | Trp | Gly Arg | Gly Cys | Ala | ||||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | Val | Ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT | CAC | ACC | AAA | GAA | GAA | AAC | GGT | CTG | GCT | CTG | AGC | CCG |
| Trp | Ile | Arg | Ser | His | Thr | Lys | Glu | Glu | Asn | Gly | Leu | Ala | Leu | Ser | Pro | |
| 355 | 360 | 365 | ||||||||||||||
| 1104 | CCG | TCT | CCG | CCG | GGT | GGT | TTC | CCG | CGT | CCG | GGT | GGT | GGT | GGT | AAC | GGT |
| Pro | Ser | Pro | Pro | Gly Gly | Phe | Pro | Arg | Pro | Gly | Gly | Gly Gly | Asn | Gly | |||
| 370 | 375 | 380 | ||||||||||||||
| 1152 | GAC | TTC | GAA | GAA | ATC | CCG | GAA | GAA | TAC | CTG | TAA | |||||
| Asp | Phe | Glu | Glu | Ile | Pro | Glu | Glu | Tyr | Leu | |||||||
| 385 | 390 |
-41CZ 288536 B6
Sekv.IDČ. 13
Délka: 1176BP, 391 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein M12 (pSJ76)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG AGC AAA ACT TGC | TAC GAA GGT AAC GGT CAC TTC TAC CGT GGT AAG | ||||||||||||||
| Met 1 | Ser Lys | Thr | Cys 5 | Tyr | Glu | Gly | Asn Gly His Phe Tyr Arg Gly Lys | |||||||||
| 10 | 15 | |||||||||||||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Mét | Gly Arg | Pro | Cys | Leu | Pro | TiP | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly | Lys | His | Asn | Tyr Cys Arg | Asn | Pro | Asp | Asn | Arg Arg | ||||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA AAA | CCG | TCT | TCT | CCG | CCG | |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp Gly | Lys | Lys | Pro | Ser | Ser | Pro | Pro | ||
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT |
| Glu | Glu | Leu | Lys | Phe | Gin | Cys | Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | |
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA AAC | CAG | CCG | TGG | TTC | |
| Lys | Ile | Ile | Gly Gly | Glu | Phe | Thr | Thr | Ile | Glu Asn | Gin | Pro | Trp | Phe | |||
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT GTT | ACC | TAC | GTT | TGC | |
| Ala | Ala | Ile | Tyr Arg Arg | His | Arg | Gly Gly | Ser Val | Thr | Tyr | Val | Cys | |||||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly Gly | Ser | Leu | Ile | Ser | Pro | Cys | Trp | Val | Ile | Ser | Ala | Thr | His | Cys | ||
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC ATC | GAC | TAC | CCG | AAA | AAA | GAA GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT | ||
| Phe | Ile | Asp | Tyr | Pro | Lys | Lys | Glu Asp Tyr | Ile | Val | Tyr | Leu | Gly Arg | ||||
| 165 | 17Q | 175 |
-42CZ 288536 B6
| 528 | TCT CGT | TTA AAC | TCT Ser | AAC ACC CAG GGT GAA ATG AAA TTC GAA GTT GAA | ||||||||||||
| Ser | Arg | Leu | Asn 180 | Asn Thr | Gin | Gly 185 | Glu Met | Lys Phe | Glu Val 190 | Glu | ||||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC | |
| Asn | Leu | Ile | Leu | His Lys | Asp | Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | ||
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA | AAA ÁTC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT | |
| Asn | Asp | Ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | Glu | Gly | Arg | Cys | Ala | |
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | Ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | Glu | Ile | Thr | Gly | Phe | Gly | Lys | Glu | |
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | Tyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Met | Thr | Val | Val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr | Tyr | Gly | Ser | |
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC | |
| Glu | Val | Thr | Thr | Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | |
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA | GTT | TGC | TCT | CTC | CAG |
| Asp | Ser | Cys | Gin | Gly | Asp | Ser | Gly | Gly | Pro | Leu | val | Cys | Ser | Leu | Gin | |
| 305 | 310 | 315 | 320 | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly Arg | Met | Thr | Leu | Thr | Gly | Ile | val | Ser | Trp | Gly Arg | Gly | Cys | Ala | |||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | Val | Ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT | CAC | ACC | AAA | GAA | GAA AAC | GGT | CTG | GCT | CTG | AGC | CCG | |
| Trp | ile | Arg | Ser | His | Thr | Lys | Glu | Glu Asn | Gly | Leu | Ala | Leu | Ser | Pro | ||
| 355 | 360 | 365 | ||||||||||||||
| 1104 | GTT | AAG | GCT | TTC | CCG | AGG | CCT | GGT | GGT | GGT | GGT | AAC | GGT | GAC | TTC | GAA |
| Val | Lys | Ala | Phe | Pro | Arg | Pro | Gly | Gly Gly Gly Asn Gly Asp | Phe | Glu | ||||||
| 370 | 375 | 380 | ||||||||||||||
| 1152 | GAA | ATC | CCG | GAA | GAA | TAC | CTG | TGA | ||||||||
| Glu | Ile | Pro | Glu | Glu | Tyr | Leu |
385 390
-43CZ 288536 B6
Sekv. ID č. 14
Délka: 1194BP, 397 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein M13 (pSJ77)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG AGC AAA ACT TGC TAC GAA GGT AAC | GGT CAC | TTC TAC CGT GGT AAG | |||||||||||||
| Met Ser Lys Thr Cys | Tyr | Glu | Gly | Asn | Gly 10 | His | Phe Tyr | Arg Gly Lys 15 | ||||||||
| 1 | 5 | |||||||||||||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly Lys | His | Asn | Tyr Cys | Arg | Asn | Pro | Asp | Asn | Arg Arg | ||||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA AAA | CCG | TCT | TCT | CCG | CCG | |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp | Gly | Lys Lys | Pro | Ser | Ser | Pro | Pro | ||
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA ACC | CTA | CGT | CCG | CGT | TTT | |
| Glu | Glu | Leu | Lys | Phe | Gin | Cys | Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | |
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA | AAC | CAG | CCG | TGG | TTC |
| Lys | Ile | Ile | Gly | Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | |
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | Ile | Tyr Arg Arg | His | Arg | Gly Gly | Ser | Val | Thr | Tyr | Val | Cys | ||||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly Gly | Ser | Leu | Ile | Ser | Pro | Cys | Trp | Val | Ile | Ser | Ala | Thr | His | Cys | ||
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC ATC | GAC | TAC | CCG | AAA AAA | GAA | GAC | TAC | ATC | OTT | TAC | CTC | GGC | CGT | ||
| Phe | ile | Asp Tyr | Pro | Lys Lys | Glu | Asp | Tyr | Ile | Val | Tyr | Leu | Gly Arg | ||||
| 165 | 170 | 175 |
-44CZ 288536 B6
| 528 | TCT CGT Ser Arg | TTA Leu | AAC TCT Asn Ser 180 | AAC ACC CAG GGT GAA ATG | AAA TTC GAA GTT GAA | |||||||||||
| Asn | Thr Gin | Gly 185 | Glu Met | Lys Phe | Glu Val Glu 190 | |||||||||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC |
| Asn | Leu | Ile | Leu | His | Lys | Asp | Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | |
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA | AAA | ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT |
| Asn | Asp | ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | Glu | Gly Arg | Cys | Ala | ||
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | Ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | Glu | Ile | Thr | Gly | Phe | Gly | Lys | Glu | |
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | Tyr | Leu | iyr | Pro | Glu | Gin | Leu | Lys | Met | Thr | Val | Val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr Tyr Gly | Ser | |||
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| Glu | Val | Thr | Thr | Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | |
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA | GTT | TGC | TCT | CTC | CAG |
| Asp | Ser | Cys | Gin | Gly | Asp | Ser | Gly | Gly | Pro | Leu | Val | Cys | Ser | Leu | Gin | |
| 305 | 310 | 315 | 320 | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly Arg | Met | Thr | Leu | Thr | Gly | Ile | Val | Ser | Trp | Gly | Arg | Gly | Cys | Ala | ||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | Val | Ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT | CAC | ACC | AAA | GAA | GAA | AAC | GGT | CTG | GCT | CTG | AGC | CCG |
| Trp | ile | Arg | Ser | His | Thr | Lys Glu | Glu | Asn | Gly | Leu | Ala Leu | Ser | Pro | |||
| 355 | 360 | 365 | ||||||||||||||
| 1104 | CCG | AGC | CCG | CCG | AGC | CCG | CCG | GGT | GGT | TTC | CCG | AGG | CCT | GGT | GGT | GGT |
| Pro | Ser | Pro | Pro | Ser | Pro | Pro | Gly Gly | Phe | Pro | Arg | Pro | Gly | Gly Gly | |||
| 370 | 375 | 380 | ||||||||||||||
| 1152 | GGT | AAC | GGT | GAC | TTC | GAA | GAA | ATC | CCG GAA | GAG | TAC | CTG | TGA | |||
| Gly | Asn | Gly Asp | Phe | Glu | Glu | Ile | Pro Glu | Glu | Tyr | Leu | ||||||
| 385 | 390 | 395 |
-45CZ 288536 B6
Sekv. IDč. 15
Délka: 1176 BP, 391 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein M14 (pSJ78)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG AGC AAA ACT | TGC TAC GAA | GGT AAC GGT CAC TTC TAC CGT GGT AAG | |||||||||||||
| Met 1 | Ser | Lys | Thr | Cys Tyr 5 | Glu | Gly Asn | Gly 10 | His Phe | Tyr Arg | Gly Lys 15 | ||||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly Lys | His | Asn | Tyr Cys Arg | Asn | Pro | Asp | Asn | Arg Arg | |||||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA | AAA | CCG | TCT | TCT | CCG | CCG |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp | Gly Lys | Lys | Pro | Ser | Ser | Pro | Pro | ||
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA ACC | CTA | CGT | CCG | CGT | TTT | |
| Glu | Glu | Leu | Lys | Phe | Gin | Cys | Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | |
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA | AAC | CAG | CCG | TGG | TTC |
| Lys | Ile | Ile | Gly | Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | |
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | Ile | Tyr Arg | Arg | His | Arg | Gly | Gly | Ser | Val | Thr | Tyr | Val | Cys | ||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly | Gly | Ser | Leu | Ile | Ser | Pro | Cys Trp | Val | Ile | Ser | Ala | Thr | His | Cys | ||
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC | ATC | GAC | TAC | CCG | AAA | AAA GAA GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT | ||
| Phe | Ile | Asp Tyr | Pro | Lys | Lys Glu Asp | Tyr | Ile | Val | Tyr | Leu | Gly Arg |
165 170 175
-46CZ 288536 B6
| 1 | ATG AGC AAA ACT TGC TAC GAA GGT AAC GGT CAC TTC TAC CGT GGT AAG | |||||||||||||||
| Met 1 | Ser Lys | Thr | Cys Tyr 5 | Glu | Gly | Asn | Gly 10 | His | Phe | Tyr | Arg Gly Lys 15 | |||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly | Lys | His | Asn | Tyr Cys | Arg | Asn | Pro | Asp | Asn | Arg Arg | |||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | Val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA | AAA | CCG | TCT | TCT | CCG | CCG |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp Gly Lys | Lys | Pro | Ser | Ser | Pro | Pro | |||
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT |
| Glu | Glu | Leu | Lys | Phe | Gin | Cys | Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | |
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA | AAC | CAG | CCG | TGG | TTC |
| Lys | Ile | Ile | Gly Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | ||
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | Ile | Tyr Arg | Arg | His | Arg | Gly Gly | Ser | Val | Thr | Tyr | val | Cys | |||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly | Gly | Ser | Leu | Ile | Ser | Pro | Cys Trp | Val | Ile | Ser | Ala | Thr | His | Cys | ||
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC | ATC | GAC | TAC | CCG | AAA | AAA | GAA GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT | |
| Phe | Ile | Asp Tyr | Pro | Lys | Lys | Glu Asp Tyr | Ile | Val | Tyr | Leu | Gly Arg | |||||
| 165 | 170 | 175 |
-47CZ 288536 B6
Sekv. IDč. 16
Délka: 1197 BP, 398 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein Ml5 (pSJ79)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG AGC AAA ACT | TGC TAC GAA | GGT AAC GGT CAC TTC TAC CGT GGT AAG | |||||||||||||
| Met 1 | Ser | Lys | Thr | Cys Tyr 5 | Glu | Gly Asn | Gly 10 | His | Phe Tyr | Arg | Gly Lys 15 | |||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly | Lys | His | Asn | Tyr | Cys Arg | Asn | Pro | Asp | Asn | Arg | Arg | ||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | Val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA | AAA | CCG | TCT | TCT | CCG | CCG |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp | Gly | Lys | Lys | Pro | Ser | Ser | Pro | Pro | |
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT |
| G1U | Glu | Leu | Lys | Phe | Gin | Cys | Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | |
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA | AAC | CAG | CCG | TGG | TTC |
| Lys | Ile | Ile | Gly | Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | |
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | ile | Tyr Arg Arg | His | Arg | Gly | Gly | Ser | Val | Thr | Tyr | Val | Cys | |||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly | Gly | Ser | Leu | Ile | Ser | Pro | Cys | Trp | Val | Ile | Ser | Ala | Thr | His | Cys | |
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC | ATC | GAC | TAC | CCG | AAA | AAA | GAA | GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT |
| Phe | Ile | Asp Tyr | Pro | Lys | Lys | Glu | Asp Tyr | Ile | Val | Tyr | Leu | Gly Arg |
165 170 175
-48CZ 288536 B6
| 528 | TCT CGT | TTA AAC | TCT Ser | AAC ACC CAG GGT GAA ATG AAA TTC GAA GTT GAA | ||||||||||||
| Ser | Arg | Leu | Asn 180 | Asn Thr | Gin | Gly 185 | Glu Met | Lys Phe | Glu Val Glu 190 | |||||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC |
| Asn | Leu | Ile 195 | Leu | His | Lys | Asp | Tyr 200 | Ser | Ala | Asp | Thr | Leu 205 | Ala | His | His | |
| 624 | AAC | GAC | ATC | GCT | CTG | CTA | AAA | ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT |
| Asn | Asp 210 | Ile | Ala | Leu | Leu | Lys 215 | Ile | Arg | Ser | Lys | Glu 220 | Gly Arg | Cys | Ala | ||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin 225 | Pro | Ser | Arg | Thr | Ile 230 | Gin | Thr | Ile | Cys | Leu 235 | Pro | Ser | Met | Tyr | Asn 240 | |
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly 245 | Thr | Ser | Cys | Glu | Ile 250 | Thr | Gly | Phe | Gly | Lys 255 | Glu | |
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp Tyr 260 | Leu | Tyr | Pro | Glu 265 | Gin | Leu | Lys | Met | Thr 270 | Val | Val | ||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile 275 | Ser | His | Arg | Glu | Cys 280 | Gin | Gin | Pro | His | Tyr 285 | Tyr Gly | Ser | ||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| Glu | Val 290 | Thr | Thr | Lys | Met | Leu 295 | Cys | Ala | Ala | Asp | Pro 300 | Gin | Trp | Lys | Thr | |
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA | GTT | TGC | TCT | CTC | CAG |
| Asp 305 | Ser | Cys | Gin | Gly | Asp 310 | Ser | Gly | Gly | Pro | Leu 315 | Val | Cys | Ser | Leu | Gin 320 | |
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly Arg | Met | Thr | Leu 325 | Thr | Gly | Ile | val | Ser 330 | Trp Gly Arg Gly | Cys 335 | Ala | |||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys 340 | Pro | Gly | Val | Tyr | Thr 345 | Arg | Val | Ser His | Phe 350 | Leu | Pro | ||
| 1056 | TGG | ATC | CGT | TCT | CAC | ACC | AAA | GAA | GAA | AAC | GGT | CTG GCT | CTG | AGC | CCG | |
| Trp | Ile | Arg 355 | Ser | His | Thr | Lys | Glu 360 | G1U | Asn | Gly | Leu | Ala 365 | Leu | Ser | Pro | |
| 1104 | CCG | AGC | CCG | CCG | AGC | CCG | CCG | GGT | GGT | TTC | GGT | CCG | AGG | CCT | GGT | GGT |
| Pro | Ser 370 | Pro | Pro | Ser | Pro | Pro 375 | Gly Gly | Phe | Gly | Pro 380 | Arg | Pro | Gly Gly | |||
| 1152 | GGT Gly | GGT Gly | AAC Asn | GGT GAC Gly Asp | TTC Phe | GAA Glu | GAA ATC Glu Ile | CCG Pro | GAA Glu | GAG Glu | TAC Tyr | CTG Leu | TGA |
385 390 395
-49CZ 288536 B6
Sekv. IDč. 17
Délka: 1185 BP, 394 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein Ml6 (pSJ81)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATC AGC AAA ACT | TGC TAC GAA | GGT AAC GGT CAC TTC TAC CGT GGT AAG | |||||||||||||
| Met 1 | Ser | Lys | Thr | Cys Tyr 5 | Glu | Gly Asn | Gly 10 | His | Phe Tyr | Arg | Gly Lys 15 | |||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly | Lys | His | Asn | Tyr Cys | Arg | Asn | Pro | Asp | Asn | Arg Arg | |||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | Val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA | AAA | CCG | TCT | TCT | CCG | CCG |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp | Gly | Lys | Lys | Pro | Ser | Ser | Pro | Pro | |
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT |
| Glu | Glu | Leu | Lys | Phe | Gin | Cys Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | ||
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA | AAC | CAG | CCG | TGG | TTC |
| Lys | Ile | Ile | Gly | Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | |
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | Ile | Tyr Arg Arg | His | Arg | Gly | Gly | Ser | Val | Thr | Tyr | val | Cys | |||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly | Gly | Ser | Leu | Ile | Ser | Pro | Cys | Trp | Val | ile | Ser | Ala | Thr | His | Cys | |
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC | ATC | GAC | TAC | CCG | AAA | AAA GAA | GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT | |
| Phe | Ile | Asp | Tyr | Pro | Lys | Lys Glu | Asp | Tyr | Ile | Val | Tyr | Leu | Gly Arg |
nn ισκ
-50CZ 288536 B6
| 528 | TCT CGT TTA AAC | TCT AAC ACC CAG GGT GAA ATG AAA TTC GAA GTT GAA | ||||||||||||||
| Ser Arg | Leu Asn 180 | Ser Asn | Thr | Gin | Gly Glu Met Lys Phe Glu Val Glu | |||||||||||
| 185 | 190 | |||||||||||||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC |
| Asn | Leu | Ile | Leu | His | Lys | Asp | Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | |
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA | AAA | ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT |
| Asn | Asp | Ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | G1U | Gly Arg | Cys | Ala | ||
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | Ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | Glu | Ile | Thr | Gly | Phe | Gly | Lys | Glu | |
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | Tyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Met | Thr | Val | Val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr Tyr Gly | Ser | |||
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| Glu | Val | Thr | Thr | Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | |
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA | GTT | TGC | TCT | CTC | CAG |
| Asp | Ser | Cys | Gin | Gly Asp | Ser | Gly | Gly | Pro | Leu | Val | Cys | Ser | Leu | Gin | ||
| 305 | 310 | 315 | 320 | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly | Arg | Met | Thr | Leu | Thr | Gly | Ile | Val | Ser | Trp | Gly Arg Gly Cys | Ala | ||||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | Val | Ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT | CAC | ACC | AAA | GAA | GAA | AAC | GGT | CTG | GCT | CTG | AGC | CCG |
| Trp | Ile | Arg | Ser | His | Thr | Lys | Glu | Glu | Asn | Gly | Leu | Ala | Leu | Ser | Pro | |
| 355 | 360 | 365 | ||||||||||||||
| 1104 | . GTT | AAG | GCT | TGG | GGA | CCG | CGG | CCG | CTG | GGT | GGT | GGT | GGT | AAC | GGT | GAC |
| Val | Lys | Ala | Trp Gly | Pro | Arg | Pro | Leu | Gly Gly Gly Gly | Asn | Gly Asp | ||||||
| 370 | 375 | 380 | ||||||||||||||
| 1152 | ! TTC | GAA | . GAA | , ATC | CCG | GAA | . GAA | . TAC | CTG | CAA TAA | ||||||
| Phe | G1U | Glu | ile | Pro | Glu | Glu | . Tyr | Leu | Gin | |||||||
| 385 | 390 |
-51CZ 288536 B6
Sekv.IDč. 18
Délka: 1185 BP, 394 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein M17 (pSJ83)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG AGC | AAA | ACT | TGC | TAC | GAA | GGT | AAC | GGT | CAC | TTC | TAC | CGT | GGT | AAG |
| Met Ser | Lys | Thr | Cys | Tyr | G1U | Gly | Asn | Gly | His | Phe | Tyr | Arg | Gly | Lys | |
| 1 | 5 | 10 | 15 | ||||||||||||
| 48 | GCT TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | |||||||||||||
| 96 | ACC GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | |||||||||||||
| 144 | CTG GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu Gly | Leu | Gly Lys | His | Asn | Tyr Cys | Arg | Asn | Pro | Asp | Asn | Arg Arg | ||||
| 50 | 55 | 60 | |||||||||||||
| 192 | CGT CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA GTT | CAG | GAA | |
| Arg Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | Val | Gin | Glu | |
| 65 | 70 | 75 | 80 | ||||||||||||
| 240 | TGC ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA | AAA | CCG | TCT | TCT | CCG | CCG |
| Cys Met | Val | His | Asp | Cys | Ala | Asp Gly | Lys | Lys | Pro | Ser | Ser | Pro | Pro | ||
| 85 | 90 | 95 | |||||||||||||
| 288 | GAA GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT |
| Glu Glu | Leu | Lys | Phe | Gin | cys Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | ||
| 100 | 105 | 110 | |||||||||||||
| 336 | AAA ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA AAC | CAG | CCG | TGG | TTC | |
| Lys Ile | Ile | Glý | Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | |
| 115 | 120 | 125 | |||||||||||||
| 384 | GCT GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala Ala | ile | Tyr Arg Arg | His | Arg | Gly Gly | Ser | Val | Thr | Tyr | Val | Cys | ||||
| 130 | 135 | 140 | |||||||||||||
| 432 | GGT GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly Gly | Ser | Leu | Ile | Ser | Pro | Cys | Trp | Val | Ile | Ser | Ala | Thr | His | Cys | |
| 145 | 150 | 155 | 160 | ||||||||||||
| 480 | TTC ATC | GAC | TAC | CCG | AAA AAA | GAA | GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT | |
| Phe Ile | Asp Tyr | Pro | Lys Lys | Glu | Asp Tyr | Ile | Val | Tyr | Leu | Gly Arg | |||||
| 165 | 170 | 175 |
-52CZ 288536 B6
| 528 | TCT CGT TTA AAC Ser Arg Leu Asn 180 | TCT AAC ACC CAG GGT GAA ATG AAA | TTC GAA GTT GAA Phe Glu Val Glu 190 | |||||||||||||
| Ser Asn Thr Gin | Gly 185 | Glu Met Lys | ||||||||||||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC |
| Asn | Leu | Ile | Leu | His | Lys | Asp Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | ||
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA AAA ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT | ||
| Asn | Asp | Ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | Glu | Gly | Arg | Cys | Ala | |
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | Ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA ATC | ACC | GGT | TTC | GGT | AAA | GAA | |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | Glu | Ile | Thr | Gly | Phe | Gly | Lys | Glu | |
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | Tyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Met | Thr | Val | Val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr | Tyr | Gly | Ser | |
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| Glu | Val | Thr | Thr | Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | |
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA GGT | GAC | TCT | GGT | GGT | CCA | CTA GTT | TGC | TCT | CTC | CAG | ||
| Asp | Ser | Cys | Gin | Gly | Asp | Ser | Gly | Gly | Pro | Leu | Val | Cys | Ser | Leu | Gin | |
| 305 | 310 | 315 | 320 | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly Arg | Met | Thr | Leu | Thr | Gly | Ile | Val | Ser | Trp | Gly Arg | Gly | Cys | Ala | |||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | Val | Ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT CAC | ACC | AAA | GAA | GAA | AAC | GGT | CTG | GCT | CTG | AGC | CCG | |
| Trp | Ile | Arg | Ser | His | Thr | Lys | Glu | Glu | Asn | Gly | Leu | Ala | Leu | Ser | Pro | |
| 355 | 360 | 365 | ||||||||||||||
| 1104 | GTT | AAG | GCT | TTC | GGA | CCG | CGG | CCG | CTG | GGT | GGT | GGT | GGT | AAC | GGT | GAC |
| Val | Lys | Ala | Phe | Gly | Pro | Arg | Pro | Leu | Gly Gly Gly Gly | Asn | Gly Asp | |||||
| 370 | 375 | 380 | ||||||||||||||
| 1152 | : ttc | GAA | GAA | . ATC | CCG | GAA | GAA | TAC | CTG | CAA TAA | ||||||
| Phe | Glu | Glu | Ile | Pro | Glu | Glu | Tyr | Leu | Gin |
385 390
-53CZ 288536 B6
Sekv.IDč. 19
Délka: 1185 BP, 394 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein Ml8 (pSJ90)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG AGC AAA ACT | TGC TAC GAA | GGT AAC GGT CAC TTC TAC CGT GGT AAG | |||||||||||||
| Met 1 | Ser | Lys | Thr | Cys Tyr 5 | Glu | Gly Asn | Gly 10 | His Phe | Tyr Arg Gly Lys 15 | |||||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly | Lys | His | Asn | Tyr | Cys | Arg | Asn | Pro | Asp | Asn | Arg Arg | ||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA | |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | Val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA | AAA | CCG | TCT | TCT | CCG | CCG |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp | Gly | Lys | Lys | Pro | Ser | Ser | Pro | Pro | |
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT |
| Glu | Glu | Leu | Lys | Phe | Gin | Cys Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | ||
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT GGT | GAG | TTC | ACC | ACC | ATC | GAA | AAC | CAG | CCG | TGG | TTC | |
| Lys | Ile | Ile | Gly Gly | Glu | Phe | Thr | Thr | Ile | G1U | Asn | Gin | Pro | Trp | Phe | ||
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | Ile | Tyr Arg Arg | His | Arg | Gly | Gly | Ser | Val | Thr | Tyr | Val | Cys | |||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT ATC | TCT | GCT | ACC | CAC | TGC | |
| Gly Gly | Ser | Leu | Ile | Ser | Pro | Cys Trp Val | Ile | Ser | Ala | Thr | His | Cys | ||||
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC ATC | GAC | TAC | CCG | AAA | AAA | GAA | GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT | |
| Phe | Ile | Asp | Tyr | Pro | Lys | Lys | Glu | Asp Tyr | Ile | Val | Tyr | Leu | Gly Arg |
165 170 175
-54CZ 288536 B6
| 528 | TCT CGT | TTA AAC | TCT Ser | AAC ACC CAG GGT GAA ATG AAA TTC GAA GTT GAA | ||||||||||||
| Ser | Arg | Leu | Asn 180 | Asn Thr | Gin | Gly 185 | Glu Met | Lys Phe | Glu Val Glu 190 | |||||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC |
| Asn | Leu | Ile | Leu | His | Lys | Asp | Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | |
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA | AAA | ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT |
| Asn | Asp | Ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | Glu | Gly Arg | Cys | Ala | ||
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | Ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | Glu | Ile | Thr | Gly | Phe | Gly | Lys | Glu | |
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | Tyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Met | Thr | Val | Val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr Tyr Gly | Ser | |||
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| Glu | Val | Thr | Thr | Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | |
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA | GTT | TGC | TCT | CTC | CAG |
| Asp | Ser | Cys | Gin | Gly | Asp | Ser | Gly | Gly | Pro | Leu | Val | Cys | Ser | Leu | Gin | |
| 305 | 310 | 315 | 320 | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly Arg | Met | Thr | Leu | Thr | Gly | Ile | Val | Ser | Trp | Gly Arg | Gly | Cys | Ala | |||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | Val | Ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT | CAC | ACC | AAA | GAA | GAA | AAC | GGT | CTG | GCT | CTG | AGC | CCG |
| Trp | Ile | Arg | Ser | His | Thr | Lys | Glu | Glu | Asn | Gly | Leu | Ala | Leu | Ser | Pro | |
| 355 | 360 | 365 | ||||||||||||||
| 1104 | GTT | CGG | GCT | TTC | GGA | CCG | CGG | CCG | CTG | GGT | GGT | GGT | GGT AAC | GGT | GAC | |
| Val | Arg | Ala | Phe | Gly | Pro | Arg | Pro | Leu | Gly | Gly | Gly | Gly Asn | Gly Asp | |||
| 370 | 375 | 380 | ||||||||||||||
| 1152 | TTC | GAA | GAA | ATC | CCG | GAA | GAA | TAC | CTG | CAA | TAA | |||||
| Phe | Glu | Glu | Ile | Pro | Glu | Glu | Tyr | Leu | Gin |
3Θ5 390
-55CZ 288536 B6
Sekv. ID č. 20
Délka: 1185 BP, 394 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein M19 (pSJ91)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG AGC AAA ACT | TGC TAC GAA | GGT AAC GGT CAC TTC TAC CGT GGT AAG | |||||||||||||
| Met 1 | Ser | Lys | Thr | Cys Tyr 5 | G1U | Gly Asn | Gly 10 | His Phe | Tyr Arg Gly Lys 15 | |||||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly | Lys | HÍS | Asn | Tyr | Cys | Arg | Asn | Pro | Asp | Asn | Arg Arg | ||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | Val | Gin | G1U | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA | AAA | CCG | TCT | TCT | CCG | CCG |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp | Gly | Lys | Lys | Pro | Ser | Ser | Pro | Pro | |
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT | |
| Glu | Glu | Leu | Lys | Phe | Gin | Cys | Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | |
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA | AAC | CAG | CCG | TGG | TTC |
| Lys | Ile | Ile | Gly | Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | |
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | Ile | Tyr Arg | Arg | HÍ3 | Arg | Gly Gly | Ser | Val | Thr | Tyr | Val | Cys | |||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly Gly | Ser | Leu | Ile | Ser | Pro | Cys | Trp | Val | Ile | Ser Ala | Thr | His | Cys | |||
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC ATC | GAC | TAC | CCG | AAA AAA GAA GAC | TAC | ATC | GTT TAC | CTC | GGC | CGT | |||||
| Phe | Ile | Asp | Tyr | Pro | Lys Lys Glu Asp | Tyr | Ile | Val Tyr | Leu | Gly Arg |
165 170 175
-56Cl 288536 B6
| 528 | TCT CGT Ser Arg | TTA Leu | AAC TCT Asn Ser 180 | AAC ACC CAG GGT GAA ATG | AAA TTC GAA GTT GAA | |||||||||||
| Asn | Thr Gin | Gly 185 | Glu Met | Lys Phe Glu 190 | Val Glu | |||||||||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC |
| Asn | Leu | Ile | Leu | His | Lys | Asp | Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | |
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA | AAA | ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT |
| Asn | Asp | Ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | Glu | Gly Arg | Cys | Ala | ||
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | Ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | Glu | Ile | Thr | Gly | Phe | Gly | Lys | Glu | |
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | Tyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Met | Thr | Val | Val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr | Tyr | Gly | Ser | |
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| Glu | Val | Thr | Thr | Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | |
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA | GTT | TGC | TCT | CTC | CAG |
| Asp | Ser | Cys | Gin | Gly | Asp | Ser | Gly | Gly | Pro | Leu | Val | Cys | Ser | Leu | Gin | |
| 305 | 310 | 315 | 320 | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly Arg | Met | Thr | Leu | Thr | Gly | Ile | Val | Ser | Trp | Gly Arg | Gly | Cys | Ala | |||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | val | Tyr | Thr | Arg | Val | Ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT | CAC | ACC | AAA | GAA | GAA | AAC | GGT | CTG | GCT | CTG | AGC | CCG |
| Trp | Ile | Arg | Ser | His | Thr | Lys | Glu | Glu | Asn | Gly | Leu | Ala | Leu | Ser | Pro | |
| 355 | 360 | 365 | ||||||||||||||
| 1104 | GTT | AAG | GCT | TAC | GGA | CCG | CGG | CCG | CTG | GGT | GGT | GGT | GGT | AAC | GGT | GAC |
| Val | Lys | Ala | Tyr Gly | Pro | Arg | Pro | Leu | Gly | Gly Gly | Gly | Asn | Gly | Asp | |||
| 370 | 375 | 380 | ||||||||||||||
| 1152 | TTC | GAA | GAA | ATC | CCG | GAA | GAA | TAC | CTG | CAA | TAA | |||||
| Phe | Glu | Glu | Ile | Pro | Glu | Glu | Tyr | Leu | Gin | |||||||
| 385 | 390 |
-57CZ 288536 B6
Sekv. ID č. 21
Délka: 1185 BP, 394 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein M20 (pSJ92)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG AGC AAA ACT TGC TAC GAA GGT AAC | GGT CAC | TTC TAC CGT GGT AAG | |||||||||||||
| Met Ser Lys Thr Cys | Tyr | Glu | Gly | Asn | Gly 10 | His | Phe Tyr | Arg Gly Lys 15 | ||||||||
| 1 | 5 | |||||||||||||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly | Lys | His | Asn | Tyr Cys | Arg | Asn | Pro | Asp | Asn | Arg Arg | |||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | Val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA | AAA | CCG | TCT | TCT | CCG | CCG |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp | Gly | Lys | Lys | Pro | Ser | Ser | Pro | Pro | |
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA AAA ACC | CTA | CGT | CCG | CGT | TTT | ||
| Glu | Glu | Leu | Lys | Phe | Gin | Cys | Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | |
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA AAC | CAG | CCG | TGG | TTC | |
| Lys | Ile | Ile | Gly Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | ||
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | Ile | Tyr Arg Arg | His | Arg | Gly | Gly | Ser | Val | Thr | Tyr | Val | Cys | |||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly | Gly | Ser | Leu | Ile | Ser | Pro | Cys | Trp | Val | Ile | Ser | Ala | Thr | His | Cys | |
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC | ATC | GAC | TAC | CCG | AAA | AAA | GAA | GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT |
| Phe | Ile | Asp Tyr | Pro | Lys | Lys | Glu | Asp Tyr | Ile | val | Tyr | Leu | Gly Arg | ||||
| 165 | 170 | 175 |
-58CZ 288536 B6
| 528 | TCT CGT TTA | AAC Asn 180 | TCT AAC ACC CAG GGT GAA ATG AAA TTC GAA GTT GAA | |||||||||||||
| Ser Arg | Leu | Ser | Asn | Thr | Gin | Gly Glu 185 | Met Lys | Phe | Glu 190 | Val | Glu | |||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC | |
| Asn | Leu | Ile | Leu | His | Lys Asp | Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | ||
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA | AAA | ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT |
| Asn | Asp | Ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | Glu | Gly Arg | Cys | Ala | ||
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | Ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | Glu | Ile | Thr | Gly | Phe | Gly | Lys | Glu | |
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | Tyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Met | Thr | Val | Val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr | Tyr | Gly | Ser | |
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| Glu | Val | Thr | Thr | Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | |
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA | GTT | TGC | TCT | CTC | CAG |
| Asp | Ser | Cys | Gin | Gly Asp | Ser | Gly | Gly | Pro | Leu | val | Cys | Ser | Leu | Gin | ||
| 305 | 310 | 315 | 320 | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly | Arg | Met | Thr | Leu | Thr | Gly | Ile | Val | Ser | Trp | Gly Arg Gly | Cys | Ala | |||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | Val | Ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT | CAC | ACC | AAA GAA | GAA | AAC | GGT | CTG | GCT | CTG | AGC | CCG | |
| Trp | Ile | Arg | Ser | His | Thr | Lys Glu | Glu | Asn | Gly | Leu | Ala | Leu | Ser | Pro | ||
| 355 | 360 | 365 | ||||||||||||||
| 1104 | GTT | GTT | GCT | TTC | GGA | CCG | CGG | CCG | CTG | GGT | GGT | GGT | GGT | AAC | GGT | GAC |
| Val | Val | Ala | Phe | Gly | Pro | Arg | Pro | Leu | Gly | Gly Gly Gly | Asn | Gly Asp | ||||
| 370 | 375 | 380 | ||||||||||||||
| 1152 | TTC | GAA | GAA | ATC | CCG | GAA GAA TAC | CTG | CAA | . TAA | |||||||
| Phe | Glu | Glu | Ile | Pro | Glu Glu Tyr | Leu | Gin |
385 390
-59CZ 288536 B6
Sekv. ID č. 22
Délka: 1182 BP, 393 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein M21 (pSJ93)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen io
| 1 | ATG AGC AAA ACT TGC TAC GAA GGT AAC | GGT CAC | TTC TAC CGT GGT AAG | |||||||||||||
| Met Ser Lys Thr Cys | Tyr | Glu | Gly | Asn | Gly 10 | His | Phe Tyr | Arg Gly Lys 15 | ||||||||
| 1 | 5 | |||||||||||||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly | Lys | His | Asn | Tyr Cys Arg | Asn | Pro | Asp | Asn | Arg Arg | ||||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | Val | Gin | Val | Gly | Leu | Lys | Pro | Leu | Val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA | AAA | CCG | TCT | TCT | CCG | CCG |
| Cys Met | Val | His | Asp | Cys | Ala | Asp Gly | Lys | Lys | Pro | Ser | Ser | Pro | Pro | |||
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT |
| Glu | Glu | Leu | Lys | Phe | Gin | Cys | Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | |
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA AAC | CAG | CCG | TGG | TTC | |
| Lys | Ile | Ile | Gly | Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | |
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | ile | Tyr Arg Arg | His | Arg | Gly Gly | Ser | Val | Thr | Tyr | Val | Cys | ||||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG GTT | ATC | TCT | GCT | ACC | CAC | TGC | |
| Gly Gly | Ser | Leu | ile | Ser | Pro | Cys | Trp Val | ile | Ser | Ala | Thr | His | Cys | |||
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC ATC | GAC | TAC | CCG | AAA | AAA | GAA | GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT | |
| Phe | ile | Asp Tyr | Pro | Lys Lys | Glu | Asp | Tyr | Ile | Val | Tyr | Leu | Gly Arg | ||||
| 165 | 170 | 175 |
-60CZ 288536 B6
| 528 | TCT CGT Ser Arg | TTA Leu | AAC TCT Asn Ser 180 | AAC ACC CAG GGT GAA ATG | AAA TTC GAA GTT GAA | |||||||||||
| Asn | Thr Gin | Gly 185 | Glu Met | Lys Phe Glu 190 | Val Glu | |||||||||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC |
| Asn | Leu | Ile | Leu | His | Lys | Asp | Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | |
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA | AAA | ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT |
| Asn | Asp | ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | Glu | Gly | Arg | Cys | Ala | |
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | Ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | Glu | Ile | Thr | Gly | Phe | Gly | Lys | G1U | |
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | Tyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Met | Thr | Val | Val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr | Tyr | Gly | Ser | |
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| Glu | Val | Thr | Thr | Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | |
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA | GTT | TGC | TCT | CTC | CAG |
| Asp | Ser | Cys | Gin | Gly Asp | Ser | Gly Gly | Pro | Leu | Val | Cys | Ser | Leu | Gin | |||
| 305 | 310 | 315 | 320 | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly Arg | Met | Thr | Leu | Thr | Gly | Ile | Val | Ser | Trp | Gly Arg | Gly | Cys | Ala | |||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | Val | ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT | CAC | ACC | AAA | GAA | GAA | AAC | GGT | CTG | GCT | CTG | AGC | CCG |
| Trp | Ile | Arg | Ser | His | Thr | Lys | Glu | Glu | Asn | Gly | Leu | Ala | Leu | Ser | Pro | |
| 355 | 360 | 365 | ||||||||||||||
| 1104 | GTT | CGG | GCT | TTC | CCG | CGG | CCG | CTG | GGT | GGT | GGT | GGT | AAC | GGT | GAC | TTC |
| Val | Arg | Ala | Phe | Pro | Arg | Pro | Leu | Gly | Gly | Gly Gly | Asn | Gly Asp | Phe | |||
| 370 | 375 | 380 | ||||||||||||||
| 1152 | GAA | GAA | ATC | CCG | GAA | GAA | TAC | CTG | CAA | TAA | ||||||
| Glu | Glu | Ile | Pro | Glu | Glu | Tyr | Leu | Gin | ||||||||
| 385 | 390 |
-61CZ 288536 B6
Sekv. ID č. 23
Délka: 1182 BP, 393 AK
Typ: sekvence nukleotidů společně s odpovídající sekvencí aminokyselin
Topologie: lineární
Druh molekuly: syntetický gen pro protein M22 (pSJ94)
Zvláštnost: N-terminální methionin byl v Escherichia coli proteolyticky odštěpen
| 1 | ATG AGC AAA ACT | TGC TAC GAA | GGT AAC GGT CAC TTC TAC CGT GGT AAG | |||||||||||||
| Met 1 | Ser | Lys | Thr | Cys Tyr 5 | Glu | Gly Asn | Gly 10 | His | Phe Tyr | Arg | Gly Lys 15 | |||||
| 48 | GCT | TCT | ACC | GAC | ACC | ATG | GGT | CGT | CCG | TGC | CTG | CCG | TGG | AAC | TCT | GCT |
| Ala | Ser | Thr | Asp | Thr | Met | Gly Arg | Pro | Cys | Leu | Pro | Trp | Asn | Ser | Ala | ||
| 20 | 25 | 30 | ||||||||||||||
| 96 | ACC | GTT | CTG | CAG | CAG | ACC | TAC | CAC | GCT | CAC | CGT | TCT | GAT | GCA | TTG | CAG |
| Thr | Val | Leu | Gin | Gin | Thr | Tyr | His | Ala | His | Arg | Ser | Asp | Ala | Leu | Gin | |
| 35 | 40 | 45 | ||||||||||||||
| 144 | CTG | GGT | CTG | GGT | AAA | CAC | AAC | TAC | TGC | CGT | AAC | CCG | GAC | AAC | CGT | CGT |
| Leu | Gly | Leu | Gly | Lys | His | Asn | Tyr | Cys | Arg | Asn | Pro | Asp | Asn | Arg Arg | ||
| 50 | 55 | 60 | ||||||||||||||
| 192 | CGT | CCG | TGG | TGC | TAC | GTT | CAG | GTT | GGT | CTG | AAA | CCG | CTA | GTT | CAG | GAA |
| Arg | Pro | Trp | Cys | Tyr | val | Gin | Val | Gly | Leu | Lys | Pro | Leu | Val | Gin | Glu | |
| 65 | 70 | 75 | 80 | |||||||||||||
| 240 | TGC | ATG | GTT | CAC | GAC | TGC | GCT | GAC | GGT | AAA | AAA | CCG | TCT | TCT | CCG | CCG |
| Cys | Met | Val | His | Asp | Cys | Ala | Asp | Gly | Lys | Lys | Pro | Ser | Ser | Pro | Pro | |
| 85 | 90 | 95 | ||||||||||||||
| 288 | GAA | GAG | CTC | AAA | TTC | CAG | TGC | GGT | CAA | AAA | ACC | CTA | CGT | CCG | CGT | TTT |
| Glu | G1U | Leu | Lys | Phe | Gin | Cys | Gly | Gin | Lys | Thr | Leu | Arg | Pro | Arg | Phe | |
| 100 | 105 | 110 | ||||||||||||||
| 336 | AAA | ATC | ATC | GGT | GGT | GAG | TTC | ACC | ACC | ATC | GAA | AAC | CAG | CCG | TGG | TTC |
| Lys | Ile | Ile | Gly Gly | Glu | Phe | Thr | Thr | Ile | Glu | Asn | Gin | Pro | Trp | Phe | ||
| 115 | 120 | 125 | ||||||||||||||
| 384 | GCT | GCT | ATC | TAC | CGT | CGT | CAC | CGT | GGT | GGT | TCT | GTT | ACC | TAC | GTT | TGC |
| Ala | Ala | Ile | Tyr | Arg | Arg | His | Arg | Gly Gly | Ser | Val | Thr | Tyr | Val | Cys | ||
| 130 | 135 | 140 | ||||||||||||||
| 432 | GGT | GGT | TCT | CTG | ATC | TCT | CCG | TGC | TGG | GTT | ATC | TCT | GCT | ACC | CAC | TGC |
| Gly | Gly | Ser | Leu | Ile | Ser | Pro | Cys | Trp | Val | Ile | Ser | Ala | Thr | His | Cys | |
| 145 | 150 | 155 | 160 | |||||||||||||
| 480 | TTC | ATC | GAC | TAC | CCG | AAA | AAA | GAA | GAC | TAC | ATC | GTT | TAC | CTC | GGC | CGT |
| Phe | Ile | Asp | Tyr | Pro | Lys | Lys | Glu | Asp | tyr | Ile | Val | Tyr | Leu | Gly Arg |
165 170 175
-62CZ 288536 B6
| 528 | TCT CGT TTA | AAC Asn 180 | TCT Ser | AAC ACC CAG GGT GAA ATG AAA TTC GAA GTT GAA | ||||||||||||
| Ser | Arg | Leu | Asn | Thr Gin Gly Glu Met Lys Phe Glu Val Glu | ||||||||||||
| 185 | 190 | |||||||||||||||
| 576 | AAC | CTG | ATC | CTG | CAC | AAA | GAC | TAC | TCT | GCT | GAC | ACC | CTG | GCT | CAC | CAC |
| Asn | Leu | Ile | Leu | His | Lys | Asp | Tyr | Ser | Ala | Asp | Thr | Leu | Ala | His | His | |
| 195 | 200 | 205 | ||||||||||||||
| 624 | AAC | GAC | ATC | GCT | CTG | CTA | AAA | ATC | CGT | TCT | AAA | GAA | GGT | CGT | TGC | GCT |
| Asn | Asp | Ile | Ala | Leu | Leu | Lys | Ile | Arg | Ser | Lys | Glu | Gly Arg | Cys | Ala | ||
| 210 | 215 | 220 | ||||||||||||||
| 672 | CAG | CCG | TCT | CGT | ACC | ATC | CAG | ACC | ATC | TGC | CTG | CCG | TCT | ATG | TAC | AAC |
| Gin | Pro | Ser | Arg | Thr | ile | Gin | Thr | Ile | Cys | Leu | Pro | Ser | Met | Tyr | Asn | |
| 225 | 230 | 235 | 240 | |||||||||||||
| 720 | GAC | CCG | CAG | TTC | GGT | ACC | TCT | TGC | GAA | ATC | ACC | GGT | TTC | GGT | AAA | GAA |
| Asp | Pro | Gin | Phe | Gly | Thr | Ser | Cys | Glu | Ile | Thr | Gly | Phe | Gly | Lys | Glu | |
| 245 | 250 | 255 | ||||||||||||||
| 768 | AAC | TCT | ACC | GAC | TAC | CTG | TAC | CCG | GAA | CAG | CTG | AAA | ATG | ACC | GTT | GTT |
| Asn | Ser | Thr | Asp | Tyr | Leu | Tyr | Pro | Glu | Gin | Leu | Lys | Met | Thr | Val | val | |
| 260 | 265 | 270 | ||||||||||||||
| 816 | AAA | CTG | ATC | TCT | CAC | CGT | GAA | TGC | CAG | CAG | CCG | CAC | TAC | TAC | GGT | TCT |
| Lys | Leu | Ile | Ser | His | Arg | Glu | Cys | Gin | Gin | Pro | His | Tyr | Tyr | Gly | Ser | |
| 275 | 280 | 285 | ||||||||||||||
| 864 | GAA | GTT | ACC | ACC | AAA | ATG | CTG | TGC | GCT | GCT | GAC | CCG | CAG | TGG | AAA | ACC |
| G1U | Val | Thr | Thr | Lys | Met | Leu | Cys | Ala | Ala | Asp | Pro | Gin | Trp | Lys | Thr | |
| 290 | 295 | 300 | ||||||||||||||
| 912 | GAC | TCT | TGC | CAA | GGT | GAC | TCT | GGT | GGT | CCA | CTA | GTT | TGC | TCT | CTC | CAG |
| Asp | Ser | Cys | Gin | Gly Asp | Ser | Gly | Gly | Pro | Leu | Val | Cys | Ser | Leu | Gin | ||
| 305 | 310 | 315 | 320 | |||||||||||||
| 960 | GGT | CGT | ATG | ACC | CTG | ACC | GGT | ATT | GTT | TCT | TGG | GGT | CGT | GGT | TGC | GCT |
| Gly Arg | Met | Thr | Leu | Thr | Gly | Ile | Val | Ser | Trp | Gly Arg | Gly Cys | Ala | ||||
| 325 | 330 | 335 | ||||||||||||||
| 1008 | CTG | AAA | GAC | AAA | CCG | GGT | GTT | TAC | ACC | CGT | GTT | TCT | CAC | TTC | CTG | CCG |
| Leu | Lys | Asp | Lys | Pro | Gly | Val | Tyr | Thr | Arg | Val | Ser | His | Phe | Leu | Pro | |
| 340 | 345 | 350 | ||||||||||||||
| 1056 | TGG | ATC | CGT | TCT | CAC | ACC | AAA | GAA | GAA | AAC | GGT | CTG | GCT | CTG | AGC | CCG |
| Trp | Ile | Arg | Ser | His | Thr | Lys | Glu | Glu | Asn | Gly | Leu | Ala | Leu | Ser | Pro | |
| 355 | 360 | 365 | ||||||||||||||
| 1104 | GTT | GTT | GCT | TTC | CCG | CGG | CCG | CTG | GGT | GGT | GGT | GGT | AAC | GGT | GAC | TTC |
| Val | Val | Ala | Phe | Pro | Arg | Pro | Leu | Gly Gly Gly Gly | Asn | Gly Asp | Phe | |||||
| 370 | 375 | 380 | ||||||||||||||
| 1152 | : GAA | . GAA | ATC | CCG | GAA | GAA | TAC | CTG | CAA TAA | |||||||
| Glu | Glu | Ile | Pro | Glu | Glu | Tyr | Leu | Gin |
385 390
Claims (17)
- PATENTOVÉ NÁROKY1. Bifunkční varianty urokinázy obecného vzorce IM4-XJ-Y, (I), ve kterémM4 značí aminokyselinovou sekvenci 47Ser až 411Leu glykosidu zbavené prourokinázy podle obr. 1,Xi značí přímou vazbu mezi M4 a Yi nebo peptid o sekvenciSer-Pro-Pro-Ser-Pro-Pro-Gly-Gly-Phe neboSer-Pro-Pro-Ser-Pro-Pro-Ser-Pro-Pro-Gly-Gly-Phe neboSer-Pro-Pro-Ser-Pro-Pro-Ser-Pro-Pro-Gly-Gly-Phe-Gly nebo peptidovou sekvenci obecného vzorce IISer-Xz-Xj-X^Xj-Xfi-Xy (II), přičemžX2 značí Pro nebo Leu,X3 značí Val nebo Pro,X4 značí Lys, Val, Arg, Gly nebo Glu,X5 značí Ala, Val, Gly, Leu nebo Ile,X6 značí Phe, Trp, Tyr nebo Val aX7 značí Gly nebo přímou vazbu mezi Xe a Yj aYi značí peptid sekvenceYz-Arg-Pro-Yj-Gly-Gly-Gly-Gly-Asn-Gly-Asp-Phe-Glu-Glu-Ile-Pro-Glu-GluTyr-Leu-Yí neboYr-Arg-Pro-Phe-Leu-Leu-Arg-Asn-Pro-Asn-Asp-Lys-Tyr-Glu-Pro-Phe-TrpGlu-Asp-Glu-Glu-Lys-Asn-Glu neboYr-Arg-Pro-Ser-Ser-Glu-Phe-Glu-Glu-Phe-Glu-Ile-Asp-Glu-Glu-Glu-Lys, přičemž-64CZ 288536 B6Y2 značí Pro nebo Val,Y3 značí Leu nebo přímou vazbu mezi Pro a Gly aY4 značí Gin nebo hydroxylovou skupinu.
- 2. Bifunkční varianty urokinázy podle nároku 1 obecného vzorce I, kdeYi značí peptid sekvenceY2-Arg-Pro-Y3-Gly-Gly-Gly-Gly-Asn-Gly-Asp-Phe-Glu-Glu-Ile-Pro-Glu-GluTyr-Leu-Y4, přičemž Y2 a Y4 mají v nároku 1 uvedený význam.
- 3. Bifunkční varianty urokinázy podle nároku 1 obecného vzorce I, kdeYi značí peptid sekvenceY2-Arg-Pro-Phe-Leu-Leu-Arg-Asn-Pro-Asn-Asp-Lys-Tyr-Glu-Pro-Phe-TrpGlu-Asp-Glu-Glu-Lys-Asn-Glu, přičemž Y2 má v nároku 1 uvedený význam.
- 4. Bifunkční varianty urokinázy podle nároku 1 a/nebo 2 obecného vzorce I, kdeX] značí peptidovou sekvenci obecného vzorce IISer-Xr-Xy-Xr-Xr-Xe-X? (II), ve kterémX2 značí Pro nebo Leu,X3 značí Val,X4 značí Lys, Val nebo Arg,X5 značí Ala, Val nebo Gly,X6 značí Phe, Trp, Tyr nebo Val aX7 značí Gly nebo přímou vazbu mezi X6 a Y].
- 5. Bifunkční varianty urokinázy podle nároku 4 obecného vzorce I, kdeX4 značí Lys nebo Val,X5 značí Ala nebo Val,X6 značí Phe, Trp nebo Tyr aX7 značí Gly nebo přímou vazbu mezi X6 a Y].
- 6. Bifunkční varianty urokinázy podle nároku 4 a/nebo 5 obecného vzorce I, kdeX7 značí přímou vazbu mezi Xe a Yi.
- 7. Bifunkční varianty urokinázy podle nároku 1 a/nebo 3 obecného vzorce I, kde Xi značí peptidovou sekvenci obecného vzorce II, ve kterémX2 značí Pro nebo Leu,X3 značí Val,X» značí Lys nebo Val,X5 značí Ala nebo Val,X6 značí Phe nebo Trp a-65CZ 288536 B6X7 značí přímou vazbu mezi Xg a Y].
- 8. Plazmidy pro použití při získávání bifunkčních variant urokinázy podle nároků 1 až 7, u kterých operon má regulovatelný promotor, Shine-Dalgamo sekvenci účinnou jako vazebné místo pro ribozomy, startovací kodon, syntetický strukturní gen se sekvencemi ID č. 1 až 24 pro bifunkční varianty urokinázy obecné vzorce I podle nároků 1 až 7 a dále od strukturního genu jeden nebo dva terminátory, přičemž tyto plazmidy jsou vhodné pro expresi bifunkčních variant urokinázy ve kmenech Escherichia coli.
- 9. Plazmidy podle nároku 8, u kterých odstup mezi Shine-Dalgamo-sekvencí a startovacím kodonem činí 6 až 12, výhodně 8 až 10 nukleotidů.
- 10. Plazmidy podle nároku 8 a/nebo 9, které jsou vybrané ze skupiny, zahrnující pSJ 69 podle obr. 2e, pSJ 76 podle obr. 2h, pSJ 77 podle obr. 2h, pSJ 78 podle obr. 2h, pSJ 79 podle obr. 2h, pSJ 81 podle obr. 2j, pSJ 83 podle obr. 21, pSJ 90 podle obr. 21, pSJ 91 podle obr. 21, pSJ 92 podle obr. 21, pSJ 93 podle obr. 21, pSJ 94 podle obr. 21, pSJ 95 podle obr. 21, pSJ 101 podle obr. 21, pSJ 102 podle obr. 21, pSJ 103 podle obr. 21, pSJ 104 podle obr. 21, pSJ 105 podle obr. 21, pSJ 106 podle obr. 2o, pSJ 109 podle obr. 21, pSJ 111 podle obr. 21, pSJ 114 podle obr. 2n a pSJ 113 podle obr. 2p.
- 11. Plazmidy podle nároku 10, které jsou vybrané ze skupiny, zahrnující pSJ 76 podle obr. 2h, pSJ 81 podle obr. 2j, pSJ 83 podle obr. 21, pSJ 90 podle obr. 21, pSJ 91 podle obr. 21, pSJ 92 podle obr. 21, pSJ 93 podle obr. 21, pSJ 94 podle obr. 21, pSJ 95 podle obr. 21, pSJ 101 podle obr. 21, pSJ 102 podle obr. 21, pSJ 103 podle obr. 21, pSJ 105 podle obr. 21, pSJ 106 podle obr. 2o, pSJ 109 podle obr. 21, pSJ 111 podle obr. 21 a pSJ 114 podle obr. 2n.
- 12. Plazmidy podle nároků 10 a/nebo 11, které jsou vybrané ze skupiny, zahrnující pSJ 76 podle obr. 2h, pSJ 81 podle obr. 2j, pSJ 83 podle obr. 21, pSJ 91 podle obr. 21, pSJ 92 podle obr. 21, pSJ 94 podle obr. 21, pSJ 95 podle obr. 21, pSJ 101 podle obr. 21, pSJ 102 podle obr. 21, pSJ 103 podle obr. 21, pSJ 105 podle obr. 21, pSJ 106 podle obr. 2o, pSJ 109 podle obr. 21, pSJ 111 podle obr. 21 a pSJ 114 podle obr. 2n.
- 13. Plazmidy podle jednoho nebo několika z nároků 10 až 12, které jsou vybrané ze skupiny, zahrnující pSJ 76 podle obr. 2h, pSJ 94 podle obr. 21, pSJ 95 podle obr. 21, pSJ 101 podle obr. 21, pSJ 102 podle obr. 21, pSJ 103 podle obr. 21, pSJ 106 podle obr. 2o, pSJ 109 podle obr. 21, pSJ 111 podle obr. 21 a pSJ 114 podle obr. 2n.
- 14. Plazmidy podle nároků 8 až 13, vyrobitelné z plazmidů pBlueskript KS II+, pUC 8 a pGR 201 podle obr. 2 a 2a až 2p.
- 15. Způsob výroby bifunkčních variant urokinázy obecného vzorce I podle nároků 1 až 7, vyznačující se tím, že se plazmidem podle nároků 8 až 13 transformuje kmen Escherichia coli, indukuje se exprese strukturního genu, vytvořený předstupeň proteinu bifunkční varianty urokinázy obecného vzorce I se z média a lýzovaných bakteriálních buněk oddělí, tento předstupeň proteinu se solubilizuje a potom se působením redox systému převede na bifunkční variantu urokinázy obecného vzorce I.
- 16. Trombolytikum, vyznačující se tím, že obsahuje jako účinnou látku bifunkční variantu urokinázy obecného vzorce I podle nároků 1 až 7.
- 17. Trombolytikum podle nároku 16 pro bolusaplikaci.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE4323754A DE4323754C1 (de) | 1993-07-15 | 1993-07-15 | Bifunktionelle Urokinasevarianten mit verbesserten fibrinolytischen Eigenschaften und thrombinhemmender Wirkung |
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| Publication Number | Publication Date |
|---|---|
| CZ170694A3 CZ170694A3 (en) | 1995-02-15 |
| CZ288536B6 true CZ288536B6 (cs) | 2001-07-11 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CZ19941706A CZ288536B6 (cs) | 1993-07-15 | 1994-07-14 | Bifunkční varianty urokinázy se zlepšenými fibrinolytickými vlastnostmi a s trombininhibičním účinkem, plazmidy pro jejich získávání, způsob jejich výroby a trombolytika tyto látky obsahující |
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|---|---|
| US (2) | US5681721A (cs) |
| EP (1) | EP0669394B1 (cs) |
| JP (1) | JPH07143878A (cs) |
| KR (1) | KR960014346A (cs) |
| CN (1) | CN1057124C (cs) |
| AT (1) | ATE189261T1 (cs) |
| AU (1) | AU679512B2 (cs) |
| CA (1) | CA2127897A1 (cs) |
| CZ (1) | CZ288536B6 (cs) |
| DE (2) | DE4323754C1 (cs) |
| DK (1) | DK0669394T3 (cs) |
| ES (1) | ES2145075T3 (cs) |
| FI (1) | FI943354L (cs) |
| GR (1) | GR3032488T3 (cs) |
| HR (1) | HRP940366B1 (cs) |
| HU (1) | HU217101B (cs) |
| IL (1) | IL109107A0 (cs) |
| NO (1) | NO941561L (cs) |
| NZ (1) | NZ260991A (cs) |
| PL (1) | PL176971B1 (cs) |
| PT (1) | PT669394E (cs) |
| RU (1) | RU2143490C1 (cs) |
| SI (1) | SI0669394T1 (cs) |
| SK (1) | SK281586B6 (cs) |
| UA (1) | UA27132C2 (cs) |
| ZA (1) | ZA944202B (cs) |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| DE4440892A1 (de) * | 1994-11-17 | 1996-05-23 | Gruenenthal Gmbh | Proteine mit fibrinolytischen und gerinnungshemmenden Eigenschaften |
| DE4442665A1 (de) * | 1994-11-30 | 1996-06-05 | Gruenenthal Gmbh | Chimäre Proteine mit fibrinolytischen und thrombinhemmenden Eigenschaften |
| CN1057125C (zh) * | 1996-04-26 | 2000-10-04 | 南京大学 | 五种尿激酶变体基因及在大肠杆菌中的表达 |
| CN1111739C (zh) * | 1997-02-04 | 2003-06-18 | 许文俊 | 凝血、纤溶功能动态测定试剂 |
| WO1999037149A1 (en) * | 1998-01-27 | 1999-07-29 | Brigham & Women's Hospital | Methods of treating cytotoxic damage |
| US7498310B1 (en) * | 1998-08-13 | 2009-03-03 | Beiersdorf Ag | Cosmetic or dermatological preparations comprising oligopeptides for lightening the skin of age marks and/or for preventing tanning of the skin, in particular tanning of the skin caused by UV radiation |
| WO2001097752A2 (en) * | 2000-06-20 | 2001-12-27 | The Trustees Of The University Of Pennsylvania | Compositions comprising urokinase for modulating muscle contractility and angiogenisis |
| US7361493B1 (en) | 2004-05-26 | 2008-04-22 | The United States Of America As Represented By The Secretary Of The Department Of Veterans Affairs | Production of urokinase in a three-dimensional cell culture |
| CN101094867B (zh) | 2004-10-19 | 2011-08-24 | 隆萨股份公司 | 用于固相肽合成的方法 |
| CN104193807B (zh) * | 2014-09-28 | 2016-06-15 | 广州贝奥吉因生物科技有限公司 | 凝血酶抑制多肽及其制备方法、应用 |
| CN105385617A (zh) * | 2015-09-23 | 2016-03-09 | 河北省科学院生物研究所 | 一株产纤溶酶的海洋放线菌株、用途以及由其制备的纤溶酶及纤溶酶的应用 |
| US11091535B2 (en) | 2016-12-16 | 2021-08-17 | The University Of Sydney | Thrombin inhibitors for treatment of stroke and related coagulative disorders |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3930944A (en) * | 1975-03-31 | 1976-01-06 | Abbott Laboratories | Urokinase production |
| DE3445517C2 (de) * | 1984-12-13 | 1993-11-18 | Ciba Geigy | Für ein Hirudin-ähnliches Protein codierende DNA-Sequenz und Verfahren zur Herstellung eines Hirudin-ähnlichen Proteins |
| US4751180A (en) * | 1985-03-28 | 1988-06-14 | Chiron Corporation | Expression using fused genes providing for protein product |
| US5002887A (en) * | 1986-01-31 | 1991-03-26 | Genetics Institute, Inc. | Truncated thrombolytic proteins |
| EP0330700B1 (en) * | 1987-08-19 | 1993-12-22 | Sagami Chemical Research Center | Human prourokinase-like polypeptide |
| DE3804600A1 (de) * | 1988-02-13 | 1989-08-24 | Basf Ag | Mischung aus einer thrombolytisch wirkenden und einer antithrombotischen substanz |
| GB8822147D0 (en) * | 1988-09-21 | 1988-10-26 | Ciba Geigy Ag | Pharmaceutically active combination |
| US5256770A (en) * | 1990-04-09 | 1993-10-26 | Schering Ag | Oxidation resistant thrombomodulin analogs |
| CA2064231A1 (en) * | 1989-07-20 | 1991-01-21 | John M. Maraganore | Combinations and methods for treating or preventing thrombotic diseases |
| GB8927722D0 (en) * | 1989-12-07 | 1990-02-07 | British Bio Technology | Proteins and nucleic acids |
| US5328898A (en) * | 1990-06-22 | 1994-07-12 | Duke University | Factor XIIIA fibrin binding fragments |
| GB9015369D0 (en) * | 1990-07-12 | 1990-08-29 | Erba Carlo Spa | Amidated fibrinolytic enzymes and their precursors,and processes for their preparation |
| US5242810A (en) * | 1990-12-07 | 1993-09-07 | Biogen, Inc. | Bifunctional inhibitors of thrombin and platelet activation |
| US5688768A (en) * | 1991-02-19 | 1997-11-18 | Cor Therapeutics, Inc. | Recombinant thrombin receptor and related pharmaceuticals |
| WO1992018139A1 (en) * | 1991-04-09 | 1992-10-29 | Brigham And Women's Hospital | Chimeric molecule with plasminogen activator activity and affinity for atherosclerotic plaques |
| US5376367A (en) * | 1991-11-22 | 1994-12-27 | Immunex Corporation | Fusion proteins comprising MGF and IL-3 |
| US5571708A (en) * | 1993-04-19 | 1996-11-05 | Bristol-Myers Squibb Company | Thrombin-activatable plasminogen activator |
| RU2061043C1 (ru) * | 1993-11-16 | 1996-05-27 | Научно-производственная фирма "Нарт" | Способ повышения устойчивости урокиназы к нагреванию |
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- 1994-07-12 RU RU94026095A patent/RU2143490C1/ru active
- 1994-07-13 CA CA002127897A patent/CA2127897A1/en not_active Abandoned
- 1994-07-13 NZ NZ260991A patent/NZ260991A/xx unknown
- 1994-07-13 KR KR1019940016792A patent/KR960014346A/ko not_active Ceased
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- 1994-07-14 JP JP6162445A patent/JPH07143878A/ja active Pending
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| PD00 | Pending as of 2000-06-30 in czech republic | ||
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