RU2829367C2 - Closed-circular dna (ccdna) and use in methods for reducing immune response associated with gene therapy or nucleic acid therapy - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology, in particular to a method of inhibiting an immune response, comprising administering an immune response inhibitor and a non-viral capsid DNA vector with covalently closed ends, containing at least one transgene, into a cell in which at least one transgene is expressed. Also disclosed are a composition, a pharmaceutical composition, a cell for expressing an immune response inhibitor and a kit for inhibiting the immune response.

EFFECT: invention is effective for inhibiting the immune response, wherein at least one transgene is expressed in a cell.

136 cl, 18 dwg, 12 tbl, 35 ex

Description

RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application No. 62/796,417, filed January 24, 2019, U.S. Provisional Application No. 62/800,303, filed February 1, 2019, U.S. Provisional Application No. 62/796,450, filed January 24, 2019, U.S. Provisional Application No. 62/800,285, filed February 1, 2019, U.S. Provisional Application No. 62/814,414, filed March 6, 2019, U.S. Provisional Application No. 62/814,424, filed March 6, 2019, and U.S. Provisional Application No. 62/857,542, filed June 5, 2019, the contents of each of which are incorporated herein by reference in their entirety.

SEQUENCE LIST

[0002] This application includes a sequence listing filed electronically in ASCII format and hereby incorporated herein by reference in its entirety. Said ASCII copy, created on January 24, 2020, is designated 131698-03320-Sequence_Listing-FINAL.txt and is 117,124 bytes in size.

AREA OF TECHNOLOGY

[0003] Embodiments of the present invention relate to the field of gene therapy, including delivery of exogenous DNA sequences to a target cell, tissue, organ or organism, as well as modifications and methods for inhibiting immune responses (e.g., innate immune responses) to said sequences.

LEVEL OF TECHNOLOGY

[0004] The goal of gene therapy is to improve clinical outcomes in patients suffering from either genetic mutations or acquired diseases caused by an aberration in the gene expression profile. Gene therapy includes the treatment or prevention of medical conditions caused by defective genes or abnormal regulation or expression, such as underexpression or overexpression, which can lead to a disorder, disease, malignancy, etc. For example, a disease or disorder caused by a defective gene can be treated, prevented, or alleviated by delivering corrective genetic material to the patient, or can be treated, prevented, or alleviated by altering or suppressing the defective gene in the patient, such as by using corrective genetic material that results in therapeutic expression of the genetic material in the patient.

[0005] The basis of gene therapy is the provision of a transcription cassette with an active gene product (sometimes called a transgene), for example, which may result in a beneficial gain-of-function effect, a detrimental loss-of-function effect, or another outcome. Such outcomes may be attributed to the expression of an activating antibody or fusion protein or an inhibitory (neutralizing) antibody or fusion protein. Gene therapy may also be used to treat disease or malignancy caused by other factors. Monogenic disorders in humans can be treated by delivering a normal gene to and expressing it in target cells. Delivery and expression of a corrective gene in a patient's target cells can be accomplished using a variety of methods, including the use of modified viruses and viral vectors for gene delivery. Among the numerous available viral-derived vectors (e.g., recombinant retrovirus, recombinant lentivirus, recombinant adenovirus, etc.), recombinant adeno-associated virus (rAAV) is gaining popularity in gene therapy as a multi-purpose vector.

[0006] Adeno-associated viruses (AAV) belong to the family Parvoviridae and, more specifically, constitute the genus Dependoparvovirus. Vectors derived from AAV (i.e., recombinant AAV (rAAV) or AAV vectors) are attractive for delivery of genetic material because (i) they are able to infect (transduce) a wide range of non-dividing and dividing cell types, including myocytes and neurons; (ii) they lack viral structural genes, which reduces host cell responses to viral infection, such as interferon-mediated responses; (iii) wild-type viruses are considered non-pathological in humans; (iv) unlike wild-type AAVs, which are able to integrate into the host cell genome, replication-defective AAV vectors lack the rep gene and typically persist as episomes, limiting the risk of insertional mutagenesis or genotoxicity; and (v) compared to other vector systems, AAV vectors are generally considered to be relatively weak immunogens and therefore do not trigger a significant immune response (see ii), thereby ensuring the stability of vector DNA and, potentially, long-term expression of therapeutic transgenes.

[0007] However, the use of AAV particles as a gene delivery vector has several serious drawbacks. One of the major drawbacks associated with rAAV is the limited viral packaging capacity of approximately 4.5 kb of heterologous DNA (Dong et al., 1996; Athanasopoulos et al., 2004; Lai et al., 2010), thereby limiting the use of AAV vectors to their ability to encode proteins smaller than 150,000 Da. A second drawback is that, due to the prevalence of wild-type AAV infection in the population, candidates for rAAV gene therapy must be screened for neutralizing antibodies that eliminate the vector from the patient. A third drawback is the immunogenicity of the capsid, which precludes re-administration to patients who have not been weaned off the initial treatment. The patient's immune system may respond to the vector, which in effect acts as a "booster" inoculation, stimulating the immune system to produce high titers of antibodies to AAV that preclude future treatment. Some recent reports have raised concerns regarding immunogenicity in high-dose situations. Another major drawback is the relatively slow onset of AAV-mediated gene expression, given that AAV single-stranded DNA must be converted to double-stranded DNA before heterologous gene expression can occur.

[0008] In addition, conventional capsid-containing AAV virions are produced by introducing a plasmid or plasmids containing the AAV genome, rep genes, and cap genes (Grimm et al., 1998). However, such capsid-containing AAV viral vectors have been found to be inefficient in transducing certain cell types and tissues, and that the capsids also induce an immune response. Accordingly, the use of adeno-associated virus (AAV) vectors for gene therapy is limited by the single administration to patients (due to the patient's immune response), the limited range of transgenic genetic material suitable for delivery in AAV vectors due to the minimal viral packaging capacity (approximately 4.5 kb), and the slow AAV-mediated gene expression.

[0009] Furthermore, mammalian immune systems include a number of mechanisms for detecting and eliminating invading pathogens and various types of aberrant cellular activities and processes that may be induced by the introduction of a viral vector or nucleic acid into a subject. For example, pattern recognition receptors (PRRs) are a class of molecules that have evolved as sensors to detect conserved molecules associated with pathogens, such as foreign nucleic acids, such as viral DNA and viral RNA, and to initiate an innate immune response. Toll-like receptors (TLRs) are a group of PRRs that detect endosomally confined nucleic acids and include TLR9 (detects dsDNA, predominantly unmethylated CpG repeats), TLR3 (detects dsRNA), and TLR7 (detects ssRNA). A second PRR system is located in the cytosol and serves to detect foreign nucleic acid, particularly double-stranded RNA, within infected cells. ¹ These PRRs, called “RIG-I-like receptors” or RLRs, include RIG-I and MDA5. The PRRs mentioned are helicases that detect structural features of RNA such as 5´-triphosphates and diphosphates, RNA replication intermediates and/or transcription products, and initiate activation of the type I interferon response. ^1,2 A third class of PRRs is triggered by cytosolic DNA, with the major intracellular DNA sensor being cGAS (cyclo-GMP-AMP synthase), which binds to DNA and activates endoplasmic reticulum (ER)-associated stimulator of interferon genes (STING), leading to activation of the type I interferon response and, in some cases, activation of ^1,4,5 other proposed cytosolic DNA sensors including absent in melanoma protein 2 (AIM2), IFN-γ-inducible protein 16 (IFI16), interferon-inducible protein X (IFIX), LRRFIP1, DHX9, DHX36, DDX41, Ku70, DNA-PKc, MRN complex (including MRE11, Rad50 and Nbs1) ^2,7 and RNA polymerase III ¹⁰ . AIM2, IFI16 and IFIX are pyrin domain and HIN200 (PYHIN) domain-containing proteins. ^2,6 Furthermore, unpaired DNA nucleotides flanking short stretches of base-paired DNA, as in the stem-loop structures of human immunodeficiency virus type 1 (HIV-1)-derived single-stranded DNA (ssDNA), have been shown to activate the type I interferon-inducing DNA sensor cGAS in a sequence-dependent manner. ^8,9 DNA structures containing unpaired guanosines flanking short (12 to 20 bp) dsDNA (Y-form DNA) had a strong stimulatory effect and specifically enhanced the enzymatic activity of cGAS. ^8,9

[0010] More recently, other intracellular microbial sensors have been identified, including NOD-like receptors (NLRs). Some NLRs also sense non-microbial danger signals and form large cytoplasmic protein complexes called inflammasomes, which are a major regulator of innate immunity and inflammation (Martinon et al., Annu. Rev. Immunol. 2009 27: 229-65).

[0011] The inflammasome is composed of NLR family receptors or AIM2 and procaspase-1. Apoptosis-associated speck-like protein containing the caspase recruitment domain (ASC) is an adaptor protein and links an NLR family member to procaspase-1. NLR family members assemble the inflammasome complex with ASC, which in turn recruits and activates caspase-1. Several members of the NLR family of proteins are involved in the formation of different inflammasomes, including NLR family pyrin domain-containing member 3 (NLRP3; also known as cyropyrin or NALP3), NLR family CARD domain-containing member 4 (NLRC4; also known as IPAF), and NLRP1. Different inflammasomes are activated by different stimuli. For example, NLRP1 is activated by lethal toxin produced by Bacillus anthracis, while NLRC4 responds to cytosolic flagellin in cells infected with Salmonella, Legionella, and Pseudomonas spp. The NLRP3 inflammasome is activated by a wide variety of stimuli, including microbial products and endogenous signals such as urate crystal, silica, amyloid fibrils, and ATP.

[0012] The NOD-like receptor (NLR) sensor component (e.g., cryopyrin (NLRP3 or NALP3)) recognizes danger signals such as damage-associated molecular patterns (DAMPs) released during tissue injury or stress (e.g., extracellular ATP, urate crystal, β-amyloid, cellular debris) and pathogen-associated molecular patterns (PAMPs). The inflammasome assembles in response to such pathogen infection or "danger" signals, which requires the interaction of the pyrin domains of cryopyrin and the adaptor component ASC, resulting in the recruitment and activation of caspase-1 (from procaspase-1) and then the maturation and release of several proinflammatory cytokines including interleukin-1β (IL-1β), IL-18, and IL-33.

[0013] In addition to NLRs, inflammasomes can be activated by members of the AIM2 family. AIM2 is characterized by the presence of a pyrin domain and a HIN DNA-binding domain and activates caspase-1 by detecting cytosolic DNA (Fernandes-Alnemri T, et al. 2009. Nature 458:509-513). Inflammasome assembly requires a preceding priming signal by TLRs, which is required to upregulate the expression of inflammasome receptors and the pro-IL-1β substrate before the second signal can initiate inflammasome complex formation (Bauernfeind FG, et al., 2009, J. Immunol. 183:787-791).

[0014] Despite their conceptual elegance, the prospect of using nucleic acid molecules for gene therapy to treat human diseases remains uncertain. The primary reason for this uncertainty lies in the apparent adverse effects associated with the host innate immune response to nucleic acid therapeutics and, thus, the way in which these materials modulate the expression of their intended targets in relation to the immune response. The current state of the art relating to the design, function, behavior, and optimization of nucleic acid molecules that can be used for clinical applications has focused primarily on: (1) antisense oligonucleotides and duplex RNAs that directly regulate gene translation and expression; (2) gene transcription-silencing RNAs that result in long-term epigenetic modifications; (3) antisense oligonucleotides that interact with and alter gene splicing structures; (4) the creation of synthetic or viral vectors that mimic the physiological functionality of the natural AAV genome or lentiviral genome; and (5) the delivery of therapeutic oligonucleotides in vivo. However, despite the advances made in the development of nucleic acid-based therapeutics, which are evident in recent clinical achievements, the field of gene therapy is still severely limited by unintended adverse events in recipients triggered by the therapeutic nucleic acids themselves.

[0015] Accordingly, there is a need in the art for a new technology that inhibits (e.g., reduces, alleviates, mitigates, prevents) an immune response to the administration of vectors or nucleic acid to a subject, thereby enabling expression of a therapeutic protein in a cell, tissue, or in the subject's body for the treatment of a wide range of diseases.

BRIEF DESCRIPTION OF THE INVENTION

[0016] The present disclosure provides methods and pharmaceutical compositions for inhibiting (i.e., reducing or suppressing) an immune response in a subject suffering from a genetic disorder and receiving gene therapy or nucleic acid therapy ("nucleic acid therapeutics" or "therapeutic nucleic acid" (TNA)). Provided herein are non-viral capsidless DNA vectors with covalently closed ends (ceDNA vectors) and inhibitors for inhibiting an immune response (e.g., an innate immune response). In some embodiments, the pharmaceutical compositions and formulations may include one or more immune response (e.g., innate immune response) inhibitors such as rapamycin and rapamycin analogs, TLR antagonists (e.g., TLR9 antagonists), cGAS antagonists, and inflammasome antagonists (e.g., any one or more of an NLRP3 inflammasome pathway inhibitor or an AIM2 inflammasome pathway inhibitor or a caspase 1 inhibitor or any combination thereof).

[0017] According to some aspects of the present disclosure, compositions and methods are provided for inhibiting (i.e., reducing or suppressing) an immune response (e.g., an innate immune response) using non-viral capsidless DNA vectors with covalently closed ends (ccDNA vectors) for expressing an inhibitor of an innate immune response from a capsidless (e.g., non-viral) DNA vector with covalently closed ends (referred to herein as a "closed-end DNA vector" or "ccDNA vector"), wherein said ccDNA vector comprises a nucleic acid sequence or codon-optimized variants thereof of an inhibitor of an immune response (e.g., an innate immune response).

[0018] According to some aspects of the present disclosure, compositions and methods are provided for inhibiting (i.e., reducing or suppressing) an immune response (e.g., an innate immune response) using non-viral capsidless DNA vectors with covalently closed ends (ccDNA vectors) for expressing rapamycin and rapamycin analogs from a capsidless (e.g., non-viral) DNA vector with covalently closed ends (referred to herein as a "closed-end DNA vector" or "ccDNA vector"), wherein the ccDNA vector comprises a nucleic acid sequence or codon-optimized variants thereof of rapamycin and rapamycin analogs. Accordingly, these ccDNA vectors can be used to produce rapamycin and rapamycin analogs to inhibit the immune system (e.g., the innate immune system).

[0019] According to some aspects of the present disclosure, compositions and methods are provided for inhibiting (i.e., reducing or suppressing) an immune response (e.g., an innate immune response) using non-viral capsidless DNA vectors with covalently closed ends (ccDNA vectors) for expressing a TLR antagonist from a capsidless (e.g., non-viral) DNA vector with covalently closed ends (referred to herein as a "closed-end DNA vector" or "ccDNA vector"), wherein the ccDNA vector comprises a nucleic acid sequence or codon-optimized variants thereof of a TLR antagonist. Accordingly, these ccDNA vectors can be used to produce a TLR antagonist for inhibiting the immune system (e.g., the innate immune system).

[0020] According to some aspects of the present disclosure, compositions and methods are provided for inhibiting (i.e., reducing or suppressing) an immune response (e.g., an innate immune response) using non-viral capsidless DNA vectors with covalently closed ends (ccDNA vectors) for expressing a cGAS antagonist from a capsidless (e.g., non-viral) DNA vector with covalently closed ends (referred to herein as a "closed-end DNA vector" or "ccDNA vector"), wherein the ccDNA vector comprises a nucleic acid sequence or codon-optimized variants thereof of a cGAS antagonist. Accordingly, these ccDNA vectors can be used to produce a cGAS antagonist for inhibiting the immune system (e.g., the innate immune system).

[0021] According to some aspects of the present disclosure, there are provided compositions and methods for inhibiting (i.e., reducing or suppressing) an immune response (e.g., an innate immune response) using non-viral capsidless DNA vectors with covalently closed ends (ccDNA vectors) for expressing an NLRP3 inflammasome pathway inhibitor or an AIM2 inflammasome pathway inhibitor or a caspase 1 inhibitor, or any combination thereof from a capsidless (e.g., non-viral) DNA vector with covalently closed ends (referred to herein as a "closed-end DNA vector" or "ccDNA vector"), wherein said ccDNA vector comprises a nucleic acid sequence, or codon-optimized variants thereof, of an NLRP3 inflammasome pathway inhibitor or an inflammasome pathway inhibitor AIM2, or a caspase 1 inhibitor, or any combination thereof. Accordingly, these ccDNA vectors can be used to produce an NLRP3 inflammasome pathway inhibitor, or an AIM2 inflammasome pathway inhibitor, or a caspase 1 inhibitor, or any combination thereof, to inhibit the immune system (e.g., the innate immune system).

[0022] In some embodiments, pharmaceutical compositions and formulations may include one or more of the immune response (e.g., innate immune response) inhibitors described herein in combination with various types of therapeutic nucleic acids (TNAs) and carriers (e.g., a lipid nanoparticle). In some embodiments, the composition further comprises an excipient or carrier. In some embodiments, the pharmaceutical composition comprises a lipid nanoparticle (LNP). In one embodiment, the LNP comprises a cationic lipid. In some embodiments, the LNP comprises polyethylene glycol (PEG). In some embodiments, the LNP comprises cholesterol.

[0023] The methods described herein typically involve the use of one or more inhibitors of an immune response (e.g., an innate immune response) (e.g., rapamycin and its analogs, TLR antagonists, cGAS antagonists) to prevent, reduce, attenuate, or even eliminate immune responses associated with the introduction of a transgene (e.g., a therapeutic nucleic acid (TNA)). The methods described herein include administering these inhibitors.

[0024] In one embodiment, the therapeutic nucleic acid is an RNA molecule or a derivative thereof. In one embodiment, the RNA molecule is an antisense oligonucleotide. In one embodiment, the antisense oligonucleotide is antisense RNA. In one embodiment, the RNA is RNA interference (RNAi).

[0025] According to one embodiment, the therapeutic nucleic acid is an mRNA molecule.

[0026] According to one embodiment, the therapeutic nucleic acid is a DNA molecule or a derivative thereof.

[0027] In one embodiment, the therapeutic nucleic acid is an antisense DNA oligonucleotide. In one embodiment, the antisense DNA oligonucleotide is a morpholine-based nucleic acid. In one embodiment, the morpholine-based nucleic acid is a phosphodiamidate morpholine oligomer (PMO).

[0028] In one embodiment, the therapeutic nucleic acid is closed-ended DNA (ccDNA). In one embodiment, the ccDNA comprises an expression cassette comprising a promoter sequence and a transgene. In one embodiment, the ccDNA comprises an expression cassette comprising a polyadenylation sequence. In one embodiment, the ccDNA comprises at least one inverted terminal repeat (ITR) flanking either the 5' or 3' end of the expression cassette. In one embodiment, the expression cassette is flanked by two ITRs, wherein the two ITRs comprise one 5' ITR and one 3' ITR. In one embodiment, the expression cassette is linked to the ITR at the 3' end (3' ITR). In one embodiment, the expression cassette is linked to the ITR at the 5' end (5' ITR). According to one embodiment, the ccDNA further comprises a spacer sequence between the 5'-ITR and the expression cassette.

[0029] In one embodiment, the ccDNA further comprises a spacer sequence between the 3'-ITR and the expression cassette. In one embodiment, the spacer sequence is at least 5 base pairs in length. In one embodiment, the spacer sequence is 5 to 200 base pairs in length. In one embodiment, the spacer sequence is 5 to 500 base pairs in length.

[0030] In one embodiment, the ITR is an ITR derived from serotype AAB. In one embodiment, AAB is selected from the group consisting of AAB1, AAB2, AAB3, AAB4, AAB5, AAB6, AAB7, AAB8, AAB9, AAB10, AAB11, and AAB12. In one embodiment, the ITR is derived from an ITR of a goose virus. In one embodiment, the ITR is derived from an ITR of a B19 virus. In one embodiment, the ITR is a wild-type ITR from a parvovirus. In one embodiment, the ITR is a mutant ITR. In one embodiment, the ccDNA comprises two mutant ITRs at both the 5' and 3' end of the expression cassette.

[0031] According to one embodiment, the ccDNA has a single-strand break or gap.

[0032] According to one embodiment, the cDNA is produced synthetically in a cell-free environment.

[0033] In one embodiment, the ccDNA is produced in a cell. In one embodiment, the ccDNA is produced in insect cells. In one embodiment, the insect cell is Sf9. In one embodiment, the ccDNA is produced in a mammalian cell. In one embodiment, the mammalian cell is a human cell line.

[0034] According to one embodiment, the therapeutic nucleic acid is a closed-ended DNA comprising at least one proteomerase target sequence at the 5' and 3' ends of its expression cassette.

[0035] According to one embodiment, the therapeutic nucleic acid is a dumbbell-shaped, closed-ended, linear duplex DNA containing two ITR hairpin structures at the 5' and 3' ends of the expression cassette.

[0036] According to one embodiment, the therapeutic nucleic acid is a DNA-based minicircle or MIDGE.

[0037] In one embodiment, the therapeutic nucleic acid is a linear DNA vector with covalently closed ends. In one embodiment, the linear DNA vector with covalently closed ends is a DNA with a minimal strand ("ministring DNA").

[0038] According to one embodiment, the therapeutic nucleic acid is doggybone DNA (dbDNA ^™ ).

[0039] According to one embodiment, the therapeutic nucleic acid is a minigene.

[0040] According to one embodiment, the therapeutic nucleic acid is a plasmid.

[0041] Accordingly, in some aspects of the present invention, there are provided methods for inhibiting or suppressing immune responses upon expression of a transgene in a cell, comprising: co-administering to the cell (1) a composition comprising a non-viral capsidless DNA vector with covalently closed ends (ccDNA vector) and (2) an inhibitor of an immune response (e.g., an innate immune response) as described herein. The ccDNA vector comprises a heterologous nucleic acid sequence encoding a transgene operably located between two different AAV inverted terminal repeat (ITR) sequences, wherein one of said ITRs comprises a functional AAV terminal resolution site and a Rep binding site, one of said ITRs comprises a deletion, insertion or substitution relative to the other ITR such that the ccDNA vector, when digested with a restriction enzyme having one recognition site on the ccDNA vector, has characteristic bands of linear and continuous DNA, compared to linear and discontinuous control DNA, when analyzed on a non-denaturing gel. As shown herein, in some embodiments, an inhibitor of an immune response (e.g., an innate immune response) is co-administered using a synthetic nanocarrier described in WO 2016/073799, the contents of which are incorporated herein by reference in their entirety. In some embodiments, the ccDNA vector is also present in a nanocarrier. In some embodiments, the one or more immune response (e.g., innate immune response) inhibitors are selected from rapamycin and rapamycin analogs, TLR antagonists (e.g., TLR9 antagonists), cGAS antagonists, and inflammasome antagonists (e.g., any one or more of an NLRP3 inflammasome pathway inhibitor or an AIM2 inflammasome pathway inhibitor or a caspase 1 inhibitor or any combination thereof). In some embodiments, the TLR9 inhibitory oligonucleotide is present on at least one of the ITRs. In some embodiments, the cGAS inhibitor is encoded by the ccDNA and is operably linked to a promoter, such as an inducible promoter. In other embodiments, the cGAS inhibitor is not encoded by the ccDNA.

[0042] Furthermore, according to one aspect of the present invention, there is provided a composition comprising (i) a non-viral capsidless DNA vector with covalently closed ends (ccDNA vector), wherein said ccDNA vector comprises a heterologous nucleic acid sequence encoding a transgene operably positioned between two different AAV inverted terminal repeat (ITR) sequences, wherein one of said ITRs comprises a functional AAV end resolution site and a Rep binding site, one of said ITRs comprises a deletion, insertion or substitution relative to the other ITR, wherein said ccDNA vector, when digested with a restriction enzyme having a single recognition site on the ccDNA vector, has characteristic bands of linear and continuous DNA, compared to linear and discontinuous control DNA, when analyzed on a non-denaturing gel, and (ii) an inhibitor of an immune response (e.g., an innate immune response) response). As shown herein, in some embodiments, the components of the composition are formulated in separate synthetic nanocarriers. In one embodiment, the components of the composition are formulated in the same synthetic nanocarrier. In some embodiments, the one or more immune response (e.g., innate immune response) inhibitors are selected from rapamycin and rapamycin analogs, TLR antagonists (e.g., TLR9 antagonists), cGAS antagonists, and inflammasome antagonists (e.g., any one or more of an NLRP3 inflammasome pathway inhibitor or an AIM2 inflammasome pathway inhibitor or a caspase 1 inhibitor or any combination thereof).

[0043] The non-viral capsidless DNA vectors described herein can be produced in susceptible host cells from an expression construct (e.g., a plasmid, bacmid, baculovirus, or integrated cell line), e.g., see the examples disclosed in International Patent Application No. PCT/US18/49996, filed September 7, 2018, or using a synthetic production method, e.g., see the examples disclosed in International Patent Application No. PCT/US19/14122, filed December 6, 2018, each of which is incorporated herein by reference in its entirety. In some embodiments, ccDNA vectors that may be used in the methods and compositions disclosed herein comprise a heterologous nucleic acid, e.g., a transgene, positioned between two inverted terminal repeat (ITR) sequences. In some embodiments, at least one of the ITRs is modified by deletion, insertion, and/or substitution compared to a wild-type ITR sequence (e.g., an AAB ITR); and at least one of the ITRs comprises a functional terminal resolution site (TRS) and a Rep binding site.

[0044] According to another aspect of the present disclosure, there is provided a method of treating a genetic disorder in a subject, said method comprising administering to said subject an effective amount of a pharmaceutical composition disclosed herein.

BRIEF DESCRIPTION OF DRAWINGS

[0045] FIG. 1 is a diagram illustrating one embodiment of a preparatory process for producing baculo-infected insect cells (BIIC) that can be used to produce a ccDNA vector in the method described in the diagram of FIG. 2. i) Two populations of naive insect cells are transfected with either a plasmid encoding a Rep protein or a plasmid producing a DNA vector; ii) the viral supernatant is collected and used to infect two new naive insect cell populations to produce BIIC-1, a DNA vector construct, and BIIC-2 (REP). BIIC refers to insect cells infected with a baculovirus. Optionally, step ii) can be repeated one or more times to produce recombinant baculovirus in larger quantities.

[0046] FIG. 2 is a diagram illustrating one embodiment for producing the ccDNA vector described herein.

[0047] FIG. 3 is a diagram illustrating one embodiment for characterizing the DNA vector (subsequent method) described herein.

[0048] FIG. 4A-FIG. 4D are schematic diagrams illustrating exemplary plasmids and plasmid components that can be used to produce the ccDNA vector disclosed herein. FIG. 4A shows an exemplary Rep plasmid, and FIG. 4B shows an exemplary TTX vector plasmid that contains a ccDNA vector template. FIG. 4C and FIG. 4D are diagrams of exemplary functional components of a DNA vector template that can be used to produce the ccDNA vectors provided herein. The transgene, which is also referred to as a nucleic acid of interest (e.g., a reporter nucleic acid such as luciferase, or, for example, a therapeutic nucleic acid), is located between two different ITRs. The modified ITR may be located on either the left side (FIG. 4C) or the right side (FIG. 4D) of the template. Additionally, the nucleic acid of interest may be operably linked to a promoter, enhancer, and termination elements. In alternative embodiments, the ITR on the left (5'-ITR) or right (3'-ITR) may be of any type. For illustrative purposes, the ITRs in the ccDNA constructs of FIG. 4C and FIG. 4D and in the Examples herein show a modified ITR (ΔITR) and a wild-type ITR (WT-ITR) and represent an example of a pair of asymmetric ITRs. However, provided herein are ccDNA vectors that comprise a heterologous nucleic acid sequence (transgene) located between any two inverted terminal repeat (ITR) sequences, wherein said ITR sequences may be a pair of asymmetric ITRs or a pair of symmetric or substantially symmetric ITRs, as those terms are defined herein. A ccDNA vector comprising an NLP disclosed herein may comprise ITR sequences that are selected from any of: (i) at least one WT-ITR and at least one modified inverted terminal repeat (mod-ITR) of AAB (e.g., asymmetric modified ITRs); (ii) two modified ITRs, wherein said pair of mod-ITRs have a different three-dimensional spatial organization relative to each other (e.g., asymmetric modified ITRs), or (iii) a pair of symmetric or substantially symmetric WT-WT-ITRs, in which each WT-ITR has the same three-dimensional spatial organization, or (iv) a pair of symmetric or substantially symmetric modified ITRs, in which each mod-ITR has the same three-dimensional spatial organization, wherein said methods according to the present disclosure may further include a delivery system, such as, but not limited to, a liposome nanoparticle-based delivery system.

[0049] FIG. 5A and FIG. 5B are drawings that illustrate one embodiment for identifying the presence of DNA vectors described herein. FIG. 5A illustrates DNA having a discontinuous structure (open DNA, e.g., a control DNA cassette isolated from a TTX vector template having open ends) and exemplary characteristic bands obtained upon cutting with a restriction endonuclease having a single recognition site on the discontinuous DNA, e.g., detection of two DNA fragments of different expected sizes (e.g., 1 kb and 2 kb) under denaturing conditions. FIG. 5B illustrates DNA having a linear and continuous closed-end structure and exemplary characteristic bands obtained upon cutting with a restriction endonuclease having a single recognition site on the linear duplex continuous DNA, e.g., detection of two DNA fragments of different sizes (e.g., 2 kb and 4 kb) under denaturing conditions, which is 2 times larger than would be expected if the DNA were discontinuous. Although the DNA is denatured, the complementary strands are covalently linked, and the resulting denatured products are single-stranded DNA that are twice the length of the corresponding discontinuous products.

[0050] FIG. 6 is an exemplary non-denaturing gel showing the presence of highly stable DNA vectors and characteristic bands confirming the presence of highly stable closed-end DNA (ccDNA vector).

[0051] FIG. 7 is a gel and standard curve for quantification for assessing DNA material obtained using the methods disclosed herein.

[0052] FIG. 8 is a Western blot analysis of FIX protein expressed from HEK293 cells containing various constructs and visualized using an anti-Factor IX antibody.

[0053] FIG. 9 is a graphical representation of the results of Example 24. Samples injected hydrodynamically show a significant increase in overall flux (e.g., luciferase expression) compared to samples injected non-hydrodynamically over the three-day study period.

[0054] FIGS. 10A and 10B represent data from the cultured THP-1 cell experiments described in the Examples, which assessed the interferon response in cells treated with a ccDNA vector and immune inhibitors. FIG. 10A shows activation of the interferon pathway in response to ccDNA in THP-1 cells with intact cGAS/STING and TLR9 pathways, but no activation in the same cells in which either pathway is disrupted. It should be noted that inclusion of either the inhibitor A151 or BX795 similarly reduces interferon pathway activation. FIG. 10B is a similar experiment showing dose-dependent inhibition of interferon induction by A151 and AS1411. In each group of columns, the 2.5 μM dose is on the left, the 1.25 μM dose is in the middle, and the 0.625 μM dose is on the right.

[0055] FIG. 11A and 11B are graphs of data obtained in Example 26. FIG. 11A shows a decrease in NF-κB induction upon introduction of ccDNA when CpGs present in the ccDNA are methylated prior to introduction into the cells. FIG. 11B also shows that inclusion of the immune inhibitor A151 reduced ccDNA-stimulated NF-κB induction to the same extent as CpG methylation in this assay.

[0056] FIG. 12A-FIG. 12C represent the results of the experiments described in Example 26. FIG. 12A and FIG. 12B are graphs of data from each of the cytokine induction assays performed on blood samples taken from mice treated with the ccDNA vector or mice treated with control LNP-poly(C), with the specific cytokine of interest shown at the top of each graph. FIG. 12C represents data from the ccDNA-driven luciferase expression assay in treated mice, showing the total flux in each group of mice throughout the study. High levels of unmethylated CpG correlated with lower total flux observed in the mice.

[0057] FIG. 13 shows total flux data obtained in the experiments described in Example 27 in mice on day 8 of neonatal development. In the study, ccDNA with high CpG content caused a decrease in flux during the assay, while ccDNA with reduced or no unmethylated CpG maintained luciferase expression. A single re-dosing modestly increased the observed expression levels in samples with minimal or no CpG content, but this sustained increase after re-dosing was not observed in samples from the high CpG group.

[0058] FIG. 14A-FIG. 14C provide results of the experiments described in Example 28. FIG. 14A and FIG. 14B are graphs of data from each of the cytokine induction assays performed on blood samples from ccDNA vector-treated STING mutant mice or control poly(C) treated samples, with the specific cytokine of interest shown at the top of each graph. With the exception of IL-18, significantly less cytokine induction was observed with low CpG ccDNA and no methylated CpG. FIG. 14C provides data from the ccDNA-driven luciferase expression assay in treated STING mutant mice, showing the full flux in each group of mice throughout the study. The results obtained again showed a correlation between high levels of unmethylated CpG in ccDNA and the observed lower total flux.

[0059] FIG. 15A and FIG. 15B show the expression of Padua FIX and FIX transgenes from the highly stable DNA vectors disclosed herein. Quantitative analysis of FIX protein levels expressed from plasmids or vectors was also assessed using the VisuLize Factor IX ELISA kit (Affinity Biologicals, #FIX-AG) following protocols provided by the supplier.

[0060] FIGS. 16A and 16B show the results of the ccDNA stability and repeat dosing study in Rag2 mice described in Example 10. FIG. 16A is a graph of the total flux versus time observed in LNP-ccDNA-Luc-treated wild-type c57bl/6 mice or Rag2 mice. FIG. 16B is a graph showing the effect of repeat dosing on luciferase transgene expression levels in Rag2 mice, with increased stable expression observed following repeat dosing (arrow indicates time of repeat dosing).

[0061] FIG. 17 shows data from the luciferase expression study from ccDNA in treated mice described in Example 29, showing the total flux in each group of mice over the course of the study. High levels of unmethylated CpG correlated with lower total flux observed in mice over time, while the use of a liver-specific promoter correlated with long-term stable expression of the transgene from the ccDNA vector for at least 77 days.

[0062] FIGS. 18A-18H show cytokine levels following administration of a ccDNA vector with pharmacological depletion of macrophages with an NLRP3 inhibitor (MCC950) or a caspase 1 inhibitor (VX765). FIG. 18A shows IFN-α levels, FIG. 18B shows IFN-γ levels showing a significant reduction in IFN-γ with the NLRP3 inhibitor MCC950 (see arrow), FIG. 18C shows IL-β levels, FIG. 18D shows IL-18 levels showing a significant reduction in IFN-γ with the NLRP3 inhibitor MCC950 (see arrow), FIG. 18E shows IL-6 levels, FIG. 18F shows IP-10 levels, FIG. 18G shows MCP-1 levels, FIG. 18H shows TNFα levels.

DETAILED DESCRIPTION OF THE INVENTION

[0063] Nucleic acid transfer vectors and nucleic acid-based therapeutic agents are promising therapeutic agents for a number of applications, such as gene expression and modulation. Viral transfer vectors may contain transgenes encoding proteins or nucleic acids. Examples of such vectors include AAV vectors, microRNAs (miRNAs), short interfering RNAs (siRNAs), and antisense oligonucleotides that bind to mutation sites in messenger RNA (such as small nuclear RNA (snRNA)). Unfortunately, the promise of these therapeutic agents has not yet been realized, largely due to cellular and humoral immune responses directed against the viral transfer vector. These immune responses include antibody, B cell, and T cell responses and are often specific for viral antigens of the viral transfer vector, such as viral capsid or envelope proteins or peptides thereof.

[0064] Currently, many potential patients have some level of pre-existing immunity against the viruses on which the viral transfer vectors are based. In fact, antibodies against viral nucleic acids (both DNA and RNA) or proteins are widely distributed in the human population. Furthermore, even if the level of pre-existing immunity is low, such as due to low immunogenicity of the viral transfer vector, such low levels may still prevent successful transduction (e.g., Jeune, et al., Human Gene Therapy Methods, 24:59-67 (2013)). Thus, even low levels of pre-existing immunity may prevent the use of a particular viral transfer vector in a patient and may require the clinician to select a viral transfer vector based on a virus of a different serotype, which may not be as effective, or even to forgo another type of therapy altogether if therapy with a different viral transfer vector is not available.

[0065] In addition, viral vectors, such as adeno-associated virus vectors, can be highly immunogenic and can induce humoral and cell-mediated immunity that can reduce efficacy, particularly with respect to repeated administration. In fact, cellular and humoral immune responses against the viral transfer vector can develop after a single administration of the viral transfer vector. Following administration of the viral transfer vector, neutralizing antibody titers can increase and remain high for several years and can reduce the efficacy of repeated administration of the viral transfer vector. Indeed, repeated administration of the viral transfer vector typically results in enhanced unwanted immune responses. In addition, CD8 ⁺ T cells specific for the viral transfer vector can emerge and eliminate transduced cells expressing the target transgene product, such as upon repeated exposure to viral nucleic acid or capsid protein similar to viral antigens. For example, it has been shown that AAV capsid nucleic acids or antigens can trigger immune-mediated destruction of hepatocytes transduced with a viral AAV transfer vector. It is believed that for many therapeutic applications, multiple rounds of administration of viral transfer vectors are necessary to obtain long-term benefits. However, the ability to do so would be severely limited, particularly if repeated administration is necessary, without the methods and compositions provided herein.

[0066] Methods and compositions are provided that provide solutions to the above-mentioned obstacles for the effective use of various nucleic acid-based therapeutics, including viral or non-viral (synthetic) vectors for transfer, and other nucleic acid-based therapeutics for treatment. The present disclosure relates to the delivery of exogenous DNA sequences to a target cell, tissue, organ or organism, as well as to modifications and methods for inhibiting (i.e., reducing or suppressing) an immune response (e.g., an innate immune response) thereto. Such modifications and methods for inhibiting (i.e., reducing or suppressing) an immune response (e.g., an innate immune response) can be used, for example, to increase the duration of transgene expression.

[0067] It has been unexpectedly discovered that the immune response (e.g., innate immune response) to a DNA transfer vector can be reduced using the methods and corresponding compositions provided herein. Accordingly, the methods and compositions can potentially improve the efficacy of treatment using viral transfer vectors and other therapeutic nucleic acid molecules and provide therapeutic benefits in the long term, even with repeated administration of the viral transfer vector or other nucleic acid-based therapeutic agents.

I. Definitions

[0068] Unless otherwise defined herein, scientific and technical terms used in connection with the present application shall have the standard meanings commonly understood by those skilled in the art to which this invention pertains. It is to be understood that the present invention is not limited to the specific methods, protocols, and reagents, etc., described herein and, as such, are subject to change. The terminology used herein is intended solely for the purpose of describing particular embodiments and is not intended to limit the scope of the present invention, which is defined solely by the claims. Definitions of common terms in immunology and molecular biology can be found in the following references: The Merck Manual of Diagnosis and Therapy, 19th edition, published by Merck Sharp & Dohme Corp., 2011 (ISBN 978-0-911910-19-3); Robert S. Porter et al. (eds.), Fields Virology, 6th ed., published by Lippincott Williams & Wilkins, Philadelphia, PA, USA (2013), Knipe, D.M. and Howley, P.M. (eds.), The Encyclopedia of Molecular Cell Biology and Molecular Medicine, published by Blackwell Science Ltd., 1999-2012 (ISBN 9783527600908); and Robert A. Meyers (ed.), Molecular Biology and Biotechnology: a Comprehensive Desk Reference, published by VCH Publishers, Inc., 1995 (ISBN 1-56081-569-8); Immunology, Werner Luttmann, published by Elsevier, 2006; Janeway's Immunobiology, Kenneth Murphy, Allan Mowat, Casey Weaver (eds.), Taylor & Francis Limited, 2014 (ISBN 0815345305, 9780815345305); Lewin's Genes XI, published by Jones & Bartlett Publishers, 2014 (ISBN-1449659055); Michael Richard Green and Joseph Sambrook, Molecular Cloning: A Laboratory Manual, 4 ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., USA (2012) (ISBN 1936113414); Davis et al., Basic Methods in Molecular Biology, Elsevier Science Publishing, Inc., New York, USA (2012) (ISBN 044460149X); Laboratory Methods in Enzymology: DNA, Jon Lorsch (ed.) Elsevier, 2013 (ISBN 0124199542); Current Protocols in Molecular Biology (CPMB), Frederick M. Ausubel (ed.), John Wiley and Sons, 2014 (ISBN047150338X, 9780471503385), Current Protocols in Protein Science (CPPS), John E. Coligan (ed.), John Wiley and Sons, Inc., 2005; and Current Protocols in Immunology (CPI) (John E. Coligan, Ada M. Kruisbeek, David H. Margulies, Ethan M. Shevach, Warren Strobe (eds.), John Wiley and Sons, Inc., 2003 (ISBN 0471142735, 9780471142737), the contents of which are incorporated herein by reference in their entirety.

[0069] As used herein, the terms "administering," "administered," and variations thereof refer to administering a composition or agent (e.g., a therapeutic nucleic acid or an immunosuppressant as described herein) to a subject, and include simultaneous and sequential administration of one or more compositions or agents. "Administering" can refer to, for example, therapeutic, pharmacokinetic, diagnostic, investigational, placebo, and experimental methods. "Administering" also includes in vitro and ex vivo treatment methods. Administration of a composition or agent to a subject is by any suitable route, including orally, pulmonarily, intranasally, parenterally (intravenously, intramuscularly, intraperitoneally, or subcutaneously), rectally, intralymphatic, intratumorally, or locally. Administration of a composition or agent to a subject is by electroporation. Administration includes self-administration and administration by another person. Administration can be by any suitable route. A suitable route of administration allows the composition or agent to perform its intended function. For example, if the appropriate route is intravenous, the composition is administered by injecting said composition or agent into a vein of the subject.

[0070] As used herein, the terms "nucleic acid therapeutic,""therapeutic nucleic acid," and "NaC" are used interchangeably and refer to any embodiment of a therapeutic agent using nucleic acids as the active component of a therapeutic agent for treating a disease or disorder. As used herein, these terms refer to RNA-based therapeutic agents and DNA-based therapeutic agents. Non-limiting examples of RNA-based therapeutic agents include mRNA, antisense RNA and oligonucleotides, ribozymes, aptamers, interfering RNA (RNAi), dsRNA Dicer substrate, short hairpin RNA (shRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA). Non-limiting examples of DNA-based therapeutics include minicircle DNA, minigene, viral DNA (e.g., lentivirus or AAV genome) or non-viral synthetic DNA vectors, closed-end linear duplex DNA (cDNA/CELiD), plasmids, bacmids, doggybone DNA vectors (dbDNA ^™ ), minimalist vector for immunologically defined gene expression (MIDGE), minimal strand non-viral DNA vector (linear covalently closed DNA vector), or dumbbell-shaped minimal DNA vector ("dumbbell DNA").

[0071] As used herein, an "effective amount" or a "therapeutically effective amount" of an active agent or therapeutic agent, such as an immunosuppressant and/or a therapeutic nucleic acid, is an amount sufficient to produce a desired effect, such as normalizing or reducing an immune response (e.g., an innate immune response) and expressing or inhibiting the expression of a target sequence compared to the level of expression detected in the absence of the therapeutic nucleic acid and/or immunosuppressant. Suitable assays for measuring the expression of a target gene or target sequence include, for example, examining protein or RNA levels using techniques known to those skilled in the art, such as dot blotting, Northern blotting, in situ hybridization, ELISA, immunoprecipitation, functional enzyme assay, and phenotypic assays known to those skilled in the art. However, dosage levels are based on a variety of factors including the type of injury, age, weight, sex, medical condition of the patient, severity of the condition, route of administration, and the particular active agent employed. Accordingly, the dosage regimen may vary considerably, but may be routinely determined by a physician using standard techniques. In addition, the terms "therapeutic amount," "therapeutically effective amounts," and "pharmaceutically effective amounts" include prophylactic or preventive amounts of the compositions of the described invention. In prophylactic or preventive applications of the described invention, the pharmaceutical compositions or medicaments are administered to a patient susceptible to or otherwise at risk of developing a disease, disorder, or condition in an amount sufficient to eliminate or reduce the risk of, lessen the severity of, or delay the onset of said disease, disorder, or condition, including the biochemical, histological, and/or behavioral symptoms of said disease, disorder, or condition, its complications, and intermediate pathological phenotypes that occur during the development of said disease, disorder, or condition. Typically, it is preferred to use a maximum dose, i.e. the highest safe dose according to some medical opinion. The terms "dose" and "dosage" are used interchangeably in this document.

[0072] As used herein, the term "therapeutic effect" refers to the effects of treatment, the results of which are assessed as desired and beneficial. A therapeutic effect may include, directly or indirectly, stopping, reducing, or eliminating the manifestation of a disease. A therapeutic effect may also include, directly or indirectly, stopping, reducing, or eliminating the progression of the manifestation of a disease.

[0073] For any therapeutic agent described herein, a therapeutically effective amount can be initially determined based on preliminary in vitro studies and/or animal models. A therapeutically effective dose can also be determined based on human data. The dose administered can be adjusted based on the relative bioavailability and potency of the compound being administered. Dose adjustment to achieve maximum efficacy based on the methods described above and other well-known methods is within the capabilities of one of ordinary skill in the art. General principles for determining therapeutic efficacy, which can be found in Chapter 1 of Goodman and Gilman, The Pharmacological Basis of Therapeutics, 10th ed., McGraw-Hill (New York) (2001), incorporated herein by reference, are summarized below.

[0074] Pharmacokinetic principles provide a basis for modifying the dosage regimen to achieve the target degree of therapeutic efficacy with a minimum of unacceptable adverse events. Additional guidance for dosage modification may be obtained in situations where plasma drug concentrations can be measured and related to the therapeutic window.

[0075] As used herein, the terms "heterologous nucleotide sequence" and "transgene" are used interchangeably and refer to a nucleic acid of interest (other than a nucleic acid encoding a capsid polypeptide) that is inserted into a ccDNA vector and can be delivered and expressed by the ccDNA vector disclosed herein.

[0076] As used herein, the terms "expression cassette" and "transcription cassette" are used interchangeably and refer to a linear region of nucleic acids that includes a transgene operably linked to one or more promoters or other regulatory sequences sufficient to direct transcription of the transgene, but that does not contain capsid coding sequences, other vector sequences, or inverted terminal repeat regions. The expression cassette may further comprise one or more cis-acting sequences (e.g., promoters, enhancers, or repressors), one or more introns, and one or more post-transcriptional regulatory elements.

[0077] The terms "polynucleotide" and "nucleic acid" as used interchangeably herein refer to a polymeric form of nucleotides of any length, whether ribonucleotides or deoxyribonucleotides. Thus, the term includes single-, double-, or multi-stranded DNA or RNA, genomic DNA, cDNA, DNA-RNA hybrids, or a polymer comprising purine and pyrimidine bases or other natural, chemically or biochemically modified, non-natural, or derivatized nucleotide bases. "Oligonucleotide" generally refers to polynucleotides comprising from about 5 to about 100 nucleotides of single- or double-stranded DNA. However, for the purposes of the present disclosure, there is no upper limit on the length of an oligonucleotide. Oligonucleotides are also known as "oligomers" or "oligos" and can be isolated from genes or chemically synthesized by methods known in the art. The terms "polynucleotide" and "nucleic acid" are to be understood as including, in the context of the embodiments described, single-stranded (such as sense or antisense) and double-stranded polynucleotides. The DNA may be in the form of, for example, antisense molecules, plasmid DNA, DNA-DNA duplexes, pre-condensed DNA, PCR products, vectors (P1, PAC, BAC, YAC, artificial chromosomes), expression cassettes, chimeric sequences, chromosomal DNA, or derivatives and combinations of these groups. DNA can be in the form of minicircle, plasmid, bacmid, minigene, minimal strand DNA (linear covalently closed DNA vector), linear closed-end duplex DNA (CELiD or ccDNA), doggybone DNA (dbDNA ^™ ), dumbbell DNA, minimalist vector for immunologically defined gene expression (MIDGE), viral vector, or non-viral vectors. RNA can be in the form of short interfering RNA (siRNA), dsRNA Dicer substrate, short hairpin RNA (shRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), mRNA, rRNA, tRNA, viral RNA (vRNA), and combinations thereof. Nucleic acids include nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, natural, and non-natural, and which have binding properties similar to those of a reference nucleic acid. Examples of such analogs and/or modified residues include, but are not limited to, phosphorothioates, phosphodiamidate-morpholine oligomer (morpholino), phosphoamidates, methylphosphonates, chiral methylphosphonates, 2'-O-methylribonucleotides, locked nucleic acid (LNA ^™ ), and peptide nucleic acids (PNA). Except where specifically limited, the term includes nucleic acids containing known analogs of natural nucleotides that have binding properties similar to those of a reference nucleic acid. Unless otherwise specified, it is also intended that a particular nucleic acid sequence also includes its conservatively modified variants (e.g., degenerate codon substitutions), alleles, orthologs, single nucleotide polymorphisms (SNPs), and complementary sequences, as well as the sequence explicitly specified.

[0078] "Nucleotides" contain the sugar deoxyribose (DNA) or ribose (RNA), a base, and a phosphate group. Nucleotides are linked together by phosphate groups.

[0079] “Bases” include purines and pyrimidines, which include, but are not limited to, the natural compounds adenine, thymine, guanine, cytosine, uracil, inosine and natural analogs, as well as synthetic derivatives of purines and pyrimidines, which include, but are not limited to, modifications that introduce new reactive groups, such as, but are not limited to, amines, alcohols, thiols, carboxylates and alkyl halides.

[0080] As used herein, the term "interfering RNA" or "RNAi" or "interfering RNA sequence" includes single-stranded RNA (e.g., mature siRNA, ssRNAi oligonucleotides, ssDNAi oligonucleotides), double-stranded RNA (i.e., duplex RNA such as siRNA, dsRNA Dicer substrate, shRNA, aiRNA, or pre-siRNA), DNA-RNA hybrid (see, e.g., PCT Publication No. WO 2004/078941), or DNA-DNA hybrid (see, e.g., PCT Publication No. WO 2004/104199) that is capable of reducing or inhibiting the expression of a target gene or sequence (e.g., by mediating the degradation of or inhibiting the translation of mRNAs that are complementary to the interfering RNA sequence) when the interfering RNA is located in the same cell as the target gene or sequence. Thus, interfering RNA refers to a single-stranded RNA that is complementary to the target mRNA sequence, or to a double-stranded RNA formed by two complementary strands or one self-complementary strand. The interfering RNA may have substantial or complete identity with the target gene or sequence, or may contain a region of mismatch (i.e., a mismatch motif). The sequence of the interfering RNA may correspond to the full-length target gene or a subsequence thereof. Preferably, the interfering RNA molecules are chemically synthesized. The disclosures of each of the above-mentioned patent documents are incorporated herein by reference in their entirety for all purposes.

[0081] The interfering RNA includes "short interfering RNA" or "siRNA", such as an interfering RNA of about 15-60, 15-50, or 15-40 (duplex) nucleotides in length, more typically about 15-30, 15-25, or 19-25 (duplex) nucleotides in length, and preferably about 20-24, 21-22, or 21-23 (duplex) nucleotides in length (e.g., each complementary sequence of a double-stranded siRNA is 15-60, 15-50, 15-40, 15-30, 15-25, or 19-25 nucleotides in length, preferably about 20-24, 21-22, or 21-23 nucleotides in length, and the double-stranded siRNA is about 15-60, 15-50, 15-40, 15-30, 15-25 or 19-25 base pairs in length, preferably about 18-22, 19-20 or 19-21 base pairs in length). The siRNA duplexes can comprise 3' overhangs having from about 1 to about 4 nucleotides or from about 2 to about 3 nucleotides and 5' phosphate ends. Examples of siRNAs include, but are not limited to, a double-stranded polynucleotide molecule assembled from two single-stranded molecules, one strand being a sense strand and the other being a complementary antisense strand; a double-stranded polynucleotide molecule assembled from a single-stranded molecule in which the sense and antisense regions are connected by a nucleic acid or non-nucleic acid linker; a double-stranded polynucleotide molecule with a hairpin secondary structure having self-complementary sense and antisense regions; and a circular single-stranded polynucleotide molecule with two or more loop structures and a stem having self-complementary sense and antisense regions, wherein the circular polynucleotide can be processed in vivo or in vitro to produce an active double-stranded siRNA molecule. As used herein, the term "siRNA" includes RNA-RNA duplexes as well as DNA-RNA hybrids (see, for example, PCT Publication No. WO 2004/078941).

[0082] As used herein, the term "nucleic acid construct" refers to a nucleic acid molecule, whether single-stranded or double-stranded, that is isolated from a natural gene or that is modified to contain nucleic acid segments by a method that would not otherwise exist in nature or is synthetic. The term "nucleic acid construct" is synonymous with the term "expression cassette" when the nucleic acid construct contains control sequences necessary for expression of a coding sequence according to the present disclosure. An "expression cassette" includes a DNA coding sequence operably linked to a promoter.

[0083] By "hybridizable" or "complementary" or "substantially complementary" is meant that a nucleic acid (e.g., RNA) includes a nucleotide sequence that allows it to non-covalently associate, i.e., form Watson-Crick base pairs and/or G/U base pairs, "renature" or "hybridize" with another nucleic acid in a sequence-specific, antiparallel manner (i.e., the nucleic acid specifically binds to the complementary nucleic acid) under appropriate conditions of temperature and ionic solution strength in vitro and/or in vivo. As is known in the art, standard Watson-Crick base pairing includes: adenine (A) pairing with thymidine (T), adenine (A) pairing with uracil (U), and guanine (G) pairing with cytosine (C). In addition, it is also known in the art that when two RNA molecules (e.g., dsRNA) hybridize, a guanine (G) base pairs with uracil (U). For example, G/U base pairing is partially responsible for the degeneracy (i.e., redundancy) of the genetic code in the case of base pairing of anti-codons of tRNA with codons in mRNA. For the purposes of the present invention, guanine (G) of the protein binding segment (dsRNA duplex) of the DNA-targeting RNA molecule of the present invention is considered complementary to uracil (U), and vice versa. Thus, when a G/U base pair can be obtained at a particular nucleotide position of the protein binding segment (dsRNA duplex) of the DNA-targeting RNA molecule according to the present invention, that position is not considered to be non-complementary, but is instead considered to be complementary.

[0084] The terms "peptide," "polypeptide," and "protein" are used interchangeably herein and refer to a polymeric form of amino acids of any length, which may include coded and non-coded amino acids, chemically or biochemically modified or derivatized amino acids, and polypeptides having modified peptide backbones.

[0085] A DNA sequence that "encodes" a particular inflammasome antagonist (e.g., any one or more of: an NLRP3 inflammasome pathway inhibitor or an AIM2 inflammasome pathway inhibitor or a caspase 1 inhibitor, or any combination thereof) is a DNA nucleic acid sequence that is transcribed into a particular RNA and/or protein. A DNA polynucleotide may encode an RNA (mRNA) that is translated into a protein, or a DNA polynucleotide may encode an RNA that is not translated into a protein (e.g., tRNA, rRNA, or DNA-targeting RNA; also referred to as "non-coding" RNA or "ncRNA").

[0086] As used herein, the term "fusion protein" refers to a polypeptide that comprises protein domains from at least two different proteins. For example, a fusion protein may comprise (i) one inflammasome antagonist (e.g., any one or more of an NLRP3 inflammasome pathway inhibitor or an AIM2 inflammasome pathway inhibitor or a caspase 1 inhibitor or any combination thereof) or a fragment thereof, and (ii) at least one gene-of-interest (GOI) protein, or alternatively, another inflammasome antagonist protein. The fusion proteins provided herein include, but are not limited to, an antibody or Fc or antigen-binding fragment of an antibody fused to an inflammasome antagonist (e.g., any one or more of an NLRP3 inflammasome pathway inhibitor or an AIM2 inflammasome pathway inhibitor or a caspase 1 inhibitor or any combination thereof), such as an extracellular domain of a receptor, a ligand, an enzyme or a peptide. The inflammasome antagonist (e.g., any one or more of an NLRP3 inflammasome pathway inhibitor or an AIM2 inflammasome pathway inhibitor or a caspase 1 inhibitor or any combination thereof) or fragment thereof that is part of a fusion protein can be a monospecific antibody or a bispecific or multispecific antibody.

[0087] As used herein, the term "genomic safe harbor gene" or "safe harbor gene" refers to a gene or loci into which a nucleic acid sequence can be inserted such that the sequence can integrate and function in a predictable manner (e.g., express a protein of interest) without significant negative consequences for endogenous gene activity or cancer promotion. In some embodiments, a safe harbor gene also is a locus or gene in which the inserted nucleic acid sequence can be expressed more efficiently and at higher levels than at a non-safe harbor site.

[0088] As used herein, the term "gene delivery" refers to the method by which foreign DNA is transferred into host cells for gene therapy applications.

[0089] As used herein, the term "terminal repeat" or "TR" includes any viral terminal repeat or any synthetic sequence that contains at least one minimal required origin of replication and a region containing a palindromic hairpin structure. The Rep binding sequence ("RBS") (also referred to as an RBE (Rep binding element)) and the terminal resolution site ("TRS") together constitute the "minimal required origin of replication" and, accordingly, a TR contains at least one RBS and at least one TRS. Each TR that is reverse complementary to one another within a defined region of a polynucleotide sequence is generally referred to as an "inverted terminal repeat" or "ITR". In the context of a virus, ITRs mediate replication, viral packaging, integration, and proviral release. As has been surprisingly found, according to the present invention, TRs that are not reverse complementary along their entire length can still perform the normal functions of ITRs, and thus, as used herein, the term ITR refers to a TR in a ccDNA genome or a ccDNA vector that is capable of mediating replication of the ccDNA vector. One of ordinary skill in the art will appreciate that more than two pairs of ITRs or asymmetric ITRs may be present in ccDNA vectors of complex configuration. An ITR may be an AAB ITR or a non-AAB ITR, or may be derived from an AAB ITR or a non-AAB ITR. For example, an ITR may be derived from a virus of the Parvoviridae family, which includes parvoviruses and dependoviruses (e.g., canine parvovirus, bovine parvovirus, murine parvovirus, porcine parvovirus, human parvovirus B-19), or the SV40 hairpin, which serves as the origin of replication of SV40, may be used as an ITR, which may be further modified by truncation, substitution, deletion, insertion, and/or addition. Viruses of the Parvoviridae family consist of two subfamilies: Parvovirinae, which infect vertebrates, and Densovirinae, which infect invertebrates. Dependoparvoviruses include the adeno-associated virus (AAV) family of viruses that are capable of replicating in vertebrate hosts, including, but not limited to, humans, primate species, bovine, canine, equine, and ovine. For convenience, in this document, the ITR located in the 5' direction (upstream) relative to the expression cassette in the ecDNA vector is referred to as the "5' ITR" or "left ITR", and the ITR located in the 3' direction (downstream) relative to the expression cassette in the ecDNA vector is referred to as the "3' ITR" or "right ITR".

[0090] "Wild-type ITR" or "WT-ITR" refers to a sequence of a naturally occurring ITR sequence in an AAV or other dependentovirus that retains, for example, Rep binding activity and Rep nicking ability. The nucleotide sequence of a WT-ITR from any AAV serotype may differ slightly from the canonical natural sequence due to degeneracy of the genetic code or drift, and thus the WT-ITR sequences contemplated for use herein include WT-ITR sequences resulting from natural variations that occur during production (e.g., replication errors).

[0091] As used herein, the term "substantially symmetrical WT-ITRs" or "a pair of substantially symmetrical WT-ITRs" refers to a pair of WT-ITRs in a single ccDNA genome or ccDNA vector that are both wild-type ITRs that have reverse complementary sequences along their entire length. For example, an ITR may be considered a wild-type sequence even if it contains one or more nucleotides that differ from the nucleotides in the canonical natural sequence, so long as the changes do not affect the properties and overall three-dimensional structure of the sequence. In some aspects, the differing nucleotides are conservative sequence changes. As one non-limiting example, a sequence having at least 95%, 96%, 97%, 98%, or 99% sequence identity to a canonical sequence (as measured, for example, using BLAST at default settings) also has a symmetrical three-dimensional spatial organization with another WT-ITR sequence such that their three-dimensional structures have the same shape in geometric space. A substantially symmetrical WT-ITR has the same A, C-C', and B-B' loops in three-dimensional space. That a substantially symmetrical WT-ITR is WT can be functionally confirmed by determining whether it has a functional Rep binding site (RBE or RBE') and a terminal resolution site (TRS) that pairs with the corresponding Rep protein. Other functions, including transgene expression under permissive conditions, may be tested, but are not required.

[0092] As used herein, the terms "modified ITR" or "mod-ITR" or "mutant ITR" are used interchangeably herein and refer to an ITR that has a mutation in at least one or more nucleotides compared to a WT-ITR from the same serotype. The mutation may result in a change in one or more of the A, C, C', B, B' regions of the ITR and may result in a change in the three-dimensional spatial organization (i.e., its three-dimensional structure in geometric space) compared to the three-dimensional spatial organization of a WT-ITR from the same serotype.

[0093] As used herein, the term "asymmetric ITRs," also referred to as "a pair of asymmetric ITRs," refers to a pair of ITRs in a single ccDNA genome or ccDNA vector that are not reverse complementary over their entire length. As one non-limiting example, an asymmetric ITR in a pair does not have a symmetric three-dimensional spatial organization with its cognate ITRs, such that their three-dimensional structures have different shapes in geometric space. That is, the asymmetric ITRs in a pair have different overall geometric structures, i.e., they have different organizations of their A, C-C', and B-B' loops in three-dimensional space (e.g., one ITR may have a short C-C' arm and/or a short B-B' arm compared to a cognate ITR). The sequence difference between the two ITRs may be due to the addition, deletion, truncation, or point mutation of one or more nucleotides. In one embodiment, one ITR of a pair of asymmetric ITRs may be a wild-type AAV ITR sequence and the other ITR is a modified ITR as defined herein (e.g., a non-wild-type ITR sequence or a synthetic sequence). In another embodiment, neither ITR of a pair of asymmetric ITRs is a wild-type AAV sequence and the two ITRs are modified ITRs that have different shapes in geometric space (i.e., different overall geometric structure). In some embodiments, one mod-ITR of a pair of asymmetric ITRs may have a short C-C' arm and the other ITR may have a different modification (e.g., a single arm or a short B-B' arm, etc.) such that they have a different three-dimensional spatial organization compared to the cognate asymmetric mod-ITR.

[0094] As used herein, the term "symmetrical ITRs" refers to a pair of ITRs in a single ccDNA genome or ccDNA vector that are mutated or modified relative to wild-type dependovirus ITR sequences and are reverse complementary throughout their entire length. Neither ITR is a wild-type AAV2 ITR sequence (i.e., they are modified ITRs, also referred to as mutant ITRs) and may differ in sequence from the wild-type ITR due to a nucleotide addition, deletion, substitution, truncation, or point mutation. For convenience, in this document, the ITR located in the 5' direction (upstream) relative to the expression cassette in the ecDNA vector is referred to as the "5' ITR" or "left ITR", and the ITR located in the 3' direction (downstream) relative to the expression cassette in the ecDNA vector is referred to as the "3' ITR" or "right ITR".

[0095] As used herein, the term "substantially symmetrical modified ITRs" or "a pair of substantially symmetrical mod-ITRs" refers to a pair of modified ITRs in a single ccDNA genome or ccDNA vector that both have reverse complementary sequences along their entire length. For example, a modified ITR may be considered substantially symmetrical even if it has some nucleotide sequences that differ from the reverse complementary sequence, so long as the changes do not affect the properties and overall shape. As one non-limiting example, a sequence is one that has at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity to a canonical sequence (as measured by BLAST using default settings) and also has a symmetrical three-dimensional spatial organization with its cognate modified ITR such that their three-dimensional structures have the same shape in geometric space. In other words, a pair of substantially symmetrical modified ITRs have loops A, C-C', and B-B' that are organized in three-dimensional space in the same way. In some embodiments, the ITRs of a mod-ITR pair may have different reverse complementary nucleotide sequences but still have the same symmetrical three-dimensional spatial organization, i.e., both ITRs have mutations that result in the same overall three-dimensional shape. For example, one ITR (e.g., a 5' ITR) in a mod-ITR pair may be from one serotype and the other ITR (e.g., a 3' ITR) may be from a different serotype, but both may have the same corresponding mutation (e.g., if the 5' ITR has a deletion in region C, then the cognate modified 3' ITR from the other serotype has a deletion at the corresponding position in region C') such that the pair of modified ITRs has the same symmetrical three-dimensional spatial organization. In such embodiments, each ITR in the pair of modified ITRs may be from different serotypes (e.g., AAB1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12), such as a combination of AAB2 and AAB6, wherein the modification in one ITR maps to the corresponding position in the cognate ITR from the other serotype. According to one embodiment, a pair of substantially symmetrical modified ITRs refers to a pair of modified ITRs (mod-ITRs), provided that the difference in nucleotide sequences between said ITRs does not affect the properties or overall shape, and they have substantially the same shape in three-dimensional space. As a non-limiting example, a mod-ITR is given that has at least 95%, 96%, 97%, 98% or 99% sequence identity to a canonical mod-ITR, determined by standard tools well known to those skilled in the art, such as BLAST (Basic Local Alignment Search Tool) or BLASTN at default settings, and also has a symmetrical three-dimensional spatial organization such that their three-dimensional structures have the same shape in geometric space. A pair of substantially symmetrical mod-ITRs have identical A, C-C' and B-B' loops in three-dimensional space, for example, if the modified ITR in a pair of substantially symmetrical mod-ITRs has a deletion of the C-C' arm, then the cognate mod-ITR has a corresponding deletion of the C-C' loop and also has a similar three-dimensional structure of the remaining A and B-B' loops, taking the same shape in geometric space as the cognate mod-ITR. The term "flanking" refers to the relative position of one nucleic acid sequence with respect to another nucleic acid sequence. Typically, in an ABC sequence, A and C flank B. The same is true for the AxBxC arrangement. Accordingly, a flanking sequence precedes or follows the flanked sequence, but does not necessarily have to be adjacent to or immediately adjacent to the flanked sequence. In one embodiment, the term "flanking" refers to the terminal repeats at each end of the linear duplex ccDNA vector. As used herein, the terms "treat," "treating," and/or "treatment" include suppressing, substantially inhibiting, slowing, or reversing the progression of a condition, substantially alleviating the clinical symptoms of a condition, or substantially preventing the onset of clinical symptoms of a condition, obtaining beneficial or targeted clinical results. Treatment also refers to achieving one or more of: (a) reducing the severity of the disorder; (b) limiting the development of symptoms characteristic of the disorder(s) being treated; (c) limiting the worsening of symptoms characteristic of the disorder(s) being treated; (d) limiting the recurrence of the disorder(s) in patients who previously had said disorder(s); and (e) limiting the recurrence of symptoms in patients who previously did not have symptoms of said disorder(s). Beneficial or targeted clinical results, such as pharmacological and/or physiological effects, include, but are not limited to, preventing the onset of a disease, disorder, or condition in a subject who may be predisposed to said disease, disorder, or condition but is not yet experiencing symptoms or exhibiting symptoms of said disease (prophylactic treatment), alleviating the symptoms of said disease, disorder, or condition, reducing the intensity of said disease, disorder, or condition, stabilizing (i.e., not worsening) said disease, disorder, or condition, preventing the spread of said disease, disorder, or condition, delaying or slowing the progression of said disease, disorder, or condition, alleviating or palliating said disease, disorder, or condition, and combinations thereof, as well as prolonging survival compared to expected survival without treatment.

[0096] As used herein, the term "increase," "amplification," "enhancement" (and similar terms) generally refers to the act of increasing, directly or indirectly, the concentration, level, function, activity, or behavior relative to the natural, expected, or average, or relative to a control state.

[0097] As used herein, the term "suppress," "reduce," "interfere with," "inhibit," and/or "reduce" (and other similar terms) generally refers to the act of decreasing, directly or indirectly, a concentration, level, function, activity, behavior relative to a natural, expected, average, or relative to a control state. By "reduce," "reduce," "reduce," or "reducing," when applied to an immune response (e.g., an immune response (e.g., an innate immune response)), with an immunosuppressant, is meant a detectable decrease in the immune response to the immunosuppressant. The amount of decrease in the immune response with an immunosuppressant can be determined relative to the level of the immune response in the presence of the immunosuppressant. The detectable reduction may be approximately 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100% or lower than the immune response detected in the presence of the immunosuppressant.

[0098] As used herein, the term "lipid" refers to a group of organic compounds that include, but are not limited to, esters of aliphatic acids and are characterized by being insoluble in water but soluble in many organic solvents. They are generally divided into at least three classes: (1) "simple lipids," which include fats and oils, as well as waxes; (2) "complex lipids," which include phospholipids and glycolipids; and (3) "lipid derivatives," such as steroids.

[0099] As used herein, the term "lipid particle" includes a lipid composition that can be used to deliver a therapeutic agent, such as a nucleic acid therapeutic agent and/or an immunosuppressant, to a target site of interest (e.g., a cell, tissue, organ, etc.). In preferred embodiments, the lipid particle of the present invention is a lipid particle containing a nucleic acid that is typically formed from a cationic lipid, a non-cationic lipid, and optionally a conjugated lipid that prevents aggregation of the particle. In other preferred embodiments, a therapeutic agent, such as a therapeutic nucleic acid, can be encapsulated in a lipid portion of the particle, which protects it from enzymatic degradation. In other preferred embodiments, an immunosuppressant can optionally be included in the lipid particles containing the nucleic acid.

[00100] As used herein, the term "lipid encapsulated" may refer to a lipid particle that provides an active agent or therapeutic agent, such as a nucleic acid (e.g., ccDNA), with complete encapsulation, partial encapsulation, or both. In a preferred embodiment, the nucleic acid is completely encapsulated within the lipid particle (e.g., to form a lipid particle containing the nucleic acid).

[00101] As used herein, the term "lipid conjugate" refers to a conjugated lipid that inhibits the aggregation of lipid particles. Such lipid conjugates include, but are not limited to, PEG-lipid conjugates such as, for example, PEG linked to dialkyloxypropyls (e.g., PEG-DAA conjugates), PEG linked to diacylglycerols (e.g., PEG-DAG conjugates), PEG linked to cholesterol, PEG linked to phosphatidylethanolamines, and PEG conjugated to ceramides (see, e.g., U.S. Patent No. 5,885,613), cationic lipids-PEG, polyoxazoline (POZ)-lipid conjugates (e.g., POZ-DAA conjugates; see, e.g., U.S. Provisional Application No. 61/294,828, filed January 13, 2010, and U.S. Provisional Application No. 61/295,140, filed January 14, 2010), polyamide oligomers (e.g., ATTA-lipid conjugates), and mixtures thereof. Additional examples of POZ-lipid conjugates are described in PCT Publication No. WO 2010/006282. PEG or POZ may be conjugated directly to the lipid or may be linked to the lipid via a linker group. Any linker group suitable for linking PEG or POZ to the lipid may be used, including, for example, non-ester linker groups and ester linker groups. Certain preferred embodiments employ non-ester linker groups such as amides or carbamates. The disclosures of each of the above-mentioned patent documents are herein incorporated by reference in their entirety for all purposes.

[00102] Representative examples of phospholipids include, but are not limited to, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyloleoylphosphatidylcholine, lysophosphatidylcholine, lysophosphatidylethanolamine, dipalmitoylphosphatidylcholine, dioleoylphosphatidylcholine, distearoylphosphatidylcholine, and dilinoleoylphosphatidylcholine. Other compounds that lack phosphorus, such as the sphingolipid and glycosphingolipid families, diacylglycerols, and β-acyloxy acids, are also included in the group designated as amphipathic lipids. In addition, the amphipathic lipids described above can be mixed with other lipids, including triglycerides and sterols.

[00103] As used herein, the term "neutral lipid" refers to any of a number of lipid molecules that exist in either an uncharged or a neutral zwitterionic form at a selected pH. At physiological pH, such lipids include, for example, diacylphosphatidylcholine, diacylphosphatidylethanolamine, ceramide, sphingomyelin, cephalin, cholesterol, cerebrosides, and diacylglycerols.

[00104] As used herein, the term "non-cationic lipid" refers to any amphipathic lipid, as well as any other neutral lipid or anionic lipid.

[00105] As used herein, the term "anionic lipid" refers to any lipid that is negatively charged at physiological pH. These lipids include, but are not limited to, phosphatidylglycerols, cardiolipins, diacylphosphatidylserines, diacylphosphatidic acids, N-dodecanoylphosphatidylethanolamines, N-succinylphosphatidylethanolamines, N-glutarylphosphatidylethanolamines, lysylphosphatidylglycerols, palmitoyloleoylphosphatidylglycerol (POPG), and other anionic modifying groups attached to neutral lipids.

[00106] As used herein, the term "hydrophobic lipid" refers to compounds having non-polar groups that include, but are not limited to, saturated and unsaturated long chain aliphatic hydrocarbon groups and such groups optionally substituted with one or more aromatic, cycloaliphatic, or heterocyclic groups. Suitable examples include, but are not limited to, diacylglycerol, dialkylglycerol, N-N-dialkylamino, 1,2-diacyloxy-3-aminopropane, and 1,2-dialkyl-3-aminopropane.

[00107] As used herein, the term "aqueous solution" refers to a composition comprising, in whole or in part, water.

[00108] As used herein, the term "organic lipid solution" refers to a composition comprising, in whole or in part, an organic solvent containing a lipid.

[00109] As used herein, the term "systemic delivery" refers to delivery of lipid particles that results in broad biodistribution of an active agent, such as interfering RNA (e.g., siRNA), in the body. Some administration techniques may result in systemic delivery of certain agents but not others. Systemic delivery means that a useful, preferably therapeutic, amount of the agent is delivered to most parts of the body. Broad biodistribution typically requires a blood lifetime such that the agent is not rapidly degraded or cleared (e.g., by first-pass organs (liver, lungs, etc.) or by rapid non-specific binding to cells) before reaching a disease site distant from the site of administration. Systemic delivery of lipid particles can be accomplished by any means known in the art, including, for example, intravenous, subcutaneous, and intraperitoneal. In a preferred embodiment, systemic delivery of lipid particles is accomplished by intravenous delivery.

[00110] As used herein, the term "local delivery" refers to the delivery of an active agent, such as an interfering RNA (e.g., siRNA), directly to a target site within the body. For example, the agent may be locally delivered by direct injection into a disease site, such as a tumor, or another target site, such as an inflammatory site, or a target organ, such as the liver, heart, pancreas, kidney, etc.

[00111] As used herein, the term "terminal repeat" or "TR" includes any viral terminal repeat or any synthetic sequence that contains at least one minimal required origin of replication and a region containing a palindromic hairpin structure. The Rep binding sequence ("RBS") (also referred to as an RBE (Rep binding element)) and the terminal resolution site ("TRS") together constitute the "minimal required origin of replication", and accordingly, a TR contains at least one RBS and at least one TRS. Each of the TRs that are reverse complementary to each other within a defined region of the polynucleotide sequence is generally referred to as an "inverted terminal repeat" or "ITR". In the context of a virus, ITRs mediate replication, viral packaging, integration, and release of the provirus. As has been surprisingly found, according to the present invention, TRs that are not reverse complementary along their entire length can still perform the normal functions of ITRs, and thus, as used herein, the term ITR refers to a TR in a ccDNA genome or a ccDNA vector that is capable of mediating replication of the ccDNA vector. One of ordinary skill in the art will appreciate that more than two pairs of ITRs or asymmetric ITRs may be present in ccDNA vectors of complex configuration. An ITR may be an AAB ITR or a non-AAB ITR, or may be derived from an AAB ITR or a non-AAB ITR. For example, an ITR may be from the Parvoviridae family, which includes parvoviruses and dependoviruses (e.g., canine parvovirus, bovine parvovirus, murine parvovirus, porcine parvovirus, human parvovirus B-19), or the SV40 hairpin, which serves as the SV40 origin of replication, may be used as an ITR, which may be further modified by truncation, substitution, deletion, insertion, and/or addition. The Parvoviridae family of viruses consists of two subfamilies: Parvovirinae, which infect vertebrates, and Densovirinae, which infect invertebrates. Dependoparvoviruses include the adeno-associated virus (AAV) family of viruses that are capable of replicating in vertebrate hosts, including, but not limited to, humans, primate species, bovine, canine, equine, and ovine. For convenience, in this document, the ITR located in the 5' direction (upstream) relative to the expression cassette in the ecDNA vector is referred to as the "5'-ITR" or "left ITR", and the ITR located in the 3' direction (downstream) relative to the expression cassette in the ecDNA vector is referred to as the "3'-ITR" or "right ITR".

[00112] "Wild-type ITR" or "WT-ITR" refers to a sequence of a native ITR sequence of AAV or another dependentovirus that retains, for example, Rep binding activity and Rep nicking ability. The nucleotide sequence of a WT-ITR from any AAV serotype may differ slightly from the canonical native sequence due to degeneracy of the genetic code or drift, and accordingly, WT-ITR sequences contemplated for use herein include WT-ITR sequences formed as a result of natural variations that occur during the production process (e.g., replication errors).

[00113] As used herein, the term "substantially symmetrical WT-ITRs" or "a pair of substantially symmetrical WT-ITRs" refers to a pair of WT-ITRs in a single ccDNA genome or ccDNA vector, in which both ITRs are wild-type ITRs that have a reverse complementary sequence along their entire length. For example, an ITR may be considered a wild-type sequence even if it has one or more nucleotides that differ from the canonical natural sequence, so long as the changes do not affect the properties and overall three-dimensional structure of the sequence. In some aspects, the differing nucleotides are conservative sequence changes. As one non-limiting example, a sequence is provided that has at least 95%, 96%, 97%, 98%, or 99% sequence identity to a canonical sequence (as measured, for example, by BLAST at default settings) and also has a symmetrical three-dimensional spatial organization relative to another WT-ITR sequence such that their three-dimensional structures have the same shape in geometric space. A substantially symmetrical WT-ITR has the same A, C-C', and B-B' loops in three-dimensional space. That a substantially symmetrical WT-ITR is WT can be functionally confirmed by determining whether it has a functional Rep binding site (RBE or RBE') and a terminal resolution site (TRS) that pairs with the corresponding Rep protein. Other functions, including transgene expression under permissive conditions, may optionally be tested.

[00114] As used herein, the terms "modified ITR" or "mod-ITR" or "mutant ITR" are used interchangeably herein and refer to an ITR that has a mutation in at least one or more nucleotides compared to a WT-ITR from the same serotype. The mutation may result in a change in one or more of the A, C, C', B, B' regions of the ITR and may result in a change in the three-dimensional spatial organization (i.e., its three-dimensional structure in geometric space) compared to the three-dimensional spatial organization of a WT-ITR from the same serotype.

[00115] As used herein, the term "asymmetric ITRs," also referred to as "asymmetric ITR pairs," refers to a pair of ITRs in a single ccDNA genome or ccDNA vector that are not reverse complementary along their entire length. As one non-limiting example, an asymmetric ITR in a pair does not have a symmetric three-dimensional spatial organization with its cognate ITR such that their three-dimensional structures have different shapes in geometric space. In other words, the asymmetric ITRs in a pair have different overall geometric structures, i.e., they have different organizations of their A, C-C´, and B-B´ loops in three-dimensional space (e.g., one ITR may have a short C-C´ arm and/or a short B-B´ arm compared to a cognate ITR). The difference in sequence between the two ITRs may be due to the addition, deletion, truncation, or point mutation of one or more nucleotides. In one embodiment, one ITR of the pair of asymmetric ITRs may be a wild-type AAV ITR sequence, and the other ITR may be a modified ITR as defined herein (e.g., a non-wild-type ITR sequence or a synthetic ITR sequence). In another embodiment, neither ITR of the pair of asymmetric ITRs is a wild-type AAV sequence, and the two ITRs are modified ITRs that have different shapes in geometric space (i.e., different overall geometric structures). In some embodiments, one mod-ITR of a pair of asymmetric ITRs may have a short C-C´ arm, and the other ITR may have a different modification (e.g., one arm or a short B-B´ arm, etc.) such that they have a different three-dimensional spatial organization compared to the cognate asymmetric mod-ITR.

[00116] As used herein, the term "symmetrical ITRs" refers to a pair of ITRs in a single ccDNA genome or ccDNA vector that are wild-type or mutated (e.g., modified relative to the wild-type) dependovirus ITR sequences and are reverse complementary along their entire length. In one non-limiting example, both ITRs are wild-type ITR sequences from AAV2. In another example, neither ITR is a wild-type AAV2 ITR sequence (i.e., they are a modified ITR, also referred to as a mutant ITR) and may differ in sequence from the wild-type ITR due to a nucleotide addition, deletion, substitution, truncation, or point mutation. For convenience, in this document, the ITR located in the 5' direction (upstream) relative to the expression cassette in the ecDNA vector is referred to as the "5' ITR" or "left ITR", and the ITR located in the 3' direction (downstream) relative to the expression cassette in the ecDNA vector is referred to as the "3' ITR" or "right ITR".

[00117] As used herein, the term "substantially symmetrical modified ITRs" or "a pair of substantially symmetrical mod-ITRs" refers to a pair of modified ITRs in a single ccDNA genome or ccDNA vector that both have reverse complementary sequences along their entire length. For example, a modified ITR may be considered substantially symmetrical even if it has some nucleotide sequences that differ from the reverse complementary sequence, so long as the changes do not affect the properties and overall shape. As one non-limiting example, a sequence is provided that has at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity to a canonical sequence (as measured by BLAST using default settings) and also has a symmetrical three-dimensional spatial organization with its cognate modified ITR such that their three-dimensional structures have the same shape in geometric space. In other words, a pair of substantially symmetrical modified ITRs have loops A, C-C', and B-B' that are organized identically in three-dimensional space. In some embodiments, the ITRs of a mod-ITR pair may have different reverse complementary nucleotide sequences but still have the same symmetrical three-dimensional spatial organization, i.e., both ITRs have mutations that result in the same overall three-dimensional shape. For example, one ITR (e.g., a 5' ITR) in a mod-ITR pair may be from one serotype and the other ITR (e.g., a 3' ITR) may be from a different serotype, but both may have the same corresponding mutation (e.g., if a 5' ITR has a deletion in region C, a cognate modified 3' ITR from a different serotype has a deletion at the corresponding position in region C') such that the pair of modified ITRs has the same symmetrical three-dimensional spatial organization. In such embodiments, each ITR in a pair of modified ITRs may be from different serotypes (e.g., AAB1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12), such as a combination of AAB2 and AAB6, wherein the modification in one ITR maps to the corresponding position in the cognate ITR from the other serotype. According to one embodiment, a pair of substantially symmetrical modified ITRs refers to a pair of modified ITRs (mod-ITRs), provided that the difference in nucleotide sequences between the ITRs does not affect the properties or overall shape, and they have substantially the same shape in three-dimensional space. As a non-limiting example, a mod-ITR is provided that has at least 95%, 96%, 97%, 98% or 99% sequence identity to a canonical mod-ITR, as determined by standard methods well known in the art, such as BLAST (Basic Local Alignment Search Tool) or BLASTN at default settings, and also has a symmetrical three-dimensional spatial organization such that their three-dimensional structure has the same shape in geometric space. A pair of substantially symmetric mod-ITRs have identical A, C-C' and B-B' loops in three-dimensional space, for example, if the modified ITR in a pair of substantially symmetric mod-ITRs has a deletion in the C-C' arm, then the cognate mod-ITR has a corresponding deletion in the C-C' loop and also has a similar three-dimensional structure of the remaining A and B-B' loops, taking the same shape in geometric space as its cognate mod-ITR.

[00118] The term "flanking" refers to the relative position of one nucleic acid sequence relative to another nucleic acid sequence. Typically, in an ABC sequence, A and C flank B. The same is true for the arrangement of AxBxC. Thus, a flanking sequence precedes or follows the sequence being flanked, but does not necessarily have to be adjacent to or immediately adjacent to the sequence being flanked. In one embodiment, the term "flanking" refers to the terminal repeats at each end of a linear duplex ccDNA vector.

[00119] As used herein, the term "ccDNA genome" refers to an expression cassette that further comprises at least one inverted terminal repeat region. The ccDNA genome may further comprise one or more spacer regions. In some embodiments, the ccDNA genome is included as an intermolecular duplex DNA polynucleotide in a plasmid or viral genome.

[00120] As used herein, the term "ccDNA spacer region" refers to an intervening sequence that separates functional elements in an ccDNA vector or ccDNA genome. In some embodiments, ccDNA spacer regions maintain two functional elements at a target distance for optimal functionality. In some embodiments, ccDNA spacer regions provide or enhance genetic stability of the ccDNA genome, such as in a plasmid or baculovirus. In some embodiments, ccDNA spacer regions facilitate pre-formed genetic manipulation of the ccDNA genome by providing convenient locations for cloning sites, etc. For example, in certain aspects, an oligonucleotide "polylinker" containing multiple restriction endonuclease sites or an out-of-open reading frame sequence designed to lack known protein (e.g., transcription factor) binding sites can be placed in the ccDNA genome to separate cis-acting factors, such as by a 6-mer, 12-mer, 18-mer, 24-mer, 48-mer, 86-mer, 176-mer insertion, etc., between the terminal resolution site and the 5' transcriptional regulatory element. Similarly, a spacer can be inserted between the polyadenylation signal sequence and the 3' terminal resolution site.

[00121] As used herein, the term "ecDNA plasmid" refers to a plasmid that contains an ecDNA genome in the form of an intermolecular duplex.

[00122] As used herein, the term "ccDNA bacmid" refers to an infectious baculovirus genome that contains the ccDNA genome in an intermolecular duplex that is capable of being replicated in E. coli as a plasmid and, accordingly, can serve as a shuttle vector for the baculovirus.

[00123] As used herein, the term "ccDNA baculovirus" refers to a baculovirus that contains a ccDNA genome as an intermolecular duplex within the baculovirus genome.

[00124] As used herein, the terms "ccDNA baculovirus-infected insect cell" and "ccDNA-BIIC" are used interchangeably and refer to an invertebrate host cell (including, but not limited to, an insect cell (e.g., an Sf9 cell)) infected with a ccDNA baculovirus.

[00125] As used herein, the term "closed-ended DNA vector" refers to a capsidless DNA vector with at least one covalently closed end, wherein at least a portion of the vector has an intramolecular duplex structure.

[00126] As used herein, the term "ccDNA" refers to capsidless, linear, double-stranded (ds), closed-ended duplex DNA for non-viral gene transfer, synthetic or otherwise. A detailed description of ccDNA is provided in international application PCT/US2017/020828, filed March 3, 2017, the contents of which are expressly incorporated herein by reference in their entirety. Certain methods for producing ccDNA containing various inverted terminal repeat (ITR) sequences and configurations using cellular methods are described in Example 1 of international applications PCT/US18/49996, filed September 7, 2018, and PCT/US2018/064242, filed December 6, 2018, each of which is expressly incorporated herein by reference in their entirety. Some methods for producing synthetic ccDNA vectors containing various ITR sequences and configurations are described, for example, in international application PCT/US2019/14122, filed January 18, 2019, the contents of which are incorporated herein by reference in their entirety.

[00127] As used herein, the terms "ccDNA vector" and "ccDNA" are used interchangeably and refer to a DNA vector with closed ends that contains at least one terminal palindrome. In some embodiments, the ccDNA contains two covalently closed ends.

[00128] As used herein, the term "ncDNA" or "nicked ccDNA" refers to DNA with closed ends that has a single-strand break or gap of 1-100 base pairs in the stem or spacer region located in the 5' direction relative to an open reading frame (e.g., a promoter and a transgene to be expressed).

[00129] As used herein, the terms "gap" and "single-strand break" are used interchangeably and refer to an interrupted portion of a synthetic DNA vector of the present invention, creating a region of single-stranded DNA in otherwise double-stranded ccDNA. A gap can be from one 1 base pair to 100 base pairs in length in one strand of duplex DNA. Typical gaps designed and created using the methods described herein and synthetic vectors produced using these methods can have, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59 or 60 base pairs in length. Exemplary gaps in the present disclosure may be from 1 base pair to 10 base pairs in length, from 1 to 20 base pairs in length, from 1 to 30 base pairs in length.

[00130] As used herein, the terms "Rep binding site," "Rep binding element," "RBE," and "RBS" are used interchangeably and refer to a binding site for a Rep protein (e.g., Rep 78 AAB or Rep 68 AAB) that, upon binding by the Rep protein, allows the Rep protein to perform its site-specific endonuclease activity on a sequence comprising the RBS. The RBS sequence and its reverse complement together form a single RBS. RBS sequences are known in the art and include, for example, 5'-GCGCGCTCGCTCGCTC-3' (SEQ ID NO: 39), which is the RBS sequence identified in AAB2. Any known RBS sequence can be used in embodiments of the present invention, including other known AAB RBS sequences and other known natural or synthetic RBS sequences. Without being bound by theory, it is believed that the nuclease domain of the Rep protein binds to the duplex nucleotide sequence GCTC and, accordingly, direct binding and stable assembly of the two known AAB Rep proteins on the duplex oligonucleotide, 5'-(GCGC)(GCTC)(GCTC)(GCTC)-3' (SEQ ID NO: 39), occurs. In addition, soluble aggregated conformers (i.e., an indefinite number of interconnected Rep proteins) dissociate and bind to oligonucleotides that contain Rep binding sites. Each Rep protein interacts with both the nucleobases and the phosphodiester backbone on each strand. The base interactions provide sequence specificity, while the phosphodiester backbone interactions are non- or less sequence specific and stabilize the protein-DNA complex.

[00131] As used herein, the terms "terminal resolution site" and "TRS" are used interchangeably herein and refer to the region in which Rep forms a tyrosine phosphodiester bond with 5'-thymidine to form a 3'-OH, which serves as a substrate for DNA extension by a cellular DNA polymerase, such as DNA polymerase delta or DNA polymerase epsilon. Alternatively, the Rep-thymidine complex can participate in a coordinated ligation reaction. In some embodiments, the TRS includes at least an unpaired thymidine. In some embodiments, the nicking efficiency of the TRS can be controlled at least in part by the distance between it and the RBS within a single molecule. If the acceptor substrate is the complement of the ITR, then the resulting product is an intramolecular duplex. TRS sequences are known in the art and include, for example, 5'-GGTTGA-3' (SEQ ID NO: 804), which is a hexanucleotide sequence identified in AAB2. Any known TRS sequence can be used in embodiments of the present invention, including other known AAB TRS sequences and other known natural or synthetic TRS sequences such as AGTT (SEQ ID NO: 085), GGTTGG (SEQ ID NO: 806), AGTTGG (SEQ ID NO: 807), AGTTGA (SEQ ID NO: 808), and other motifs such as RRTTRR (SEQ ID NO: 809).

[00132] As used herein, the terms "sense" and "antisense" refer to the orientation of a structural element on a polynucleotide. Sense and antisense versions of an element are inversely complementary to each other.

[00133] As used herein, the terms "synthetic AAV vector" and "synthetic production of an AAV vector" refer to an AAV vector and synthetic methods for producing it in a completely cell-free environment.

[00134] As used herein, "reporters" refer to proteins that can be used to provide detectable indicators. Reporters typically generate a measurable signal, such as fluorescence, color, or luminescence. Sequences encoding reporter proteins encode proteins whose presence in a cell or organism is easily observed. For example, fluorescent proteins cause a cell to fluoresce when excited by light of a specific wavelength, luciferases cause a cell to catalyze a reaction that generates light, and enzymes such as β-galactosidase convert a substrate into a colored product. Exemplary reporter polypeptides that can be used for experimental or diagnostic purposes include, but are not limited to, β-lactamase, β-galactosidase (LacZ), alkaline phosphatase (AP), thymidine kinase (TK), green fluorescent protein (GFP) and other fluorescent proteins, chloramphenicol acetyltransferase (CAT), luciferase, and others well known in the art.

[00135] As used herein, the term "effector protein" refers to a polypeptide that provides a detectable indicator, either, for example, as a reporter polypeptide, or more preferably, as a cell-killing polypeptide, for example, a toxin, or an agent that confers sensitivity to killing by a selected agent or in the absence thereof. Effector proteins include any protein or peptide that directly targets or damages host cell DNA and/or RNA. For example, effector proteins can include, but are not limited to, a restriction endonuclease that targets a host cell DNA sequence (whether a genomic or extrachromosomal element), a protease that cleaves a target polypeptide necessary for cell survival, a DNA gyrase inhibitor, and a ribonuclease-type toxin. In some embodiments, expression of an effector protein controlled by a synthetic biological circuit described herein may participate as a factor in another synthetic biological circuit, which increases the range and complexity of the sensitivity of the biological circuit system.

[00136] Transcriptional regulators refer to transcriptional activators and repressors that either activate or repress transcription of a gene of interest, such as an inflammasome antagonist (e.g., an inhibitor of one or more of the NLRP3 and/or AIM2 inflammasome pathway or a caspase inhibitor 1). Promoters are regions of nucleic acid that initiate transcription of a particular gene. Transcriptional activators typically bind in the vicinity of transcriptional promoters and recruit RNA polymerase to directly initiate transcription. Repressors bind to transcriptional promoters and sterically hinder RNA polymerase from initiating transcription. Other transcriptional regulators can serve as either activators or repressors depending on their binding site and cellular and environmental conditions. Non-limiting examples of classes of transcriptional regulators include, but are not limited to, homeodomain proteins, zinc finger proteins, forkhead proteins, and leucine zipper proteins.

[00137] As used herein, a "repressor protein" or "inducer protein" is a protein that binds to a regulatory sequence element and represses or activates, respectively, transcription of sequences operably linked to said regulatory sequence element. Preferred repressor and inducer proteins described herein are responsive to the presence or absence of at least one input agent or environmental factor. Preferred proteins described herein are modular in form and comprise, for example, separable DNA-binding and input agent-binding or responsive elements or domains.

[00138] As used herein, the term "carrier" includes any and all possible solvents, dispersion media, bases, coatings, diluents, antibacterial and antifungal agents, isotonic and absorption delaying agents, buffers, carrier solutions, suspensions, colloids, and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Auxiliary active ingredients may also be included in the compositions. The expression "pharmaceutically acceptable" refers to molecular entities and compositions that do not result in a toxic, allergic, or similar undesirable reaction when administered to a host.

[00139] As used herein, the term "input agent responsive domain" is a domain of a transcription factor that binds or otherwise responds to a condition or input agent in a manner that renders the DNA-binding fusion domain attached thereto responsive to the presence of said condition or input agent. In one embodiment, the presence of said condition or input agent results in a conformational change in the input agent responsive domain, or in the protein to which it is fused, that modifies the transcription-modulating activity of the transcription factor.

[00140] The term "in vivo" refers to assays or methods that are performed in or within an organism, such as a multicellular animal. In some aspects described herein, a method or use is considered to be performed "in vivo" if a single-celled organism, such as a bacterium, is used. The term "ex vivo" refers to methods and uses that are performed using a living cell with an intact membrane that is outside of a multicellular animal or plant, such as explants, cultured cells, including primary cells and cell lines, transformed cell lines, and extracted tissues or cells, including blood cells, among others. The term "in vitro" refers to assays and methods that do not require a cell with an intact membrane, such as cell extracts, and may relate to the administration of a programmable synthetic biological circuit in a cell-free system, such as an environment that does not contain cells or cell systems, such as cell extracts.

[00141] As used herein, the term "promoter" refers to any nucleic acid sequence that regulates the expression of another nucleic acid sequence by directing the transcription of said nucleic acid sequence, which may be a heterologous target gene encoding a protein or RNA. Promoters may be constitutive, inducible, repressible, tissue-specific, or any combination thereof. A promoter is a control region of a nucleic acid sequence that controls the initiation and rate of transcription of the remainder of the nucleic acid sequence. A promoter may also contain genetic elements to which regulatory proteins and molecules, such as RNA polymerase and other transcription factors, may bind. In some embodiments of the aspects described herein, a promoter may direct the expression of a transcription factor that regulates the expression of the promoter itself. The promoter sequence may contain a transcription initiation site as well as protein binding domains responsible for binding RNA polymerase. Eukaryotic promoters will often, but not always, contain "TATA" boxes and "CAT" boxes. A variety of promoters, including inducible promoters, may be used to drive expression of transgenes in the ccDNA vectors disclosed herein. The promoter sequence may be bordered at its 3' end by a transcription initiation site and extend upstream (in the 5' direction) to include the minimum number of bases or elements necessary to initiate transcription at levels detectable above background.

[00142] As used herein, the term "enhancer" refers to a cis-acting regulatory sequence (e.g., 50-1500 base pairs) that binds one or more proteins (e.g., activator proteins or a transcription factor) to enhance transcriptional activation of a nucleic acid sequence. Enhancers can be located up to 1,000,000 base pairs upstream of the start site of the gene they regulate or downstream of the start site of the gene they regulate. An enhancer can be located within an intronic region or in an exonic region of an unrelated gene.

[00143] A promoter may be said to control the expression or control the transcription of a nucleic acid sequence that it regulates. The terms "operably linked," "operably positioned," "operably linked," "under the control of," and "under transcriptional control" indicate that the promoter is in the correct functional position and/or orientation relative to the nucleic acid sequence that it regulates to control the initiation of transcription and/or expression of said sequence. As used herein, an "inverted promoter" refers to a promoter in which the nucleic acid sequence is arranged in the reverse orientation such that the strand that was the sense strand becomes the non-coding strand, and vice versa. Inverted promoter sequences may be used in various embodiments to control the state of a switch. Additionally, in various embodiments, a promoter may be used in combination with an enhancer.

[00144] A promoter may be a promoter that is naturally associated with a gene or sequence, and may also be obtained by isolating non-coding 5' sequences located 5' relative to a coding segment and/or exon of a particular gene or sequence. Such a promoter may be referred to as "endogenous." Similarly, in some embodiments, an enhancer may be an enhancer that is naturally associated with a nucleic acid sequence and located either 3' or 5' relative to said sequence.

[00145] In some embodiments, the coding segment of the nucleic acid is placed under the control of a "recombinant promoter" or a "heterologous promoter," both of which terms refer to a promoter that is not normally associated with the encoded nucleic acid sequence to which the promoter is operably linked in its natural environment. A recombinant or heterologous enhancer refers to an enhancer that is not normally associated with a particular nucleic acid sequence in its natural environment. Such promoters or enhancers may include promoters or enhancers of other genes; promoters or enhancers isolated from any other prokaryotic, viral, or eukaryotic cell; and synthetic promoters or enhancers that are not "naturally occurring," i.e., contain different elements of different transcriptional regulatory regions and/or expression-altering mutations introduced using genetic engineering techniques known in the art. In addition to producing promoter and enhancer nucleic acid sequences synthetically, promoter sequences may be produced using recombinant cloning and/or nucleic acid amplification technology, including PCR, as applied to the synthetic biological circuits and modules disclosed herein (see, e.g., U.S. Patent No. 4,683,202, U.S. Patent No. 5,928,906, each of which is incorporated herein by reference). Additionally, the use of control sequences that direct transcription and/or expression of the sequences in non-nuclear organelles such as mitochondria, chloroplasts, etc. is also contemplated.

[00146] As described herein, an "inducible promoter" is a promoter that is characterized by initiation or enhancement of transcriptional activity in the presence of, under the influence of, or in contact with an inducer or inducing agent. An "inducer" or "inducing agent" as defined herein may be endogenous or is typically an exogenous compound or protein that is administered in a manner that provides its activity in inducing the transcriptional activity of an inducible promoter. In some embodiments, the inducer or inducing agent, i.e., chemical, compound, or protein, may itself be produced by transcription or expression of a nucleic acid sequence (i.e., the inducer may be an inducer protein expressed by another component or module), which itself may be under the control of an inducible promoter. In some embodiments, an inducible promoter is induced in the absence of certain agents, such as a repressor. Examples of inducible promoters include, but are not limited to, tetracycline-sensitive, metallothione-sensitive, ecdysone-sensitive promoters, mammalian viral promoters (e.g., adenovirus late promoter and murine mammary tumor virus long terminal repeat (MMTV-LTR) promoter) and other steroid-sensitive promoters, rapamycin-sensitive promoters, etc.

[00147] The terms "DNA regulatory sequences," "control elements," and "regulatory elements," as used interchangeably herein, refer to transcriptional and translational control sequences, such as promoters, enhancers, polyadenylation signals, terminators, protein destruction signals, and the like, that provide for and/or regulate the transcription of a non-coding sequence (e.g., a DNA targeting RNA) or a coding sequence (e.g., a site-directed modifying polypeptide or a Cas9/Csn1 polypeptide) and/or regulate the translation of the encoded polypeptide.

[00148] The term "operably linked" refers to a placement in close proximity wherein the components so described are in a relationship that allows them to function in their intended manner. For example, a promoter is operably linked to a coding sequence if the promoter affects its transcription or expression. An "expression cassette" includes a heterologous DNA sequence that is operably linked to a promoter or other regulatory sequence sufficient to direct transcription of a transgene in a ccDNA vector. Suitable promoters include, for example, tissue-specific promoters. Promoters may also be derived from AAV.

[00149] As used herein, the term "subject" refers to a human or animal that is being treated, including prophylactic treatment, using a ccDNA vector according to the present invention. Typically, the animal is a vertebrate animal, such as, but not limited to, a primate, a rodent, a pet, or a game animal. Primates include, but are not limited to, chimpanzees, cynomolgus monkeys, spider monkeys, and macaques, such as rhesus monkeys. Rodents include mice, rats, marmots, ferrets, rabbits, and hamsters. Domestic and game animals include, but are not limited to, cows, horses, pigs, deer, bison, buffalo, feline species such as a domestic cat, canine species such as a dog, fox, wolf, bird species such as a chicken, emu, ostrich, and fish such as trout, catfish, and salmon. In certain embodiments of the aspects described herein, the subject is a mammal, such as a primate or a human. The subject may be male or female. Furthermore, the subject may be an infant or a child. In some embodiments, the subject may be a newborn or unborn subject, such as the subject is in utero. Preferably, the subject is a mammal. The mammal may be a human, a non-human primate, a mouse, a rat, a dog, a cat, a horse, or a cow, but these examples are not limiting. Mammals other than humans can be advantageously used as subjects in animal models of diseases and disorders. In addition, the methods and compositions described herein can be used with domesticated animals and/or companion animals. The human subject can be of any age, gender, race, or ethnicity, such as Caucasian (white), Asian, African, Black, African American, African European, Hispanic, Middle Eastern, etc. In some embodiments, the subject can be a patient or other subject in a clinical setting. In some embodiments, the subject is already undergoing a course of treatment. In some embodiments, the subject is an embryo, fetus, neonate, infant, child, adolescent, or adult. In some embodiments, the subject is a human fetus, human neonate, human infant, human child, human adolescent, or adult. In some embodiments, the subject is an animal embryo or a non-human embryo or a non-human primate embryo. In some embodiments, the subject is a human embryo.

[00150] As used herein, the term "host cell" includes any cell type that is susceptible to transformation, transfection, transduction, etc., using a nucleic acid construct or ccDNA expression vector of the present disclosure. As non-limiting examples, the host cell can be an isolated primary cell, pluripotent stem cells, CD34 ⁺ cells, induced pluripotent stem cells, or any of a number of immortalized cell lines (e.g., HepG2 cells). Alternatively, the host cell can be a cell in situ or in vivo in a tissue, organ, or organism.

[00151] The term "exogenous" refers to a substance present in a cell other than its natural source. As used herein, the term "exogenous" may refer to a nucleic acid (e.g., a nucleic acid encoding a polypeptide) or a polypeptide that has been introduced by a process involving human action into a biological system, such as a cell or organism, in which it is not normally found, and introduction of the nucleic acid or polypeptide into such a cell or organism is desirable. Alternatively, "exogenous" may refer to a nucleic acid or polypeptide that has been introduced by a process involving human action into a biological system, such as a cell or organism, in which it is found in relatively low amounts, and an increase in the amount of the nucleic acid or polypeptide in the cell or organism is desirable, such as to obtain ectopic expression or levels. In contrast, the term "endogenous" refers to a substance that is native to a biological system or cell.

[00152] The term "sequence identity" refers to the relatedness between two nucleotide sequences. For the purposes of the present disclosure, the degree of sequence identity between two deoxyribonucleotide sequences is determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, supra) as implemented in the Needle program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, supra), preferably version 3.0.0 or later. The optional parameters used are a gap opening penalty of 10, a gap extension penalty of 0.5, and an EDNAFULL substitution matrix (EMBOSS version NCBI NUC4.4). The Needle output value, called the "longest identical fragment length" (obtained using the "-nobrief" option), is used as the percent identity and is calculated as follows: (Identical deoxyribonucleotides×100)/(Alignment length - Total number of gaps in alignment). The alignment length is preferably at least 10 nucleotides, preferably at least 25 nucleotides, more preferably at least 50 nucleotides, and most preferably at least 100 nucleotides.

[00153] As used herein, the term "homology" or "homologous" is defined as the percentage of nucleotide residues that are identical to the nucleotide residues in a corresponding sequence on a target chromosome after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percentage of sequence identity. Alignment for the purpose of determining the percentage of nucleotide sequence homology can be achieved in a variety of ways known in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, ClustalW2, or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms necessary to achieve maximum alignment over the entire length of the sequences being compared. In some embodiments, a nucleic acid sequence (e.g., a DNA sequence), such as a homology arm, is considered "homologous" when the sequence is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more identical to a corresponding native or unedited nucleic acid sequence (e.g., genomic sequence) of the host cell.

[00154] As used herein, the term "heterologous" refers to a nucleotide or polypeptide sequence that is not found in a native nucleic acid or protein, respectively. A heterologous nucleic acid sequence may be linked to a native nucleic acid sequence (or a variant thereof) (e.g., by genetic engineering) to produce a chimeric nucleotide sequence encoding a chimeric polypeptide. A heterologous nucleic acid sequence may be linked to a variant polypeptide (e.g., by genetic engineering) to produce a nucleotide sequence encoding a fusion variant polypeptide.

[00155] A "vector" or "expression vector" is a replicon, such as a plasmid, bacmid, phage, virus, virion, or cosmid, to which another segment of DNA, i.e., an "insert," can be attached to cause replication of the attached segment in a cell. A vector can be a nucleic acid construct designed for delivery into a host cell or for transfer between different host cells. As used herein, a vector can be viral or non-viral in origin and/or final form, however, for the purpose of the present disclosure, a "vector" generally refers to a ccDNA vector as used herein. The term "vector" includes any genetic element that is capable of replication when associated with appropriate control elements and that can transfer gene sequences into cells. In some embodiments, a vector can be an expression vector or a recombinant vector.

[00156] As used herein, the term "expression vector" refers to a vector that directs the expression of RNA or a polypeptide from sequences linked to transcriptional regulatory sequences on the vector. The expressed sequences will often, but not necessarily, be heterologous with respect to the cell. An expression vector may contain additional elements, for example, an expression vector may have two replication systems, allowing it to be maintained in two organisms, for example, human cells for expression and a prokaryotic host for cloning and amplification. The term "expression" refers to cellular processes involved in the production of RNA and proteins and, where appropriate, the secretion of proteins, including, where applicable, but not limited to, for example, transcription, transcript processing, translation, and protein folding, modification, and processing. "Expression products" include RNA transcribed from a gene and polypeptides produced by translation of mRNA transcribed from a gene. The term "gene" means a nucleic acid sequence that is transcribed (DNA) into RNA in vitro or in vivo when operably linked to appropriate regulatory sequences. A gene may, but need not, include regions preceding and following the coding region, such as the 5'-untranslated (5'-UTR) or "leader" sequences and the 3'-UTR or "trailer" sequences, as well as intervening sequences (introns) between individual coding segments (exons).

[00157] By "recombinant vector" is meant a vector that includes a heterologous nucleic acid sequence or "transgene" that is capable of expression in vivo. It is to be understood that the vectors described herein, in some embodiments, can be combined with other suitable compositions and therapies. In some embodiments, the vector is episomal. Use of a suitable episomal vector provides a means of maintaining the nucleotide of interest in a subject in multi-copy extrachromosomal DNA, thereby eliminating potential chromosomal integration effects.

[00158] As used herein, the term "genetic disease" refers to a disease caused in whole or in part, directly or indirectly, by one or more abnormalities in the genome, in particular a condition that is present from birth. The abnormality may be a mutation, insertion, or deletion. The abnormality may affect the coding sequence of a gene or its regulatory sequence. Genetic disorder may include but is not limited to DMD (Duchenne muscular dystrophy), hemophilia, cystic fibrosis, Huntington's chorea, familial hypercholesterolemia (LDL receptor defect), hepatoblastoma, Wilson's disease, congenital hepatic porphyria, inherited disorders of liver metabolism, Lesch-Nyhan syndrome, sickle cell anemia, thalassemia, xeroderma pigmentosum, Fanconi anemia, retinitis pigmentosa, ataxia-telangiectasia, Bloom syndrome, retinoblastoma, and Tay-Sachs disease.

[00159] As used herein, the term "inhibitory polynucleotide" refers to a DNA or RNA molecule that reduces or prevents the expression (transcription or translation) of a second (target) polynucleotide. Inhibitory polynucleotides include antisense polynucleotides, ribozymes, and external targeting sequences. The term "inhibitory polynucleotide" further includes DNA and RNA molecules, such as RNAi, that encode existing inhibitory molecules, such as DNA molecules that encode ribozymes.

[00160] As used herein, "gene silencing" or "silenced gene" with respect to the activity of an RNAi molecule, such as an siRNA or miRNA, refers to a reduction in the mRNA level of a target gene (e.g., NLRP3, AIM2, or caspase-1 mRNA) in a cell by at least about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 99%, about 100% of the mRNA level detected in the cell without the presence of the siRNA or RNA interference molecule. According to one preferred embodiment, mRNA levels are reduced by at least about 70%, about 80%, about 90%, about 95%, about 99%, about 100%.

[00161] As used herein, the term "RNAi" refers to any type of interfering RNA, including, but not limited to, siRNAi, shRNAi, endogenous microRNA, and artificial microRNA. For example, it includes sequences previously identified as siRNAs, regardless of the subsequent RNA processing mechanism (i.e., although siRNAs are believed to have a specific processing mode in vivo that results in mRNA cleavage, such sequences may be included in vectors between the flanking sequences described herein). The term "RNAi" may include both RNAi molecules that silence a gene and RNAi effector molecules that activate gene expression. By way of example only, in some embodiments, RNAi agents that serve to inhibit or suppress a gene can be used in the methods, kits, and compositions disclosed herein, such as to inhibit an immune response (e.g., an innate immune response).

[00162] As used herein, the term "comprising" or "comprises" is used to refer to compositions, methods, and their respective component(s) that are essential to the said method or composition, but may include non-specified elements, whether essential or not.

[00163] As used herein, the term "consisting essentially of" refers to elements necessary for a particular embodiment. The term allows for the presence of elements that do not substantially affect the basic and novel or functional characteristic(s) of the specified embodiment. The use of the word "comprising" indicates inclusion, not limitation.

[00164] The term "consisting of" refers to the compositions, methods, and their respective components described herein that do not include any elements not listed in the specified embodiment description.

[00165] As used herein, the term "consisting essentially of" refers to elements that are necessary for a particular embodiment. The term allows for the presence of additional elements that do not materially affect the basic and novel or functional characteristic(s) of the specified embodiment of the present invention.

[00166] In the present specification and the appended claims, singular terms (respectively, "a," "an," and "the" in the original English language text) include their plural equivalents unless the context clearly dictates otherwise. Accordingly, for example, reference to "a method" includes one or more methods and/or steps of the type described herein and/or those that will be understood by those skilled in the art after reading this disclosure, and the like. Likewise, the term "or" is intended to include "and" unless the context clearly dictates otherwise. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. As used herein, "for example" (the abbreviation "e.g." in the original English language text, derived from the Latin expression "exempli gratia") indicates a non-limiting example. Accordingly, “for example” is synonymous with the term “for instance”.

[00167] Except in the operating examples or where otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used herein are to be understood as being modified by the term "about" in all instances. The term "about" when used in connection with percentages may mean ±1%. The present invention is further explained in more detail in the following examples, but the scope of the present invention is not limited thereto.

[00168] The groups of alternative elements or embodiments of the present invention disclosed herein are not to be construed as limitations. Each member of a group may be referred to and claimed individually or in any combination with other members of the group or other elements disclosed herein. One or more members of a group may be included or excluded from a group for reasons of convenience and/or patentability. In any such inclusion or exclusion, the specification herein is deemed to contain the group modified so that it satisfies the condition of a written description of all Markush groups used in the appended claims.

[00169] In some embodiments of any of the aspects, the disclosure described herein does not relate to a method of cloning humans, methods of modifying the genetic identity of a human germline, uses of human embryos for industrial or commercial purposes, or methods of modifying the genetic identity of animals that may cause them suffering without any significant medical benefit to a human or animal, as well as animals obtained using such methods.

[00170] Definitions of other terms are provided herein in describing various aspects of the present invention.

II. Nucleic acids

[00171] Nucleic acids are large, highly charged molecules that are rapidly degraded and eliminated from the body and generally have poor pharmacological properties because they are recognized as foreign to the body and become a target of the immune response (e.g., the innate immune response). Therefore, certain nucleic acids, such as therapeutic nucleic acids or nucleic acids used for research purposes (e.g., antisense oligonucleotide or viral vectors), can often trigger immune responses in vivo. The present disclosure provides pharmaceutical compositions and methods that can alleviate, reduce, or eliminate such immune responses and increase the efficacy of nucleic acids by increasing expression levels by maximizing the stability of the nucleic acid in a state of reduced immunological reactivity in a recipient subject. This can also minimize any potential adverse events that may lead to organ damage or other toxicity during gene therapy. Many of the compositions and methods provided herein relate to administering a specific inhibitor of an immune response (e.g., an innate immune response) in combination with a nucleic acid (e.g., a therapeutic nucleic acid or a nucleic acid used for research purposes), which reduces the immune response (e.g., an innate immune response) triggered by the presence of the nucleic acid.

[00172] Immunogenic/immunostimulatory nucleic acids can include both deoxyribonucleic acids and ribonucleic acids. For deoxyribonucleic acids (DNA), a particular sequence or motif has been shown to induce immune stimulation in mammals. These sequences or motifs include, but are not limited to, CpG motifs, pyrimidine-rich sequences, and palindromic sequences. CpG motifs in deoxyribonucleic acid are often recognized by endosomal Toll-like receptor 9 (TLR-9), which in turn triggers both the innate immune stimulation pathway and the adaptive immune stimulation pathway. Certain immunostimulatory ribonucleic acid (RNA) sequences bind to Toll-like receptor 6 and 7 (TLR-6 and TLR-7) and are thought to activate a proinflammatory response via the immune response (e.g., the innate immune response). In addition, double-stranded RNA can often be immunostimulatory due to its binding to TLR-3. Thus, foreign nucleic acid molecules of pathogenic or therapeutic origin can be highly immunogenic in vivo.

[00173] The present document provides for the characterization and development of nucleic acid molecules for potential therapeutic use in combination with antagonists of the immune response (e.g., the innate immune response). In some embodiments, chemical modification of oligonucleotides is described, where appropriate, to alter and improve in vivo properties (delivery, stability, lifetime, folding, target specificity) as well as their biological function and mechanism that directly correlate with the therapeutic application.

[00174] Exemplary therapeutic nucleic acids of the present invention that may be immunostimulatory and require the use of the immunosuppressants disclosed herein may include, but are not limited to, minigenes, plasmids, minicircles, short interfering RNA (siRNA), microRNA (miRNA), antisense oligonucleotides (ASO), ribozymes, closed-ended double-stranded DNA (e.g., ccDNA, CELiD, linear covalently closed DNA ("minimal strand"), doggybone (dbDNA ^™ ), closed-ended protelomeric DNA, or dumbbell linear DNA), dsRNA substrate Dicer, short hairpin RNA (shRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), mRNA, tRNA, rRNA, and viral DNA vectors, viral RNA vector; and any combination thereof.

[00175] siRNA or miRNA, which can negatively regulate intracellular levels of specific proteins through a process called RNA interference (RNAi), are also provided by the present invention as nucleic acid therapeutic agents. Upon introduction of siRNA or miRNA into the cytoplasm of a host cell, said double-stranded RNA constructs bind to a protein called RISC. The RISC complex removes the sense strand of the siRNA or miRNA. The RISC complex, when combined with complementary mRNA, cleaves the mRNA and releases the cleaved strands. RNAi is accomplished by inducing specific degradation of mRNA, resulting in negative regulation of the corresponding protein.

[00176] Antisense oligonucleotides (ASOs) and ribozymes that inhibit the translation of mRNA into protein may be nucleic acid therapeutics. In the case of antisense constructs, these single-stranded deoxynucleic acids have a sequence complementary to the mRNA sequence of the target protein and are capable of binding to the mRNA via Watson-Crick base pairing. This binding prevents translation of the target mRNA and/or triggers degradation of the mRNA transcript by RNase H. As a result, the antisense oligonucleotide has increased specificity of action (i.e., down-regulation of a specific disease-associated protein).

[00177] According to any of the methods provided herein, the therapeutic nucleic acid may be a therapeutic RNA. The therapeutic RNA may be an mRNA translation inhibitor, an RNA interference (RNAi) agent, a catalytically active RNA molecule (ribozyme), a transfer RNA (tRNA), or an RNA that binds an mRNA transcript (ASO), a protein, or another molecular ligand (aptamer). According to any of the methods provided herein, the RNAi agent may be a double-stranded RNA, a single-stranded RNA, a microRNA, a short interfering RNA, a short hairpin RNA, or a triplex-forming oligonucleotide.

[00178] In some embodiments, the therapeutic nucleic acid is a double-stranded DNA with closed ends, such as ccDNA. In some embodiments, the expression and/or production of the therapeutic protein in the cell occurs from a non-viral DNA vector, such as a ccDNA vector. A distinct advantage of ccDNA vectors for expressing a therapeutic protein compared to conventional AAV vectors and even lentiviral vectors is that there is no size limitation for the heterologous nucleic acid sequences encoding the target protein. Thus, even a large therapeutic protein can be expressed from a single ccDNA vector. Thus, ccDNA vectors can be used to express a therapeutic protein in a subject in need thereof.

[00179] In general, a ccDNA vector for expressing a therapeutic protein disclosed herein comprises, in 5' to 3' direction: a first inverted terminal repeat (ITR) of an adeno-associated virus (AAV), a nucleotide sequence of interest (e.g., an expression cassette described herein), and a second AAV ITR. The ITR sequences are selected from any of: (i) at least one WT-ITR and at least one modified AAV inverted terminal repeat (mod-ITR) (e.g., asymmetric modified ITRs); (ii) two modified ITRs, wherein the pair of mod-ITRs has a different three-dimensional spatial organization relative to each other (e.g., asymmetric modified ITRs), or (iii) a pair of symmetric or substantially symmetric WT-WT-ITRs, in which each of the WT-ITRs has the same three-dimensional spatial organization, or (iv) a pair of symmetric or substantially symmetric modified ITRs, in which each of the mod-ITRs has the same three-dimensional spatial organization.

III. cDNA vectors

[00180] According to some aspects of the present disclosure, non-viral capsidless DNA vectors with covalently closed ends (ccDNA vector) are provided that are administered in combination with rapamycin or rapamycin analogs. In some embodiments, rapamycin or a rapamycin analog is present in a supersaturated amount in a synthetic nanocarrier as described in WO 2016/073799. In some embodiments, the ccDNA vector is also present in the same nanocarrier.

[00181] According to some aspects of the present disclosure, non-viral capsidless DNA vectors with covalently closed ends (ccDNA) are provided that are administered in combination with one or more TLR9 antagonists. Also provided herein are ccDNA constructs comprising sequences encoding, among other things, one or more TLR9 inhibitory oligonucleotides.

[00182] According to some aspects of the present disclosure, non-viral capsidless DNA vectors with covalently closed ends (ccDNA) are provided that are administered in combination with one or more cGAS antagonists. Also provided herein are ccDNA constructs comprising sequences encoding, among other things, one or more RNAs or proteins that inhibit cGAS.

[00183] According to some aspects of the present disclosure, non-viral capsidless DNA vectors with covalently closed ends (ccDNA) are provided that are administered in combination with one or more inflammasome antagonists (e.g., any one or more of an NLRP3 inflammasome pathway inhibitor or an AIM2 inflammasome pathway inhibitor or a caspase 1 inhibitor or any combination thereof). Also provided herein are ccDNA constructs comprising sequences encoding, among other things, one or more inflammasome antagonists (e.g., any one or more of an NLRP3 inflammasome pathway inhibitor or an AIM2 inflammasome pathway inhibitor or a caspase 1 inhibitor or any combination thereof).

[00184] As will be appreciated, the cDNA vector technologies described herein can be adapted to any level of complexity or can be used modularly, wherein the expression of various components of an immune response inhibitor (e.g., an innate immune response), such as those described herein, can be controlled independently. For example, it is intended that the ccDNA vector technologies developed herein may be as simple as using a single ccDNA vector to express a single heterologous gene sequence (e.g., a single inhibitor of an immune response (e.g., an innate immune response), such as those described herein, for example, c), or may be as complex as using multiple ccDNA vectors, wherein each vector expresses multiple inhibitors of an immune response (e.g., an innate immune response), such as those described herein, for example, or a nucleic acid sequence encoding one or more inhibitors of an immune response (e.g., an innate immune response), such as those described herein, and, for example, associated cofactors or accessory proteins, each independently controlled by different promoters.

[00185] In one embodiment, a single ccDNA vector can be used to express a single component of an inflammasome antagonist (e.g., any one or more of an NLRP3 inflammasome pathway inhibitor or an AIM2 inflammasome pathway inhibitor or a caspase 1 inhibitor or any combination thereof). Alternatively, a single ccDNA vector can be used to express multiple components (e.g., at least 2), such as expressing two or more NLRP3 inflammasome pathway inhibitors and/or two or more AIM2 inflammasome pathway inhibitors and/or two or more caspase 1 inhibitors or any combination thereof under the control of a single promoter (e.g., a strong promoter), optionally using an IRES sequence(s) to ensure proper expression of each of the components, e.g., cofactors or accessory proteins.

[00186] Also provided herein, in another embodiment, is a single ccDNA vector comprising at least two inserts, wherein expression of each insert is under the control of its own promoter. Promoters may include multiple copies of the same promoter, multiple different promoters, or any combination thereof. One skilled in the art will appreciate that it is often desirable to express multiple inflammasome antagonists at different expression levels, allowing control of the stoichiometry of the individual components expressed to ensure efficient expression and, in the case of a protein, folding and combining of the protein in the cell.

[00187] In some embodiments, the synthetic ccDNA is produced by excision from a double-stranded DNA molecule. Synthetic production of ccDNA vectors is described in Examples 2-6 of international application PCT/US19/14122, filed January 18, 2019, which is incorporated herein by reference in its entirety. One exemplary method for producing a ccDNA vector using a synthetic method involves excision of a double-stranded DNA molecule. In general, a ccDNA vector can be produced using a double-stranded DNA construct, for example, see FIGS. 7A-8E of PCT/US19/14122. In some embodiments, the double-stranded DNA construct is a ccDNA plasmid, for example, see FIG. 6 in international patent application PCT/US2018/064242, filed on December 6, 2018.

[00188] In some embodiments, the construct for producing a ccDNA vector comprises a regulatory switch as described herein.

[00189] Another exemplary method for producing a ccDNA vector using a synthetic method that involves assembling various oligonucleotides is provided in Example 3 of PCT/US19/14122, wherein the ccDNA vector is produced by synthesizing a 5' oligonucleotide and a 3' ITR oligonucleotide and ligating said ITR oligonucleotides into a double-stranded polynucleotide containing an expression cassette. FIG. 11B of PCT/US19/14122 provides an exemplary method for ligating a 5' ITR oligonucleotide and a 3' ITR oligonucleotide into a double-stranded polynucleotide containing an expression cassette.

[00190] An exemplary method for producing a ccDNA vector using the synthetic method set forth in Example 4 of PCT/US19/14122 utilizes a single-stranded linear DNA comprising two sense ITRs that flank a sense sequence of an expression cassette and are covalently attached to two antisense ITRs that flank the antisense sequence of the expression cassette, and the ends of said single-stranded linear DNA are then ligated to produce a single-stranded molecule with closed ends. One non-limiting example includes synthesizing and/or producing a single-stranded DNA molecule, annealing portions of said molecule to produce a single linear DNA molecule that has one or more base-paired secondary structure regions, and then ligating the free 5' and 3' ends together to produce a closed single-stranded molecule.

[00191] One skilled in the art may envision additional variations of cDNA vector technologies or adapt them from protein production methods using standard vectors.

[00192] Non-viral capsidless DNA vectors can be produced in susceptible host cells from an expression construct (e.g., a plasmid, bacmid, baculovirus, or integrated cell line), e.g., see the examples disclosed in International Patent Application No. PCT/US18/49996, filed September 7, 2018, or using a synthetic production method, e.g., see the examples disclosed in International Patent Application No. PCT/US19/14122, filed December 6, 2018, each of which is incorporated herein by reference in its entirety. In some embodiments, ccDNA vectors that may be used in the methods and compositions disclosed herein comprise a heterologous nucleic acid, e.g., a transgene, located between two inverted terminal repeat (ITR) sequences. In some embodiments, at least one of the ITRs is modified by a deletion, insertion, and/or substitution relative to a wild-type ITR sequence (e.g., an AAB ITR); and at least one of the ITRs comprises a functional terminal resolution site (TRS) and a Rep binding site. In one embodiment, at least one of the ITRs has at least one polynucleotide deletion, insertion, or substitution relative to the corresponding AAB ITR (e.g., SEQ ID NO: 1 or SEQ ID NO: 51 for wild-type AAV2) to induce replication of the vector DNA in a host cell in the presence of the Rep protein. As discussed above, any ITR can be used. For illustrative purposes, the ITRs in the ccDNA constructs in Table 1A are a modified ITR and a WT ITR. However, provided herein are ccDNA vectors that comprise a heterologous nucleic acid sequence (transgene) located between two inverted terminal repeat (ITR) sequences, wherein said ITR sequences may be a pair of asymmetric ITRs or a pair of symmetric or substantially symmetric ITRs, as those terms are defined herein. A ccDNA vector comprising a nuclear localization signal (NLS) disclosed herein may comprise ITR sequences that are selected from any of: (i) at least one WT-ITR and at least one modified inverted terminal repeat (mod-ITR) of AAB (e.g., asymmetric modified ITRs); (ii) two modified ITRs, wherein said pair of mod-ITRs have a different three-dimensional spatial organization relative to each other (e.g., asymmetric modified ITRs), or (iii) a pair of symmetric or substantially symmetric WT-WT-ITRs, in which each WT-ITR has the same three-dimensional spatial organization, or (iv) a pair of symmetric or substantially symmetric modified ITRs, in which each mod-ITR has the same three-dimensional spatial organization, wherein said methods according to the present disclosure may further include a delivery system, such as, but not limited to, a liposome nanoparticle-based delivery system.

[00193] In some embodiments, the methods and compositions described herein relate to the use of an inhibitor of an immune response (e.g., an innate immune response) disclosed herein for co-administration with any ccDNA vector, including, but not limited to, a ccDNA vector comprising asymmetric ITRs disclosed in International Patent Application PCT/US18/49996, filed September 7, 2018 (see, e.g., Examples 1-4); The ccDNA vector for gene editing disclosed in International Patent Application PCT/US18/64242, filed December 6, 2018 (see, for example, Examples 1-7), or the ccDNA vector for producing antibodies or fusion proteins disclosed in International Patent Application PCT/US19/18016, filed February 14, 2019 (see, for example, Examples 1-4), or the ccDNA vector for controlled expression of a transgene disclosed in International Patent Application PCT/US19/18927, filed February 22, 2019, each of which is incorporated herein by reference in its entirety. In some embodiments, it is also contemplated that the methods and compositions described herein using an inhibitor of an immune response (e.g., an innate immune response) disclosed herein can be used with a ccDNA vector produced by a synthetic process, such as a ccDNA vector produced in a cell-free or insect-free system for producing ccDNA, as disclosed in International Application No. PCT/US19/14122, filed January 18, 2019, which is incorporated herein by reference in its entirety.

[00194] The ccDNA vector is preferably duplex or self-complementary, at least over a portion of the molecule, e.g., the transgene. The ccDNA vector has covalently closed ends and, accordingly, is preferably resistant to exonuclease (e.g., Exo I or Exo III) digestion for more than an hour at 37°C. The presence of the Rep protein in host cells (e.g., insect cells or mammalian cells) facilitates replication of the ccDNA vector polynucleotide template, which has a modified ITR that induces the production of a non-viral capsidless DNA vector with covalently closed ends. The covalently capped molecule continues to accumulate in susceptible cells by replication and is preferably sufficiently stable over time in the presence of the Rep protein under standard replication conditions, such as to accumulate at yields of at least 1 pg/cell, preferably at least 2 pg/cell, preferably at least 3 pg/cell, more preferably at least 4 pg/cell, even more preferably at least 5 pg/cell.

[00195] In particular, in one embodiment, DNA vectors are prepared by providing cells (e.g., insect cells or mammalian cells, e.g., 293 cells, etc.) carrying a vector polynucleotide template (e.g., an expression construct) that comprises two different ITRs (e.g., the AAB ITR) and a nucleotide sequence of interest (heterologous nucleic acid, expression cassette) located between the ITRs, wherein at least one of said ITRs is a modified ITR that comprises an insertion, substitution, or deletion relative to the other ITR. The vector polynucleotide template described herein comprises at least one functional ITR that comprises a Rep binding site (RBS; e.g., 5'-GCGCGCTCGCTCGCTC-3' for AAB2) and a functional terminal resolution site (TRS; e.g., 5'-AGTT). The cells do not express viral capsid proteins and the vector polynucleotide matrix lacks sequences encoding the viral capsid.

[00196] In the presence of Rep, a vector polynucleotide template having at least one modified ITR is replicated to produce a ccDNA vector. The production of the ccDNA vector occurs in two steps: first, excision ("extraction") of the template from the vector backbone (e.g., plasmid, bacmid, genome, etc.) using Rep proteins, and second, Rep-mediated replication of the excised vector genome. The Rep proteins and Rep binding sites of the various AAV serotypes are well known to those skilled in the art. One skilled in the art will appreciate the need to select a Rep protein from a serotype that binds to and replicates a functional ITR.

[00197] Cells harboring a vector polynucleotide either already contain Rep (e.g., an inducible rep cell line) or are transduced with a vector that contains Rep and then grown under conditions that allow replication and release of the ccDNA vector. The ccDNA vector DNA is then collected and isolated from the cells. The presence of capsidless non-viral ccDNA vector DNA can be confirmed by digesting the vector DNA isolated from the cells with a restriction enzyme that has a single recognition site on the vector DNA and analyzing the digested DNA material on a non-denaturing gel to confirm the presence of characteristic bands of linear and continuous DNA, as compared to linear and discontinuous DNA. For example, FIG. 6 is a gel confirming the production of a ccDNA vector from several TTX plasmid constructs using one embodiment for producing such vectors described in the Examples. The ccDNA vector is confirmed by a characteristic band profile in the gel, as discussed with respect to Figure 4D. FIG. 5A and FIG. 5B are drawings illustrating one embodiment for identifying the presence of closed-ended ccDNA vectors produced by the methods of the present invention.

[00198] The vector polynucleotide expression template (e.g., a TTX plasmid, bacmid, etc.) and/or ii) the polynucleotide that encodes Rep can be introduced into the cells using any means well known to those skilled in the art, including, but not limited to, transfection (e.g., calcium phosphate, a nanoparticle, or a liposome) or introduction via viral vectors, e.g., HSV or baculovirus. For example, the vector polynucleotide expression template used to produce the ccDNA vectors of the present invention can be a plasmid (e.g., TTX plasmids, e.g., see FIG. 4B), a bacmid (e.g., a TTX bacmid), and/or a baculovirus (e.g., a TTX baculovirus). In one embodiment, the TTX plasmid comprises a restriction cloning site (e.g., SEQ ID NO: 7) operably located between the ITRs into which a heterologous nucleic acid (e.g., an expression cassette containing a reporter gene or a therapeutic nucleic acid) can be inserted.

[00199] According to one preferred embodiment, the host cells used to produce the ccDNA vectors described herein are insect cells. According to another preferred embodiment, a baculovirus is used to deliver both a polynucleotide that encodes a Rep protein and a polynucleotide template for expressing a non-viral DNA vector construct for the ccDNA vector. Examples of such methods for producing and isolating ccDNA vectors are described in FIGS. 1-33.

[00200] According to another aspect of the present invention, there are provided host cell lines having a DNA vector polynucleotide expression template (ccDNA vector template) described herein stably integrated into their own genome for use in the production of a non-viral DNA vector. Methods for producing such cell lines are described in Lee, L. et al. (2013) Plos One 8(8): e69879, which is incorporated herein by reference in its entirety. Preferably, the Rep protein (e.g., as described in Example 1) is added to the host cells at a multiplicity of infection (MOI) of 3. In one embodiment, the host cell line is an invertebrate cell line, preferably an Sf9 insect cell line. If the host cell line is a mammalian cell line, preferably 293 cells, said cell lines can contain a stably integrated vector polynucleotide template, and a second vector, such as a herpes virus, can be used to introduce the Rep protein into the cells, which allows for excision and amplification of the ccDNA vector in the presence of Rep.

[00201] Preferably, the ccDNA comprises one or more functional ITR polynucleotide sequences that include a Rep binding site (RBS; 5'-GCGCGCTCGCTCGCTC-3' for AAB2, SEQ ID NO: 39) and a terminal resolution site (TRS; 5'-AGTT).

[00202] Capsidless cDNA vectors can be derived from expression constructs (e.g., TTX plasmids, TTX bacmid, TTX baculovirus) that further include a defined combination of cis-regulatory elements, such as the WHP post-transcriptional regulatory element (WPRE) and bovine growth hormone poly(A) (BGH). Suitable expression cassettes for use in expression constructs are free from packaging constraints imposed by the viral capsid. Expression cassettes according to the present disclosure include a promoter that can influence overall expression levels as well as cell specificity. In the case of transgene expression, they can include a highly active immediate early promoter of viral origin. Expression cassettes may comprise a tissue-specific eukaryotic promoter to restrict expression of the transgene to specific cell types and to reduce toxic effects and immune responses caused by unregulated ectopic expression. In some embodiments, the expression cassette may comprise a synthetic regulatory element, such as a CAG promoter (SEQ ID NO: 3). The CAG promoter comprises (i) a cytomegalovirus (CMV) early enhancer element (e.g., SEQ ID NO: 309), (ii) a promoter, first exon, and first intron of the chicken beta-actin gene, and (iii) a splice acceptor of the rabbit beta-globin gene. Alternatively, for example, the expression cassette may comprise an alpha-1 antitrypsin (AAT) promoter (e.g., SEQ ID NO: 4), a liver-specific (LP1) promoter (e.g., SEQ ID NO: 5), or a HAAT promoter (e.g., SEQ ID NO: 135), or a human elongation factor-1-alpha (EF1-α) promoter (SEQ ID NO: 6), or a fragment of EF1-α (SEQ ID NO: 66), or a MND promoter (SEQ ID NO: 70). In some embodiments, the expression cassette comprises one or more constitutive promoters, such as a retroviral promoter from the Rous sarcoma virus (RSV) LTR (optionally with an RSV enhancer), a cytomegalovirus (CMV) immediate early promoter (optionally with a CMV enhancer), and the like. Alternatively, an inducible promoter, a transgene native promoter, a tissue-specific promoter, or various promoters known in the art may be used. In one embodiment, an endogenous or native promoter is used in the expression cassette for the gene coding sequence.

[00203] Inducible gene editing using ccDNA vectors can be accomplished using methods described in, for example, Dow et al. Nat Biotechnol 33:390-394 (2015); Zetsche et al. Nat Biotechnol 33:139-42 (2015); Davis et al. Nat Chem Biol 11:316-318 (2015); Polstein et al. Nat Chem Biol 11:198-200 (2015); and/or Kawano et al. Nat Commun 6:6256 (2015), the contents of each of which are herein incorporated by reference in their entirety. The expression cassettes may also include a post-transcriptional element, in particular the North American woodchuck hepatitis virus (WHP) post-transcriptional regulatory element (WPRE) (SEQ ID NO: 72), to enhance expression of the transgene. Other post-transcriptional processing elements may be used, such as the herpes simplex virus or hepatitis B virus (HBV) thymidine kinase gene post-transcriptional element. The expression cassettes may include a polyadenylation sequence known in the art or a variant thereof, such as a natural sequence derived from the bovine BGHpA sequence or the SV40pA viral sequence (e.g., SEQ ID NO: 10), or a synthetic sequence. Some expression cassettes may also include the SV40 5' late poly(A) signal enhancer (USE) sequence. USE can be used in combination with SV40pA or heterologous poly(A) signal.

The time for collecting and accumulating the DNA vectors described herein from the cells can be selected and optimized to achieve high yields of DNA vectors. For example, the time of collection can be selected taking into account cell viability, cell morphology, cell proliferation, etc. Typically, the cells can be collected after a sufficient time has passed after baculovirus infection to obtain DNA vectors (e.g., TTX vectors), but before the majority of the cells begin to die due to viral toxicity. DNA vectors can be isolated, for example, using plasmid purification kits, such as the Endo-Free ^™ Plasmid kits from Qiagen. Other methods developed for isolating plasmids can also be adapted for DNA vectors. Generally, any method for purifying nucleic acid known in the art can be used.

Regulatory sequences and effectors

[00204] In embodiments, the ccDNA vector comprises a second nucleotide sequence (e.g., a regulatory sequence) in addition to one or more nucleotide sequences encoding a therapeutic protein. In embodiments, the gene regulatory sequence is operably linked to a nucleotide sequence encoding a therapeutic protein. In embodiments, the regulatory sequence is suitable for controlling the expression of a therapeutic protein in a host cell. In embodiments, the regulatory sequence includes a suitable promoter sequence capable of directing transcription of a gene operably linked to the promoter sequence, such as a nucleotide sequence encoding a therapeutic protein according to the present disclosure. In embodiments, the second nucleotide sequence includes an intron sequence linked to the 5' end of the nucleotide sequence encoding the therapeutic protein. In embodiments, the enhancer sequence is provided in the 5' direction relative to the promoter to enhance the efficiency of the promoter. According to embodiments, the regulatory sequence comprises an enhancer and a promoter, wherein said second nucleotide sequence comprises an intron sequence in the 5' direction relative to the nucleotide sequence encoding the therapeutic protein, wherein said intron comprises one or more cleavage sites for a nuclease, and wherein said promoter is operably linked to the nucleotide sequence encoding the nuclease. According to embodiments, the regulatory sequence used is native to the coding sequence in the vector.

[00205] Promoters: Suitable promoters, including those described above, may be from viruses and may accordingly be referred to as viral promoters, or they may be from any organism, including prokaryotic or eukaryotic organisms. Suitable promoters may be used to drive expression by any RNA polymerase (e.g., pol I, pol II, pol III). Exemplary promoters include, but are not limited to, the SV40 early promoter, the promoter from the mouse mammary tumor virus long terminal repeat (LTR); the adenovirus large late promoter (Ad MLP); herpes simplex virus (HSV) promoter, cytomegalovirus (CMV) promoter such as CMV immediate early promoter region (CMVIE), Rous sarcoma virus (RSV) promoter, human small nuclear RNA U6 promoter (U6, e.g., SEQ ID NO: 18) (Miyagishi et al., Nature Biotechnology 20, 497-500 (2002)), improved U6 promoter (e.g., Xia et al., Nucleic Acids Res. 2003 Sep. 1; 31(17)), human H1 promoter (H1) (e.g., SEQ ID NO: 19), CAG promoter, human alpha-1-antitrypsin (HAAT) promoter (e.g., SEQ ID NO: 135), etc. In embodiments, the promoters are altered at their intron-containing 3' end to include one or more nuclease cleavage sites. In some embodiments, the DNA containing the nuclease cleavage site(s) is foreign to the promoter DNA.

[00206] A promoter may contain one or more specific transcriptional regulatory sequences to further enhance expression and/or to spatially and/or temporally alter the expression of said promoter. A promoter may also contain distal enhancer or repressor elements, which may be located up to several thousand base pairs from the transcription start site. A promoter may be derived from sources including viruses, bacteria, fungi, plants, insects, and animals. A promoter may regulate expression of a gene component constitutively or differentially with respect to the cell, tissue, or organ where expression occurs, or with respect to the developmental stage at which expression occurs, or in response to external stimuli such as physiological stresses, pathogens, metal ions, or inducing agents. Representative examples of promoters include the bacteriophage T7 promoter, the bacteriophage T3 promoter, the SP6 promoter, the lac operator/promoter, the tac promoter, the SV40 late promoter, the SV40 early promoter, the RSV-LTR promoter, the CMV immediate early promoter, the SV40 early promoter, or the SV40 late promoter and CMV immediate early promoter, as well as the promoters listed below. Such promoters and/or enhancers can be used to express any gene of interest, such as gene editing molecules, donor sequences, therapeutic proteins, etc. For example, a vector can comprise a promoter that is operably linked to a nucleic acid sequence encoding a therapeutic protein. The promoter operably linked to the therapeutic protein encoding sequence may be a promoter from simian virus 40 (SV40), a murine mammary tumor virus (MMTV) promoter, a human immunodeficiency virus (HIV) promoter such as the bovine immunodeficiency virus (BIV) long terminal repeat (LTR) promoter, a Moloney virus promoter, an avian leukemia virus (ALV) promoter, a cytomegalovirus (CMV) promoter such as the CMV immediate early promoter, an Epstein-Barr virus (EBV) promoter, or a Rous sarcoma virus (RSV) promoter. The promoter may also be a promoter from a human gene such as human ubiquitin C (hUbC), human actin, human myosin, human hemoglobin, human muscle creatine, or human metallothionein. The promoter may also be a tissue-specific promoter, such as a liver-specific promoter, for example, human alpha-1-antitrypsin (HAAT), natural or synthetic. In one embodiment, delivery to the liver can be achieved using specific targeting of endogenous ApoE by a composition comprising a ccDNA vector, which is directed to hepatocytes by a low-density lipoprotein (LDL) receptor present on the surface of the hepatocyte.

[00207] According to one embodiment, the promoter used is a native promoter of a gene encoding a therapeutic protein. Promoters and other regulatory sequences of corresponding genes encoding therapeutic proteins are known and have been characterized. The promoter region used may further include one or more additional regulatory sequences (e.g., native), such as enhancers.

[00208] Non-limiting examples of suitable promoters for use in accordance with the present invention include the CAG promoter, for example (SEQ ID NO: 3), the HAAT promoter (SEQ ID NO: 135), the human EF1-α promoter (SEQ ID NO: 6) or a fragment of the EF1-α promoter (SEQ ID NO: 66), and the rat EF1-α promoter (SEQ ID NO: 310).

[00209] Enhancers: In some embodiments, a ccDNA expressing an inflammasome antagonist (e.g., an inhibitor of one or more of the NLRP3 and/or AIM2 inflammasome pathway, or a caspase 1 inhibitor) comprises one or more enhancers. In some embodiments, the enhancer sequence is located 5' relative to the promoter sequence. In some embodiments, the enhancer sequence is located 3' relative to the promoter sequence. Exemplary enhancers are listed in Table 1 herein.

Table 1. Approximate enhancer sequences

Description Length Tissue specificity CG Contents Subsequence Cytomegalovirus enhancer 518 Constitutive 22 TCAATATTGGCCATTAGCCATATTATTCATTGGTTATATAGCATAAATCAATATTGGCTATTGGCCATTGCATACGTTGTATCTATATCATAATATGTACATTTATATTGGCTCATGTCCAATATGACC GCCATGTTGGCATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAAC GACCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCAT ATGCCAAGTCCGCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTACGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGG Liver-specific enhancer of human apolipoprotein E/C-I 777 Liver 13 AGGCTCAGAGGCACACAGGAGTTTCTGGGCTCACCCTGCCCCCTTCCAACCCCTCAGTTCCCATCCTCCAGCAGCTGTTTGTGTGCTGCCTCTGAAGTCCACACTGAACAAACTTCAGCCTACTCATGTCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACC TTGGAGCTGGGGCAGAGGTCAGAGACCTCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGTCCGGGTTCAAAACCACTTGCTGGGTGGGGAGTCGTCAGTAAGTGGCTATGCCCCGACCCCGAAGCC TGTTTCCCCATCTGTACAATGGAAATGATAAAGACGCCCATCTGATAGGGTTTTTGTGGCAAATAAACATTTGGTTTTTTTGTTTTGTTTTGTTTTGTTTTTTGAGATGGAGGTTTGCTCTGTCGCCCAGGCTGGAGTGCAGTGACACAATCTCATCTCACCACAACCTTCCCCTGCCTCAGCCTCCCAAGTAG CTGGGATTACAAGCATGTGCCACCACACCTGGCTAATTTTCTATTTTTAGTAGAGACGGGTTTCTCCATGTTGGTCAGCCTCAGCCTCCCAAGTAACTGGGATTACAGGCCTGTGCCACCACACCCGGCTAATTTTTTCTATTTTTGACAGGGACGGGGTTTCACCATGTTGGTCAGGCTGGTCTAGAGGTACCG Mouse CMV enhancer without CpG 427 Constitutive 0 GAGTCAATGGGAAAAACCCATTGGAGCCAAGTACACTGACTCAATAGGGACTTTCCATTGGGTTTTGCCCAGTACATAAGGTCAATAGGGGGTGAGTCAACAGGAAAGTCCCATTGGAGCCAAGTACATTGAGTCAATAGGGACTTTCCAATGGGTTTTGCCCAGTACATAAGGTCAATGGGAGGTAAGCCAATGGGTTTTTCCCATTACTGA CATGTATACTGAGTCATTAGGGACTTTCCAATGGGTTTTGCCCAGTACATAAGGTCAATAGGGGTGAATCAACAGGAAAGTCCCATTGGAGCCAAGTACACTGAGTCAATAGGGACTTTCCATTGGGTTTTGCCCAGTACAAAAGGTCAATAGGGGGTGAGTCAATGGGTTTTTCCCATTATTGGCACATACATAAGGTCAATAGGGGTGACTA Enhancer HS-CRM8 SERP 83 Liver 4 CGGGGGAGGCTGCTGGTGAATATTAACCAAGGTCACCCCAGTTATCGGAGGAGCAAACAGGGGCTAAGTCCACACGCGTGGTA Liver-specific enhancer of human apolipoprotein E/C-I 777 Liver 12 AGGCTCAGAGGCACACAGGAGTTTCTGGGCTCACCCTGCCCCCTTCCAACCCCTCAGTTCCCATCCTCCAGCAGCTGTTTGTGTGCTGCCTCTGAAGTCCACACTGAACAAACTTCAGCCTACTCATGTCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACC TTGGAGCTGGGGCAGAGGTCAGAGACCTCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGTCCGGGTTCAAAACCACTTGCTGGGTGGGGAGTCGTCAGTAAGTGGCTATGCCCCGACCCCGAAGCC TGTTTCCCCATCTGTACAATGGAAATGATAAAGACGCCCATCTGATAGGGTTTTTGTGGCAAATAAACATTTGGTTTTTTTGTTTTGTTTTGTTTTGTTTTTTGAGATGGAGGTTTGCTCTGTCGCCCAGGCTGGAGTGCAGTGACACAATCTCATCTCACCACAACCTTCCCCTGCCTCAGCCTCCCAAGTAG CTGGGATTACAAGCATGTGCCACCACACCTGGCTAATTTTCTATTTTTAGTAGAGACGGGTTTCTCCATGTTGGTCAGCCTCAGCCTCCCAAGTAACTGGGATTACAGGCCTGTGCCACCACACCCGGCTAATTTTTTCTATTTTTGACAGGGACGGGGTTTCACCATGTTGGTCAGGCTGGTCTAGAGGTACTG APOe/c-1 enhancer 34 bp and AAT X-region 32 bp. 66 Liver 1 GTTTGCTGCTTGCAATGTTTGCCCATTTTAGGGTGGACACAGGACGCTGTGGTTTCTGAGCCAGGG hAPO-HCR insulator and enhancer sequence 212 Liver 4 GGAGGGGTGGAGTCGTGACCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGG hAPO-HCR enhancer derived from SPK9001 330 Liver 4 AGGCTCAGAGGCACACAGGAGTTTCTGGGCTCACCCTGCCCCCTTCCAACCCCTCAGTTCCCATCCTCCAGCAGCTGTTTGTGTGCTGCCTCTGAAGTCCACACTGAACAAACTTCAGCCTACTCATGTCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGC AAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGGTACCCGGG hAPO-HCR enhancer 194 Liver 3 CCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTTTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGG Invivogen SV40 Enhancer 240 Constitutive 0 GGGCCTGAAATAACCTCTGAAAGAGGAACTTGGTTAGGTACCTTCTGAGGCTGAAAGAACCAGCTGTGGAATGTGTGTCAGTTAGGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGA AGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCAGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCATAGTCCCACTA HS-CRM8 SERP enhancer with all spacers/restriction sites removed 73 Liver 2 CGGGGGAGGCTGCTGGTGAATATTAACCAAGGTCACCCCAGTTATCGGAGGAGCAAACAGGGGCTAAGTCCAC Enhancer alpha mic/bik 100 Liver 0 AGGTTAATTTTTAAAAAGCAGTCAAAAGTCCAAGTGGCCCTTGGCAGCATTTACTCTCTCTGTTTGCTCTGGTTAATAATCTCAGGAGCACAAAACATTCC Human CMV enhancer without CpG v2 296 Constitutive 0 GTTACATAACTTATGGTAAATGGCCTGCCTGGCTGACTGCCCAATGACCCCTGCCCAATGATGTCAATAATGATGTATGTTCCCATGTAATGCCAATAGGGACTTTCCATTGATGTCAATGGGTGGAGTATTTATGGTAACTGCCCAC TTGGCAGTACATCAAGTGTATCATATGCCAAGTATGCCCCCTATTGATGTCAATGATGGTAAATGGCCTGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTATGTATTAGTCATTGCTATTA Enhancer SV40 235 Constitutive 1 GGCCTGAAATAACCTCTGAAAGAGGAACTTGGTTAGGTACCTTCTGAGGCGGAAAGAACCAGCTGTGGAATGTGTGTCAGTTAGGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCA GAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCAGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCATAGTCCC

[00210] 5'-UTR and Intron Sequences: In some embodiments, the ccDNA vector comprises a 5'-UTR sequence and/or an intron sequence that is located 3' relative to the 5'-ITR sequence. In some embodiments, the 5'-UTR is located 5' relative to a transgene, such as a sequence encoding an inflammasome antagonist (e.g., an inhibitor of one or more of the NLRP3 and/or AIM2 inflammasome pathway, or an inhibitor of caspase 1). Exemplary 5'-UTR sequences are listed in Table 2A.

Table 2A: Approximate 5´-UTR and intron sequences

Description Length Source CG Contents Subsequence Synthetic 5´-UTR element consisting of 5´-UTR/intron of chicken B-actin and intron and 1st exon of rabbit B-globin 1127 137 GGAGTCGCTGCGACGCTGCCTTCGCCCCGTGCCCCGCTCCGCCGCCGCCTCGCGCCGCCCGCCCCGGCTCTGACTGACCGCGTTACTCCCACAGGTGAGCGGGCGGGACGGCCCTTCTCCTCCGGGCTGTAATTAGCGCT TGGTTTAATGACGGCTTGTTTCTTTTCTGTGGCTGCGTGAAAGCCTTGAGGGGCTCCGGGAGGGCCCTTTGTGCGGGGGGGAGCGGCTCGGGGGGTGCGTGCGTGTGTGTGTGCGTGGGGAGCGCCGCGTGCGGCCCGCGC TGCCCGGCGGCTGTGAGCGCTGCGGGCGCGGCGCGGGGCTTTGTGCGCTCCGCAGTGTGCGCGAGGGGAGCGCGGCCGGGGGCGGTGCCCCGCGGTGCGGGGGGGGCTGCGAGGGGAACAAAGGCTGCGTGCGGGGTGTGT GCGTGGGGGGGTGAGCAGGGGGTGTGGGCGCGGCGGTCGGGCTGTAACCCCCCCCTGCACCCCCCTCCCCGAGTTGCTGAGCACGGCCCGGCTTCGGGTGCGGGGCTCCGTACGGGGCGTGGCGCGGGGCTCGCCGTGCCG GGCGGGGGGTGGCGGCAGGTGGGGGTGCCGGGCGGGGCGGGGCCGCCTCGGGCCGGGGAGGGCTCGGGGGAGGGGCGCGGCGGCCCCCGGAGCGCCGGCGGCTGTCGAGGCGCGGCGAGCCGCAGCCATTGCCTTTTATGG TAATCGTGCGAGAGGGCGCAGGGACTTCCTTTGTCCCAAATCTGTGCGGAGCCGAAATCTGGGAGGCGCCGCCGCACCCCCTCTAGCGGGCGCGGGGCGAAGCGGTGCGGCGCCGGCAGGAAGGAAATGGGCGGGGAGGGC CTTCGTGCGTCGCCGCGCCGCCGTCCCCTTCTCCCTCCAGCCTCGGGGCTGTCCGCGGGGGGACGGCTGCCTTCGGGGGGGACGGGGCAGGGCGGGGTTCGGCTTCTGGCGTGTGACCGGCGGCTCTAGAGCCTCTGCT AACCATGTTTTAGCCTTCTTCTTTTTCCTACAGCTCCTGGGCAACGTGCTGGTTATTGTGCTGTCTCATCATTTGTCGACAGAATTCCTCGAAGATCCGAAGGGGTTCAAGCTTGGCATTCCGGTACTGTTGGTAAAGCCA Modified SV40 intron 93 0 CTCTAAGGTAAATATAAAATTTTTAAGTGTATAATGTGTTAAACTACTGATTCTAATTGTTTCTCTCTTTTAGATTCCAACCTTTGGAACTGA 5´-UTR of hAAT immediately before the ORF (3´ CGGA may represent a spacer/restriction enzyme cleavage site and was included in the sequence) 54 1 GCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATCCGGA Synthetic intron of the CET promoter set 173 0 CTGCCTTCTCCCTCCTGTGAGTTTGGTAAGTCACTGACTGTCTATGCCTGGGAAAGGGTGGGCAGGAGATGGGGCAGTGCAGGAAAAGTGGCACTATGAACCCTGCAGCCCTAGACAATTGTACTAACCTTTCTTCTCTTTCCTCTCCTGACAGGTTGGTGTACAGTAGCTTCC Small mouse virus (MVM) intron 91 0 AAGAGGTAAGGGTTTAAGGGATGGTTGGTTGGTGGGGTATTAATGTTTAATTACCTGGAGCACCTGCCTGAAATCACTTTTTTTCAGGTTG 5'-UTR hAAT 54 0 GCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATAATTA 5´-UTR hAAT in combination with modSV40 intron 147 1 GCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATCCGGACTCTAAGGTAAATATAAAATTTTTAAGTGTATAATGTGTTAAACTACTGATTCTAATTGTTTCTCTCTTTTAGATTCCAACCTTTGGAACTGA 5´-UTR hAAT (3´ TAATTA may represent a spacer/restriction enzyme cleavage site and was included in the sequence) in combination with the modSV40 intron 147 0 GCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATAATTACTCTAAGGTAAATATAAAATTTTTAAGTGTATAATGTGTTAAACTACTGATTCTAATTGTTTCTCTCTTTTAGATTCCAACCTTTGGAACTGA 42 bp of the 5´-UTR of AAT, derived from BMN270, includes the Kozak sequence 48 https://www.ncbi.nlm.nih.gov/pubmed/29292164 1 TCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATCGCCACC Intron/enhancer from EF1-α 128 US 2017/0216408 6 GCTAGCAGGTAAGTGCCGTGTGTGGTTCCCGCGGGCCTGGCCTCTTTACGGGTTATGGCCCTTGCGTGCCTTGAATTACTGACACTGACATCCACTTTTTCTTTTTCTCCACAGGTTTAAAACGCCACC Synthetic SBR intron derived from Sangamo CRMSBS2-Intron3 - includes Kozak sequence 98 WO 2017074526 2 AAGAGGTAAGGGTTTAAGTTATCGTTAGTTCGTGCACCATTAATGTTTAATTACCTGGAGCACCTGCCTGAAATCATTTTTTTTTCAGGTTGGCTAGT Endogenous hFVIII 5´-UTR 172 NG_011403.1 0 GCTTAGTGCTGAGCACATCCAGTGGGTAAAGTTCCTTAAAATGCTCTGCAAAGAAATTGGGACTTTTCATTAAATCAGAAATTTTACTTTTTTCCCCTCCTGGGAGCTAAAGATATTTTAGAGAAGAATTAACCTTTTGCTTCTCCAGTTGAACATTTGTAGCAATAAGTCA 5´-UTR hAAT + modSV40 + Kozak sequence 160 http://www.bloodjournal.org/content/early/2005/12/01/blood-2005-10-4035?sso-checked=true 1 GCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATCCGGACTCTAAGGTAAATATAAAATTTTTAAGTGTATAATGTGTTAAACTACTGATTCTAATTGTTTCTCTCTTTTAGATTCCAACCTTTGGAACTGAATTCTAGACCACC 5´-UTR hFIX and Kozak sequence 29 US20160375110 0 ACCACTTTCACAATCTGCTAGCAAAGGTT Chimeric intron 133 U47119.2 2 GTAAGTATCAAGGTTACAAGACAGGTTTAAGGAGACCAATAGAAACTGGGCTTGTCGAGACAGAGAAGACTCTTGCGTTTCTGATAGGCACCTATTGGTCTTACTGACATCCACTTTGCCTTTCTCTCCACAG Large fragment of the 5´-UTR of human alpha-1-antitrypsin (AAT) 341 9 TGGGCAGGAACTGGGCACTGTGCCCAGGGCATGCACTGCCTCCACGCAGCAACCCTCAGAGTCCTGAGCTGAACCAAGAAGGAGGAGGGGGTCGGGCCTCCGAGGAAGGCCTAGCCGCTGCTGCTGCCAGGAATTCCAGGTTGGAGGGGCGGCAACCTCCTGCCAGCCTT CAGGCCACTCTCCTGTGCCTGCCAGAAGAGACAGAGCTTGAGGAGAGCTTGAGGAGAGCAGGAAAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATCGACA 5pUTR 316 US9644216 6 TCTAGAGAAGCTTTATTGCGGTAGTTTATCACAGTTAAATTGCTAACGCAGTCAGTGCTTCTGACACAACAGTCTCGAACTTAAGCTGCAGTGACTCTCTTAAGGTAGCCTTGCAGAAGTTGGTCGTGAGGCACTGGGCAGGTAAGTATCAAGGTTAC AAGACAGGTTTAAGGAGACCAATAGAAACTGGGCTTGTCGAGACAGAGAAGACTCTTGCGTTTCTGATAGGCACCTATTGGTCTTACTGACATCCACTTTGCCTTTCTCCACAGGTGTCCACTCCCAGTTCAATTACAGCTCTTAAGGCCCTGCAG 5pUTR human ABCB4 cDNA (variant A, predominant isoform) 76 NM_000443 8 CAAAGTCCAGGCCCCTCTGCTGCAGCGCCCGCGTCCAGAGGCCCTGCCAGACACGCGCGAGGTTCGAGGCTGAG 5pUTR human ABCB11 cDNA 127 NM_003742 2 AGAATGATGAAAACCGAGGTTGGAAAAGGTTGTGAAACCTTTTTAACTCTCCACAGTGGAGTCCATTATTTCCTCTGGCTTCCTCAAATTCATATTCACAGGGTCGTTGGCTGTGGGTTGCAATTACC 5pUTR human glucose-6-phosphatase 80 NM_000151.3 0 ATAGCAGAGCAATCACCACCAAGCCTGGAATAACTGCAAGGGCTCTGCTGACATCTTCCTGAGGTGCCAAGGAAATGAGG 5pUTR MCK derived from rAAVirh74.MCK GALGT2. Contains 53 bp of endogenous mouse MCK exon 1 (untranslated), 16S/19S SV40 late splicing signals, 5pUTR derived from pCMVB plasmid. 208 https://patentimages.storage.googleapis.com/4f/8a/d6/b915c650f5eeb5/WO2017049031A1.pdf 8 GGGTCACCACCACCTCCACAGCACAGACAGACACTCAGGAGCCAGCCAGCCAGGTAAGTTTAGTCTTTTTGTCTTTTATTTCAGGTCCCGGATCCGGTGGTGGTGCAAATCAAAGAACTGCTCCTCAGTGGATGTTGCCTTTACTTCTAGGCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCTGCGGAATTGTACCCGCGGCCGCG CpG-free 5´-UTR synthetic intron (SI 126) 159 0 AAGCTTCTGCCTTCTCCCTCCTGTGAGTTTGGTAAGTCACTGACTGTCTATGCCTGGGAAAGGGTGGGCAGGAGATGGGGCAGTGCAGGAAAAGTGGCACTATGAACCCTGCAGCCCTAGACAATTGTACTAACCTTCTTCTCTTTCCTCTCCTGACAG 5'-UTR of the human cytochrome b-245 alpha chain (CYBA) gene 36 (NM_000101.4) 5 CGCGCTAGCAGTGTCCCAGCCGGGTTCGTGTCGCC 5'-UTR of the human 2,4-dienoyl-CoA reductase 1 gene (DECR1) 141 (NM_001330575.1) 14 ACGCCGCCTGGGTCCCAGTCCCCGTCCCATCCCCCGGCGGCCTAGGCAGCGTTTCCAGCCCCGAGAACTTTGTTCTTTTTGTCCCGCCCCCTGCGCCCAACCGCCTGCGCCGCCTTCCGGCCCGAGTTCTGGAGACTCAAC 5'-UTR of the human glial maturation factor gamma (GMFG) gene 110 (NM_001301008.1) 4 GTTGGATGAAACCTTCCTCCTACTGCACAGCCCGCCCCCCTACAGCCCCGGTCCCCACGCCTAGAAGACAGCGGAACTAAGAAAAGAAGAGGCCTGTGGACAGAACAATC 5´-UTR of human late endosomal/lysosomal adaptor, activator of MAPK and MTOR 2 (LAMTOR2) 164 (NM_001145264.1) 13 GGTGGGGCGGGGTTGAGTCGGAACCACAATAGCCAGGCGAAGAAACTACAACTCCCAGGGCGTCCCGGAGCAGGCCAACGGGACTACGGGAAGCAGCGGGCAGCGGCCCGCGGGAGGCACCTCGGAGATCTGGGTGCAAAAGCCCAGGGTTAGGAACCGTAGGC 5´-UTR of human myosin light chain 6B (MYL6B) 127 (NM_002475.4) 8 GGCCACCGGAATTAACCCTTCAGGGCTGGGGGCCGCGCTATGCCCCGCCCCCTCCCCAGCCCCAGACACGGACCCCGCAGGAGATGGGTGCCCCCATCCGCACACTGTCCTTTGGCCACCGGACATC Large fragment of the 5´-UTR of human alpha-1-antitrypsin (AAT) 341 9 TGGGCAGGAACTGGGCACTGTGCCCAGGGCATGCACTGCCTCCACGCAGCAACCCTCAGAGTCCTGAGCTGAACCAAGAAGGAGGAGGGGGTCGGGCCTCCGAGGAAGGCCTAGCCGCTGCTGCTGCCAGGAATTCCAGGTTGGAGGGGCGGCAACCTCCTGCCAGCCTTCAGGCCACTCTCCTGTGCCTGCCAGAAGACAGAGCTTGAGGAGAGCTTGAGGAGAGCAGGAAAGCCTCCCCCGTT GCCCCTCTGGATTCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATCGACA

[00211] 3'-UTR sequences: In some embodiments, the ccDNA vector comprises a 3'-UTR sequence that is located 5' relative to a 3'-ITR sequence. In some embodiments, the 3'-UTR is located 3' from a transgene, such as a sequence encoding an inflammasome antagonist (e.g., an inhibitor of one or more of the NLRP3 and/or AIM2 inflammasome pathway, or an inhibitor of caspase 1). Exemplary 3'-UTR sequences are listed in Table 2B.

Table 2B: Approximate 3´-UTR and intron sequences

Description Length Source CG Contents Subsequence WHP post-transcriptional response element 581 20 GAGCATCTTACCGCCATTTATTCCCATATTTGTTCTGTTTTTCTTGATTTGGGTATACATTTAAATGTTAATAAAACAAAATGGTGGGGCAATCATTTACATTTTTAGGGATATGTAATTACTAGTTCAGGTGTATTGCCACAAGACAAACATGTTAAGAAACTTTCCCGTTATTTACGCTCTGTTCCTGTTAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGATATTCTTAACTATGTTGCTC CTTTTACGCTGTGTGGATATGCTGCTTTATAGCCTCT GTATCTAGCTATTGCTTCCCGTACGGCTTTCGTTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTTAGAGGAGTTGTGGCCCGTTGTCCGTCAACGTGGCGTGGTGTGCTCTGTGTTTGCTGACGCAACCCCCACTGGCTGGGGCATTGCCACCACCTGTCAACTCCTTTCTGGGACTTTCGCTTTCCCCCTCCCGATCGCCACGGCAGAACTCATCGCCGCCTGCCTTGCCCG CTGCTGGACAGGGGCTAGGTTGCTGGGCACTGATAATTCCGTGGTGTTGTC Triplet repeat of the mir-142 binding site 77 1 TCCATAAAGTAGGAAACACTACACGATTCCATAAAGTAGGAAACACTACATCACTCCATAAAGTAGGAAACACTACA 3´-UTR of hFIX and poly(A) spacer derived from SPK9001 88 US2016/0375110 0 TGAAAGATGGATTTCCAAGGTTAATTCATTGGAATTGAAAATTAACAGAGATCTAGAGCTGAATTCCTGCAGCCAGGGGGATCAGCCT 3pUTR of human hemoglobin beta chain (HBB) 395 1 TAAAATACAGCATAGCAAAACTTTAACCTCCAAATCAAGCCTCTACTTGAATCCTTTTCTGAGGGATGAATAAGGCATAGGCATCAGGGGCTGTTGCCAATGTGCATTAGCTGTTTGCAGCCTCACCTTCTTTCATGGAGTTTAAGATATAGTGTATTTTCCCAAGGTTTGAACTAGCTCTTCATTTCTTTATGTTTTAAATGCACTGACCTCCCACATTCCCTTTTTAGTAAAATATTCAGAAATAATTT AAATACATCATTGCAATGAAAATAAATGTTTTTTATTAGGCAGAATCCAGATGCTCAAGGCCCTTCATAATATCCCCCAGTTTAGTAGTTGGACTTAGGGAACAAAGGAACCTTTAATAGAAATTGGACAGCAAGAAAGCGAGC S/MAR interferon-beta (scaffold/matrix-associated region) 800 0 AGTCAATATGTTCACCCCAAAAAAGCTGTTTGTTAACTTGCCAACCTCATTCTAAAATGTATATAGAAGCCCAAAAGACAATAACAAAAATATTCTTGTAGAACAAAATGGGAAAGAATGTTCCACTAAATATCAAGATTTAGAGCAAAGCATGAGATGTGTGGGGATAGACAGTGAGGCTGATAAAATAGAGTAGAGCTCAGAAACAGACCCATTGATATATGTAAGTGACCTATGAAAAAAATGGCATTTTA CAATGGGAAAATGATGGTCTTTTTCTTTTTAGAAAAACAGGGAAATATATTTATATGTAAAAAATAAAAGGGAACCCATATGTCATACCATACACACAAAAAAATTCCAGTGAATTATAAGTCTAAATGGAGAAGGCAAAACT TTAAATCTTTTAGAAAATAATATAGAAGCATGCCATCAAGACTTCAGTGTAGAGAAAAATTTCTTATGACTCAAAGTCCTAACCACAAAGAAAAGATTGTTAATTAGATTGCATGAATATTAAGACTTATTTTTAAAATTAAAAAACCATTAAGAAAAGTCAGGCCATAGAATGACAGAAAATATTTGCAACACCCCAGTAAAGAGAATTGTAATATGCAGATTATAAAAAGAAGTCTTACAAATCAGTAAAAATAAAACTAG ACAAAAATTTGAACAGATGAAAGAGAAACTCTAAATAATCATTACACATGAGAAACTCAATCTCAGAAATCAGAGAACTATCATTGCATATACACTAAATTAGAGAAATATTAAAAGGCTAAGTAACATCTGTGGC MAR beta-globulin (matrix associated region) 407 0 AATTATCTCTAAGGCATGTGAACTGGCTGTCTTGGTTTTCATCTGTACTTCATCTGCTACCTCTGTGACCTGAAACATATTTATAATTCCATTAAGCTGTGCATATGATAGATTTATCATATGTATTTTCCTTAAAGGATTTTTGTAAGAACTAATTGAATTGATACCTGTAAAGTCTTTATCACACTACCCAATAAATAATAAATCTCTTTGTTCAGCTCTCTGTTTCTATAAATATGTACCAGTT TTATTGTTTTTAGTGGTAGTGATTTTATTCTCTTTCTATATATATACACACACATGTGTGTGCATTCATAAATATATACAATTTTTATGAATAAAAAATTATTAGCAATCAATATTGAAAACCACTGATTTTTGTTTATGTGAGCAAACAGCAGATTAAAAG Human albumin 3'-UTR sequence 186 1 CATCACATTTAAAAGCATCTCAGCCTACCATGAGAATAAGAGAAAGAAAATGAAGATCAAAAGCTTATTCATCTGTTTTTCTTTTTCGTTGGTGTAAAGCCAACACCCTGTCTAAAAAACATAAATTTCTTTAATCATTTTGCCTCTTTTCTCTGTGCTTCAATTAATAAAAAAATGGAAAGAATCT CpG-minimized 3pUTR from HBB 395 0 TAAAATACAGCATAGCAAAACTTTAACCTCCAAATCAAGCCTCTACTTGAATCCTTTTCTGAGGGATGAATAAGGCATAGGCATCAGGGGCTGTTGCCAATGTGCATTAGCTGTTTGCAGCCTCACCTTCTTTCATGGAGTTTAAGATATAGTGTATTTTCCCAAGGTTTGAACTAGCTCTTCATTTCTTTATGTTTTAAATGCACTGACCTCCCACATTCCCTTTTTAGTAAAATATTCAGAAATAATTT AAATACATCATTGCAATGAAAATAAATGTTTTTTATTAGGCAGAATCCAGATGCTCAAGGCCCTTCATAATATCCCCCAGTTTAGTAGTTGGACTTAGGGAACAAAGGAACCTTTAATAGAAATTGGACAGCAAGAAAGCCAGC WHP post-transcriptional response element. 3'-cytosine is missing. 580 20 GAGCATCTTACCGCCATTTATTCCCATATTTGTTCTGTTTTTCTTGATTTGGGTATACATTTAAATGTTAATAAAACAAAATGGTGGGGCAATCATTTACATTTTTAGGGATATGTAATTACTAGTTCAGGTGTATTGCCACAAGACAAACATGTTAAGAAACTTTCCCGTTATTTACGCTCTGTTCCTGTTAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGATATTCTTAACTATGTTGCTC CTTTTACGCTGTGTGGATATGCTGCTTTATAGCCTCT GTATCTAGCTATTGCTTCCCGTACGGCTTTCGTTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTTAGAGGAGTTGTGGCCCGTTGTCCGTCAACGTGGCGTGGTGTGCTCTGTGTTTGCTGACGCAACCCCCACTGGCTGGGGCATTGCCACCACCTGTCAACTCCTTTCTGGGACTTTCGCTTTCCCCCTCCCGATCGCCACGGCAGAACTCATCGCCGCCTGCCTTGCCCG CTGCTGGACAGGGGCTAGGTTGCTGGGCACTGATAATTCCGTGGTGTTGT 3'-UTR of the human cytochrome b-245 alpha chain (CYBA) gene 64 (NM_000101.4) 5 CCTCGCCCCGGACCTGCCCTCCCGCCAGGTGCACCCACCTGCAATAAATGCAGCGAAGCCGGGA Shortened WPRE3 sequence with minimal gamma and alpha elements 247 WPRE3 see https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3975461/ 10 GATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAAATCCTGGTTAGTTCTTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGG 3pUTR of human hemoglobin beta chain (HBB) 144 1 AAATACATCATTGCAATGAAAATAAATGTTTTTTATTAGGCAGAATCCAGATGCTCAAGGCCCTTCATAATATCCCCCAGTTTAGTAGTTGGACTTAGGGAACAAAGGAACCTTTAATAGAAATTGGACAGCAAGAAAGCGAGC First 62 bp of 3pUTR WPRE element 62 1 GAGCATCTTACCGCCATTTATTCCCATATTTGTTCTGTTTTTCTTGATTTGGGTATACATTT

[00212] Polyadenylation sequences: A sequence encoding a polyadenylation sequence can be included in a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response), as described herein, to stabilize mRNA expressed from the ccDNA vector, as well as to facilitate nuclear export and translation. In one embodiment, the ccDNA vector does not include a polyadenylation sequence. In other embodiments, a ccDNA vector for expressing an inflammasome antagonist includes at least 1, at least 2, at least 3, at least 4, at least 5, at least 10, at least 15, at least 20, at least 25, at least 30, at least 40, at least 45, at least 50 or more adenine dinucleotides. In some embodiments, the polyadenylation sequence comprises about 43 nucleotides, about 40-50 nucleotides, about 40-55 nucleotides, about 45-50 nucleotides, about 35-50 nucleotides, or any range in between.

[00213] The expression cassettes may include any polyadenylation sequence known in the art, or a variant thereof. In some embodiments, the polyadenylation (polyA) sequence is selected from any of the sequences listed in Table 3. Other polyA sequences commonly known in the art may also be used, including, for example, but not limited to, the natural sequence isolated from bovine BGHpA (e.g., SEQ ID NO: 9) or SV40 polyA (SV40pA) (e.g., SEQ ID NO: 10), or a synthetic sequence. Some expression cassettes may also include the SV40 5' enhancer late polyA signal (USE) sequence. In some embodiments, the USE sequence may be used in combination with SV40pA or a heterologous polyA signal. The poly(A) sequences are located in the 3' direction relative to the transgene encoding the inflammasome antagonist.

[00214] Expression cassettes may also include a post-transcriptional element to enhance expression of the transgene. In some embodiments, a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) (e.g., SEQ ID NO: 72) is used to enhance expression of the transgene. Other post-transcriptional processing elements may be used, such as a post-transcriptional element from the herpes simplex virus or hepatitis B virus (HBV) thymidine kinase gene. Secretory sequences may be linked to the transgenes, such as the VH-02 and VK-A26 sequences, such as SEQ ID NO: 950 and SEQ ID NO: 951.

Table 3: Approximate poly(A) sequences

Description Length Source CG Contents Subsequence Terminator and polyadenylation sequence of bovine growth hormone. 225 3 TGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCGGTGGGCTCTATGGC Synthetic poly(A) derived from BMN270 49 https://www.ncbi.nlm.nih.gov/pubmed/29292164 0 AATAAAAGATCTTTATTTTCATTAGATCTGTGTGTTGGTTTTTTGTGTG Synthetic poly(A) derived from SPK8011 54 US2017/0216408 2 GCGGCCGCAATAAAAGATCAGAGCTCTAGAGATCTGTGTGTTGGTTTTTTGTGT Synthetic poly(A) and insulator sequence derived from Sangamo_CRMSBS2-Intron3 74 WO2017074526 2 GGATCCAATAAAATATCTTTATTTTCATTACATCTGTGTGTTGGTTTTTTGTGTGTTTTCCTGTAACGATCGGG SV40 late poly(A) and 3´-insulator sequences derived from Nathwani hFIX 143 http://www.bloodjournal.org/content/early/2005/12/01/blood-2005-10-4035?sso-checked=true 1 CTCGATGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTTAACAACAACAATTGCATTCATTTTATGTTTCAGGTTCAGGGGGAGGTGTGGGAGGTTTTTTAAACTAGT Poly(A) from bGH derived from SPK9001 228 US2016/0375110 0 CTACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCTTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCACATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCAGTGGGCTCTATGG CpG-free poly(A) SV40 222 0 CAGACATGATAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTTAACAACAACAATTGCATTCATTTTATGTTTCAGGTTCAGGGGGAGATGTGGGAGGTTTTTTAAAGCAAGTAAAACCTCTACAAATGTGGTA Late poly(A) SV40 226 0 CCAGACATGATAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTTAACAACAACAATTGCATTCATTTTATGTTTCAGGTTCAGGGGGAGGTGTGGGAGGTTTTTTAAAGCAAGTAAAACCTCTACAAATGTGGTATGG Poly(A) C60pAC30HSL containing the poly(A) A64 sequence and the stem-loop sequence of histone C30 129 0 GTTAACAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAATGCATCCCCCCCCCCCCCCCCCCCCCCCCCCCCAAAGGCTCTTTTCAGAGCCACCA Poly(A) used in J. Chou's glucose-6-phosphatase (G6Pase) constructs containing SV40 poly(A) 232 US9644216 4 GCGGCCGCGGGGATCCAGACATGATAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTTAACAACAACAATTGCATTCATTTTATGTTTCAGGTTCAGGGGGAGGTGTGGGAGGTTTTTTAGTCGACCATGCTGGGGAGAGATCT SV40 polyadenylation signal 135 0 GATCCAGACATGATAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTT Herpes virus thymidine kinase polyadenylation signal 49 4 CGGCAATAAAAAGACAGAATAAAACGCACGGGTGTTGGGTCGTTTGTTC SV40 late polyadenylation signal 226 0 CCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGG Sequence of the 3´-UTR and terminator/poly(A) of human albumin 416 2 CATCACATTTAAAAGCATCTCAGCCTACCATGAGAATAAGAGAAAGAAAATGAAGATCAAAAGCTTATTCATCTGTTTTTCTTTTTCGTTGGTGTAAAGCCAACACCCTGTCTAAAAAACATAAATTTCTTTAATCATTTTGCCTCTTTTCTCTGTGCTTCAATTAATAAAAAAATGGAAAGAATCTAATAGAGTGGTACAGCACTGTT ATTTTTCAAAGATGTGTTGCTATCCTGAAAATTCTGTAGGTTCTGTGGAAGTTCCAGTGTTCTCTCTTATTCCACTTCGGTAGAGGATTTCTAGTTTCTTGTGGGCTAATTAAATAAATCATTAATACTCTTCTAAGTTATGGATTATAAACATTCAAAATAATATTTTGACATTATGATAATTCTGAATAAAAGAACAAAAACCATG Sequence of the 3´-UTR and terminator/poly(A) of human albumin 415 2 ATCACATTTAAAAGCATCTCAGCCTACCATGAGAATAAGAGAAAGAAAATGAAGATCAAAAGCTTATTCATCTGTTTTTCTTTTTCGTTGGTGTAAAGCCAACACCCTGTCTAAAAAACATAAATTTCTTTAATCATTTTGCCTCTTTTCTCTGTGCTTCAATTAATAAAAAAATGGAAAGAATCTAATAGAGTGGTACAGCACTGTT ATTTTTCAAAGATGTGTTGCTATCCTGAAAATTCTGTAGGTTCTGTGGAAGTTCCAGTGTTCTCTCTTATTCCACTTCGGTAGAGGATTTCTAGTTTCTTGTGGGCTAATTAAATAAATCATTAATACTCTTCTAAGTTATGGATTATAAACATTCAAAATAATATTTTGACATTATGATAATTCTGAATAAAAGAACAAAAACCATG CpG-free short poly(A) SV40 122 0 TAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTT CpG-free short poly(A) SV40 133 0 TGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTTAACAACAACAATTGCATTCATTTTATGTTTCAGGTTCAGGGGGAGGTGTGGGAGGTTTTTTAAA

[00215] According to one embodiment, the vector polynucleotide (ccDNA vector) comprises a pair of two different ITRs selected from the group consisting of: SEQ ID NO: 1 and SEQ ID NO: 52; and SEQ ID NO: 2 and SEQ ID NO: 51. According to one embodiment of each of these aspects, the vector polynucleotide or non-viral capsidless DNA vectors with covalently closed ends comprise a pair of ITRs selected from the group consisting of: SEQ ID NO: 101 and SEQ ID NO: 102; SEQ ID NO: 103 and SEQ ID NO: 104, SEQ ID NO: 105 and SEQ ID NO: 106; SEQ ID NO: 107 and SEQ ID NO: 108; SEQ ID NO: 109 and SEQ ID NO: 110; SEQ ID NO: 111 and SEQ ID NO: 112; SEQ ID NO:113 and SEQ ID NO:114; and SEQ ID NO:115 and SEQ ID NO:116. In some embodiments, the ccDNA vectors do not have an ITR that comprises any sequence selected from SEQ ID NO:500-529.

[00216] The timing of collection and accumulation of the DNA vectors described herein from cells can be selected and optimized to achieve high yields of ccDNA vectors. For example, the timing of collection can be selected based on cell viability, cell morphology, cell expansion, etc. In one embodiment, the cells are cultured under sufficient conditions and harvested a sufficient time after baculovirus infection to produce DNA vectors (e.g., TTX vectors), but before the majority of the cells begin to die due to viral toxicity. The DNA vectors can be isolated using plasmid purification kits, such as endotoxin-free plasmid isolation kits from Qiagen. Other methods developed for isolating plasmids can also be adapted for DNA vectors. Generally, any method for purifying nucleic acids can be used.

[00217] DNA vectors can be purified using any DNA purification methods known to those skilled in the art. In one embodiment, ccDNA vectors are purified as DNA molecules. In another embodiment, ccDNA vectors are purified as exosomes or microparticles.

[00218] In one embodiment, a capsidless non-viral DNA vector comprises or is derived from a plasmid comprising a polynucleotide template comprising, in the specified order: the first inverted terminal repeat (ITR) of an adeno-associated virus (AAV), a nucleotide sequence of interest (e.g., an exogenous DNA expression cassette), and a modified AAV ITR, wherein said template nucleic acid molecule lacks the ability to encode an AAV capsid protein. In a further embodiment, the template nucleic acid of the present invention lacks sequences encoding a viral capsid protein (i.e., lacks AAV capsid genes as well as capsid genes of other viruses). Furthermore, in a particular embodiment, the template nucleic acid molecule also lacks sequences encoding an AAV Rep protein. Accordingly, in a preferred embodiment, the nucleic acid molecule of the present invention lacks both the functional AAV cap and AAV rep genes.

[00219] In one embodiment, a ccDNA vector may include an ITR structure that is mutated relative to the wild-type AAB2 ITR disclosed herein, but still retains a functional portion of the RBE, trs, and RBE'. In some embodiments, ccDNA vectors do not have an ITR that comprises any sequence selected from SEQ ID NO: 500-529.

[00220] In some embodiments, a transgene encoding an inflammasome antagonist (e.g., any one or more of an NLRP3 inflammasome pathway inhibitor or an AIM2 inflammasome pathway inhibitor or a caspase 1 inhibitor or any combination thereof) may also encode a secretory sequence such that the inflammasome antagonist is targeted to the Golgi apparatus and the endoplasmic reticulum (ER), where the inflammasome antagonist (e.g., any one or more of an NLRP3 inflammasome pathway inhibitor or an AIM2 inflammasome pathway inhibitor or a caspase 1 inhibitor or any combination thereof) will be folded into the correct conformation by chaperone molecules upon its passage through the ER and release from the cell. Exemplary secretory sequences include, but are not limited to, VH-02 (SEQ ID NO: 950) and VK-A26 (SEQ ID NO: 951) and the Igκ signal sequence, as well as the Gluc secretory signal that ensures the secretion of the tagged protein from the cytosol, the TMD-ST secretory sequence that directs the tagged protein to the Golgi apparatus.

[00221] Nuclear Localization Sequences: In some embodiments, a ccDNA vector for expressing, e.g., an inhibitor of an immune response (e.g., an innate immune response) comprises one or more nuclear localization sequences (NLS), e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more NLSs. In some embodiments, the one or more NLSs are located at or near the amino terminus, at or near the carboxy terminus, or a combination thereof (e.g., one or more NLSs at the amino terminus and/or one or more NLSs at the carboxy terminus). If more than one NLS is present, each NLS can be selected independently of the others such that one NLS is present in more than one copy and/or in combination with one or more other NLSs present in one or more copies. Non-limiting examples of NLSs are shown in Table 4.

Table 4: Nuclear localization signals

SOURCE SUBSEQUENCE SEQ ID NO. Large T-antigen of SV40 virus PKKKRKV (coded CCCAAGAAGAAGAGGAAGGTG) 315 Nucleoplasmin KRPAATKKAGQAKKKK 316 C-myc PAAKRVKLD 317 RQRRNELKRSP 318 hRNPA1 M9 NQSSNFGPMKGGNFGGRSSGPYGGGGQYFAKPRNQGGY 319 IBB domain importin-alpha RMRIZFKNKGKDTAELRRRRVEVSVELRKAKKDEQILKRRNV 320 Myoma T protein VSRKRPRP 321 PPKKARED 322 p53 human PQPKKKPL 323 c-abl IV mouse SALIKKKKKMAP 324 NS1 influenza virus DRLRR 325 PKQKKRK 326 Hepatitis delta virus antigen RKLKKKIKKL 327 Mouse Mx1 protein REKKKFLKRR 328 Human poly(ADP-ribose) polymerase KRKGDEVDGVDEVAKKKSKK 329 Glucocorticoid steroid hormone receptor (human) RKCLQAGMNLEARKTKK 330

[00222] Regulatory switches: A molecular regulatory switch is a switch that generates a measurable change in state in response to a signal. Regulatory switches can also be used to fine-tune the expression of an immune response inhibitor (e.g., an innate immune response) described herein such that the immune response inhibitor is expressed in a desired manner, including, but not limited to, expressing the immune response inhibitor at a target expression level or in a target amount, or, alternatively, in the presence or absence of a particular signal, including a cellular signaling event. For example, as described herein, expression of an immune response inhibitor from a ccDNA vector can be turned on or off upon the occurrence of a particular condition. In some embodiments, the switch is an ON/OFF switch that is designed to start or stop (i.e., terminate) the expression of an inhibitor of an immune response (e.g., an innate immune response) in the ccDNA vector in a controlled and regulated manner. In some embodiments, the switch may include a kill switch that may instruct a cell containing the ccDNA vector to initiate programmed cell death upon activation of the switch. Exemplary regulatory switches contemplated for use in a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response) may be used to regulate transgene expression and are described in more detail in International Application No. PCT/US18/49996, which is incorporated herein by reference in its entirety.

(i) Binary Regulatory Switches

[00223] In some embodiments, a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response) comprises a regulatory switch that can serve to controllably modulate the expression of an inflammasome antagonist. For example, an expression cassette located between the ITRs of the ccDNA vector can further comprise a regulatory region, such as a promoter, cis-element, repressor, enhancer, etc., that is operably linked to a nucleic acid sequence encoding an inhibitor of an immune response (e.g., an innate immune response), wherein the regulatory region is regulated by one or more cofactors or exogenous agents. As a non-limiting example, regulatory regions can be modulated by small molecule switches or inducible or repressible promoters. Non-limiting examples of inducible promoters are hormone-inducible or metal-inducible promoters. Other exemplary inducible promoter/enhancer elements include, but are not limited to, the RU486 inducible promoter, the ecdysone inducible promoter, the rapamycin inducible promoter, and the metallothionein promoter.

(ii) Small molecule regulatory switches

[00224] Various small molecule regulatory switches known in the art are known in the art and can be combined with an immune response (e.g., innate immune response) inhibitor disclosed herein to produce a regulatory switch-controlled ccDNA vector. In some embodiments, the regulatory switch can be selected from any one or a combination of the following: an orthogonal ligand/nuclear receptor pair, such as a retinoid receptor/LG335 variant and GRQCIMFI, together with an artificial promoter controlling expression of an operably linked transgene, as disclosed in Taylor et al. BMC Biotechnology 10 (2010): 15; engineered steroid receptors, such as a modified progesterone receptor with a truncated C-terminus that cannot bind progesterone but binds RU486 (mifepristone) (U.S. Patent No. 5,364,791); the Drosophila ecdysone receptor and its ecdysteroid ligands (Saez, et al., PNAS, 97(26)(2000), 14512-14517); or the switch controlled by the antibiotic trimethoprim (TMP), as disclosed in Sando R 3 ^rd ; Nat Methods. 2013, 10(11):1085-8. In some embodiments, the regulatory switch for controlling the transgene or expressed by the ccDNA vector is a prodrug activating switch, such as that disclosed in U.S. Patents 8,771,679 and 6,339,070.

(iii) Regulatory switches "with access code"

[00225] In some embodiments, the regulatory switch may be an "access code switch" or an "access code circuit." Access code switches allow for fine-tuning of the control of expression of a transgene from a ccDNA vector when specific conditions occur, i.e., a combination of conditions must be present for expression and/or repression of the transgene to occur. For example, at least conditions A and B must be present for expression of the transgene to occur. The regulatory switch may be any number of conditions, such as at least 2, or at least 3, or at least 4, or at least 5, or at least 6, or at least 7 or more conditions, that must be present for expression of the transgene to occur. In some embodiments, at least 2 conditions must be present (e.g., conditions A, B), and in some embodiments, at least 3 conditions must be present (e.g., A, B, and C, or A, B, and D). By way of example only, condition A, B, and C are required for gene expression from a ccDNA containing a regulatory switch with the access code "ABC". Conditions A, B, and C may be as follows; condition A is the presence of a condition or disease, condition B is a hormonal response, and condition C is a response to transgene expression. For example, if the transgene edits a defective EPO gene, then condition A is the presence of chronic kidney disease (CKD), condition B is met when hypoxic conditions are present in the subject's kidneys, condition C is the defective recruitment of erythropoietin-producing cells (EPCs) to the kidneys; or alternatively, defective activation of HIF-2. When oxygen levels rise or the target EPO level is reached, the transgene is turned off again until conditions 3 occur that turn it on again.

[00226] In some embodiments, a regulatory switch with an access code or "access code circuit" provided for use in a ccDNA vector comprises hybrid transcription factors (TFs) to expand the range and complexity of environmental signals used to determine biocontainment conditions. Unlike a deadman switch, which triggers cell death in the presence of a predetermined condition, an "access code circuit" allows for cell survival or transgene expression in the presence of a specific "access code" and can be easily reprogrammed to allow transgene expression and/or cell survival only in the presence of a predetermined environmental condition or access code.

[00227] Any and all combinations of the regulatory switches disclosed herein, such as small molecule switches, nucleic acid-based switches, small molecule-nucleic acid-based hybrid switches, transgene post-transcriptional regulatory switches, post-translational regulation, radiation-controlled switches, hypoxia-mediated switches, and other regulatory switches known to those skilled in the art disclosed herein, can be used in the access code regulatory switch as disclosed herein. Regulatory switches contemplated for use are also discussed in the review article by Kis et al., J R Soc Interface. 12: 20141000 (2015) and are summarized in Table 1 of the Kis article. According to some embodiments, the control switch for use in the access code system may be selected from any of the switches or combinations thereof disclosed in Table 11 of international patent application PCT/US18/49996, which is incorporated herein by reference in its entirety.

(iv) Nucleic acid-based regulatory switches for control of transgene expression

[00228] In some embodiments, a regulatory switch for controlling the expression of an inhibitor of an immune response (e.g., an innate immune response) by a ccDNA is based on a nucleic acid-based control mechanism. Examples of nucleic acid-based control mechanisms are known in the art and are contemplated for use. For example, such mechanisms include riboswitches such as those disclosed, for example, in US 2009/0305253, US2008/0269258, US2017/0204477, WO2018026762A1, US Patent 9,222,093, and European Patent Application EP288071, and are also disclosed in the review article by Villa JK et al. Microbiol Spectr. 2018 May;6(3). Also included are metabolite-sensitive transcriptional biosensors, such as those disclosed in WO2018/075486 and WO2017/147585. Other mechanisms known in the art that are contemplated for use include transgene silencing using a siRNA or RNAi molecule (e.g., miR, shRNA). For example, a ccDNA vector may comprise a regulatory switch encoding an RNAi molecule that is complementary to a portion of a transgene expressed by the ccDNA vector. When such RNAi is expressed, even if the ccDNA vector expresses a transgene (e.g., an inflammasome antagonist (e.g., an inhibitor of one or more of the NLRP3 and/or AIM2 inflammasome pathway, or a caspase 1 inhibitor), it will be suppressed by the complementary RNAi molecule, and if RNAi is not expressed when the transgene (e.g., an inflammasome antagonist) is expressed by the ccDNA vector, the transgene will not be suppressed by RNAi.

[00229] In some embodiments, the regulatory switch is a tissue-specific self-inactivating regulatory switch, such as disclosed in US2002/0022018, whereby the regulatory switch intentionally turns off a transgene (e.g., an inflammasome antagonist) at a site where expression of the transgene might otherwise be detrimental. In some embodiments, the regulatory switch is a recombinase-reversible gene expression system, such as disclosed in US2014/0127162 and US Patent 8,324,436.

(v) Post-transcriptional and post-translational regulatory switches.

[00230] In some embodiments, the regulatory switch for controlling expression of an inhibitor of an immune response (e.g., an innate immune response) by a ccDNA vector is a post-transcriptional modification system. For example, such a regulatory switch can be a tetracycline- or theophylline-sensitive riboswitch-aptazyme disclosed in US2018/0119156, GB201107768, WO2001/064956A3, EP 2707487, and Beilstein et al., ACS Synth. Biol., 2015, 4 (5), pp. 526-534; Zhong et al., Elife. 2016 Nov 2; 5. pii: e18858. In some embodiments, it is envisaged that one of ordinary skill in the art may encode both a transgene and an inhibitory siRNA that contains a ligand-responsive aptamer (negative switch), resulting in a ligand-responsive positive switch.

(vi) Other example regulatory switches

[00231] Any known regulatory switch can be used in a ccDNA vector to control expression of an inhibitor of an immune response (e.g., an innate immune response) by the ccDNA vector, including switches triggered by environmental changes. Additional examples include, but are not limited to; the BOC method according to Suzuki et al., Scientific Reports 8; 10051 (2018); an extension of the genetic code and a non-physiological amino acid; radiation-controlled or ultrasound-controlled positive/negative (on/off) switches (see, e.g., Scott S. et al., Gene Ther. 2000 Jul; 7 (13): 1121-5; U.S. Patents 5,612,318; 5,571,797; 5,770,581; 5,817,636; WO1999/025385A1. In some embodiments, the regulatory switch is controlled by an implantable system, e.g., as disclosed in U.S. Patent 7,840,263; US2007/0190028A1, in which gene expression is controlled by one or more forms of energy, including electromagnetic energy, that activates promoters operably linked to a transgene in a ccDNA vector.

[00232] In some embodiments, a regulatory switch contemplated for use in a ccDNA vector is a hypoxia-mediated or stress-activated switch, such as those disclosed in WO1999060142A2, US Patents 5,834,306; 6,218,179; 6,709,858; US2015/0322410; Greco et al., (2004) Targeted Cancer Therapies 9, S368, as well as FROG, TOAD, and NRSE elements and conditionally inducible suppressor elements, including hypoxia response elements (HREs), inflammatory response elements (IREs), and shear stress-activated elements (SSAEs), such as those disclosed in U.S. Patent 9,394,526. Such an embodiment can be used to "turn on" transgene expression from a ccDNA vector following ischemia or in ischemic tissues and/or tumors.

(vii). "Emergency switches"

[00233] Other embodiments described herein relate to a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response) described herein comprising a kill switch. The "kill switch" disclosed herein allows for the cell containing the ccDNA vector to be destroyed or subjected to programmed cell death as a method for permanently removing the administered ccDNA vector from the subject's system. One of skill in the art will appreciate that the use of "kill switches" in ccDNA vectors for expressing an inhibitor of an immune response (e.g., an innate immune response) will typically involve targeting the ccDNA vector to a limited number of cells whose loss is acceptable to the subject or to a cell type whose apoptosis is desirable (e.g., cancer cells). In all aspects, the "kill switch" disclosed herein is designed to ensure rapid and reliable killing of a cell containing the ccDNA vector in the absence of an input survival signal or other predetermined condition. In other words, the "kill switch" encoded by the ccDNA inflammasome antagonist expression vector described herein can limit the survival of a cell containing the ccDNA vector to an environment defined by specific input signals. Such "kill switches" serve to provide a biological containment function should it be necessary to remove the ccDNA inflammasome antagonist expression vector from a subject or to ensure that the encoded inflammasome antagonist is not expressed from it.

[00234] Other "kill switches" known to one of ordinary skill in the art are contemplated for use in the ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response) disclosed herein, such as those disclosed in US2010/0175141; US2013/0009799; US2011/0172826; US2013/0109568, as well as the "kill switches" disclosed in Jusiak et al., Reviews in Cell Biology and Molecular Medicine; 2014; 1-56; Kobayashi et al., PNAS, 2004; 101; 8419-9; Marchisio et al., Int. Journal of Biochem and Cell Biol., 2011; 43; 310-319; and Reinshagen et al., Science Translational Medicine, 2018, 11.

[00235] Accordingly, in some embodiments, a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response) may comprise a "kill switch" nucleic acid construct that comprises a nucleic acid encoding an effector toxin or a reporter protein, wherein expression of the effector toxin (e.g., a death protein) or reporter protein is controlled by a predetermined condition. For example, the predetermined condition may be the presence of an environmental agent, such as, for example, an exogenous agent, without which the cell will by default express the effector toxin (e.g., a death protein) and will be destroyed. In alternative embodiments, the predetermined condition is the presence of two or more environmental agents, such that the cell will only survive when supplied with two or more of the required exogenous agents, and without any of them, the cell containing the ccDNA vector is destroyed.

[00236] In some embodiments, a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response) is modified to include a "kill switch" for the destruction of cells containing said ccDNA vector to effectively stop the expression of a transgene expressed by said ccDNA vector (e.g., expression of an inflammasome antagonist) in vivo. In particular, the ccDNA vector is further genetically modified to express a switch protein that does not function in mammalian cells under normal physiological conditions. Cells expressing the switch protein will only be destroyed upon administration of a drug or exposure to an environmental condition that specifically targets said switch protein, which will stop expression of the therapeutic protein or peptide. For example, it has been reported that cells expressing HSV thymidine kinase can be killed by administration of drugs such as ganciclovir and cytosine deaminase. See, e.g., Dey and Evans, “Suicide Gene Therapy by Herpes Simplex Virus-1 Thymidine Kinase (HSV-TK),” in Targets in Gene Therapy, You (ed.) (2011); and Beltinger et al., Proc. Natl. Acad. Sci. USA 96(15):8699–8704 (1999). In some embodiments, the ccDNA vector may contain an siRNA-based “kill switch” called DISE (Death Induced by Survival gene Elimination) (Murmann et al., Oncotarget. 2017; 8:84643–84658. Induction of DISE in ovarian cancer cells in vivo).

[00237] According to another embodiment, the inhibitor of an immune response (e.g., an innate immune response) expressed from ccDNA vectors further comprises an additional functionality, such as fluorescence, enzymatic activity, a secretion signal, or an immune cell activator.

[00238] In some embodiments, a ccDNA encoding an inhibitor of an immune response (e.g., an innate immune response) may further comprise a linker domain, for example. As used herein, the term "linker domain" refers to an oligo- or polypeptide region of about 2 to 100 amino acids in length that links together any of the domains/regions of an inflammasome antagonist described herein. In some embodiments, linkers may include or be composed of flexible residues, such as glycine and serine, such that adjacent protein domains may move freely relative to one another. Longer linkers may be used when it is desirable to ensure that two adjacent domains do not sterically interfere with one another. Linkers may be cleavable or non-cleavable. Examples of cleavable linkers include 2A linkers (e.g., T2A), 2A-like linkers, or functional equivalents thereof, and combinations thereof. The linker may be a T2A linker region derived from Thosea asigna virus.

IV. Method for obtaining ccDNA vector

A. Obtaining in the general case

[00239] Certain methods for producing a ccDNA vector for expressing, for example, an inhibitor of an immune response (e.g., an innate immune response) comprising a pair of asymmetric ITRs or a pair of symmetric ITRs as defined herein are described in Section IV of international application PCT/US18/49996, filed September 7, 2018, which is incorporated herein by reference in its entirety. In some embodiments, a ccDNA vector for expressing an inflammasome antagonist disclosed herein can be produced using insect cells as described herein. In alternative embodiments, the ccDNA vector for expressing an inflammasome antagonist disclosed herein can be produced synthetically and, in some embodiments, cell-free, as disclosed in International Application PCT/US19/14122, filed January 18, 2019, which is incorporated herein by reference in its entirety.

[00240] As described herein, in one embodiment, a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response), for example, can be produced, for example, by a method comprising the steps of: a) incubating a population of host cells (e.g., insect cells) carrying a polynucleotide template of an expression construct (e.g., a ccDNA plasmid, ccDNA bacmid, and/or ccDNA baculovirus) lacking viral capsid encoding sequences in the presence of a Rep protein under effective conditions and for a period of time sufficient to induce production of said ccDNA vector in the host cells, and wherein said host cells lack viral capsid encoding sequences; and b) collecting and isolating the ccDNA vector from the host cells. The presence of the Rep protein induces replication of the vector polynucleotide with the modified ITR to produce the ccDNA vector in the host cell. However, viral particles (e.g., AAV virions) are not expressed. Thus, there is no size limitation such as that naturally present in AAV vectors or other viral vectors.

[00241] The presence of a ccDNA vector isolated from host cells can be confirmed by digesting DNA isolated from the host cell with a restriction enzyme that has a single recognition site on the ccDNA vector and analyzing the digested DNA material on a non-denaturing gel to confirm the presence of characteristic bands of linear and continuous DNA versus linear and discontinuous DNA.

[00242] According to another aspect of the present invention, there is provided the use of host cell lines having a stably integrated DNA expression vector polynucleotide template (cDNA template) into their own genome in the production of a non-viral DNA vector, for example as described in Lee, L. et al. (2013) Plos One 8(8): e69879. Preferably, Rep is added to the host cells at an MOI of about 3. When the host cell line is a mammalian cell line, for example HEK293 cells, the cell lines can have a stably integrated vector polynucleotide template, and a second vector, such as a herpes virus, can be used to introduce the Rep protein into the cells, allowing excision and amplification of the ccDNA in the presence of Rep and a helper virus.

[00243] In one embodiment, the host cells used to produce ccDNA vectors for expressing an inhibitor of an immune response (e.g., an innate immune response), such as described herein, are insect cells, and a baculovirus is used to deliver both the polynucleotide that encodes the Rep protein and the polynucleotide template of the non-viral DNA expression construct of the ccDNA vector, such as described in FIGS. 4A-4D and in Example 1. In some embodiments, the host cell is modified to express the Rep protein.

[00244] The ccDNA vector is then harvested and isolated from the host cells. The timing of harvesting and accumulation of the ccDNA vectors described herein from the cells can be selected and optimized to achieve high yields of ccDNA vectors. For example, the timing of harvesting can be selected based on cell viability, cell morphology, cell expansion, etc. In one embodiment, the cells are cultured under sufficient conditions and harvested a sufficient time after baculovirus infection to obtain ccDNA vectors, but before the majority of the cells begin to die due to baculovirus toxicity. The DNA vectors can be isolated using plasmid purification kits, such as the Endo-Free Plasmid Kits from Qiagen. Other methods developed for isolating plasmids can also be adapted for DNA vectors. Generally, any method for purifying nucleic acids can be used.

[00245] DNA vectors can be purified by any DNA purification methods known to those skilled in the art. In one embodiment, ccDNA vectors are purified as DNA molecules. In another embodiment, ccDNA vectors are purified as exosomes or microparticles.

The presence of a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response) can be confirmed by digesting vector DNA isolated from cells with a restriction enzyme having a single recognition site on the vector DNA and analyzing both the digested and undigested DNA material using gel electrophoresis to confirm the presence of characteristic bands of linear and continuous DNA versus linear and discontinuous DNA. FIG. 4C and FIG. 4D illustrate one embodiment for identifying the presence of closed-ended ccDNA vectors produced by the methods described herein.

B. cDNA plasmid

[00246] A ccDNA plasmid is a plasmid used to subsequently produce a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response), for example. In some embodiments, a ccDNA plasmid can be constructed using known techniques to provide at least the following elements as operably linked components in the direction of transcription: (1) a modified 5'-ITR sequence; (2) an expression cassette comprising a cis-regulatory element, such as a promoter, an inducible promoter, a regulatory switch, enhancers, and the like; and (3) a modified 3'-ITR sequence, wherein the 3'-ITR sequence is symmetrical with respect to the 5'-ITR sequence. In some embodiments, the expression cassette flanked by the ITR comprises a cloning site for the introduction of an exogenous sequence. The expression cassette replaces the rep and cap coding regions of the AAV genomes.

[00247] In one aspect, a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response) is, for example, derived from a plasmid, referred to herein as a "ccDNA plasmid," encoding, in this order: the first inverted terminal repeat (ITR) of an adeno-associated virus (AAV), an expression cassette containing a transgene, and a mutated or modified ITR of AAV, wherein said ccDNA plasmid lacks sequences encoding an AAV capsid protein. According to alternative embodiments, the ccDNA plasmid encodes, in the specified order: a first (or 5') modified or mutated AAV ITR, an expression cassette containing a transgene, and a second (or 3') modified AAV ITR, wherein said ccDNA plasmid lacks sequences encoding an AAV capsid protein, and wherein said 5' and 3' ITRs are symmetrical relative to each other. According to alternative embodiments, the ccDNA plasmid encodes, in the specified order: a first (or 5') modified or mutated AAV ITR, an expression cassette containing a transgene, and a second (or 3') mutated or modified AAV ITR, wherein said ccDNA plasmid lacks sequences encoding an AAV capsid protein, and wherein said modified 5' and 3' ITRs have the same modifications (i.e., they are reverse complementary or symmetrical relative to each other).

[00248] According to a further embodiment, the ccDNA plasmid system lacks sequences encoding a viral capsid protein (i.e., lacks AAV capsid genes, as well as capsid genes from other viruses). Furthermore, according to a particular embodiment, the ccDNA plasmid also lacks sequences encoding an AAV Rep protein. Accordingly, in a preferred embodiment, the ccDNA plasmid lacks functional AAV cap and AAV rep genes, GG-3′ for AAV2, and a variable palindromic hairpin sequence.

[00249] The ccDNA plasmid of the present invention can be produced using natural nucleotide sequences from the genomes of any of the AAV serotypes, which are well known in the art. In one embodiment, the ccDNA plasmid backbone is derived from the genome of AAB1, AAB2, AAB3, AAB4, AAB5, AAB5, AAB7, AAB8, AAB9, AAB10, AAB11, AAB12, AABrh8, AABrh10, AAB-DJ, and AAB-DJ8. For example, NCBI: NC002077; NC001401; NC001729; NC001829; NC006152; NC006260; NC006261; from the Kotin and Smith Index of Viruses ("The Springer Index of Viruses"), accessible at a link maintained by Springer (web address: oesys.springer.de/viruses/database/mkchapter.asp?virID=42.04.) (note: references to a URL or database refer to the contents of the linked URL or database as of the actual filing date of this application). In a particular embodiment, the ccDNA plasmid backbone is derived from the AAV2 genome. In another particular embodiment, the ccDNA plasmid backbone is a synthetic backbone constructed by genetic engineering to include at its 5' and 3' ends ITRs derived from one of said AAV genomes.

[00250] The ccDNA plasmid may optionally include a selectable or selective marker for use in generating a cell line producing the ccDNA vector. In one embodiment, the selectable marker may be inserted after (i.e., in the 3' direction) the 3'-ITR sequence. In another embodiment, the selectable marker may be inserted before (i.e., in the 5' direction) the 5'-ITR sequence. Suitable selectable markers include, for example, markers that confer drug resistance. Selectable markers may be, for example, blasticidin S, kanamycin, geneticin, and the like resistance genes. In a preferred embodiment, the selectable marker for selection for drug sensitivity is a blasticidin S resistance gene.

[00251] An exemplary ccDNA vector (e.g., pAAV0) for expressing an inflammasome antagonist (e.g., an inhibitor of one or more of the NLRP3 and/or AIM2 inflammasome pathway, or an inhibitor of caspase 1) is derived from a pAAV plasmid. A method for producing a rAAV vector can comprise: (a) delivering a pAAV plasmid as described above to a host cell, wherein both the host cell and the plasmid lack genes encoding a capsid protein, (b) culturing the host cell under conditions that allow production of the ccDNA genome; and (c) harvesting the cells and isolating the AAV genome produced by the cells.

C. Approximate method for obtaining ccDNA vectors from ccDNA plasmids

[00252] The present document also provides methods for producing capsid-less ccDNA vectors for expressing an inhibitor of an immune response (e.g., an innate immune response), in particular a method with a sufficiently high yield to provide sufficient vector for in vivo experiments.

[00253] In some embodiments, a method for producing a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response), for example, comprises the following steps: (1) introducing a nucleic acid construct comprising an expression cassette and two symmetric ITR sequences into a host cell (e.g., Sf9 cells), (2) optionally creating a clonal cell line, such as using a selectable marker present on a plasmid, (3) introducing a Rep-encoding gene (either by transfection or infection with a baculovirus carrying said gene) into said insect cell; and (4) harvesting the cells and purifying the ccDNA vector. The nucleic acid construct comprising the expression cassette and two ITR sequences described above for producing the ccDNA vector may be in the form of a ccDNA plasmid or a bacmid, or a baculovirus produced using the ccDNA plasmid as described below. The nucleic acid construct may be introduced into the host cell by transfection, viral transduction, stable integration, or other methods known in the art.

D. Cell lines:

[00254] Host cell lines used in producing a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response) can, for example, include insect cell lines derived from Spodoptera frugiperda, such as Sf9, Sf21, or Trichoplusia ni cells, or cell lines from other invertebrates, vertebrates, or other eukaryotes, including mammalian cells. Other cell lines known to those of ordinary skill in the art can also be used, such as HEK293, Huh-7, HeLa, HepG2, HeplA, 911, CHO, COS, MeWo, NIH3T3, A549, HT1 180, monocytes, and mature and immature dendritic cells. Host cell lines can be transfected to stably express the ccDNA plasmid to produce a high-yield ccDNA vector.

[00255] The ccDNA plasmids can be introduced into Sf9 cells by transient transfection using reagents (e.g., liposomal, calcium phosphate) or physical means (e.g., electroporation) known in the art. Alternatively, stable Sf9 cell lines can be generated with the ccDNA plasmid stably integrated into their genomes. Such stable cell lines can be generated by incorporating a selectable marker into the ccDNA plasmid as described above. If the ccDNA plasmid used to transfect the cell line includes a selectable marker, such as for an antibiotic, cells that have been transfected with the ccDNA plasmid and have integrated the ccDNA plasmid DNA into their genome can be selected by adding the antibiotic to the cell culture media. Resistant cell clones can then be isolated using single cell dilution or colony transfer techniques and expanded.

E. Isolation and purification of ccDNA vectors:

[00256] Examples of a method for producing and isolating ccDNA vectors are described in FIGS. 1-7 and in specific examples below. ccDNA vectors for expressing an inhibitor of an immune response (e.g., an innate immune response), such as those disclosed herein, can be produced from producer cells expressing the AAV Rep protein(s) further transformed with a ccDNA plasmid, a ccDNA bacmid, or a ccDNA baculovirus. Plasmids that can be used to produce ccDNA vectors include plasmids that encode an inflammasome antagonist or plasmids that encode one or more REP proteins.

[00257] According to one aspect, the polynucleotide encodes a Rep AAB protein (Rep 78 or 68) delivered to the producer cell in a plasmid (Rep plasmid), bacmid (Rep bacmid), or baculovirus (Rep baculovirus). The Rep plasmid, Rep bacmid, and Rep baculovirus can be produced using the methods described above.

[00258] Described herein are methods for producing a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response), for example. Expression constructs used to produce a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response) described herein can be a plasmid (e.g., ccDNA plasmids), a bacmid (e.g., a ccDNA bacmid), and/or a baculovirus (e.g., a ccDNA baculovirus). By way of example only, a ccDNA vector can be produced from cells coinfected with a ccDNA baculovirus and a Rep baculovirus. Rep proteins produced from the Rep baculovirus can replicate the ccDNA baculovirus to produce ccDNA vectors. Alternatively, ccDNA vectors for expression of the inflammasome antagonist can be generated from cells stably transfected with a construct containing the AAV Rep protein coding sequence (Rep78/52) delivered in Rep plasmids, Rep bacmids, or Rep baculovirus. ccDNA baculovirus can be transiently transfected into cells, replicated using the Rep protein, and can produce ccDNA vectors.

[00259] A bacmid (e.g., a ccDNA bacmid) can be transfected into susceptible insect cells, such as Sf9 cells, Sf21 cells, Tni (Trichoplusia ni) cells, High Five cells, and produce a ccDNA baculovirus, which is a recombinant baculovirus that includes sequences containing symmetric ITRs and an expression cassette. The insect cells can be re-infected with the ccDNA baculovirus to produce a further generation of recombinant baculovirus. Optionally, the step can be repeated one or more times to produce more recombinant baculovirus.

[00260] The timing of collection and accumulation of the ccDNA vectors for expression of an inhibitor of an immune response (e.g., an innate immune response) described herein from cells can be selected and optimized to achieve high yields of ccDNA vectors. For example, the collection time can be selected based on cell viability, cell morphology, cell expansion, etc. Typically, cells can be collected after a period of time following baculovirus infection sufficient to obtain ccDNA vectors (e.g., ccDNA vectors), but before the majority of cells have died due to viral toxicity. ccDNA vectors can be isolated from Sf9 cells using plasmid purification kits, such as the Qiagen ENDO-FREE PLASMID® kits. Other methods developed for plasmid isolation can also be adapted for ccDNA vectors. In general, any known methods in the art for purifying nucleic acids, as well as commercially available DNA extraction kits, can be used.

[00261] Alternatively, purification can be accomplished by alkaline lysis of the cell pellet, centrifugation of the resulting lysate, and chromatographic separation. As one non-limiting example, this method can be accomplished by loading the supernatant onto an ion exchange column (e.g., SARTOBIND Q®) that retains nucleic acids, followed by elution (e.g., with 1.2 M NaCl) and further chromatographic purification on a gel filtration column (e.g., 6 Fast Flow GE). The capsidless AAV vector is then isolated, for example, by precipitation.

[00262] In some embodiments, ccDNA vectors for expressing an inhibitor of an immune response (e.g., an innate immune response) can also be purified in the form of exosomes or microparticles. It is known in the art that many cell types release not only soluble proteins but also complex protein/nucleic acid cargoes by shedding membrane microvesicles (Cocucci et al., 2009; EP 10306226.1). Such vesicles include microvesicles (also called microparticles) and exosomes (also called nanovesicles); both contain proteins and RNA as cargo. Microvesicles are formed by direct budding of the plasma membrane, while exosomes are released into the extracellular environment by fusion of multivesicular endosomes with the plasma membrane. Accordingly, ccDNA vector-containing microvesicles and/or exosomes can be isolated from cells transduced with ccDNA plasmid or bacmid or baculovirus produced using ccDNA plasmid.

[00263] Microvesicles can be isolated by filtration or ultracentrifugation of the culture medium at 20,000×g, and exosomes at 100,000×g. The optimal duration of ultracentrifugation can be determined experimentally and depends on the specific cell type from which the vesicles are isolated. Preferably, the culture medium is first clarified by low-speed centrifugation (e.g., at 2000×g for 5-20 minutes) and centrifugally concentrated using, for example, an AMICON® spin column (Millipore®, Watford, UK). Microvesicles and exosomes can be further purified by FACS or MACS using specific antibodies that recognize specific surface antigens present on the microvesicles and exosomes. Other methods for purifying microvesicles and exosomes include, but are not limited to, immunoprecipitation, affinity chromatography, filtration, and magnetic beads coated with specific antibodies or aptamers. After purification, the vesicles are washed, for example, with phosphate-buffered saline (PBS). One advantage of using microvesicles or exosomes to deliver ccDNA-containing vesicles is that such vesicles can be targeted to different cell types by incorporating into their membranes proteins recognized by specific receptors on the corresponding cell types. (See also EP 10306226)

[00264] Another aspect of the present invention pertains to methods for purifying ccDNA vectors from host cell lines that have stably integrated the ccDNA construct into their own genome. In one embodiment, ccDNA vectors are purified as DNA molecules. In another embodiment, ccDNA vectors are purified as exosomes or microparticles.

[00265] FIG. 5 of international application PCT/US18/49996 shows a gel confirming the production of cDNA from several cDNA plasmid constructs using the method described in the Examples. The cDNA is confirmed based on the characteristic band pattern on the gel (see FIG. 5A).

V. Pharmaceutical compositions and formulations

[00266] The present invention provides pharmaceutical compositions and formulations comprising a therapeutic nucleic acid and one or more inhibitors of an immune response (e.g., an innate immune response) as described herein. In some embodiments, a pharmaceutical composition comprising a therapeutic nucleic acid and one or more inhibitors of an immune response (e.g., an innate immune response) may include a pharmaceutically acceptable excipient or carrier. In some embodiments, the pharmaceutical composition comprises a closed-ended DNA vector, such as a ccDNA vector as described herein, and rapamycin or a rapamycin analog, and a pharmaceutically acceptable carrier or diluent. In some embodiments, the pharmaceutical composition comprises a closed-ended DNA vector, such as a ccDNA vector as described herein, and a TLR inhibitor (e.g., a TLR9 inhibitor), and a pharmaceutically acceptable carrier or diluent. In some embodiments, the pharmaceutical composition comprises a closed-ended DNA vector, such as a ccDNA vector as described herein, and a cGAS inhibitor, and a pharmaceutically acceptable carrier or diluent. In some embodiments, the pharmaceutical composition comprises a closed-ended DNA vector, such as a ccDNA vector as described herein, and an inflammasome antagonist (e.g., any one or more of an NLRP3 inflammasome pathway inhibitor or an AIM2 inflammasome pathway inhibitor or a caspase 1 inhibitor or any combination thereof), and a pharmaceutically acceptable carrier or diluent.

[00267] The DNA vectors disclosed herein can be included in pharmaceutical compositions suitable for administration to a subject for in vivo delivery to the subject's cells, tissues, or organs, including, in some embodiments, pharmaceutical compositions comprising the inhibitors of an immune response (e.g., an innate immune response) described herein. Typically, a pharmaceutical composition comprises the ccDNA vectors disclosed herein and a pharmaceutically acceptable carrier. For example, the TTX vectors of the present invention can be included in a pharmaceutical composition suitable for a targeted therapeutic administration route (e.g., parenteral administration). Passive tissue transduction by high-pressure intravenous or intra-arterial infusion, as well as intracellular injection such as intranuclear microinjection or intracytoplasmic injection, is also contemplated. Pharmaceutical compositions for therapeutic purposes can be prepared in the form of a solution, microemulsion, dispersion, liposomes or other ordered structure suitable for high concentration of the TTX vector. Sterile injection solutions can be prepared by incorporating the TTX vector compound in the required amount in an appropriate buffer with one or a combination of ingredients listed above, if necessary, followed by sterilization by filtration.

[00268] Pharmaceutically active compositions comprising a TTX vector can be formulated to deliver a transgene in nucleic acid to recipient cells, resulting in therapeutic expression of said transgene therein. The composition can also include a pharmaceutically acceptable carrier.

[00269] The compositions and vectors provided herein can be used to deliver a transgene for a variety of purposes. In some embodiments, the transgene encodes a protein or functional RNA that is intended for use in research purposes, such as to create a somatic transgenic animal model carrying the transgene, such as to study the function of the transgene product. In another example, the transgene encodes a protein or functional RNA that is intended for use in creating an animal model of a disease. In some embodiments, the transgene encodes one or more peptides, polypeptides, or proteins that can be used to treat or prevent pathological conditions in a mammalian subject. The transgene can be transferred to a patient (e.g., expressed in the patient) in an amount sufficient to treat a disease associated with decreased expression, lack of expression, or dysfunction of the gene. In some embodiments, the transgene is a gene editing molecule (e.g., a nuclease). In certain embodiments, the nuclease is a CRISPR-associated nuclease (Cas nuclease).

[00270] In some embodiments, the pharmaceutically active compositions described herein can be administered in combination with an antihistamine or a steroid. In some embodiments, the antihistamine or steroid is administered in the same composition as the pharmaceutically active compositions described herein. In some embodiments, the antihistamine or steroid is administered in a separate composition than the pharmaceutically active compositions described herein. In some embodiments, the antihistamine or steroid is administered concurrently with the pharmaceutically active composition. In some embodiments, the antihistamine or steroid is administered sequentially with the pharmaceutically active composition. Any antihistamine known in the art can be used in the embodiments herein. According to some embodiments, the antihistamine agent is one or more of ompheniramine, buclizine, chlorpheniramine, cinnarizine, clemastine, cyclizine, cyproheptadine, diphenhydramine, diphenylpyralin, doxylamine, meclozine, pheniramine, promethazine, triprolidine, acrivastine, astemizole, cetirizine, desloratadine, fexofenadine, levocetirizine, loratadine, mizolastine, terfenadine, a pharmaceutically acceptable salt thereof, or a combination thereof. Any steroid known in the art can be used in the embodiments herein. According to some embodiments, the steroid is one or more of at least one of fluoxymesterone, mesterolone, methandrostenolone, nandrolone undecanoate, nandrolone cyplonate, oxandrolone, oxymetholone, nandrolone hexyloxyphenylpropionate, testosterone, prednisone, cortisol, cortisone, prednisolone, dexamethasone, betamethasone, triamcinolone, beclomethasone, fludrocortisone, deoxycorticosterone, aldosterone, and stanozolol.

[00271] Pharmaceutical compositions for therapeutic purposes generally must be sterile and stable under the conditions of manufacture and storage. Sterile injectable solutions can be prepared by incorporating the ccDNA vector compound in the required amount in an appropriate buffer with one or a combination of ingredients listed above, if necessary, followed by filtered sterilization.

Single dosage form

[00272] According to some embodiments, the pharmaceutical compositions may be presented as a unit dosage form. The unit dosage form is typically adapted to one or more specific routes of administration of the pharmaceutical composition. In some embodiments, the unit dosage form is adapted to be administered by inhalation. In some embodiments, the unit dosage form is adapted to be administered by a vaporizer. In some embodiments, the unit dosage form is adapted to be administered by a nebulizer. In some embodiments, the unit dosage form is adapted to be administered by an aerosol generator. In some embodiments, the unit dosage form is adapted to be administered orally, buccally, or sublingually. In some embodiments, the unit dosage form is adapted to be administered intravenously, intramuscularly, or subcutaneously. In some embodiments, the unit dosage form is adapted to be administered intrathecally or intracerebroventricularly. In some embodiments, the pharmaceutical composition is formulated for topical administration. The amount of active ingredient that can be combined with a carrier material to produce a unit dosage form will typically be the amount of the compound that produces a therapeutic effect.

IV. Administration and Dosage

[00273] The disclosure provided herein describes methods for preventing, reducing, or eliminating an adverse immune response (e.g., an innate immune response) in a subject (e.g., a human subject) by administering to said subject at least one inhibitor of an immune response (e.g., an innate immune response) described herein and a nucleic acid (e.g., a therapeutic nucleic acid, a nucleic acid used for research purposes), wherein said administrations of the inhibitor of an immune response (e.g., an innate immune response) and the administration of the nucleic acid are timed so as to provide modulation of the immune response (e.g., an innate immune response) when the administration of the two agents is provided as a combination. The two agents may be administered simultaneously in a single formulation, at the same time in different formulations, or they may be administered separately at different times.

[00274] According to one embodiment, the expressed inhibitor of an immune response (e.g., an innate immune response) disclosed herein does not elicit an immune system response, but rather suppresses the innate immune system in a subject by at least 10% or 20% or 30% or 40% or 50% or 60% or 70% or 80% or 90% or 95% or 98% or 99% or 100% compared to not administering a ccDNA vector expressing the inhibitor.

[00275] The technology described herein is generally directed to methods of administering to a subject closed-ended DNA vectors together with one or more inhibitors of an immune response (e.g., an innate immune response) selected from one or more or a combination of rapamycin or rapamycin analogs, TLR inhibitors (e.g., TLR9), cGAS inhibitors, and one or more inflammasome antagonists (e.g., any one or more of an NLRP3 inflammasome pathway inhibitor or an AIM2 inflammasome pathway inhibitor or a caspase 1 inhibitor or any combination thereof) described herein. In some embodiments, the closed-ended DNA vector includes, but is not limited to, the ccDNA vectors disclosed herein and mRNA, antisense RNA and oligonucleotide, ribozymes, aptamer, interfering RNA (RNAi), dsRNA Dicer substrate, short hairpin RNA (shRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), minicircle DNA, minigene, viral DNA (e.g., lentivirus or AAV genome) or non-viral synthetic DNA vectors, closed-ended linear duplex DNA (ccDNA/CELiD), plasmids, bacmids, doggybone DNA vectors (dbDNA ^™ ), minimalist vector for immunologically defined gene expression (MIDGE), minimal strand non-viral DNA vector (linear covalently closed DNA vector) or a dumbbell-shaped minimal DNA vector ("dumbbell DNA"). (See, for example, WO2010/0086626, the contents of which are incorporated herein by reference in their entirety). According to some embodiments, the innate immune response inhibitors and nucleic acids may be administered to a subject or patient in any combination. For example, one or more immune response inhibitors (e.g., innate immune response) may be administered. In some embodiments, an immune response inhibitor (e.g., an innate immune response) described herein and nucleic acids (e.g., a minicircle, minigene, minimal strand covalently closed DNA, doggybone DNA (dbDNA ^™ ), dumbbell DNA, closed-end linear duplex DNA (ccDNA and CELiD), plasmid-based circular vector, antisense oligonucleotide (ASO), RNAi, siRNA, mRNA, etc.) are administered to a subject or patient. In some embodiments, rapamycin or rapamycin analogs, one or more TLR9 inhibitors, and nucleic acids are administered to a subject or patient. In some embodiments, rapamycin or rapamycin analogs, one or more cGAS inhibitors, and nucleic acids are administered to a subject or patient. In some embodiments, the subject or patient is administered rapamycin or rapamycin analogs, one or more inflammasome antagonists, and nucleic acids. In some embodiments, the subject or patient is administered rapamycin or rapamycin analogs, one or more TLR9 inhibitors, one or more cGAS inhibitors, and nucleic acids. In some embodiments, the subject or patient is administered rapamycin or rapamycin analogs, one or more TLR9 inhibitors, one or more inflammasome antagonists, and nucleic acids. In some embodiments, the subject or patient is administered one or more TLR9 inhibitors, one or more cGAS inhibitors, and a ccDNA vector comprising nucleic acids. In some embodiments, the subject or patient is administered one or more TLR9 inhibitors, one or more cGAS inhibitors, one or more inflammasome antagonists, and nucleic acids. In some embodiments, the subject or patient is administered rapamycin or rapamycin analogs, one or more TLR9 inhibitors, one or more cGAS inhibitors, one or more inflammasome antagonists, and nucleic acids.

[00276] In some embodiments, one or more inhibitors of an immune response (e.g., an innate immune response) and one or more nucleic acids (e.g., a minicircle, a minigene, a minimal strand of covalently closed DNA, a doggybone DNA (dbDNA ^™ ), a dumbbell DNA, a closed-end linear duplex DNA (ccDNA and CELiD), a plasmid-based circular vector, an antisense oligonucleotide (ASO), RNAi, siRNA, mRNA, etc.) can be administered to a subject simultaneously. For example, a method can include administering to a subject an inhibitor of an immune response (e.g., an innate immune response) and a nucleic acid therapeutic agent as two separate formulations, but simultaneously. In another example, the method may include simultaneously administering to a subject an inhibitor of an immune response (e.g., an innate immune response) and a therapeutic nucleic acid in the same formulation at the same time.

[00277] In some embodiments, one or more inhibitors of an immune response (e.g., an innate immune response) and one or more nucleic acids (e.g., a minicircle, a minigene, a minimal strand of covalently closed DNA, a doggybone DNA (dbDNA ^™ ), a dumbbell DNA, a closed-end linear duplex DNA (ccDNA and CELiD), a plasmid-based circular vector, an antisense oligonucleotide (ASO), RNAi, siRNA, mRNA, etc.) may be sequentially administered to a subject. For example, an inhibitor of an immune response (e.g., an innate immune response) may be administered prior to the administration of a therapeutic nucleic acid.

[00278] In cases of sequential administration, there may be a delay period between the administration of one or more inhibitors of the immune response (e.g., innate immune response) and the TNC. For example, an inhibitor of an immune response (e.g., an innate immune response) can be administered hours, days, or weeks before administration of the TNC (e.g., at least 30 minutes, at least 1 hour, at least 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, at least 8 hours, at least 9 hours, at least 10 hours, at least 11 hours, at least 12 hours, at least 13 hours, at least 14 hours, at least 15 hours, at least 16 hours, at least 17 hours, at least 18 hours, at least 19 hours, at least 20 hours, at least 21 hours, at least 22 hours, at least 23 hours, at least at least 24 hours, at least about 2 days, at least about 3 days, at least about 4 days, at least about 5 days, at least about 6 days, at least about 1 week, at least about 2 weeks, at least about 3 weeks, and at least about 4 weeks prior to administration of the nucleic acid). In some embodiments, the immune response (e.g., innate immune response) inhibitor can be administered about thirty (30) minutes prior to administration of the TNA. In some embodiments, the immune response (e.g., innate immune response) inhibitor can be administered about one (1) hour prior to administration of the nucleic acid. In some embodiments, the immune response (e.g., innate immune response) inhibitor can be administered about two (2) hours prior to administration of the nucleic acid. In some embodiments, an inhibitor of an immune response (e.g., an innate immune response) can be administered about three (3) hours before administration of the nucleic acid. In some embodiments, an inhibitor of an immune response (e.g., an innate immune response) can be administered about four (4) hours before administration of the nucleic acid. In some embodiments, an inhibitor of an immune response (e.g., an innate immune response) can be administered about five (5) hours before administration of the nucleic acid. In some embodiments, an inhibitor of an immune response (e.g., an innate immune response) can be administered about six (6) hours before administration of the nucleic acid. In some embodiments, an inhibitor of an immune response (e.g., an innate immune response) can be administered about seven (7) hours before administration of the nucleic acid. In some embodiments, an inhibitor of an immune response (e.g., an innate immune response) can be administered about eight (8) hours before administration of the nucleic acid. In some embodiments, an inhibitor of an immune response (e.g., an innate immune response) may be administered about nine (9) hours before administration of the nucleic acid. In some embodiments, an inhibitor of an immune response (e.g., an innate immune response) may be administered about ten (10) hours before administration of the nucleic acid.

[00279] In one embodiment, an inhibitor of an immune response (e.g., an innate immune response) is administered no more than about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, or 24 hours before administration of the nucleic acid. In some embodiments, the inhibitor of an immune response (e.g., an innate immune response) may be administered no more than about 1 day, about 2 days, about 3 days, about 4 days, about 6 days, or about 7 days prior to administration of the nucleic acid.

[00280] In some embodiments, an inhibitor of an immune response (e.g., an innate immune response) can be administered about 30 minutes, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, or 24 hours after administration of the nucleic acid. In some embodiments, the inhibitor of an immune response (e.g., an innate immune response) may be administered about 1 day, about 2 days, about 3 days, about 4 days, about 6 days, or about 7 days after administration of the nucleic acid.

[00281] According to one embodiment, the inhibitor of an immune response (e.g., an innate immune response) is administered no more than about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, or 24 hours after administration of the nucleic acid. In some embodiments, the inhibitor of an immune response (e.g., an innate immune response) may be administered no more than about 1 day, about 2 days, about 3 days, about 4 days, about 6 days, or about 7 days after administration of the nucleic acid.

[00282] In some embodiments, one or more inhibitors of an immune response (e.g., an innate immune response) can be administered multiple times before, simultaneously with, and/or after administration of the nucleic acid.

[00283] In some embodiments, the nucleic acid (e.g., the ccDNA vector) can be administered in a single dose or in multiple doses. In some embodiments, more than one dose can be administered to the subject. Multiple doses can be administered as needed since the ccDNA vector does not elicit an immune response from the host against the capsid due to the absence of a viral capsid. In some embodiments, the number of doses administered can be, for example, 2 to 10 or more doses, such as 2, 3, 4, 5, 6, 7, 8, 9, 10, or more.

[00284] In some embodiments, the nucleic acid may be administered and re-dosed multiple times in combination with one or more immune response (e.g., innate immune response) inhibitors disclosed herein. For example, the therapeutic nucleic acid may be administered on day 0 with one or more immune response inhibitors administered before, after, or concurrently with the administration of the nucleic acid in a first dosing regimen. After the initial treatment on Day 0, the second dosing (re-dose) may be given at about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, or about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, or about 1 year, about 2 years, about 3 years, about 4 years, about 5 years, about 6 years, about 7 years, about 8 years, about 9 years, about 10 years, about 11 years, about 12 years, about 13 years, about 14 years, about 15 years, about 16 years, about 17 years, about 18 years, about 19 years, about 20 years, about 21 years, about 22 years, about 23 years, about 24 years, about 25 years, about 26 years, about 27 years, about 28 years, about 29 years, about 30 years, about 31 years, about 32 years, about 33 years, about 34 years, about 35 years, about 36 years, about 37 years, about 38 years, about 39 years, about 40 years, about 41 years, about 42 years, about 43 years, about 44 years, about 45 years, about 46 years, about 47 years, about 48 years, about 49 years, or about 50 years after the initial nucleic acid treatment, preferably with one or more immune response inhibitors (e.g., innate immune response) disclosed herein.

[00285] According to some embodiments, repeated dosing of a nucleic acid results in an increase in expression of the nucleic acid. According to some embodiments, the increase in expression of the nucleic acid after repeated dosing, compared to the expression of the nucleic acid after the first dose, is from about 0.5 times to about 10 times, from about 1 times to about 5 times, from about 1 times to about 2 times, or about 0.5 times, about 1 times, about 2 times, about 3 times, about 4 times, about 5 times, about 6 times, about 7 times, about 8 times, about 9 times, or about 10 times higher after repeated dosing of the nucleic acid.

[00286] Typically, the dosage will vary depending on the specific characteristics of the cDNA vector, the expression efficiency, and the age, condition, and gender of the patient. The dosage can be determined by one skilled in the art and, unlike conventional AAV vectors, can also be adjusted by the individual physician in the event of any complication, since cDNA vectors do not contain immune-activating capsid proteins that prevent repeated dosing.

[00287] According to some embodiments, more than one administration (e.g., two, three, four or more administrations) of a nucleic acid (e.g., a ccDNA vector) for protein expression as disclosed herein can be used to achieve a target level of gene expression over various time periods, such as daily, weekly, monthly, annually, etc.

[00288] According to any of the embodiments disclosed herein, the nucleic acid can be a therapeutic nucleic acid.

Therapeutic effect

[00289] The efficacy of a ccDNA vector expressing an inhibitor of an immune response (e.g., an innate immune response) disclosed herein to suppress or attenuate the innate immune system can be determined by a skilled clinician. However, a treatment is considered an "effective treatment," as that term is used herein, if any or all signs or symptoms of the innate immune system are reduced and/or altered in a beneficial manner, or if other clinically accepted symptoms or markers of the disease are improved or alleviated, such as by at least 10%, following treatment with a ccDNA vector encoding an inhibitor of an immune response (e.g., an innate immune response) disclosed herein. Exemplary markers and symptoms are discussed in the Examples herein. Efficacy can also be measured by the absence of deterioration in the individual, as assessed by stabilization of the disease or the need for medical interventions (i.e., the progression of the disease is stopped or at least slowed). Methods for measuring these parameters are known to those skilled in the art and/or described herein. Treatment includes any treatment of a disease in an individual or animal (some non-limiting examples include a human or a mammal) and includes: (1) inhibiting a disease, such as stopping or slowing the progression of a disease or disorder; or (2) alleviating a disease, such as causing a regression of symptoms; and (3) preventing or reducing the likelihood of developing a disease, or preventing secondary diseases/disorders associated with said disease, such as liver or kidney failure. An effective amount for treating a disease means an amount that, when administered to a mammal in need thereof, is sufficient to provide effective treatment, as that term is defined herein, of that disease.

[00290] The effectiveness of the agent can be determined by assessing physical indicators that are specific to the disease. Standard methods for analyzing disease indicators are known in the art. For example, physical indicators for the innate immune system include, for example, but are not limited to, soluble CD14 (sCD14) and IL-18, IL-22 in plasma or blood, inflammasome proteins such as AIM2, NLRP3, NLRP1, ASC and caspase-1 in cerebrospinal fluid or blood, activation of cytokine pathways can be used as a functional indicator of NLRP3 and/or AIM2 inflammasome pathway activation or caspase 1 activation, and includes biomarkers such as, but are not limited to: interleukin (IL)-1β, IL-6, IL-8, IL-18, interferon (IFN)-γ, interferon (IFN)-α, monocyte chemoattractant protein (MCP)-1 and/or tumor necrosis factor (TNF)-α.

[00291] In one embodiment, the ccDNA vector comprises a nucleic acid sequence for expressing an inhibitor of an immune response (e.g., an innate immune response) disclosed herein, e.g., which is functional for suppressing the innate immune system. In a preferred embodiment, the inhibitor of an immune response (e.g., an innate immune response) disclosed herein, e.g., as disclosed herein, does not elicit an immune system reaction, but rather suppresses or weakens the immune system in a subject.

[00292] Pharmaceutical compositions for therapeutic purposes generally must be sterile and stable under the conditions of manufacture and storage. The composition can be prepared as a solution, microemulsion, dispersion, liposomes or other ordered structure suitable for high concentration of a closed-end DNA vector, such as an ecDNA vector. Sterile injectable solutions can be prepared by incorporating the ecDNA vector in the required amount in an appropriate buffer with one or a combination of ingredients listed above, if necessary, followed by sterilization by filtration.

[00293] The closed-ended DNA vector, including the ccDNA vector, and the inhibitor of an immune response (e.g., an innate immune response) disclosed herein can be included in a pharmaceutical composition suitable for topical, systemic, intra-amniotic, intrathecal, intracranial, intra-arterial, intravenous, intralymphatic, intraperitoneal, subcutaneous, tracheal, interstitial (e.g., intramuscular, intracardiac, intrahepatic, intrarenal, intracerebral), intrathecal, intravesical, conjunctival (e.g., extraorbital, intraorbital, retroorbital, intraretinal, subretinal, choroidal, subchoroidal, intrastromal, intracameral, and intravitreal), intracochlear, and mucosal (e.g., oral, rectal, nasal) administration. Passive tissue transduction by high-pressure intravenous or intra-arterial infusion, as well as intracellular injection such as intranuclear microinjection or intracytoplasmic injection, is also envisaged.

[00294] In some aspects, the methods provided herein involve delivering one or more closed-ended DNA vectors, including a ccDNA vector, and an inhibitor of an immune response (e.g., an innate immune response) described herein to a host cell. Also provided herein are cells produced by such methods and organisms (such as animals, plants, or fungi) containing or derived from such cells. Methods of delivering nucleic acids can include lipofection, nucleofection, microinjection, biolistics, liposomes, immunoliposomes, polycation or lipid:nucleic acid conjugates, naked DNA, and agent-enhanced DNA uptake. Lipofection is described, for example, in U.S. Patent Nos. 5,049,386, 4,946,787, and 4,897,355, and lipofection reagents are sold commercially (e.g., Transfectam ^™ and Lipofectin ^™ ). Delivery can be to cells (e.g., in vitro or ex vivo administration) or to target tissues (e.g., in vivo administration).

[00295] Various techniques and methods for delivering nucleic acids to cells are known in the art. For example, a closed-ended DNA vector, including a ccDNA vector, and rapamycin or a rapamycin analog described herein can be formulated as lipid nanoparticles (LNPs), lipidoids, liposomes, lipid nanoparticles, lipoplexes, or core-shell nanoparticles. Typically, LNPs are comprised of nucleic acid molecules (e.g., ccDNA), one or more ionizable or cationic lipids (or salts thereof), one or more non-ionic or neutral lipids (e.g., a phospholipid), a molecule that prevents aggregation (e.g., PEG or a PEG-lipid conjugate), and optionally a sterol (e.g., cholesterol).

[00296] Another method for delivering a closed-ended DNA vector, including a ccDNA vector, and an inhibitor of an immune response (e.g., an innate immune response) described herein into a cell is to conjugate the nucleic acid to a ligand that is internalized by the cell. For example, the ligand can bind a receptor on the cell surface and be internalized by endocytosis. The ligand can be covalently linked to a nucleotide in the nucleic acid. Exemplary conjugates for delivering nucleic acids into a cell are described, for example, in WO2015/006740, WO2014/025805, WO2012/037254, WO2009/082606, WO2009/073809, WO2009/018332, WO2006/112872, WO2004/090108, WO2004/091515, and WO2017/177326.

[00297] The nucleic acids and closed-ended DNA vector, including the ccDNA vector, described herein can also be delivered into a cell by transfection. Suitable transfection methods include, but are not limited to, lipid-mediated transfection, cationic polymer-mediated transfection, or calcium phosphate precipitation. Transfection reagents are well known in the art and include, but are not limited to, TurboFect Transfection Reagent (Thermo Fisher Scientific®), Pro-Ject Reagent (Thermo Fisher Scientific®), TRANSPASS ^™ P Protein Transfection Reagent (New England Biolabs®), CHARIOT ^™ Protein Delivery Reagent (Active Motif), PROTEOJUICE ^™ Protein Transfection Reagent (EMD Millipore®), 293fectin, LIPOFECTAMINE ^™ 2000, LIPOFECTAMINE ^™ 3000 (Thermo Fisher Scientific®), LIPOFECTAMINE ^™ (Thermo Fisher Scientific®), LIPOFECTIN ^™ (Thermo Fisher Scientific®), DMRIE-C, CELLFECTIN ^™ (Thermo Fisher Scientific®), OLIGOFECTAMINE ^™ (Thermo Fisher Scientific®), LIPOFECTACE ^™ , FUGENE ^™ (Roche®, Basel, Switzerland), FUGENE ^™ HD (Roche®), TRANSFECTAM ^™ (Promega®, Madison, WI), TFX-10 ^™ (Promega®), TFX-20 ^™ (Promega®), TFX-50 ^™ (Promega®), TRANSFECTIN ^™ (BioRad®, Hercules, CA), SILENTFECT ^™ (Bio-Rad®), Effectene ^™ (Qiagen®, Valencia, CA), DC-chol (Avanti Polar Lipids), GENEPORTER ^™ (Gene Therapy Systems®, San Diego, CA), DHARMAFECT 1 ^™ (Dharmacon®, Lafayette, CO), DHARMAFECT 2 ^™ (Dharmacon®), DHARMAFECT 3 ^™ (Dharmacon®), DHARMAFECT 4 ^™ (Dharmacon®), ESCORT ^™ III (Sigma®, St. Louis, MO), and ESCORT ^™ IV (Sigma Chemical Co.). Nucleic acids such as ccDNA can also be delivered into the cell using microfluidic methods known to those skilled in the art.

[00298] The closed-ended DNA vector, including the ccDNA vector, and the inhibitor of an immune response (e.g., an innate immune response) described herein can also be administered directly into the body to transduce cells in vivo. Administration can be by any of the routes commonly used to bring the molecule into maximum contact with blood or tissue cells, including, but not limited to, injection, infusion, topical application, and electroporation. Suitable methods for administering such nucleic acids are available and well known to those skilled in the art, and although more than one route can be used to administer a particular composition, a particular route can often provide a faster and more effective response than another route of administration.

[00299] Methods of administering a closed-ended DNA vector, including a ccDNA vector, and an innate immune response inhibitor described herein may include delivery to hematopoietic stem cells, such as by methods described, for example, in U.S. Patent No. 5,928,638.

[00300] A closed-ended DNA vector, including a ccDNA vector, and an inhibitor of an immune response (e.g., an innate immune response) described herein can be added to liposomes for delivery to a target cell or organ in a subject. Liposomes are vesicles that have at least one lipid bilayer. Liposomes are typically used as vehicles for delivering drugs/therapeutics in the context of pharmaceutical development. They work by fusing with the cell membrane and rearranging its lipid structure to deliver a drug or active pharmaceutical ingredient (API). Liposomal compositions for such delivery are comprised of phospholipids, in particular compounds containing a phosphatidylcholine group, but said compositions can also include other lipids. Exemplary liposomes and liposomal formulations are disclosed in international application PCT/US2018/050042, filed September 7, 2018, and in international application PCT/US2018/064242, filed December 6, 2018, for example, see the section entitled “Pharmaceutical Formulations.”

[00301] Various delivery methods known in the art, or modifications thereof, can be used to deliver a closed-ended DNA vector, including a ccDNA vector, and an inhibitor of an immune response (e.g., an innate immune response) described herein in vitro or in vivo. For example, in some embodiments, ccDNA vectors are delivered by creating a transient opening in a cell membrane using mechanical, electrical, ultrasound, hydrodynamic, or laser energy so as to facilitate entry of DNA into target cells. For example, a ccDNA vector can be delivered by transiently rupturing a cell membrane by forcing a cell through a channel of limited size, or by other methods known in the art. In some cases, a ccDNA vector is separately directly administered by injecting naked DNA into skin, thymus, cardiac muscle, skeletal muscle, or liver cells. In some cases, a ccDNA vector is delivered using a gene gun. Spherical gold or tungsten particles (1-3 µm in diameter) coated with capsidless AAV vectors can be accelerated to high speeds using compressed gas to penetrate target tissue cells.

[00302] Compositions comprising a closed-ended DNA vector, including a ccDNA vector, and rapamycin or a rapamycin analog as described herein and a pharmaceutically acceptable carrier are specifically provided herein. In some embodiments, the ccDNA vector is formulated with a lipid delivery system, such as liposomes, as described herein. In some embodiments, such compositions are administered by any route desired by the skilled artisan. The compositions can be administered to a subject by a variety of routes, including oral, parenteral, sublingual, transdermal, rectal, transmucosal, topical, inhalational, buccal, intrapleural, intravenous, intraarterial, intraperitoneal, subcutaneous, intramuscular, intranasal, intrathecal, and intraarticular, or combinations thereof. For veterinary use, the composition may be administered in a reasonably acceptable formulation in accordance with normal veterinary practice. The veterinarian can readily determine the dosage regimen and route of administration that are most appropriate for a particular animal. The compositions may be administered using conventional syringes, needle-free injection devices, "microparticle bombardment gene guns" or other physical methods such as electroporation ("EP"), hydrodynamic methods or the use of ultrasound.

[00303] In some cases, a closed-ended DNA vector, including a ccDNA vector, and one or more immune response inhibitors (e.g., innate immune response) described herein are delivered by hydrodynamic injection, which is a simple and highly efficient method for direct intracellular delivery of any water-soluble compounds and particles to internal organs and skeletal muscles throughout the limb.

[00304] In some cases, the closed-ended DNA vector, including the ccDNA vector, and one or more inhibitors of an immune response (e.g., an innate immune response) described herein are delivered using ultrasound by creating nanoscopic pores in a membrane to facilitate intracellular delivery of DNA particles to cells of internal organs or tumors, so the size and concentration of the closed-ended DNA vector are of great importance to the efficiency of the system. In some cases, the closed-ended DNA vectors, including the ccDNA vector, and one or more inhibitors of an immune response (e.g., an innate immune response) described herein are delivered using magnetofection using magnetic fields to concentrate particles containing nucleic acid in target cells.

[00305] In some cases, chemical delivery systems may be used, such as those using nanoscale complexes that involve compaction of negatively charged nucleic acid with polycationic nanoscale particles, such as cationic liposomes/micelles or cationic polymers. Cationic lipids used in this delivery method include, but are not limited to, monovalent cationic lipids, polyvalent cationic lipids, guanidine-containing compounds, cholesterol derivative compounds, cationic polymers (e.g., polyethyleneimine, poly-L-lysine, protamine, other cationic polymers), and lipid-polymer hybrids.

A. Exosomes:

[00306] In some embodiments, a closed-ended DNA vector, including a ccDNA vector, and one or more inhibitors of an immune response (e.g., an innate immune response) described herein are delivered packaged in an exosome. Exosomes are small membrane vesicles of endocytic origin that are released into the extracellular environment following fusion of multivesicular bodies with the plasma membrane. Their surface consists of the lipid bilayer of the cellular membrane of the donor cell, they contain cytosol from the cell that produced the exosome, and they display membrane proteins of the parent cell on the surface. Exosomes are produced by a variety of cell types, including epithelial cells, B and T lymphocytes, mast cells (MC), and dendritic cells (DC). According to some embodiments, exosomes with a diameter of 10 nm to 1 μm, 20 nm to 500 nm, 30 nm to 250 nm, 50 nm to 100 nm are provided for use. Exosomes can be isolated for delivery to target cells either using their donor cells or by introducing specific nucleic acids into them. Various approaches known in the art can be used to obtain exosomes containing capsid-less AAV vectors according to the present invention.

B. Microparticles/nanoparticles:

[00307] In some embodiments, a closed-ended DNA vector, including a ccDNA vector, and rapamycin or a rapamycin analog described herein are delivered via a lipid nanoparticle. Typically, the lipid nanoparticles comprise an ionizable aminolipid (e.g., heptatriaconta-6,9,28,31-tetraen-19-yl-4-(dimethylamino)butanoate, DLin-MC3-DMA, phosphatidylcholine (1,2-distearoyl-sn-glycero-3-phosphocholine, DSPC), cholesterol, and a shell lipid (polyethylene glycol-dimyristoylglycerol, PEG-DMG), e.g., as described by Tam et al. (2013). Advances in Lipid Nanoparticles for siRNA delivery. Pharmaceuticals 5(3): 498-507. In some embodiments, the lipid nanoparticle has an average diameter of from about 10 to about 1000 nm. In some embodiments, the lipid nanoparticle has a diameter of less than 300 nm. In some embodiments the lipid nanoparticle has a diameter of from about 10 to about 300 nm. In some embodiments, the lipid nanoparticle has a diameter of less than 200 nm. In some embodiments, the lipid nanoparticle has a diameter of from about 25 to about 200 nm. In some other embodiments, the lipid particles comprising the therapeutic nucleic acid and/or the immunosuppressant typically have an average diameter of from about 20 nm to about 100 nm, from 30 nm to about 150 nm, from about 40 nm to about 150 nm, from about 50 nm to about 150 nm, from about 60 nm to about 130 nm, from about 70 nm to about 110 nm, from about 70 nm to about 100 nm, from about 80 nm to about 100 nm, from about 90 nm to about 100 nm, from about 70 nm to about 90 nm, from about 80 nm to about 90 nm, from about 70 nm to about 80 nm, or about 30 nm, 35 nm, 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 85 nm, 90 nm, 95 nm, 100 nm, 105 nm, 110 nm, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm or 150 nm to ensure efficient delivery. Lipid particles containing a nucleic acid and a method for producing the same are disclosed, for example, in PCT/US18/50042, U.S. Patent Publication Nos. 20040142025 and 20070042031, the disclosures of which are incorporated herein by reference in their entireties for all purposes. In some embodiments, a lipid nanoparticle preparation (e.g., a composition comprising a plurality of lipid nanoparticles) has a size distribution in which the average size (e.g., diameter) is from about 70 nm to about 200 nm, and more typically the average size is about 100 nm or less.

[00308] According to some embodiments, a liquid pharmaceutical composition comprising a nucleic acid (e.g., a therapeutic nucleic acid, a nucleic acid used for research purposes) and/or an inhibitor of an immune response (e.g., an innate immune response) according to the present invention can be prepared as a lipid particle formulation. In some embodiments, the lipid particle comprising the nucleic acid can be formed from a cationic lipid. In some other embodiments, the lipid particle comprising the nucleic acid can be formed from a non-cationic lipid. According to a preferred embodiment, the lipid particle according to the present invention is a lipid particle containing a nucleic acid, which is formed from a cationic lipid containing a nucleic acid selected from the group consisting of mRNA, antisense RNA and oligonucleotide, ribozymes, aptamer, interfering RNA (RNAi), dsRNA substrate Dicer, short hairpin RNA (shRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), minicircle DNA, minigene, viral DNA (e.g. lentivirus or AAV genome) or non-viral synthetic DNA vectors, closed-end linear duplex DNA (cDNA/CELiD), plasmids, bacmids, doggybone DNA vectors (dbDNA™), minimalist vector for immunologically defined gene expression (MIDGE), a non-viral minimal strand DNA vector (linear, covalently closed DNA vector) or a dumbbell-shaped minimal DNA vector ("dumbbell DNA").

[00309] Various lipid nanoparticles known in the art can be used to deliver a closed-ended DNA vector, including the ccDNA vector described herein. For example, various delivery methods using lipid nanoparticles are described in U.S. Patent Nos. 9,404,127, 9,006,417, and 9,518,272.

[00310] In some embodiments, a closed-ended DNA vector, including a ccDNA vector, and one or more inhibitors of an immune response (e.g., an innate immune response) described herein are delivered using a gold nanoparticle. Typically, the nucleic acid can be covalently linked to the gold nanoparticle or non-covalently linked to the gold nanoparticle (e.g., linked by a charge-charge interaction), such as described by Ding et al. (2014). Gold Nanoparticles for Nucleic Acid Delivery. Mol. Ther. 22(6); 1075-1083. In some embodiments, gold nanoparticle-nucleic acid conjugates are prepared using methods described, for example, in U.S. Patent No. 6,812,334.

C. Conjugates

[00311] In some embodiments, a closed-ended DNA vector, including a ccDNA vector, and one or more inhibitors of an immune response (e.g., an innate immune response) described herein, as disclosed herein, are conjugated (e.g., covalently linked) to an agent that enhances cellular uptake. An "agent that enhances cellular uptake" is a molecule that facilitates the transport of a nucleic acid across a lipid membrane. For example, a nucleic acid can be conjugated to a lipophilic compound (e.g., cholesterol, tocopherol, etc.), a cell penetrating peptide (CPP) (e.g., penetratin, TAT, Syn1B, etc.), and polyamines (e.g., spermine). Additional examples of agents that enhance cellular uptake are disclosed, for example, in Winkler (2013). Oligonucleotide conjugates for therapeutic applications. Ther. Deliv. 4(7); 791-809.

[00312] In some embodiments, a closed-ended DNA vector, including a ccDNA vector, and one or more inhibitors of an immune response (e.g., an innate immune response) described herein, as disclosed herein, are conjugated to a polymer (e.g., a polymer molecule) or a folate molecule (e.g., a folic acid molecule). In general, delivery of nucleic acids conjugated to polymers is known in the art, such as described in WO 2000/34343 and WO 2008/022309. In some embodiments, a ccDNA vector disclosed herein is conjugated to a polyamide polymer, such as described in U.S. Patent No. 8,987,377. In some embodiments, the nucleic acid described in the present disclosure is conjugated to a folic acid molecule as described in U.S. Patent No. 8,507,455.

[00313] In some embodiments, a closed-ended DNA vector, including a ccDNA vector, and rapamycin or a rapamycin analog described herein, as disclosed herein, are conjugated to a carbohydrate, such as described in U.S. Patent No. 8,450,467.

[00314] In some embodiments, the lipid nanoparticles may be conjugated with other groups to prevent aggregation. Such lipid conjugates include, but are not limited to, PEG-lipid conjugates such as, for example, PEG linked to dialkyloxypropyls (e.g., PEG-DAA conjugates), PEG linked to diacylglycerols (e.g., PEG-DAG conjugates), PEG linked to cholesterol, PEG linked to phosphatidylethanolamines, and PEG conjugated to ceramides (see, e.g., U.S. Patent No. 5,885,613), cationic lipids-PEG, polyoxazoline (POZ)-lipid conjugates (e.g., POZ-DAA conjugates; see, e.g., U.S. Provisional Application No. 61/294,828, filed January 13, 2010, and U.S. Provisional Application No. 61/295,140, filed January 14, 2010), polyamide oligomers (e.g., ATTA-lipid conjugates), and mixtures thereof. Additional examples of POZ-lipid conjugates are described in PCT Publication No. WO 2010/006282. PEG or POZ may be conjugated directly to the lipid or may be linked to the lipid via a linker group. Any linker group suitable for linking PEG or POZ to the lipid may be used, including, for example, non-ester linker groups and ester linker groups. Certain preferred embodiments employ non-ester linker groups such as amides or carbamates. The disclosures of each of the above-mentioned patent documents are herein incorporated by reference in their entirety for all purposes.

D. Nanocapsule

[00315] Alternatively, nanocapsule formulations of a closed-ended DNA vector, including a ccDNA vector, and rapamycin or a rapamycin analog described herein can be used, as disclosed herein. Typically, nanocapsules can entrap substances in a stable and reproducible manner. To avoid side effects due to intracellular polymer overload, such ultrafine particles (about 0.1 μm in size) should be designed using polymers that are degradable in vivo. The use of biodegradable polyalkyl cyanoacrylate nanoparticles that meet such requirements is contemplated.

E. Liposomes

[00316] A closed-ended DNA vector, including a ccDNA vector, and one or more inhibitors of an immune response (e.g., an innate immune response) described herein can be added to liposomes for delivery to a target cell or organ in a subject. Liposomes are vesicles that have at least one lipid bilayer. Liposomes are typically used as vehicles for delivering drugs/therapeutics in the context of pharmaceutical development. They work by fusing with the cell membrane and rearranging its lipid structure to deliver a drug or active pharmaceutical ingredient (API). Liposomal compositions for such delivery are comprised of phospholipids, in particular compounds containing a phosphatidylcholine group, but said compositions can also include other lipids.

[00317] The preparation and use of liposomes are generally known to those skilled in the art. Liposomes with improved serum stability and circulating half-life have been developed (U.S. Patent No. 5,741,516). In addition, various methods for producing liposomes and liposome-like preparations as potential drug carriers have been described (U.S. Patents Nos. 5,567,434; 5,552,157; 5,565,213; 5,738,868; and 5,795,587).

F. Exemplary compositions of liposomes and lipid nanoparticles (LNPs)

[00318] A closed-ended DNA vector, including a ccDNA vector, and one or more inhibitors of an immune response (e.g., an innate immune response) described herein can be added to liposomes for delivery to a cell, such as a cell in which expression of a transgene is desired. Liposomes are vesicles that have at least one lipid bilayer. Liposomes are typically used as vehicles for delivering drugs/therapeutics in the context of pharmaceutical development. They work by fusing with the cell membrane and rearranging its lipid structure to deliver a drug or active pharmaceutical ingredient (API). Liposomal compositions for such delivery are comprised of phospholipids, in particular compounds containing a phosphatidylcholine group, but said compositions can also include other lipids.

[00319] Lipid nanoparticles (LNPs) containing ccDNA are disclosed in international application PCT/US2018/050042, filed September 7, 2018, and international application PCT/US2018/064242, filed December 6, 2018, each of which is incorporated herein by reference in its entirety, and are provided for use in the methods and compositions disclosed herein.

[00320] According to some aspects of the present disclosure, a liposomal formulation is provided that includes one or more compounds with a polyethylene glycol (PEG) functional group (so-called "PEGylated compounds"), which can reduce the immunogenicity/antigenicity of the compound(s), provide their hydrophilicity and hydrophobicity, and reduce the frequency of dosing. In other cases, the liposomal formulation simply includes a polyethylene glycol (PEG) polymer as an additional component. In such aspects, the molecular weight of the PEG or PEG functional group can be from 62 Da to about 5000 Da.

[00321] According to some aspects of the present disclosure, a liposomal formulation is provided that delivers an API with a sustained or controlled release profile over a period of hours to weeks. In some related aspects, the liposomal formulation can comprise aqueous chambers that are bounded by lipid bilayers. In other related aspects, the liposomal formulation encapsulates the API with components that undergo a physical transition at elevated temperature that releases the API over a period of hours to weeks.

[00322] In some aspects, the liposomal formulation comprises sphingomyelin and one or more lipids disclosed herein. In some aspects, the liposomal formulation comprises optisomes.

[00323] According to some aspects of the present disclosure, a liposomal formulation is provided that includes one or more lipids selected from: N-(carbonylmethoxypolyethyleneglycol 2000)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine, sodium salt, (distearoyl-sn-glycerophosphoethanolamine), MPEG (methoxypolyethyleneglycol)-conjugated lipid, HSPC (hydrogenated soy phosphatidylcholine); PEG (polyethyleneglycol); DSPE (distearoyl-sn-glycerophosphoethanolamine); DSPC (distearoylphosphatidylcholine); DOPC (dioleoylphosphatidylcholine); DPPG (dipalmitoylphosphatidylglycerol); EPC (egg phosphatidylcholine); DOPS (dioleoylphosphatidylserine); POPC (palmitoyloleoylphosphatidylcholine); SM (sphingomyelin); MPEG (methoxypolyethyleneglycol); DMPC (dimyristoylphosphatidylcholine); DMPG (dimyristoylphosphatidylglycerol); DSPG (distearoylphosphatidylglycerol); DEPC (dierucoylphosphatidylcholine); DOPE (dioleoyl-sn-glycerophosphoethanolamine), cholesterol sulfate (CS), dipalmitoylphosphatidylglycerol (DPPG), DOPC (dioleoyl-sn-glycerophosphatidylcholine), or any combination thereof.

[00324] According to some aspects of the present disclosure, there is provided a liposomal formulation comprising a phospholipid, cholesterol, and a PEGylated lipid in a molar ratio of 56:38:5. According to some aspects, the total lipid content of the liposomal formulation is 2-16 mg/mL. According to some aspects of the present disclosure, there is provided a liposomal formulation comprising a lipid comprising a phosphatidylcholine functional group, a lipid comprising an ethanolamine functional group, and a PEGylated lipid. According to some aspects of the present disclosure, there is provided a liposomal formulation comprising a lipid comprising a phosphatidylcholine functional group, a lipid comprising an ethanolamine functional group, and a PEGylated lipid, in a molar ratio of 3:0.015:2, respectively. According to some aspects of the present disclosure, there is provided a liposomal formulation comprising a lipid comprising a phosphatidylcholine functional group, cholesterol, and a PEGylated lipid. According to some aspects of the present disclosure, there is provided a liposomal formulation comprising a lipid comprising a phosphatidylcholine functional group and cholesterol. According to some aspects, the pegylated lipid is PEG-2000-DSPE. According to some aspects of the present disclosure, there is provided a liposomal formulation comprising DPPG, soy PC, MPEG-DSPE lipid conjugate and cholesterol.

[00325] According to some aspects of the present disclosure, there is provided a liposomal formulation comprising one or more lipids comprising a phosphatidylcholine functional group and one or more lipids comprising an ethanolamine functional group. According to some aspects of the present disclosure, there is provided a liposomal formulation comprising one or more of: lipids comprising a phosphatidylcholine functional group, lipids comprising an ethanolamine functional group, and sterols, such as cholesterol. According to some aspects, the liposomal formulation comprises DOPC/DEPC; and DOPE.

[00326] According to some aspects of the present disclosure, a liposomal formulation is provided further comprising one or more pharmaceutical excipients, such as sucrose and/or glycine.

[00327] According to some aspects of the present disclosure, a liposomal formulation with a unilamellar or multilamellar structure is provided. According to some aspects of the present disclosure, a liposomal formulation is provided that comprises multivesicular particles and/or foam-based particles. According to some aspects of the present disclosure, a liposomal formulation is provided that has a relative size larger than conventional nanoparticles and is from about 150 to 250 nm. According to some aspects, the liposomal formulation is a lyophilized powder.

[00328] According to some aspects of the present disclosure, there is provided a liposomal formulation that is prepared and loaded with the ccDNA vectors disclosed or described herein by adding a weak base to a mixture containing isolated ccDNA outside the liposome. Such addition increases the pH outside the liposome to about 7.3 and stimulates entry of the API into the liposome. According to some aspects of the present disclosure, there is provided a liposomal formulation having an acidic pH inside the liposome. In such cases, the pH inside the liposome can be 4-6.9, and more preferably pH 6.5. According to other aspects of the present disclosure, there is provided a liposomal formulation prepared using intraliposomal drug stabilization technology. In such cases, polymeric or non-polymeric highly charged anions and intraliposomal entrapment agents are used, for example, sucrose polyphosphate or octasulfate.

[00329] According to some aspects of the present disclosure, there is provided a lipid nanoparticle comprising a DNA vector, including an eDNA vector as described herein, and an ionizable lipid. For example, a lipid nanoparticle formulation that is prepared and loaded with eDNA obtained by the method disclosed in international application PCT/US2018/050042, filed September 7, 2018, which is incorporated herein. This can be accomplished by vigorously mixing ethanol solutions of lipids with an aqueous solution of eDNA at a low pH that protonates the ionizable lipid and provides favorable energetic characteristics for eDNA/lipid association and particle nucleation. The particles can be further stabilized by dilution with water and removal of the organic solvent. The particles can be concentrated to a target level.

[00330] Typically, the lipid particles are prepared at a total lipid to ccDNA ratio (weight or mass ratio) of from about 10:1 to 30:1. In some embodiments, the lipid to ccDNA ratio (weight ratio; w/w ratio) can range from about 1:1 to about 25:1, from about 10:1 to about 14:1, from about 3:1 to about 15:1, from about 4:1 to about 10:1, from about 5:1 to about 9:1, or from about 6:1 to about 9:1. In accordance with some embodiments of any of the aspects or embodiments herein, the composition has a total lipid to ccDNA ratio of about 15:1. In accordance with some embodiments of any of the aspects or embodiments herein, the composition has a total lipid to ccDNA ratio of about 30:1. According to some embodiments of any of the aspects or embodiments herein, the composition has a total lipid to ccDNA ratio of about 40:1. According to some embodiments of any of the aspects or embodiments herein, the composition has a total lipid to ccDNA ratio of about 50:1. The amounts of lipid and ccDNA can be adjusted to provide a target N/P ratio, such as an N/P ratio of 3, 4, 5, 6, 7, 8, 9, 10 or higher. Typically, the total lipid content of the lipid particles can be in the range of about 5 mg/mL to about 30 mg/mL.

[00331] An ionizable lipid is typically used to condense a nucleic acid cargo, such as ccDNA, at low pH and to promote membrane association and fusogenicity. Typically, ionizable lipids are lipids that contain at least one amino group that is positively charged or becomes protonated under acidic conditions, such as at pH 6.5 or below. Ionizable lipids are also referred to herein as cationic lipids.

[00332] Exemplary ionizable lipids are described in International PCT Patent Publications WO2015/095340, WO2015/199952, WO2018/011633, WO2017/049245, WO2015/061467, WO2012/040184, WO2012/000104, WO2015/074085, WO2016/081029, WO2017/004143, WO2017/075531, WO2017/117528, WO2011/022460, WO2013/148541, WO2013/116126, WO2011/153120, WO2012/044638, WO2012/054365, WO2011/090965, WO2013/016058, WO2012/162210, WO2008/042973, WO2010/129709, WO2010/144740, WO2012/099755, WO2013/049328, WO2013/086322, WO2013/086373, WO2011/071860, WO2009/132131, WO2010/048536, WO2010/088537, WO2010/054401, WO2010/054406, WO2010/054405, WO2010/054384, WO2012/016184, WO2009/086558, WO2010/042877, WO2011/000106, WO2011/000107, WO2005/120152, WO2011/141705, WO2013/126803, WO2006/007712, WO2011/038160, WO2005/121348, WO2011/066651, WO2009/127060, WO2011/141704, WO2006/069782, WO2012/031043, WO2013/006825, WO2013/033563, WO2013/089151, WO2017/099823, WO2015/095346 and WO2013/086354, and US Patent Publications US2016/0311759, US2015/0376115, US2016/0151284, US2017/0210697, US2015/0140070, US2013/0178541, US2013/0303587, US2015/0141678, US2015/0239926, US2016/0376224, US2017/0119904, US2012/0149894, US2015/0057373, US2013/0090372, US2013/0274523, US2013/0274504, US2013/0274504, US2009/0023673, US2012/0128760, US2010/0324120, US2014/0200257, US2015/0203446, US2018/0005363, US2014/0308304, US2013/0338210, US2012/0101148, US2012/0027796, US2012/0058144, US2013/0323269, US2011/0117125, US2011/0256175, US2012/0202871, US2011/0076335, US2006/0083780, US2013/0123338, US2015/0064242, US2006/0051405, US2013/0065939, US2006/0008910, US2003/0022649, US2010/0130588, US2013/0116307, US2010/0062967, US2013/0202684, US2014/0141070, US2014/0255472, US2014/0039032, US2018/0028664, US2016/0317458 and US2013/0195920, the entire contents of which are incorporated herein by reference.

[00333] According to some embodiments, the ionizable lipid is MC3 (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl-4-(dimethylamino)butanoate (DLin-MC3-DMA or MC3), which has the following structure:

.

[00334] The lipid DLin-MC3-DMA is described in Jayaraman et al., Angew. Chem. Int. Ed Engl. (2012), 51(34): 8529-8533, the contents of which are incorporated herein by reference in their entirety.

[00335] In some embodiments, the ionizable lipid is the ATX-002 lipid described in WO 2015/074085, the contents of which are incorporated herein by reference in their entirety.

[00336] In some embodiments, the ionizable lipid is (13Z,16Z)-N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine (Compound 32), described in WO 2012/040184, the contents of which are incorporated herein by reference in their entirety.

[00337] In some embodiments, the ionizable lipid is Compound 6 or Compound 22, described in WO 2015/199952, the contents of which are incorporated herein by reference in their entirety.

[00338] Without limitation, the ionizable lipid can comprise 20-90% (mol) of the total lipids present in the lipid nanoparticle. For example, the molar content of the ionizable lipid can be 20-70% (mol), 30-60% (mol), or 40-50% (mol) of the total lipids present in the lipid nanoparticle. In some embodiments, the ionizable lipid comprises from about 50 mol % to about 90 mol % of the total lipids present in the lipid nanoparticle.

[00339] According to some aspects, the lipid nanoparticle may further comprise a non-cationic lipid. Non-ionic lipids include amphipathic lipids, neutral lipids, and anionic lipids. Accordingly, the non-cationic lipid may be a neutral uncharged, zwitterionic, or anionic lipid. Non-cationic lipids are typically used to enhance fusogenicity.

[00340] Exemplary non-cationic lipids contemplated for use in methods and compositions comprising a DNA vector, including the ccDNA vector described herein, are described in international application PCT/US2018/050042, filed September 7, 2018, and PCT/US2018/064242, filed December 6, 2018, which are incorporated herein in their entireties.

[00341] Exemplary non-cationic lipids are described in International Patent Publication WO 2017/099823 and United States Patent Publication US 2018/0028664, the contents of which are incorporated herein by reference in their entirety.

[00342] The non-cationic lipid may comprise 0-30% (mol) of the total lipids present in the lipid nanoparticle. For example, the non-cationic lipid comprises 5-20% (mol) or 10-15% (mol) of the total lipids present in the lipid nanoparticle. In various embodiments, the molar ratio of ionizable lipid to neutral lipid ranges from about 2:1 to about 8:1.

[00343] In some embodiments, the lipid nanoparticles do not contain any phospholipids. In some aspects, the lipid nanoparticle may further comprise a component, such as a sterol, to ensure membrane integrity.

[00344] One exemplary sterol that is suitable for use in a lipid nanoparticle is cholesterol and its derivatives. Exemplary cholesterol derivatives are described in International Patent Application WO 2009/127060 and US Patent Publication US 2010/0130588, the contents of which are incorporated herein by reference in their entirety.

[00345] A membrane integrity component, such as a sterol, may comprise 0-50% (mol) of the total lipid present in the lipid nanoparticle. In some embodiments, such a component comprises 20-50% (mol) 30-40% (mol) of the total lipid content of the lipid nanoparticle.

[00346] In some aspects, the lipid nanoparticle may further comprise polyethyleneglycol (PEG) or a conjugated lipid molecule. These are typically used to inhibit aggregation of the lipid nanoparticles and/or to provide steric stabilization. Exemplary conjugated lipids include, but are not limited to, PEG-lipid conjugates, polyoxazoline (POZ)-lipid conjugates, polyamide-lipid conjugates (such as ATTA-lipid conjugates), cationic group-polymer-lipid (CPL) conjugates, and mixtures thereof. In some embodiments, the conjugated lipid molecule is a PEG-lipid conjugate, such as a methoxypolyethyleneglycol-conjugated lipid. Exemplary PEG-lipid conjugates include, but are not limited to, PEG-diacylglycerol (DAG) (such as 1-(monomethoxypolyethyleneglycol)-2,3-dimyristoylglycerol (PEG-DMG)), PEG-dialkyloxypropyl (DAA), PEG-phospholipid, PEG-ceramide (Cer), pegylated phosphatidylethanolamine (PEG-PE), PEG-succinate diacylglycerol (PEGS-DAG) (such as 4-O-(2',3'-di(tetradecanoyloxy)propyl-1-O)-(w-methoxy(polyethoxy)ethyl)butanedioate (PEG-S-DMG)), PEG-dialkoxypropylcarbam, sodium salt of N-(carbonylmethoxypolyethyleneglycol) 2000)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine or a mixture thereof. Additional exemplary PEG-lipid conjugates are described, for example, in US 5,885,613, US 6,287,591, US 2003/0077829, US 2003/0077829, US 2005/0175682, US 2008/0020058, US 2011/0117125, US 2010/0130588, US 2016/0376224 and US 2017/0119904, the contents of which are incorporated herein by reference in their entirety.

[00347] In some embodiments, the PEG lipid is a compound as disclosed in US 2018/0028664, the contents of which are incorporated herein by reference in their entirety.

[00348] In some embodiments, the PEG lipid is disclosed in US20150376115 or US2016/0376224, the contents of which are incorporated herein by reference in their entirety.

[00349] The PEG-DAA conjugate can be, for example, PEG-dilauryloxypropyl, PEG-dimyristyloxypropyl, PEG-dipalmityloxypropyl, or PEG-distearyloxypropyl. The PEG lipid may be one or more of PEG-DMG, PEG-dilaurylglycerol, PEG-dipalmitoylglycerol, PEG-disterylglycerol, PEG-dilaurylglycamide, PEG-dimyristylglycamide, PEG-dipalmitoylglycamide, PEG-disterylglycamide, PEG-cholesterol (1-[8'-(cholest-5-ene-3[beta]-oxy)carboxamido-3',6'-dioxaoctanoyl]carbamoyl-[omega]-methylpolyethyleneglycol, PEG-DMB (3,4-ditetradecoxylbenzyl-[omega]-methylpolyethyleneglycol ether), and 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-[methoxypolyethyleneglycol-2000]. In some examples, the PEG lipid may be selected from the group consisting of PEG-DMG, 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-[methoxypolyethyleneglycol-2000].

[00350] Lipids conjugated to a molecule other than PEG may also be used in place of the PEG lipid. For example, polyoxazoline (POZ)-lipid conjugates, polyamide-lipid conjugates (such as ATTA-lipid conjugates), and cationic group-polymer-lipid (CPL) conjugates may be used in place of or in conjunction with the PEG lipid. Exemplary conjugated lipids, i.e., PEG-lipids, (POZ)-lipid conjugates, ATTA-lipid conjugates and cationic polymer-lipids are described in International Patent Application Publications WO 1996/010392, WO 1998/051278, WO 2002/087541, WO 2005/026372, WO 2008/147438, WO 2009/086558, WO 2012/000104, WO 2017/117528, WO 2017/099823, WO 2015/199952, WO 2017/004143, WO 2015/095346, WO 2012/000104, WO 2012/000104 and WO 2010/006282, US Patent Application Publications US 2003/0077829, US 2005/0175682, US 2008/0020058, US 2011/0117125, US 2013/0303587, US 2018/0028664, US 2015/0376115, US 2016/0376224, US 2016/0317458, US 2013/0303587, US 2013/0303587 and US 20110123453, and US Patents US 5,885,613, US 6,287,591, US 6,320,017 and US 6,586,559, the contents of which are incorporated herein by reference in their entirety.

[00351] In some embodiments, one or more additional compounds may be a therapeutic agent. The therapeutic agent may be selected from any class suitable for use in therapeutic purposes. In other words, the therapeutic agent may be selected from any class suitable for use in therapeutic purposes. In other words, the therapeutic agent may be selected in accordance with the therapeutic goal and the target biological effect. For example, if the ccDNA in the LNP can be used to treat cancer, the additional compound can be an anti-cancer agent (e.g., a chemotherapeutic agent, a targeted cancer therapy (including, but not limited to, a small molecule, an antibody, or an antibody-drug conjugate). According to another example, if the ccDNA-containing LNP can be used to treat an infection, the additional compound can be an antimicrobial agent (e.g., an antibiotic or an antiviral compound). According to yet another example, if the ccDNA-containing LNP can be used to treat an immune disease or disorder, the additional compound can be a compound that modulates an immune response (e.g., an immunosuppressant, an immunostimulatory compound, or a compound that modulates one or more specific immune pathways). In some embodiments, other mixtures of different lipid nanoparticles containing other compounds, such as ccDNA encoding another protein or another compound, such as a therapeutic agent, can be used in the compositions and methods of the present invention.

[00352] In some embodiments, the additional compound is an immunomodulatory agent. For example, the additional compound is an immunosuppressant. In some embodiments, the additional compound is an immunostimulatory agent.

[00353] Also provided herein is a pharmaceutical composition comprising a lipid nanoparticle encapsulated ccDNA vector and rapamycin or a rapamycin analog as described herein and a pharmaceutically acceptable carrier or excipient. Also provided herein is a pharmaceutical composition comprising a lipid nanoparticle encapsulated ccDNA vector and a pharmaceutically acceptable carrier or excipient, wherein said rapamycin or rapamycin analog is co-administered to a subject in another composition as described herein.

[00354] According to some aspects of the present invention, a lipid nanoparticle formulation is provided that further comprises one or more pharmaceutical excipients. According to some embodiments, the lipid nanoparticle formulation further comprises sucrose, Tris, trehalose, and/or glycine.

[00355] A closed-ended DNA vector, including a ccDNA vector, and optionally one or more inhibitors of an immune response (e.g., an innate immune response) described herein can be complexed with a lipid portion (portion) of a particle or can be encapsulated in a lipid portion (position) of a lipid nanoparticle. In some embodiments, a DNA vector, including a ccDNA vector described herein, can be completely encapsulated in a lipid portion of a lipid nanoparticle, which protects it from degradation by a nuclease, such as in an aqueous solution. In some embodiments, a DNA vector in a lipid nanoparticle, including a ccDNA vector described herein, is not substantially degraded after exposure of the lipid nanoparticle to a nuclease at 37°C for at least about 20, 30, 45, or 60 minutes. In some embodiments, the ccDNA in the lipid nanoparticle is not substantially degraded after incubating the particle in serum at 37°C for at least about 30, 45, or 60 minutes, or at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, or 36 hours.

[00356] In certain embodiments, the lipid nanoparticles are substantially non-toxic to a subject, such as a mammal, such as a human. In some aspects, the lipid nanoparticle formulation is a lyophilized powder.

[00357] In some embodiments, the lipid nanoparticles are solid core particles that have at least one lipid bilayer. In other embodiments, the lipid nanoparticles have a non-bilayer structure, i.e., have a non-lamellar (i.e., non-bilayer) morphology. Without limitation, non-bilayer morphology can include, for example, three-dimensional tubes, rods, cubic symmetry particles, etc. For example, the morphology of the lipid nanoparticles (lamellar versus non-lamellar) can be readily assessed and characterized using, for example, cryo-TEM (cryo-transmission electron microscopy) analysis, as described in US2010/0130588, the contents of which are incorporated herein by reference in their entirety.

[00358] According to some other embodiments, lipid nanoparticles having a non-lamellar morphology are electron dense. According to some aspects of the present disclosure, a lipid nanoparticle is provided that is either unilamellar or multilamellar in structure. According to some aspects of the present disclosure, a lipid nanoparticle composition is provided that comprises multivesicular particles and/or foam-based particles.

By controlling the composition and concentration of the lipid components, the rate at which the lipid conjugate is released from the lipid particle can be controlled, and in turn, the rate at which the lipid nanoparticle becomes fusogenic. In addition, other variables, including, for example, pH, temperature, or ionic strength, can be used to change and/or control the rate at which the lipid nanoparticle becomes fusogenic. Other methods that can be used to control the rate at which the lipid nanoparticle becomes fusogenic will be apparent to those of ordinary skill in the art upon review of this disclosure. It will also be appreciated that the size of the lipid particles can be controlled by controlling the composition and concentration of the lipid conjugate. The pKa of cationic lipids in the formulations can be compared to the efficiency of LNPs for nucleic acid delivery (see Jayaraman et al, Angewandte Chemie, International Edition (2012), 51(34), 8529-8533; Semple et al, Nature Biotechnology 28, 172-176 (2010), both of which are incorporated herein by reference in their entireties). The preferred pKa range is from ~5 to ~7. The pKa of each cationic lipid in the lipid nanoparticles can be determined using an assay based on 2-(p-toluidino)-6-naphthalenesulfonic acid (TNS) fluorescence.

VI. Inhibitors of the immune response

[00359] Provided herein are immune response inhibitors. In accordance with embodiments, the immune response inhibitors are innate immune response inhibitors.

Rapamycin or rapamycin analogs

[00360] According to some aspects of the present disclosure, non-viral capsidless DNA vectors with covalently closed ends (ccDNA vector) are provided that are administered in combination with rapamycin or rapamycin analogs. In some embodiments, rapamycin or a rapamycin analog is present in a supersaturated amount in a synthetic nanocarrier as described in WO 2016/073799. In some embodiments, the ccDNA vector is also present in the same nanocarrier.

[00361] In some embodiments of the compositions and methods described herein, rapamycin or a rapamycin analog is co-administered to the subject with a ccDNA vector. In some embodiments of the compositions and methods described herein, the ccDNA vector and rapamycin or a rapamycin analog are co-administered in a single formulation. In some embodiments of the compositions and methods described herein, rapamycin or a rapamycin analog is present at a supersaturated concentration in a synthetic nanocarrier as described in WO 2016/073799. In some embodiments, the ccDNA vector is also present in the same nanocarrier. In some embodiments, the ccDNA vector formulated with a lipid nanoparticle is also present in the same nanocarrier.

[00362] In some embodiments, the rapamycin analog is any of the rapamycin analogs known in the art, such as any of the rapamycin analogs described in U.S. Patent 5,138,051 or WO 2017/040341, the contents of each of which are incorporated herein by reference in their entirety.

[00363] In some embodiments, the rapamycin analog is a compound of Formula I, shown below:

[00364] In some embodiments, the rapamycin analog is a compound of Formula II wherein the configuration of the substituents at C-33 of Formula I is the R-configuration, as shown below:

[00365] In some embodiments, the rapamycin analog is a compound of Formula III, as shown below:

or a pharmaceutically acceptable salt thereof, wherein: R ¹ is OH or OCH ₃ R ² is H or FR ³ is H, OH or OCH ₃ ; and R ⁴ is OH or OCH ₃ .

[00366] In some embodiments, the rapamycin analog is a compound of Formula III in pure chiral form as a single diastereomer of Formula IV, as shown below:

[00367] In some embodiments, the rapamycin analog is a compound of Formula III in pure chiral form as a single diastereomer of Formula V, as shown below:

[00368] In some embodiments, the rapamycin analog is a compound of Formula III in pure chiral form as a single diastereomer of Formula VI, as shown below:

[00369] In some embodiments, the rapamycin analog is a compound of Formula III in pure chiral form as a single diastereomer of Formula VII, as shown below:

[00370] In some embodiments, the rapamycin analog is a compound of Formula III in pure chiral form as a single diastereomer of Formula VIII, as shown below:

[00371] In some embodiments, the rapamycin analog is a compound of Formula IX, as shown below:

or a pharmaceutically acceptable salt thereof, wherein R ² is H or F, R ³ is OH or OCH ₃ ; and R ⁴ is OCH ₃ or OH. In certain embodiments, R ⁴ is OCH ₃ . In certain embodiments, R4 is OCH ₃ , R2 is F, and R ³ is OCH3. In certain embodiments, R ⁴ is OCH ₃ , R ² is H, and R ³ is OH. In certain embodiments, R ² is H, R ₃ is H, and R ⁴ is OH. In various embodiments, the compounds of Formula IX are provided as a racemic mixture.

[00372] Accordingly, in some embodiments, the rapamycin analog is selected from any of Formulas I-IX or a derivative thereof.

[00373] In some embodiments, rapamycin or a rapamycin analog is delivered or administered using a synthetic nanocarrier as described in WO 2016/073799, incorporated herein by reference in its entirety.

[00374] As described in WO 2016/073799, the concentration of rapamycin in the formulation during the preparation of the synthetic nanocarrier, relative to the solubility limit of rapamycin in said formulation, can significantly affect the ability of the resulting synthetic nanocarriers to induce immune tolerance. In addition, the nature of the dispersion of rapamycin in the synthetic nanocarriers can affect the ability to initially sterilize by filtration of the resulting synthetic nanocarriers. Accordingly, in some embodiments, synthetic nanocarriers created under conditions that result in a concentration of rapamycin exceeding its solubility in the resulting nanocarrier suspension are used in the compositions and methods described herein. Such synthetic nanocarriers can provide longer-lasting immune tolerance and can be initially sterilized by filtration.

[00375] According to some embodiments, the ccDNA vector is administered in conjunction with a composition comprising synthetic nanocarriers comprising a hydrophobic polyester carrier material and rapamycin or a rapamycin analog, wherein said rapamycin or rapamycin analog is present in the synthetic nanocarriers in a stable, supersaturated amount that is less than 50% (w/w) based on the weight of rapamycin or rapamycin analog relative to the weight of the hydrophobic polyester carrier material.

[00376] According to one embodiment of any of the compositions or methods provided herein, the mass values are recipe mass values of materials that are combined during the manufacture of the synthetic nanocarriers. According to one embodiment of any of the compositions or methods provided herein, the mass values are mass values of materials in the resulting synthetic nanocarrier composition.

[00377] In some embodiments of any of the compositions or methods provided herein, rapamycin or a rapamycin analog is present in a stable, supersaturated amount that is less than 45% (w/w). In one embodiment of any of the compositions or methods provided herein, rapamycin or a rapamycin analog is present in a stable, supersaturated amount that is less than 40% (w/w). In one embodiment of any of the compositions or methods provided herein, rapamycin or a rapamycin analog is present in a stable, supersaturated amount that is less than 35% (w/w). In one embodiment of any of the compositions or methods provided herein, rapamycin or a rapamycin analog is present in a stable, supersaturated amount that is less than 30% (w/w). In one embodiment of any of the compositions or methods provided herein, rapamycin or a rapamycin analog is present in a stable, supersaturated amount that is less than 25% (w/w). In one embodiment of any of the compositions or methods provided herein, rapamycin or a rapamycin analog is present in a stable, supersaturated amount that is less than 20% (w/w). In one embodiment of any of the compositions or methods provided herein, rapamycin or a rapamycin analog is present in a stable, supersaturated amount that is less than 15% (w/w). In one embodiment of any of the compositions or methods provided herein, rapamycin or a rapamycin analog is present in a stable, supersaturated amount that is less than 10% (w/w). According to one embodiment of any of the compositions or methods provided herein, rapamycin or a rapamycin analog is present in a stable, supersaturated amount that is greater than 7% (w/w).

[00378] According to one embodiment of any of the compositions or methods provided herein, the hydrophobic polyester carrier material comprises PLA, PLG, PLGA, or polycaprolactone. According to one embodiment of any of the compositions or methods provided herein, the hydrophobic polyester carrier material further comprises PLA-PEG, PLGA-PEG, or PCL-PEG.

[00379] According to one embodiment of any of the compositions or methods provided herein, the amount of hydrophobic polyester carrier material in the synthetic nanocarriers is 5-95% (w/w) hydrophobic polyester carrier material/total solids. According to one embodiment of any of the compositions or methods provided herein, the amount of hydrophobic polyester carrier material in the synthetic nanocarriers is 60-95% (w/w) hydrophobic polyester carrier material/total solids.

[00380] According to one embodiment of any of the compositions or methods provided herein, the synthetic nanocarriers further comprise a nonionic surfactant with an HLB value of less than or equal to 10. According to one embodiment of any of the compositions or methods provided herein, the nonionic surfactant with an HLB value of less than or equal to 10 comprises a sorbitan ester, an aliphatic alcohol, an aliphatic acid ester, an ethoxylated aliphatic alcohol, a poloxamer, an aliphatic acid, cholesterol, a cholesterol derivative, or a bile acid or salt. According to one embodiment of any of the compositions or methods provided herein, the nonionic surfactant with an HLB value of less than or equal to 10 comprises SPAN 40, SPAN 20, oleyl alcohol, stearyl alcohol, isopropyl palmitate, glycerol monostearate, BRIJ 52, BRIJ 93, pluronic acid P-123, pluronic acid L-31, palmitic acid, dodecanoic acid, glycerol tripalmitate, or glycerol trilinoleate. According to one embodiment of any of the compositions or methods provided herein, the nonionic surfactant with an HLB value of less than or equal to 10 is SPAN 40.

[00381] According to one embodiment of any of the compositions or methods provided herein, a nonionic surfactant with an HLB value of less than or equal to 10 is encapsulated in the synthetic nanocarriers, present on the surface of the synthetic nanocarriers, or both. According to one embodiment of any of the compositions or methods provided herein, the amount of the nonionic surfactant with an HLB value of less than or equal to 10 is >0.1 but <15% (w/w) of the nonionic surfactant with an HLB value of less than or equal to 10/hydrophobic polyester carrier material. In one embodiment of any of the compositions or methods provided herein, the amount of nonionic surfactant with an HLB value of less than or equal to 10 is >1 but <13% (w/w) of nonionic surfactant with an HLB value of less than or equal to 10/hydrophobic polyester carrier material. In one embodiment of any of the compositions or methods provided herein, the amount of nonionic surfactant with an HLB value of less than or equal to 10 is >1 but <9% (w/w) of nonionic surfactant with an HLB value of less than or equal to 10/hydrophobic polyester carrier material.

[00382] According to one embodiment of any of the compositions or methods provided herein, the composition may initially be sterilized by filtration through a filter with a pore size of 0.22 microns.

[00383] In one embodiment of any of the compositions or methods provided herein, the average particle size distribution obtained using dynamic light scattering of synthetic nanocarriers is greater than 120 nm in diameter. In one embodiment of any of the compositions or methods provided herein, the diameter is greater than 150 nm. In one embodiment of any of the compositions or methods provided herein, the diameter is greater than 200 nm. In one embodiment of any of the compositions or methods provided herein, the diameter is greater than 250 nm. In one embodiment of any of the compositions or methods provided herein, the diameter is less than 300 nm. In one embodiment of any of the compositions or methods provided herein, the diameter is less than 250 nm. In one embodiment of any of the compositions or methods provided herein, the diameter is less than 200 nm.

[00384] According to one embodiment of any of the compositions or methods provided herein, rapamycin or a rapamycin analog is encapsulated in synthetic nanocarriers.

[00385] According to one embodiment of any of the compositions or methods provided herein, the composition further comprises a pharmaceutically acceptable carrier.

[00386] In one embodiment of any of the compositions or methods provided herein, the rapamycin or rapamycin analog is present in a supersaturated amount that is at least 1% greater than the saturation limit of rapamycin or the rapamycin analog in the hydrophobic polyester carrier material. In one embodiment of any of the compositions or methods provided herein, the rapamycin or rapamycin analog is present in a supersaturated amount that is at least 5% greater than the saturation limit of rapamycin or the rapamycin analog in the hydrophobic polyester carrier material. In one embodiment of any of the compositions or methods provided herein, the rapamycin or rapamycin analog is present in a supersaturated amount that is at least 10% greater than the saturation limit of rapamycin or the rapamycin analog in the hydrophobic polyester carrier material. In one embodiment of any of the compositions or methods provided herein, the rapamycin or rapamycin analog is present in a supersaturated amount that is at least 15% greater than the saturation limit of rapamycin or rapamycin analog in the hydrophobic polyester carrier material. In one embodiment of any of the compositions or methods provided herein, the rapamycin or rapamycin analog is present in a supersaturated amount that is at least 20% greater than the saturation limit of rapamycin or rapamycin analog in the hydrophobic polyester carrier material. In one embodiment of any of the compositions or methods provided herein, the rapamycin or rapamycin analog is present in a supersaturated amount that is at least 25% greater than the saturation limit of rapamycin or rapamycin analog in the hydrophobic polyester carrier material. According to one embodiment of any of the compositions or methods provided herein, rapamycin or a rapamycin analog is present in a supersaturated amount that is at least 30% greater than the saturation limit of rapamycin or a rapamycin analog in the hydrophobic polyester carrier material.

[00387] In another embodiment of any of the compositions or methods provided herein, the amount of rapamycin or rapamycin analog exceeds the saturation limit by at least 1%. In another embodiment, the amount of rapamycin or rapamycin analog exceeds the saturation limit by at least 5%. In another embodiment, the amount of rapamycin or rapamycin analog exceeds the saturation limit by at least 10%. In another embodiment, the amount of rapamycin or rapamycin analog exceeds the saturation limit by at least 15%. In another embodiment, the amount of rapamycin or rapamycin analog exceeds the saturation limit by at least 20%. In another embodiment, the amount of rapamycin or rapamycin analog exceeds the saturation limit by at least 25%. In another embodiment, the amount of rapamycin or rapamycin analog exceeds the saturation limit by at least 30%.

cGAS inhibitors

[00388] According to some aspects of the present disclosure, non-viral capsidless DNA vectors with covalently closed ends (ccDNA) are provided that are administered in combination with one or more cGAS antagonists. Also provided herein are ccDNA constructs comprising sequences encoding, among other things, one or more RNAs or proteins that inhibit cGAS.

[00389] cGAS is another class of cytosolic DNA-triggered PRR that binds to DNA and activates ER-associated stimulator of interferon genes (STING). This leads to activation of the type I interferon response and, in some cases, activation of other putative cytosolic DNA sensors, including absent in melanoma protein 2 (AIM2), IFN-γ-inducible protein 16 (IFI16), interferon-inducible protein X (IFIX), LRRFIP1, DHX9, DHX36, DDX41, Ku70, DNA-PKc, MRN complex (including MRE11, Rad50, and Nbs1) and RNA polymerase III. AIM2, IFI16, and IFIX are proteins containing the pyrin domain and the HIN200 (PYHIN) domain. Furthermore, it was shown that unpaired DNA nucleotides flanking short stretches of base-paired DNA, as in the stem-loop structures of single-stranded DNA (ssDNA) derived from human immunodeficiency virus type 1 (HIV-1), activated the type I interferon-induced DNA sensor cGAS in a sequence-dependent manner. DNA structures containing unpaired guanosines flanking short (12 to 20 bp) dsDNA (Y-form DNA) had a strong stimulatory effect and specifically enhanced the enzymatic activity of cGAS

[00390] cGAS directly binds DNA through interactions with the sugar-phosphate backbone of both DNA strands (S.R. Paluden. Microbiology and Molecular Biology Reviews. 2015. 79(2): 225). This causes a conformational change in the enzyme that allows the nucleotide substrates ATP and GTP to access the active site, leading to the synthesis of cGAMP (A. Dempsey and A.G. Bowie, Virology 2015 May, 0: 146-152). cGAMP then binds STING, leading to the production of type I interferon (A. Dempsey and A.G. Bowie, Virology 2015 May, 0: 146-152). Importantly, cGAS contacts dsDNA only through the phosphate backbone of DNA, resulting in sequence-independent sensitivity (A. Dempsey and A.G. Bowie, Virology 2015 May, 0: 146–152). It has also been shown that cGAS can be activated by unpaired DNA nucleotides, in particular guanosines flanking short base-paired regions of DNA of 12–20 bp, as in the stem-loop structures of single-stranded DNA (ssDNA) derived from human immunodeficiency virus type 1 (HIV-1) (M.H. Christnesen and S.R. Paluden. Cellular and Molecular Immunology. 2017. 14:4–13; A-M Herzner et al., 2015. Nature Immunology).

[00391] Accordingly, structural features of cDNA important for innate immune activation by PRRs include, but are not limited to, modified AAV inverted terminal repeat (ITR) sequences, including the Rep binding site (RBS) and the terminal resolution site (TRS); hairpin sequences within the ITR; a CG-rich RBS sequence; lack of DNA methylation; and a linear duplex DNA structure with flanking ITRs that may contain, for example, single-stranded DNA in the form of a loop.

[00392] In some embodiments of the compositions and methods described herein, the cGAS inhibitor is administered to the subject in conjunction with the ccDNA. In some embodiments of the compositions and methods described herein, wherein the cGAS inhibitor is an RNA or protein sequence, the ccDNA encodes an RNA or protein inhibitor of cGAS.

[00393] In some embodiments, the cGAS inhibitor is an antimalarial drug (J. An et al., J. Immunol. March 27, 2015). In some embodiments, the antimalarial drug is an aminoquinoline or aminoacridine antimalarial drug (J. An et al., J. Immunol. March 27, 2015). In some embodiments, the antimalarial drug is selected from quinacrine (QC), 9-amino-6-chloro-2-methoxyacridine (AMCA), hydroxychloroquine (HCQ), and chloroquine (CQ) (J. An et al., J. Immunol. March 27, 2015).

[00394] In some embodiments, the cGAS inhibitor is a small molecule compound that binds to the catalytic pocket of cGAS (J. Vincent et al., Nature Communications, 8:750). In some embodiments, the small molecule compound that binds to the catalytic pocket of cGAS is selected from RU166365, RU281332, RU320521, RU320519, RU320461, RU320462, RU320520, RU320467, and RU320582 (J. Vincent et al., Nature Communications, 8:750). In some embodiments, the small molecule compound that binds to the catalytic pocket of cGAS is RU320521 (J. Vincent et al., Nature Communications, 8:750). In some embodiments, the small molecule compound that binds to the catalytic pocket of cGAS is selected from compound 15, compound 16, compound 17, compound 18, compound 19, and PF-06928215 (J. Vincent et al., Nature Communications, 8:750; PLOS ONE. September 21, 2017). In some embodiments, the small molecule compound that binds to the catalytic pocket of cGAS is PF-06928215 (PLOS ONE. September 21, 2017).

[00395] In some embodiments, the cGAS inhibitor is any of the small molecule compounds described in US 20160068560, the contents of which are incorporated herein by reference in their entirety.

[00396] According to some embodiments of the compositions and methods described herein, the cGAS inhibitor is encoded by ccDNA administered to the subject (including, for example, subsequent delivery of ccDNA). According to some embodiments of the compositions and methods described herein, the cGAS inhibitor encoded by the ccDNA administered to the subject is the Kaposi's sarcoma-associated herpesvirus ORF52 protein having the amino acid sequence MAAPRGRPKKDLTMEDLTAKISQLTVENRELRKALGSTADPRDRPLTATEKEAQLTATVGALSAAAAKKIEARVRTIFSKVVTQKQVDDALKGLSLRIDVCMSDGGTAKPPPGANNRRRRGASTTRAGVDD (SEQ ID NO: 882), or a variant thereof that inhibits cGAS (M.H. Christnesen and S.R. Paluden. Cellular and Molecular Immunology. 2017. 14:4-13). According to some embodiments of the compositions and methods described herein, the cGAS inhibitor encoded by the ccDNA administered to the subject is an ortholog of ORF52 of a gammaherpesvirus.

[00397] According to some embodiments of the compositions and methods described herein, the cGAS inhibitor encoded by the ccDNA administered to the subject is the cytoplasmic isoform of LANA (latency-associated nuclear antigen) of Kaposi's sarcoma-associated herpesvirus, also referred to herein as the "cytoplasmic isoform of LANA", or a variant thereof that inhibits cGAS (Zhang G. et al., Proc Natl Acad Sci U S A. 2016 Feb 23;113(8):E1034-43). LANA or ORF73 has a sequence of the following 1129 amino acids: MAPPGMRLRSGRSTGAPLTRGSCRKRNRSPERCDLGDDLHLQPRRKHVADSVDGRECGPHTLPIPGSPTVFTSGLPAFVSSPTLPVAPIPSPAPATPLPPPALLPPVTTSSSPIPPSHPVSPGTTDTHSPSPALPPTQSPE SSQRPPLSSPTGRPDSSTPMRPPPSQTTPPHSPTTPPPEPPSKSSPDSLAPSTLRSLRKRRLSSPQGPSTLNPICQSPPVSPPRCDFANRSVYPPWATESPIYVGSSSDGDTPPRQPPTSPISIGSSSPSEGSWGDDTAM LVLLAEIAEEASKNEKECSENNQAGEDNGDNEISKESQVDKDDNDNKDDEEEQETDEEDEEEDDEEDDEEDDEEDDEEDDEEDDEEDDEEEDEEEDEEEDEEEDEEEEEEDDDDEDNEDEEDDEEEDKKEDEEDGGDGNK TLSIQSSQQQQEPQQQEPQQQEPQQQEPQQQEPQQQEPQQQEPQQQEPQQREPQQREPQQREPQQREPQQREPQQREPQQREPQQREPQQREPQQREPQQREPQQREPQQQEPQQQEPQQQEPQQQEPQQQEPQQQEPQQQ EPQQQEPQQQEPQQQEPQQQEPQQQEPQQQDEQQQDEQQQDEQQQDEQQQDEQQQDEQQQDEQQQDEQEQQDEQQQDEQQQQDEQEQQEEQEQQEEQQQDEQQQDEQQQDEQQQDEQEQQDEQQQDEQQQQDEQEQQEEQE QQEEQEQQEEQEQQEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQEQELEEVEEQEQEQEEQELEEVEE QEQEQEEQEEQELEEVEEQEEQELEEVEEQEEQELEEVEEQEQQGVEQQEQETVEEPIILHGSSSEDEMEVDYPVVSTHEQIASSPPGDNTPDDDPQPGPSREYRYVLRTSPPHRPGVRMRRVPVTHPKKPHPRYQQPPVP YRQIDDCPAKARPQHIFYRRFLGKDGRRDPKCQWKFAVIFWGNDPYGLKKLSQAFQFGGVKAGPVSCLPHPGPDQSPITYCVYVYCQNKDTSKKVQMARLAWEASHPLAGNLQSSIVKFKKPLPLTQPGENQGPGDSPQEMT (SEQ ID NO: 883).

[00398] A non-limiting example of a truncated cytoplasmic isoform of LANA for use with the ccDNAs described herein is LANAΔ161 or SEQ ID NO: 532 (which lacks amino acids 161-1162 of SEQ ID NO: 884).

[00399] In some embodiments of the compositions and methods described herein, the cGAS inhibitor is an antibody or antigen-binding fragment that binds cGAS. In some embodiments of the compositions and methods described herein, the antibody or antigen-binding fragment that binds cGAS is encoded by ccDNA.

[00400] In some embodiments of the compositions and methods described herein, the cGAS inhibitor is an RNA-based cGAS inhibitor, such as a siRNA specific for cGAS. In some embodiments of the compositions and methods described herein, the RNA-based cGAS inhibitor is encoded by ccDNA.

[00401] According to some embodiments of the compositions and methods described herein, the cGAS inhibitor is an miRNA-based cGAS inhibitor, such as miR-25 (GGCCAGTGTTGAGAGGCGGAGACTTGGGCAATTGCTGGACGCTGCCCTGGGCATTGCACTTGTCTCGGTCTGACAGTGCCGGCC; SEQ ID NO: 885) and miR-93 (CTGGGGGCTCCAAAGTGCTGTTCGTGCAGGTAGTGTGATTACCCAACCTACTGCTGAGCTAGCACTTCCCGAGCCCCCGG; SEQ ID NO: 886) ¹¹ . MiR-25 and miR-93 are believed to target nuclear receptor coactivator 3 (NCOA3), an epigenetic factor that maintains basal levels of cGAS expression, leading to cGAS repression (Wu et al. 2017. Nat. Cell Biol. 19(10):1286-1296). In some embodiments of the compositions and methods described herein, the miRNA-based cGAS inhibitor is encoded by ccDNA.

TLR inhibitors

[00402] According to some aspects of the present disclosure, non-viral capsidless DNA vectors with covalently closed ends (ccDNA) are provided that are administered in combination with one or more TLR antagonists. Also provided herein are ccDNA constructs comprising sequences encoding, among other things, one or more TLR inhibitory oligonucleotides. According to some aspects of the present disclosure, non-viral capsidless DNA vectors with covalently closed ends (ccDNA) are provided that are administered in combination with one or more TLR9 antagonists. Also provided herein are ccDNA constructs comprising sequences encoding, among other things, one or more TLR9 inhibitory oligonucleotides.

[00403] In some embodiments, the TLR9 inhibitor is a small molecule antagonist. In another embodiment, the TLR9 inhibitor is an anti-TLR9 antibody. In some embodiments, the anti-TLR9 antibody is a monoclonal antibody. In some embodiments of the compositions and methods described herein, one or more terminal structural elements of the ccDNA, such as ITR sequences, comprise a TLR9 inhibitory oligonucleotide sequence.

[00404] In some embodiments of the compositions and methods described herein, the TLR9 inhibitory oligonucleotide has one or more of the following features: (i) three consecutive G nucleotides at the 3' end; (ii) a CC(T) triplet at the 5' end; and (iii) the distance between the 5'-CC(T) and the subsequent GGG triplet is optimally 3-5 nucleotides in length. In some embodiments, the TLR9 inhibitory oligonucleotide has the sequence 5'CCTN(3-5)G(3-5)RR3' (SEQ ID NO: 887). In some embodiments, the TLR9 inhibitory oligonucleotide does not have an intrastrand and/or interstrand Hoogsteen hydrogen bond between adjacent Gs.

[00405] According to some embodiments of the compositions and methods described herein, the TLR9 inhibitory oligonucleotide is a class G TLR9 inhibitory oligonucleotide having G4 stacking characteristics and comprises multiple G3 triplets or G4 tetrads, such as an inhibitory oligonucleotide comprising TTAGGGn (SEQ ID NO: 888). Non-limiting examples of such a TLR9 inhibitory class G oligonucleotide include ODN-2088 (TCCTGGCGGGGAAGT, SEQ ID NO: 889), ODN-2114 (TCCTGGAGGGGAAGT, SEQ ID NO: 890), poly(G) (GGGGGGGGGGGGGGGGGGGGGG, SEQ ID NO: 891), ODN-A151 (TTAGGGTTAGGGTTAGGGTTAGGG, SEQ ID NO: 892), G-ODN (CTCCTATTGGGGGTTTCCTAT, SEQ ID NO: 893), as well as IRS-869 (TCCTGGAGGGGTTGT, SEQ ID NO: 894) and AS1411 (GGTGGTGGTGGTGGTTGTGGTGGTGGTGG, SEQ ID NO: 903).

[00406] According to some embodiments of the compositions and methods described herein, the TLR9 inhibitory oligonucleotide is a Class R TLR9 inhibitory oligonucleotide having characteristics including palindromicity and/or the presence of short 5' or 3' overhangs, such as an INH-1 inhibitory oligonucleotide. Non-limiting examples of such a Class R TLR9 inhibitory oligonucleotide include INH-1 (CCTGGATGGGAATTCCCATCCAGG, SEQ ID NO: 895), INH-4 (TTCCCATCCAGGCCTGGATGGGAA, SEQ ID NO: 896), and IRS-661 (TGCTTGCAAGCTTGCAAGCA, SEQ ID NO: 897).

[00407] According to some embodiments of the compositions and methods described herein, the TLR9 inhibitory oligonucleotide is a class B TLR9 inhibitory oligonucleotide having linear characteristics and a 5'-CC(T)→GGG-3' motif, such as the INH-18 inhibitory oligonucleotide. Non-limiting examples of such a class B TLR9 inhibitory oligonucleotide include ODN-2088 (TCCTGGCGGGGAAGT, SEQ ID NO: 889), ODN-2114 (TCCTGGAGGGGAAGT, SEQ ID NO: 890), 4024 (TCCTGGATGGGAAGT, SEQ ID NO: 898), 4084F (CCTGGATGGGAA, SEQ ID NO: 899), INH-13 (CTTACCGCTGCACCTGGATGGGAA, SEQ ID NO: 900), INH-18 (CCTGGATGGGAACTTACCGCTGCA, SEQ ID NO: 901), G-ODN (CTCCTATTGGGGGTTTCCTAT, SEQ ID NO: 893), IRS-869 (TCCTGGAGGGTTGT, SEQ ID NO: 864), IRS-954 TGCTCCTGGAGGGGTTGT, SEQ ID NO: 902) and AS1411 (GGTGGTGGTGGTTGTGGTGGTGGTGG, SEQ ID NO: 903).

[00408] According to some embodiments of the compositions and methods described herein, a coding sequence encoded by a ccDNA, such as a transgene sequence, is modified such that CpG dinucleotides located in a codon triplet for a selected amino acid are replaced by a codon triplet for the same amino acid that lacks a CpG dinucleotide.

[00409] In some embodiments of the compositions and methods described herein, wherein the TLR9 inhibitor is an RNA or protein sequence, the ccDNA encodes an RNA or protein inhibitor of TLR9. In some embodiments of the compositions and methods described herein, the TLR9 inhibitor is an antibody or antigen-binding fragment that binds TLR9. In some embodiments of the compositions and methods described herein, the antibody or antigen-binding fragment that binds TLR9 is encoded by ccDNA.

[00410] According to some embodiments of the compositions and methods described herein, a TLR9 inhibitor is administered to the subject in conjunction with the ccDNA. Non-limiting examples of TLR9 inhibitors can be found in "Classification, Mechanisms of Action, and Therapeutic Applications of Inhibitory Oligonucleotides for Toll-Like Receptors (TLR) 7 and 9," P.S. Lenert, Mediators of Inflammation, Vol. 2010, 986596; US20150203850; and US2017026800, the contents of each of which are incorporated herein by reference in their entireties.

[00411] Accordingly, in some embodiments of the compositions and methods described herein, the TLR9 inhibitor is administered to the subject in conjunction with the ccDNA.

[00412] In some embodiments of the compositions and methods described herein, the TLR9 inhibitor is encoded in cis by the ccDNA administered to the subject (including, for example, subsequent delivery of the ccDNA). In some embodiments of the compositions and methods described herein, the TLR9 inhibitor is administered in trans by the ccDNA administered to the subject.

[00413] According to some embodiments of the compositions and methods described herein, the TLR9 inhibitor is an oligonucleotide that inhibits TLR9.

[00414] In some embodiments of the compositions and methods described herein, the TLR9 inhibitory oligonucleotide has one or more of the following features: (i) three consecutive G nucleotides at the 3' end; (ii) a CC(T) triplet at the 5' end; and (iii) the distance between the 5'CC(T) and the subsequent GGG triplet is optimally 3-5 nucleotides in length. In some embodiments, the TLR9 inhibitory oligonucleotide has the sequence 5'CCTN(3-5)G(3-5)RR3' (SEQ ID NO: 887). In some embodiments, the TLR9 inhibitory oligonucleotide does not have an intrastrand and/or interstrand Hoogsteen hydrogen bond between adjacent Gs.

[00415] In some embodiments of the compositions and methods described herein, the TLR9 inhibitor is an antibody or antigen-binding fragment that binds TLR9. In some embodiments of the compositions and methods described herein, the antibody or antigen-binding fragment that binds TLR9 is encoded by ccDNA.

[00416] According to some embodiments of the compositions and methods described herein, the TLR9 inhibitor is an endosomal acidification inhibitor, such as chloroquine.

Inflammasome antagonists

NLRP3 inflammasome pathway inhibitors:

[00417] In some embodiments, the inflammasome antagonist inhibits NLRP3. The term "NLRP3", also referred to as cryopyrin, refers to the NOD-like receptor family pyrin domain-containing inflammasome 3 or NACHT, LRR, and PYD domain-containing protein 3 (NALP3), also known as cryopyrin, cold-inducible autoinflammatory syndrome 1 (CIAS1), caterpillar-like receptor 1.1 (CLR1.1), or pyrin domain-containing Apafl-like protein 1 (PYPAF1). NALP3 is also known by other names: NLRP3 PYD-NACHT-NAD-LRR NALP3 Cias1, Pypaf1, Mmig1 PYD-NACHT-NAD-LRR. NLRP3 is a component of a polyprotein oligomer consisting of NLRP3 protein, ASC (apoptosis-associated speck-like protein containing CARD), and procaspase 1.

[00418] NLRP3 inhibitors contemplated for use in the methods and compositions described herein are disclosed in Shao, Bo-Zong, et al., “NLRP3 inflammasome and its inhibitors: a review.” Frontiers in pharmacology 6 (2015): 262, and Wang et al., Lab investigation, 2017, 97; 922-934, which are incorporated herein by reference in their entireties.

[00419] In some embodiments, the NLRP3 inflammasome inhibitor is MCC950 or a functional derivative thereof. MCC950 has the formula:

and is a potent and selective inhibitor of NLRP3. MCC950 blocks the release of IL-1β induced by NLRP3 activators such as ATP, MSU, and nigericin by preventing the oligomerization of the inflammasome adaptor protein ASC (apoptosis-associated speck-like protein containing CARD) (Coll RC. et al., 2015. A small-molecule inhibitor of the NLRP3 inflammasome for the treatment of inflammatory diseases. Nature Med 21(3), 248–255.; Guo H. et al., 2015. Inflammasomes: mechanism of action, role in disease, and therapeutics. Nat Med. 21(7):677–87; Ren, Honglei, et al. “Selective NLRP3 (Pyrin Domain-Containing Protein 3) Inflammasome Inhibitor Reduces Brain Injury After Intracerebral Hemorrhage." Stroke (2017): STROKEAHA-117.).

[00420] In some embodiments, the NLRP3 inflammasome inhibitor is Bay11-7082, which has the following structure:

and have been reported to selectively inhibit NLRP3 inflammasome activity in macrophages, independent of their inhibitory effect on NF-κB activity (Juliana C. et al, 2010. Anti-inflammatory Compounds Parthenolide and Bay11-7082 Are Direct Inhibitors of the Inflammasome. J. Biol Chem. 285(13): 9792–9802].

[00421] In some embodiments, the NLRP3 inflammasome inhibitor is glibenclamide (also known as glyburide), which has the following structure:

which blocks the maturation of caspase-1 and pro-IL-1β by inhibiting K ⁺ efflux (Laliberte RE et al., 1999. ATP treatment of human monocytes promotes caspase-1 maturation and externalization. J Biol Chem. 274(52):36944-51). Glibenclamide also potently blocks the activation of the NRLP3 inflammasome induced by PAMPs, DAMPs, and crystalline substances (Lamkanfi M et al., 2009. Glyburide inhibits the Cryopyrin/Nalp3 inflammasome. J. Cell Biol., 187: 61 - 70; Dostert C et al., 2009. Malarial hemozoin is a Nalp3 inflammasome activating danger signal. PLoS One. 4(8): e6510).

[00422] In some embodiments, the NLRP3 inflammasome inhibitor is isoliquiritigenin (also known as ILG), which has the following structure:

which is a chalcone-type flavonoid isolated from licorice root (Glycyrrhiza uralensis) and has been reported to inhibit NLRP3-activated ASC oligomerization (Honda H. et al., 2014. Isoliquiritigenin is a potent inhibitor of NLRP3 inflammasome activation and diet-induced adipose tissue inflammation. J Leukoc Biol. 96(6):1087-100.). NLRP3-dependent IL-1β production was inhibited by low concentrations of isoliquiritigenin (1 to 10 μM), indicating that isoliquiritigenin can block the NLRP3 inflammasome at both the priming and activation stages.

[00423] In some embodiments, the NLRP3 inflammasome inhibitor is 6673-34-0; (5-chloro-2-methoxy-N-[2-(4-sulfamoylphenyl)ethyl]benzamide)), which is disclosed in US20160052876, which is incorporated herein by reference in its entirety. In some embodiments, the NLRP3 inflammasome inhibitor is any of the small molecule compounds described in US20160052876, the disclosure of which is incorporated herein by reference in its entirety.

[00424] In some embodiments, the NLRP3 inflammasome inhibitor is a cysteinyl leukotriene receptor antagonist disclosed in Ozaki et al., 2015; Coll et al., 2011; Haerter et al., 2009 and U.S. Patent 7,498,460, which are each incorporated by reference in their entireties. The cysteinyl leukotriene receptor antagonist has been reported to inhibit both the NLRP3 inflammasome and AIM2-induced IL-1 processing by preventing ASC oligomerization and also appears to have other functions in innate immune responses other than its role as an adaptor for inflammasome formation (Ozaki et al., 2015).

[00425] In some embodiments, small molecule inhibitors targeting NLRP3 and AIM2 have been characterized and widely described in (Ozaki et al., 2015). The vast majority of these are pharmacological inhibitors that have been repurposed to target the inflammasome (Guo et al., 2015) and include: parthenolide (Juliana et al., 2010), Bay 11-708 (Juliana et al., 2010), CRID3 (Coll et al., 2011), auranofin (Isakov et al., 2014), isoliquiritigenin (Honda et al., 2014), 3,4-methylenedioxy-*-nitrostyrene (He et al., 2014), cyclopentenone prostaglandin 15d-PJ2 (Maier et al., 2015), and 25-hydroxycholesterol (25-HC) (Reboldi et al., 2014). Moreover, type I interferon has also been shown to suppress inflammasome activation through a poorly understood mechanism (Guarda et al., 2011). However, it has recently been demonstrated that the IFN-stimulated gene product, cholesterol 25-hydroxylase (Ch25h), antagonizes both IL1b transcription and activation of the NLRP3, NLRC4, and AIM2 inflammasomes, suggesting that Ch25h has broad inhibitory activity across multiple inflammasomes (Reboldi et al., 2014).

[00426] The coding sequence of NLRP3 is provided under NCBI accession numbers NM _- 004895.1 (SEQ ID NO: 530), NM _- 183395 (SEQ ID NO: 531), NM _- 001079821 (SEQ ID NO: 532), NM _- 001127461 (SEQ ID NO: 533), and NM _- 001127462 (SEQ ID NO: 534). In this case, the translation initiation codon of NLRP3 is preferably a codon located 6 nucleotides after the translation initiation codon described in each of these NCBI accession numbers. Examples of a mutant NLRP3 gene include an NLRP3 gene in which an adenine at position 1709, counted from the translation initiation codon (in the case of the coding region shown under the NCBI accession numbers, position 1715, counted from the translation initiation codon) is guanine, a cytosine at position 1043 (position 1049 in the coding region shown under the NCBI accession numbers), counted from the translation initiation codon, is thymine, or a guanine at position 587 (position 593 in the coding region shown under the NCBI accession numbers), counted from the translation initiation codon, is adenine. NLRP3 is preferably a variant in which the nucleotide at position 1079 is mutated to guanine. As one skilled in the art will appreciate, there may be variants of the NLRP3 gene that encode a functionally equivalent NLRP3 that retains the function, at least in part, of activating caspase-1 and/or stimulating the maturation of inflammatory cytokines such as interleukin 1β and interleukin 18. Such a functionally equivalent NLRP3 may, accordingly, include amino acid substitutions, deletions, or additions that do not eliminate activity.

[00427] In some embodiments of the compositions and methods described herein, the NLRP3 inflammasome inhibitor is an RNA-based NLRP3 inhibitor (RNAi), such as an siRNA specific for NLRP3. In some embodiments of the compositions and methods described herein, the RNA-based NLRP3 inhibitor is encoded by ccDNA. The NLRP3 siRNA may be commercially available, such as SI03060323 (Qiagen®).

[00428] In some embodiments, the NLRP3 inhibitor is RNAi encoded in ccDNA. For the avoidance of doubt, the amino acid sequence of the human NLRP3 protein is NM _- 004895.1 (SEQ ID NO: 539) and is provided below:

MKMASTRCKLARYLEDLEDVDLKKFKMHLEDYPPQKGCIPLPRG QTEKADHVDLATLMIDFNGEEKAWAMAVWIFAAINRRDLYEKAKRDEPKWGSDNARVS NPTVICQEDSIEEEWMGLLEYLSRISICKMKKDYRKKYRKYVRSRFQCIEDRNARLGE SVSLNKRYTRLRLIKEHRSQQEREQELLAIGKTKTCESPVSPIKMELLFDPDDEHSEP VHTVVFQGAAGIGKTILARKMMLDWASGTLYQDRFDYLFYIHCREVSLVTQRSLGDLI MSCCPDPNPPIHKIVRKPSRILFLMDGFDELQGAFDEHIGPLCTDWQKAERGDILLSS LIRKKLLPEASLLITTRPVALEKLQHLLDHPRHVEILGFSEAKRKEYFFKYFSDEAQA RAAFSLIQENEVLFTMCFIPLVCWIVCTGLKQQMESGKSLAQTSKTTTAVYVFFLSSL LQPRGGSQEHGLCAHLWGLCSLAADGIWNQKILFEESDLRNHGLQKADVSAFLRMNLF QKEVDCEKFYSFIHMTFQEFFAAMYYLLEEEKEGRTNVPGSRLKLPSRDVTVLLENYG KFEKGYLIFVVRFLFGLVNQERTSYLEKKLSCKISQQIRLELLKWIEVKAKAKKLQIQ PSQLELFYCLYEMQEEDFVQRAMDYFPKIEINLSTRMDHMVSSFCIENCHRVESLSLG FLHNMPKEEEEEEKEGRHLDMVQCVLPSSSHAACSHGLVNSHLTSSFCRGLFSVLSTS QSLTELDLSDNSLGDPGMRVLCETLQHPGCNIRRLWLGRCGLSHECCFDISLVLSSNQ KLVELDLSDNALGDFGIRLLCVGLKHLLCNLKKLWLVSCCLTSACCQDLASVLSTSHS LTRLYVGENALGDSGVAILCEKAKNPQCNLQKLGLVNSGLTSVCCSALSSVLSTNQNL THLYLRGNTLGDKGIKLLCEGLLHPDCKLQVLELDNCNLTSHCCWDLSTLLTSSQSLR KLSLGNNDLGDLGVMMFCEVLKQQSCLLQNLGLSEMYFNYETKSALETLQEEKPELTV

VFEPSW (SEQ ID NO: 539)

[00429] The human NLRP3 protein is encoded by the NLRP3 gene, which comprises the nucleic acid sequences NM _- 004895.1 (SEQ ID NO: 530), NM _- 183395 (SEQ ID NO: 531), NM _- 001079821 (SEQ ID NO: 532), NM _- 001127461 (SEQ ID NO: 533), and NM _- 001127462 (SEQ ID NO: 534), and the human NLRP3 protein has the amino acid sequence NM _- 004895 (SEQ ID NO: 539).

[00430] NLRP3 inhibitors further include antisense polynucleotides that can be used to inhibit transcription of the NLRP3 gene and, therefore, activation of the NLRP3 inflammasome. Polynucleotides complementary to a segment of a polynucleotide encoding NLRP3 (e.g., a polynucleotide as set forth in SEQ ID NOs: 530-534) are designed to bind to mRNA encoding NLRP3 and to inhibit translation of such mRNA. Antisense polynucleotides can be encoded by a ccDNA vector as disclosed herein and can optionally be operably linked to a tissue-specific or inducible promoter as disclosed herein.

[00431] Inhibition of NLRP3 mRNA can be performed using gene silencing RNAi molecules according to methods commonly known to those skilled in the art. For example, oligonucleotide duplexes of gene silencing siRNAs specifically targeting human NLRP3 (NM _- 004895.1) can be easily used to suppress the expression of NLRP3. Targeting NLRP3 mRNA can be successfully performed using siRNA; and other siRNA molecules can be easily prepared by those skilled in the art based on the known sequence of the target mRNA. Accordingly, to avoid any doubt, a person of ordinary skill in the art can design nucleic acid inhibitors, such as RNAi agents (silencing RNAs), for the nucleic acid sequence of NM _- 004895.1, which is shown below:

1 gtagatgagg aaactgaagt tgaggaatag tgaagagttt gtccaatgtc atagccccgt

61 aatcaacggg acaaaaattt tcttgctgat gggtcaagat ggcatcgtga agtggttgtt

121 caccgtaaac tgtaatacaa tcctgtttat ggatttgttt gcatattttt ccctccatag

181 ggaaaccttt cttccatggc tcaggacaca ctcctggatc gagccaacag gagaactttc

241 tggtaagcat ttggctaact tttttttttt tgagatggag tcttgctgtg tcgcctaggc

301 tggagtgcag tggcgtgatc ttggctcact gcagcctcca cttcccgggt tcaatcaatt

361 ctcctacctc aacttcctga gtagctggga ttacaggcgc ccgccaccac acccggctca

421 tttttgtact tttagtagag acacagtttt gccatgttgg ccaggctggt cttgaattcc

481 tcagctcagg tgatctgcct gccttggcct ctcaaagtgc tgggattaca ggcgtgagcc

541 actgtgcccg gccttggcta acttttcaaa attaaagatt ttgacttgtt acagtcatgt

601 gacatttttt tctttctgtt tgctgagttt ttgataattt atatctctca aagtggagac

661 tttaaaaaag actcatccgt gtgccgtgtt cactgcctgg tatcttagtg tggaccgaag

721 cctaaggacc ctgaaaacag ctgcagatga agatggcaag cacccgctgc aagctggcca

781 ggtacctgga ggacctggag gatgtggact tgaagaaatt taagatgcac ttagaggact

841 atcctcccca gaagggctgc atccccctcc cgaggggtca gacagagaag gcagaccatg

901 tggatctagc cacgctaatg atcgacttca atggggagga gaaggcgtgg gccatggccg

961 tgtggatctt cgctgcgatc aacaggag acctttatga gaaagcaaaa agagatgagc

1021 cgaagtgggg ttcagataat gcacgtgttt cgaatcccac tgtgatatgc caggaagaca

1081 gcattgaaga ggagtggatg ggtttactgg agtacctttc gagaatctct atttgtaaaa

1141 tgaagaaaga ttaccgtaag aagtacagaa agtacgtgag aagcagattc cagtgcattg

1201 aagacaggaa tgcccgtctg ggtgagagtg tgagcctcaa caaacgctac acacgactgc

1261 gtctcatcaa ggagcaccgg agccagcagg agagggagca ggagcttctg gccatcggca

1321 agaccaagac gtgtgagagc cccgtgagtc ccattaagat ggagttgctg tttgaccccg

1381 atgatgagca ttctgagcct gtgcacaccg tggtgttcca gggggcggca gggattggga

1441 aaacaatcct ggccaggaag atgatgttgg actgggcgtc ggggacactc taccaagaca

1501 ggtttgacta tctgttctat atccactgtc gggaggtgag ccttgtgaca cagaggagcc

1561 tggggggacct gatcatgagc tgctgccccg acccaaaccc acccatccac aagatcgtga

1621 gaaaaccctc cagaatcctc ttcctcatgg acggcttcga tgagctgcaa ggtgcctttg

1681 acgagcacat aggaccgctc tgcactgact ggcagaaggc cgagcgggga gacattctcc

1741 tgagcagcct catcagaaag aagctgcttc ccgaggcctc tctgctcatc accacgagac

1801 ctgtggccct ggagaaactg cagcacttgc tggaccatcc tcggcatgtg gagatcctgg

1861 gtttctccga ggccaaaagg aaagagtact tcttcaagta cttctctgat gaggcccaag

1921 ccagggcagc cttcagtctg attcaggaga acgaggtcct cttcaccatg tgcttcatcc

1981 ccctggtctg ctggatcgtg tgcactggac tgaaacagca gatggagagt ggcaagagcc

2041 ttgcccagac atccaagacc accaccgcgg tgtacgtctt cttcctttcc agtttgctgc

2101 agccccgggg agggagccag gagcacggcc tctgcgccca cctctggggg ctctgctctt

2161 tggctgcaga tggaatctgg aaccagaaaa tcctgtttga ggagtccgac ctcaggaatc

2221 atggactgca gaaggcggat gtgtctgctt tcctgaggat gaacctgttc caaaaggaag

2281 tggactgcga gaagttctac agcttcatcc acatgacttt ccaggagttc tttgccgcca

2341 tgtactacct gctggaagag gaaaaggaag gaaggacgaa cgttccaggg agtcgtttga

2401 agcttcccag ccgagacgtg acagtccttc tggaaaacta tggcaaattc gaaaaggggt

2461 atttgatttt tgttgtacgt ttcctctttg gcctggtaaa ccaggagagg acctcctact

2521 tggagaagaa attaagttgc aagatctctc agcaaatcag gctggagctg ctgaaatgga

2581 ttgaagtgaa agccaaagct aaaaagctgc agatccagcc cagccagctg gaattgttct

2641 actgtttgta cgagatgcag gaggaggact tcgtgcaaag ggccatggac tatttcccca

2701 agattgagat caatctctcc accagaatgg accacatggt ttcttccttt tgcattgaga

2761 actgtcatcg ggtggagtca ctgtccctgg ggtttctcca taacatgccc aaggaggaag

2821 aggaggagga aaaggaaggc cgacaccttg atatggtgca gtgtgtcctc ccaagctcct

2881 ctcatgctgc ctgttctcat ggattggtga acagccacct cacttccagt ttttgccggg

2941 gcctcttttc agttctgagc accagccaga gtctaactga attggacctc agtgacaatt

3001 ctctggggga cccagggatg agagtgttgt gtgaaacgct ccagcatcct ggctgtaaca

3061 ttcggagatt gtggttgggg cgctgtggcc tctcgcatga gtgctgcttc gacatctcct

3121 tggtcctcag cagcaaccag aagctggtgg agctggacct gagtgacaac gccctcggtg

3181 acttcggaat cagacttctg tgtgtgggac tgaagcacct gttgtgcaat ctgaagaagc

3241 tctggttggt cagctgctgc ctcacatcag catgttgtca ggatcttgca tcagtattga

3301 gcaccagcca ttccctgacc agactctatg tgggggagaa tgccttggga gactcaggag

3361 tcgcaatttt atgtgaaaaa gccaagaatc cacagtgtaa cctgcagaaa ctggggttgg

3421 tgaattctgg ccttacgtca gtctgttgtt cagctttgtc ctcggtactc agcactaatc

3481 agaatctcac gcacctttac ctgcgaggca acactctcgg agacaagggg atcaaactac

3541 tctgtgaggg actcttgcac cccgactgca agcttcaggt gttggaatta gacaactgca

3601 acctcacgtc acactgctgc tgggatcttt ccacacttct gacctccagc cagagcctgc

3661 gaaagctgag cctgggcaac aatgacctgg gcgacctggg ggtcatgatg ttctgtgaag

3721 tgctgaaaca gcagagctgc ctcctgcaga acctggggtt gtctgaaatg tatttcaatt

3781 atgagacaaa aagtgcgtta gaaacacttc aagaagaaaa gcctgagctg accgtcgtct

3841 ttgagccttc ttggtaggag tggaaacggg gctgccagac gccagtgttc tccggtccct

3901 ccagctgggg gccctcaggt ggagagagct gcgatccatc caggccaaga ccacagctct

3961 gtgatccttc cggtggagtg tcggagaaga gagcttgccg acgatgcctt cctgtgcaga

4021 gcttgggcat ctcctttacg ccagggtgag gaagacacca ggacaatgac agcatcgggt

4081 gttgttgtca tcacagcgcc tcagttagag gatgttcctc ttggtgacct catgtaatta

4141 gctcattcaa taaagcactt tctttatttt tctcttctct gtctaacttt ctttttccta

4201 tcttttttct tctttgttct gtttactttt gctcatatca tcattcccgc tatctttcta

4261 ttaactgacc ataacacaga actagttgac tatatattat gttgaaattt tatggcagct

4321 atttatttat ttaaattttt tgtaacagtt ttgttttcta ataagaaaaa tccatgcttt

4381 ttgtagctgg ttgaaaattc aggaatatgt aaaacttttt ggtatttaat taaattgatt

ccttttctta attttaaaaa aaaaaaaaaa (SEQ ID NO: 530)

[00432] In some embodiments, the NLRP3 inflammasome inhibitor is an siRNA, thereby inhibiting NLRP3 inflammasome mRNA. In some embodiments, the NLRP3 inflammasome inhibitor is GUGCAUUGAAGACAGGAAUTT (SEQ ID NO: 540) (Wang et al., Laboratory Invest. (2017) 97: 922-934, which is incorporated herein by reference in its entirety), which inhibits the expression of human NLRP3 or a fragment or homolog thereof that is at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90% identical thereto. In some embodiments, the NLRP3 inflammasome inhibitor is a commercially available siRNA, such as that available from Santa Cruz® (catalog number sc-40327).

[00433] In some embodiments, the NLRP3 inflammasome inhibitor is an RNAi that is complementary to an RNAi target sequence in human NM_001079821.2, NCBI 114548 (NLRP3) gene. The RNAi agent that inhibits NLRP3 can be a nucleic acid that is complementary to 17-21 consecutive bases of SEQ ID NO: 541-551 shown in Table 5A.

[00434] Table 5A: RNAi target sequences for NLRP3 inhibition:

Target sequence SEQ ID NO: Clone ID GGCTGTAACATTCGGAGATTG 541 TRCN0000419896 TCATCATTCCCGCTATCTTTC 542 TRCN0000420883 CCGTAAGAAGTACAGAAAGTA 543 TRCN0000062723 GAGACTCAGGAGTCGCAATTT 544 TRCN0000431574 CCTCATGTAATTAGCTCATTC 545 TRCN0000427726 GTGGATCTAGCCACGCTAATG 546 TRCN0000432208 CCACAGTGTAACCTGCAGAAA 547 TRCN0000062725 CCAGCCAGAGTCTAACTGAAT 548 TRCN0000062724 GCGTTAGAAACACTTCAAGAA 549 TRCN0000062726 GCTGGAATTGTTCTACTGTTT 550 TRCN0000062727 CCACATGACTTTCCAGGAGTT 551 TRCN0000101069

[00435] In some embodiments, the NLRP3 inflammasome inhibitor is an siRNA agent. Exemplary siRNA sequences that inhibit NLRP3 are shown in Table 5B.

[00436] Table 5B: Exemplary siRNAs that inhibit NLRP3

Clone ID Target sequence Forward and reverse oligonucleotide sequences TRCN0000419896 GGCTGTAACATTCGGAGATTG (SEQ ID NO: 552) Direct: CCGGGGCTGTAACATTCGGAGATTGCTCGAGCAATCTCCGAATGTTACAGCCTTTTTG (SEQ ID NO: 553) Reverse: AATTCAAAAAGGCTGTAACATTCGGAGATTGCTCGAGCAATCTCCGAATGTTACAGCC (SEQ ID NO: 554) TRCN0000420883 TCATCATTCCCGCTATCTTTC (SEQ ID NO: 555) Direct:CCGGTCATCATTCCCGCTATCTTTCCTCGAGGAAAGATAGCGGGAATGATGATTTTTG (SEQ ID NO: 556) Reverse: AATTCAAAATCATCATTCCCGCTATCTTTCCTCGAGGAAAGATAGCGGGAATGATGA (SEQ ID NO: 557) TRCN0000062723 CCGTAAGAAGTACAGAAAGTA (SEQ ID NO: 558) Direct: CCGGCCGTAAGAAGTACAGAAAGTACTCGAGTACTTTCTGTACTTCTTACGGTTTTTG (SEQ ID NO: 559) Reverse: AATTCAAAAACCGTAAGAAGTACAGAAAGTACTCGAGTACTTTCTGTACTTCTTACGG (SEQ ID NO: 560) TRCN0000431574 GAGACTCAGGAGTCGCAATTT (SEQ ID NO: 561) Direct: CCGGGAGACTCAGGAGTCGCAATTTCTCGAGAAATTGCGACTCCTGAGTCTCTTTTTG (SEQ ID NO: 562) Reverse: AATTCAAAAGAGACTCAGGAGTCGCAATTTCTCGAGAAATTGCGACTCCTGAGTCTC (SEQ ID NO: 563) TRCN0000427726 CCTCATGTAATTAGCTCATTC (SEQ ID NO: 564) Direct: CCGGCCTCATGTAATTAGCTCATTCCTCGAGGAATGAGCTAATTACATGAGGTTTTTG (SEQ ID NO: 565) Reverse: AATTCAAAACCTCATGTAATTAGCTCATTCCTCGAGGAATGAGCTAATTACATGAGG (SEQ ID NO: 566) TRCN0000432208 GTGGATCTAGCCACGCTAATG (SEQ ID NO: 567) Direct: CCGGGTGGATCTAGCCACGCTAATGCTCGAGCATTAGCGTGGCTAGATCCACTTTTTG (SEQ ID NO: 568) Reverse: AATTCAAAAAGTGGATCTAGCCACGCTAATGCTCGAGCATTAGCGTGGCTAGATCCAC (SEQ ID NO: 569) TRCN0000062725 CCACAGTGTAACCTGCAGAAA (SEQ ID NO: 570) Direct: CCGGCCACAGTGTAACCTGCAGAAACTCGAGTTTCTGCAGGTTACACTGTGGTTTTTG (SEQ ID NO: 570) Reverse: AATTCAAAAACCACAGTGTAACCTGCAGAAACTCGAGTTTCTGCAGGTTACACTGTGG (SEQ ID NO: 571) TRCN0000062724 CCAGCCAGAGTCTAACTGAAT (SEQ ID NO: 572) Direct: CCGGCCAGCCAGAGTCTAACTGAATCTCGAGATTCAGTTAGACTCTGGCTGGTTTTTG (SEQ ID NO: 573) Reverse: AATTCAAAAACCAGCCAGAGTCTAACTGAATCTCGAGATTCAGTTAGACTCTGGCTGG (SEQ ID NO: 574) TRCN0000062726 GCGTTAGAAACACTTCAAGAA (SEQ ID NO: 575) Direct: CCGGGCGTTAGAAACACTTCAAGAACTCGAGTTCTTGAAGTGTTTCTAACGCTTTTTG (SEQ ID NO: 576) Reverse: AATTCAAAAAGCGTTAGAAACACTTCAAGAACTCGAGTTCTTGAAGTGTTTCTAACGC (SEQ ID NO: 577) TRCN0000062727 GCTGGAATTGTTCTACTGTTT (SEQ ID NO: 578) Direct: CCGGGCTGGAATTGTTCTACTGTTTCTCGAGAAACAGTAGAACAATTCCAGCTTTTTG (SEQ ID NO: 579) Reverse: AATTCAAAAAGCTGGAATTGTTCTACTGTTTCTCGAGAAACAGTAGAACAATTCCAGC (SEQ ID NO: 580) TRCN0000101069 CCACATGACTTTCCAGGAGTT (SEQ ID NO: 581) Direct: CCGGCCACATGACTTTCCAGGAGTTCTCGAGAACTCCTGGAAAGTCATGTGGTTTTTG (SEQ ID NO: 582) Reverse: AATTCAAAACCACATGACTTTCCAGGAGTTCTCGAGAACTCCTGGAAAGTCATGTGG (SEQ ID NO: 583) TRCN0000191875 GAAAGCCAAAGCTAAGAAGTA (SEQ ID NO: 584) Direct: CCGGGAAAGCCAAAGCTAAGAAGTACTCGAGTACTTCTTAGCTTTGGCTTTCTTTTTG (SEQ ID NO: 585) Reverse: AATTCAAAAAGAAAGCCAAAGCTAAGAAGTACTCGAGTACTTCTTAGCTTTGGCTTTC (SEQ ID NO: 586)

[00437] In some embodiments, the NLRP3 inflammasome inhibitor is a miRNA (miR) that inhibits NLRP3 expression or an agonist of a miR that inhibits NLRP3 expression. Exemplary miRs that inhibit NLRP3 are miR-9 and miR-223.

[00438] miR-9 inhibits NLRP3 inflammasome activation (Wang, Yue, et al. "MicroRNA-9 inhibits NLRP3 inflammasome activation in human atherosclerosis inflammation cell models through the JAK1/STAT signaling pathway." Cellular Physiology and Biochemistry 41.4 (2017): 1555-1571.). Accordingly, pre-miR-9 or miR-9 can be used to inhibit NLRP3. The sequence of mature miR-9 (MIMAT0000441) is 5´-UCU UUG GUU AUC U AG CUG UAU GA-3´ (SEQ ID NO: 587). hsa-miR-9-5p (UCUUUGGUUAUCUAGCUGUAUGA) (SEQ ID NO: 588). In some embodiments, the NLRP3 inflammasome inhibitor is the miR-9 agonist SQ22538 (SQ; 9-(tetrahydro-2-furanyl)-9H-purin-6-amine), which has been reported to increase miR-9 expression (Ham, Onju, et al. “Small molecule-mediated induction of miR-9 suppressed vascular smooth muscle cell proliferation and neointima formation after balloon injury.” Oncotarget 8.55 (2017): 93360.). The formula for SQ22538 is shown below:

[00439] miR-223 inhibits NLRP3 inflammasome activity. (Bauernfeind, Franz, et al. “NLRP3 inflammasome activity is negatively controlled by miR-223.” The Journal of Immunology 189.8 (2012): 4175–4181; Feng, Zunyong, et al. “Ly6G+ neutrophil-derived miR-223 inhibits the NLRP3 inflammasome in mitochondrial DAMP-induced acute lung injury.” Cell death & disease 8.11 (2017): e3170). miR-223 can be synthesized as mmu-miR-223. At least one or 2 or 3 or 4 blocks of sequence complementary to miR-223 (5'-TGGGGTATTTGACAAACTGACA-3' (SEQ ID NO: 589) can be used to inhibit NLRP3. cbn-mir-233 MI0024890 has the sequence: UCGCCCAUCCCGUUGUUCCAAUAUUCCAACAACAAGUGAUUAUUGAGCAAUGCGCAUGUGCGG (SEQ ID NO: 590); cbr-mir-233 MI0000530 has the sequence: AAGCAUUUUUCUGUCCCGCGCAUCCCUUUGUUCCAAUAUUCAAACCAGUAGAAAGAUUAUUGAGCAAUGCGCAUGUGCGGACAGAUUGAAUAGCUG (SEQ ID NO: 591); cel-mir-233 MI0000308 has the sequence: AUAUAGCAUCUUUCUGUCUCGCCCAUCCCGUUGCUCCAAUAUUCUAACAACAAGUGAUUAUUGAGCAAUGCGCAUGUGCGGGAUAGACUGAUGGCUGC (SEQ ID NO: 592); crm-mir-233 MI0011059 has the sequence: UGAAGCGUCUCUGUCCCGCUCAUCCUGUUGUUCCAAUAUUCCAACAGCCCAGUGAUUAUUGAGCAAUGCGCAUGUGCGGGACAGAUUGUAUGCUGCCAU (SEQ ID NO: 593).

[00440] In some embodiments, the NLRP3 inflammasome inhibitor is an anti-miRNA (anti-miR) that inhibits the expression of a miR that suppresses the expression or function of NLRP3. Exemplary anti-miRs are anti-miR-22 and anti-miR-33. miR22 has been demonstrated to maintain NLRP3 expression (Li, S., et al., "MiR-22 sustains NLRP3 expression and attenuates H. pylori-induced gastric carcinogenesis." Oncogene 37.7 (2018): 884.). The mature sequence of miR-22 is hsa-miR-22 (hsa-miR-22-5p MIMAT000449) and is as follows: AGUUCUUCAGUGGCAAGCUUUA (SEQ ID NO: 594) with the stem-loop sequence as follows: hsa-mir-22 MI0000078 has the sequence: GGCUGAGCCGCAGUAGUUCUUCAGUGGCAAGCUUUAUGUCCUGACCCAGCUAAAGCUGCCAGUUGAAGAACUGUUGCCCUCUGCC (SEQ ID NO: 595).

[00441] miR-33 has been reported to positively regulate NLRP3 mRNA and protein expression and caspase-1 activity in primary macrophages (Xie, Qingyun, et al. “MicroRNA - 33 regulates the NLRP3 inflammasome signaling pathway in macrophages.” Molecular medicine reports 17.2 (2018): 3318–3327). The mature sequence of miR-33 is mmu-miR-33-5p or MIMAT0000667; and is: GUGCAUUGUAGUUGCAUUGCA (SEQ ID NO: 596); the stem-loop sequence is as follows: mmu-mir-33 MI0000707: CUGUGGUGCAUUGUAGUUGCAUUGCAUGUUCUGGCAAUACCUGUGCAAUGUUUCCACAGUGCAUCACGG (SEQ ID NO: 597).

[00442] Accordingly, in some embodiments, the NLRP3 inhibitor is an anti-miR-22 that is complementary to at least a portion, such as 15-25 units of SEQ ID NO: 594 or SEQ ID NO: 595, or an anti-miR-33 that is complementary to at least a portion, such as 15-21 units of SEQ ID NO: 596 or SEQ ID NO: 597.

[00443] In some embodiments of the compositions and methods described herein, the NLRP3 inflammasome inhibitor is an anti-human NLRP3 antibody (catalog number AF6789) from R&D Systems (Minneapolis, MN). In some embodiments, the NLRP3 inhibitor antibody is encoded by ccDNA.

In some embodiments of the compositions and methods described herein, the NLRP3 inhibitor is an antibody or antigen-binding fragment that binds NLRP3. In some embodiments of the compositions and methods described herein, the antibody or antigen-binding fragment that binds NLRP3 is encoded by ccDNA.

An NLRP3 inflammasome inhibitor refers to compounds that inhibit or at least reduce the activity of the inflammasome, including glyburide and its functionally equivalent precursors or derivatives, caspase-1 inhibitors, adenosine monophosphate-activated protein kinase (AMPK) activators, and P2X7 inhibitors. Inhibition of the NLRP3 inflammasome can be achieved by a single compound or a combination of compounds that inhibit the inflammasome or caspase-1 but do not result in changes in the enzymatic activity of cytochrome P450 (cyp), including the cyp, 3A4, 2C9, and 2C19 isoforms, which may adversely affect statin metabolism and thereby reduce the bioavailability of statins.

AIM2 inflammasome pathway inhibitors

[00444] In some embodiments, an inflammasome antagonist inhibits AIM2. AIM2, alternatively known as PISA, is a 343 amino acid polypeptide (see Genbank Accession No. AF024714.1; RefSeq Accession No. NP_004824.1) (SEQ ID NO: 598). AIM2 is a member of the IFI20X/IF116 family and is known to be expressed in the spleen, small intestine, peripheral blood leukocytes, and testes. AIM2 contains a PYD domain that is involved in interaction with ASC, as well as a HIN200 domain that is involved in interaction with dsDNA. AIM2 is thought to play a role in oncogenic reversal and may control cell proliferation. AIM2 expression is induced by interferon gamma.

[00445] In some embodiments of the compositions and methods described herein, the AIM2 inhibitor is an antibody or antigen-binding fragment that binds AIM2. In some embodiments of the compositions and methods described herein, the antibody or antigen-binding fragment that binds NLRP3 is encoded by ccDNA. AIM2 inhibitors are disclosed in Farshchian et al., Oncotarget 2017; 8(28); 45825-45836, which is incorporated herein by reference in its entirety.

[00446] In some embodiments, the AIM2 inflammasome inhibitor is a monoclonal antibody to human ASC (clone 23-4, MBL, Nagoya, Japan) that is reported to interfere with PYD from ASC. In some embodiments, the AIM2 inflammasome inhibitor is an anti-human AIM2 antibody (catalog #8055) (Cell Signaling Technology® (Beverly, MA). In some embodiments, the AIM2 inflammasome inhibitor is an endogenous AIM2 inhibitor, such as the pyrin-containing proteins recently described (Khare et al., 2014; de Almeida et al., 2015) or the cathelicidin antimicrobial peptides reported by Schauber and colleagues (Dombrowski et al., 2011). In some embodiments, the AIM2 inflammasome inhibitor is any of the compounds disclosed in Miriam Canavase's short review, "the duality of AIM2 inflammasome: A focus on its role in autoimmunity and Skin diseases. Am. J. Pharm &Toxicology; 2016).

[00447] In some embodiments, the AIM2 inflammasome inhibitor is P202, which is a tetramer of p202 and has been reported to reduce AIM2 activation and prevent dsDNA-dependent clustering of ASC and activation of the AIM2 inflammasome (Fernandes-Alnemri, Teresa, et al. "The AIM2 inflammasome is critical for innate immunity to Francisella tularensis." Nature immunology 11.5 (2010): 385; Yin, Qian, et al. "Molecular mechanism for p202-mediated specific inhibition of AIM2 inflammasome activation." Cell reports 4.2 (2013): 327-339). In some embodiments of the compositions and methods described herein, P202 is encoded by ccDNA.

[00448] In some embodiments, the AIM2 inflammasome inhibitor is any of the small molecule compounds described in WO2017138586A or US2013/0158100A1, the contents of each of which are incorporated herein by reference in their entirety.

[00449] In some embodiments of the compositions and methods described herein, the AIM2 inhibitor is an RNA-based AIM2 inhibitor, such as an siRNA specific for AIM2. In some embodiments of the compositions and methods described herein, the RNA-based AIM2 inhibitor is encoded by a ccDNA. The human AIM2 protein is encoded by the AIM2 gene comprising the nucleic acid sequence NM_004833.2 (SEQ ID NO: 600), and the human AIM2 protein has the amino acid sequence NP_004824.1 (SEQ ID NO: 598). AIM2 inhibitors further include antisense polynucleotides that can be used to inhibit transcription of the AIM2 gene and, accordingly, activation of the AIM2 inflammasome. Polynucleotides complementary to a segment of a polynucleotide encoding AIM2 (e.g., a polynucleotide as set forth in SEQ ID NO: 600) are designed to bind to and inhibit translation of an mRNA encoding AIM2. Antisense polynucleotides may be encoded by a ccDNA vector as disclosed herein and may optionally be operably linked to a tissue-specific or inducible promoter as disclosed herein. Inhibition of AIM2 mRNA can be accomplished using gene silencing RNAi molecules according to methods commonly known to those skilled in the art. For example, oligonucleotide duplexes of gene silencing siRNAs specifically targeting human AIM2 (NM_004833.2) can be readily used to inhibit expression of AIM2. Targeting of AIM2 mRNA can be advantageously accomplished using siRNA; and other siRNA molecules can be easily obtained by those skilled in the art based on the known sequence of the target mRNA. Accordingly, to avoid any doubt, one of ordinary skill in the art can develop nucleic acid-based inhibitors, such as RNAi (RNA silencing) agents, for the nucleic acid sequence NM_004833.2, which is shown below:

1 atagacattt tcttctgtgg ctgctagtga gaacccaaac cagctcagcc aattagagct

61 ccagttgtca ctcctaccca cactgggcct gggggtgaag ggaagtgttt attaggggta

121 catgtgaagc cgtccagaag tgtcagagtc tttgtagctt tgaaagtcac ctaggttatt

181 tgggcatgct ctcctgagtc ctctgctagt taagctctct gaaaagaagg tggcagaccc

241 ggtttgctga tcgccccagg gatcaggagg ctgatcccaa agttgtcaga tggagagtaa

301 atacaaggag atactcttgc taacaggcct ggataacatc actgatgagg aactggatag

361 gtttaagttc tttctttcag acgagtttaa tattgccaca ggcaaactac atactgcaaa

421 cagaatacaa gtagctacct tgatgattca aaatgctggg gcggtgtctg cagtgatgaa

481 gaccattcgt atttttcaga agttgaatta tatgcttttg gcaaaacgtc ttcaggagga

541 gaaggagaaa gttgataagc aatacaaatc ggtaacaaaa ccaaagccac taagtcaagc

601 tgaaatgagt cctgctgcat ctgcagccat cagaaatgat gtcgcaaagc aacgtgctgc

661 accaaaagtc tctcctcatg ttaagcctga acagaaacag atggtggccc agcaggaatc

721 tatcagagaa gggtttcaga agcgctgttt gccagttatg gtactgaaag caaagaagcc

781 cttcacgttt gagacccaag aaggcaagca ggagatgttt catgctacag tggctacaga

841 aaaggaattc ttctttgtaa aagtttttaa tacactgctg aaagataaat tcattccaaa

901 gagaataatt ataatagcaa gatattatcg gcacagtggt ttcttagagg taaatagcgc

961 ctcacgtgtg ttagatgctg aatctgacca aaaggttaat gtcccgctga acattatcag

1021 aaaagctggt gaaaccccga agatcaacac gcttcaaact cagccccttg gaacaattgt

1081 gaatggtttg tttgtagtcc agaaggtaac agaaaagaag aaaaacatat tatttgacct

1141 aagtgacaac actgggaaaa tggaagtact gggggttaga aacgaggaca caatgaaatg

1201 taaggaagga gataaggttc gacttacatt cttcacactg tcaaaaaatg gagaaaaact

1261 acagctgaca tctggagttc atagcaccat aaaggttatt aaggccaaaa aaaaaacata

1321 gagaagtaaa aaggaccaat tcaagccaac tggtctaagc agcatttaat tgaagaatat

1381 gtgatacagc ctcttcaatc agattgtaag ttacctgaaa gctgcagttc acaggctcct

1441 ctctccacca aattaggata gaataattgc tggataaaca aattcagaat atcaacagat

gatcacaata aacatctgtt tctcattcaa aaaaaaaaaa aaaaaaaaaa aaaaaaaa

(SEQ ID NO: 600)

In some embodiments, the AIM2 inflammasome inhibitor is an siRNA, thereby inhibiting AIM2 inflammasome mRNA. In some embodiments, the AIM2 inflammasome inhibitor is 5'-CCCGAAGATCAACACGCTTCA-3' (SEQ ID NO: 601) or 5'-AAAGGTTAATGTCCCGCTGAA-3' (SEQ ID NO: 665) (both from Farshchian et al. Oncotarget (2017) 8: 45825-45836), which inhibits the expression of human AIM2 or a fragment or homolog thereof that is at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90% identical thereto.

[00450] In some embodiments of the compositions and methods described herein, the AIM2 inflammasome inhibitor is an RNA-based AIM2 inhibitor, such as an siRNA specific for AIM2. In some embodiments of the compositions and methods described herein, the RNA-based AIM2 inhibitor is encoded by ccDNA. AIM2 siRNA may be commercially available, for example, SI04261432 (Qiagen®); or RCN0000096104 (#1), TRCN0000096105 (#2), TRCN0000096106 (#3) from OpenBiosystems® (Huntsville, AL).

[00451] In some embodiments, the AIM2 inflammasome inhibitor is A151 (5'-TTAGGGTTAGGGTTAGGGTTAGGG-3' (SEQ ID NO: 602) or C151 (5'-TTCAAATTCAAATTCAAATTCAAA-3' (SEQ ID NO: 603), which is synthesized with a phosphorothioate (PO) backbone. A151 (also referred to as ODN TTAGGG) is a synthetic oligonucleotide (ODN) containing 4 repeats of the TTAGGG immunosuppressive motif (SEQ ID NO: 604), commonly found in mammalian telomeric DNA (Steinhagen F. et al., 2017. Suppressive oligodeoxynucleotides containing TTAGGG motifs inhibit cGAS activation in human monocytes. Eur J Immunol). A151 blocks AIM2 inflammasome activation in response to cytosolic dsDNA but requires a phosphorothioate (PO) backbone (Kaminsji et al., J Immunol 2013; 191:3876–3883, Synthetic Oligodeoxynucleotides Containing Suppressive TTAGGG Motifs Inhibit AIM2 Inflammasome Activation; Eichholz K. et al., 2016. Immune-Complexed Adenovirus Induce AIM2-Mediated Pyroptosis in Human Dendritic Cells. PLoS Pathog. 12(9): e1005871). In some embodiments, the AIM2 inflammasome inhibitor is A151 (5′-TTAGGGTTAGGGTTAGGGTTAGGG-3′ (SEQ ID NO: 602) or at least one TTAGGG repeat (SEQ ID NO: 604), each with a phosphorothioate (PO) backbone. In some embodiments, the AIM2 inflammasome inhibitor is A151 (5´-TTAGGGTTAGGGTTAGGGTTAGGG-3´ (SEQ ID NO: 602) or at least one TTAGGG repeat (SEQ ID NO: 604), which does not have a phosphodiester (PE) backbone. In some embodiments of the compositions and methods described herein, the AIM2 inflammasome inhibitor is encoded by a ccDNA administered to a subject (including, for example, subsequent delivery (ecDNA). According to some embodiments of the compositions and methods described herein, the AIM2 inflammasome inhibitor encoded by the ecDNA administered to the subject is A151 (SEQ ID NO: 602).

[00452] In some embodiments, the AIM2 inflammasome inhibitor is an RNAi that is complementary to an RNAi target sequence in human NM_001348247.1 (SEQ ID NO: 566), NCBI gene 9447 (AIM2). The RNAi agent that inhibits AIM2 can be a nucleic acid complementary to 17-21 consecutive bases of SEQ ID NO: 605-610, shown in Table 5C.

[00453] Table 5C: RNAi target sequences for AIM2 inhibition:

Target sequence SEQ ID NO: Clone ID AGCCACTAAGTCAAGCTGAAA 605 TRCN0000107503 CCAACTGGTCTAAGCAGCATT 606 TRCN0000107500 GAAACGAGGACACAATGAAAT 607 TRCN0000413154 GCCACTAAGTCAAGCTGAAAT 608 TRCN0000107502 CTGGAGTTCATAGCACCATAA 609 TRCN0000107504 CCCGCTGAACATTATCAGAAA 610 TRCN0000107501

[00454] In some embodiments, the AIM2 inflammasome inhibitor is an siRNA agent. Exemplary siRNA sequences that inhibit AIM2 are shown in Table 5D.

[00455] Table 5D: Exemplary siRNAs that inhibit AIM2

Clone ID Target sequence Forward and reverse oligonucleotide sequence TRCN0000107503 AGCCACTAAGTCAAGCTGAAA (SEQ ID NO: 666) Direct: CCGGAGCCACTAAGTCAAGCTGAAACTCGAGTTTCAGCTTGACTTAGTGGCTTTTTTG (SEQ ID NO: 667) Reverse: AATTCAAAAAAGCCACTAAGTCAAGCTGAAACTCGAGTTTCAGCTTGACTTAGTGGCT (SEQ ID NO:668) TRCN0000107500 CCAACTGGTCTAAGCAGCATT (SEQ ID NO: 669) Direct: CCGGCCAACTGGTCTAAGCAGCATTCTCGAGAATGCTGCTTAGACCAGTTGGTTTTTG (SEQ ID NO: 670) Reverse: AATTCAAAACCAACTGGTCTAAGCAGCATTCTCGAGAATGCTGCTTAGACCAGTTGG (SEQ ID NO: 671) TRCN0000413154 GAAACGAGGACACAATGAAAT (SEQ ID NO: 672) Direct: CCGGGAAACGAGGACACAATGAAATCTCGAGATTTCATTGTGTCCTCGTTTCTTTTTG (SEQ ID NO: 673) Reverse: AATTCAAAAGAAACGAGGACACAATGAAATCTCGAGATTTCATTGTGTCCTCGTTTC (SEQ ID NO: 674) TRCN0000107502 GCCACTAAGTCAAGCTGAAAT (SEQ ID NO: 675) Direct: CCGGGCCACTAAGTCAAGCTGAAATCTCGAGATTTCAGCTTGACTTAGTGGCTTTTTG (SEQ ID NO: 676) Reverse: AATTCAAAAAGCCACTAAGTCAAGCTGAAATCTCGAGATTTCAGCTTGACTTAGTGGC (SEQ ID NO:677) TRCN0000107504 CTGGAGTTCATAGCACCATAA (SEQ ID NO: 678) Direct: CCGGCTGGAGTTCATAGCACCATAACTCGAGTTATGGTGCTATGAACTCCAGTTTTTG (SEQ ID NO: 679) Reverse: AATTCAAAAACTGGAGTTCATAGCACCATAACTCGAGTTATGGTGCTATGAACTCCAG (SEQ ID NO: 680) TRCN0000107501 CCCGCTGAACATTATCAGAAA (SEQ ID NO: 681) Direct: CCGGCCCGCTGAACATTATCAGAAACTCGAGTTTCTGATAATGTTCAGCGGGTTTTTG (SEQ ID NO:682) Reverse: AATTCAAAACCCGCTGAACATTATCAGAAACTCGAGTTTCTGATAATGTTCAGCGGG (SEQ ID NO: 683) TRCN0000107503 AGCCACTAAGTCAAGCTGAAA (SEQ ID NO: 684) Direct: CCGGAGCCACTAAGTCAAGCTGAAACTCGAGTTTCAGCTTGACTTAGTGGCTTTTTTG (SEQ ID NO: 685) Reverse: AATTCAAAAAAGCCACTAAGTCAAGCTGAAACTCGAGTTTCAGCTTGACTTAGTGGCT (SEQ ID NO: 686) TRCN0000107500 CCAACTGGTCTAAGCAGCATT (SEQ ID NO: 687) Direct: CCGGCCAACTGGTCTAAGCAGCATTCTCGAGAATGCTGCTTAGACCAGTTGGTTTTTG (SEQ ID NO: 688) Reverse: AATTCAAAACCAACTGGTCTAAGCAGCATTCTCGAGAATGCTGCTTAGACCAGTTGG (SEQ ID NO: 689) TRCN0000413154 GAAACGAGGACACAATGAAAT (SEQ ID NO: 690) Direct: CCGGGAAACGAGGACACAATGAAATCTCGAGATTTCATTGTGTCCTCGTTTCTTTTTG (SEQ ID NO: 691) Reverse: AATTCAAAAGAAACGAGGACACAATGAAATCTCGAGATTTCATTGTGTCCTCGTTTC (SEQ ID NO: 692) TRCN0000107502 GCCACTAAGTCAAGCTGAAAT (SEQ ID NO: 693) Direct: CCGGGCCACTAAGTCAAGCTGAAATCTCGAGATTTCAGCTTGACTTAGTGGCTTTTTG (SEQ ID NO: 694) Reverse: AATTCAAAAAGCCACTAAGTCAAGCTGAAATCTCGAGATTTCAGCTTGACTTAGTGGC (SEQ ID NO: 695) TRCN0000107504 CTGGAGTTCATAGCACCATAA (SEQ ID NO: 696) Direct: CCGGCTGGAGTTCATAGCACCATAACTCGAGTTATGGTGCTATGAACTCCAGTTTTTG (SEQ ID NO: 697) Reverse: AATTCAAAAACTGGAGTTCATAGCACCATAACTCGAGTTATGGTGCTATGAACTCCAG (SEQ ID NO: 698) TRCN0000107501 CCCGCTGAACATTATCAGAAA (SEQ ID NO: 699) Direct: CCGGCCCGCTGAACATTATCAGAAACTCGAGTTTCTGATAATGTTCAGCGGGTTTTTG (SEQ ID NO: 700) Reverse: AATTCAAAACCCGCTGAACATTATCAGAAACTCGAGTTTCTGATAATGTTCAGCGGG (SEQ ID NO: 701) TRCN0000107503 AGCCACTAAGTCAAGCTGAAA (SEQ ID NO: 702) Direct: CCGGAGCCACTAAGTCAAGCTGAAACTCGAGTTTCAGCTTGACTTAGTGGCTTTTTTG (SEQ ID NO: 703) Reverse: AATTCAAAAAAGCCACTAAGTCAAGCTGAAACTCGAGTTTCAGCTTGACTTAGTGGCT (SEQ ID NO: 704) TRCN0000107500 CCAACTGGTCTAAGCAGCATT (SEQ ID NO: 705) Direct: CCGGCCAACTGGTCTAAGCAGCATTCTCGAGAATGCTGCTTAGACCAGTTGGTTTTTG (SEQ ID NO: 706) Reverse: AATTCAAAACCAACTGGTCTAAGCAGCATTCTCGAGAATGCTGCTTAGACCAGTTGG (SEQ ID NO: 707) TRCN0000413154 GAAACGAGGACACAATGAAAT (SEQ ID NO: 708) Direct: CCGGGAAACGAGGACACAATGAAATCTCGAGATTTCATTGTGTCCTCGTTTCTTTTTG (SEQ ID NO: 709) Reverse: AATTCAAAAGAAACGAGGACACAATGAAATCTCGAGATTTCATTGTGTCCTCGTTTC (SEQ ID NO:710) TRCN0000107502 GCCACTAAGTCAAGCTGAAAT (SEQ ID NO: 711) Direct: CCGGGCCACTAAGTCAAGCTGAAATCTCGAGATTTCAGCTTGACTTAGTGGCTTTTTG (SEQ ID NO: 712) Reverse: AATTCAAAAAGCCACTAAGTCAAGCTGAAATCTCGAGATTTCAGCTTGACTTAGTGGC (SEQ ID NO: 713) TRCN0000107504 CTGGAGTTCATAGCACCATAA (SEQ ID NO:714) Direct: CCGGCTGGAGTTCATAGCACCATAACTCGAGTTATGGTGCTATGAACTCCAGTTTTTG (SEQ ID NO: 715) Reverse: AATTCAAAAACTGGAGTTCATAGCACCATAACTCGAGTTATGGTGCTATGAACTCCAG (SEQ ID NO: 716) TRCN0000107501 CCCGCTGAACATTATCAGAAA (SEQ ID NO: 717) Direct: CCGGCCCGCTGAACATTATCAGAAACTCGAGTTTCTGATAATGTTCAGCGGGTTTTTG (SEQ ID NO: 718) Reverse: AATTCAAAACCCGCTGAACATTATCAGAAACTCGAGTTTCTGATAATGTTCAGCGGG (SEQ ID NO: 719) TRCN0000107503 AGCCACTAAGTCAAGCTGAAA (SEQ ID NO: 720) Direct: CCGGAGCCACTAAGTCAAGCTGAAACTCGAGTTTCAGCTTGACTTAGTGGCTTTTTTG (SEQ ID NO: 721) Reverse: AATTCAAAAAAGCCACTAAGTCAAGCTGAAACTCGAGTTTCAGCTTGACTTAGTGGCT (SEQ ID NO: 722) TRCN0000107500 CCAACTGGTCTAAGCAGCATT (SEQ ID NO: 723) Direct: CCGGCCAACTGGTCTAAGCAGCATTCTCGAGAATGCTGCTTAGACCAGTTGGTTTTTG (SEQ ID NO: 724) Reverse: AATTCAAAACCAACTGGTCTAAGCAGCATTCTCGAGAATGCTGCTTAGACCAGTTGG (SEQ ID NO: 725) TRCN0000413154 GAAACGAGGACACAATGAAAT (SEQ ID NO: 726) Direct: CCGGGAAACGAGGACACAATGAAATCTCGAGATTTCATTGTGTCCTCGTTTCTTTTTG (SEQ ID NO: 727) Reverse: AATTCAAAAGAAACGAGGACACAATGAAATCTCGAGATTTCATTGTGTCCTCGTTTC (SEQ ID NO: 728) TRCN0000107502 GCCACTAAGTCAAGCTGAAAT (SEQ ID NO: 729) Direct: CCGGGCCACTAAGTCAAGCTGAAATCTCGAGATTTCAGCTTGACTTAGTGGCTTTTTG (SEQ ID NO: 730) Reverse: AATTCAAAAAGCCACTAAGTCAAGCTGAAATCTCGAGATTTCAGCTTGACTTAGTGGC (SEQ ID NO: 731) TRCN0000107504 CTGGAGTTCATAGCACCATAA (SEQ ID NO: 732) Direct: CCGGCTGGAGTTCATAGCACCATAACTCGAGTTATGGTGCTATGAACTCCAGTTTTTG (SEQ ID NO: 733) Reverse: AATTCAAAAACTGGAGTTCATAGCACCATAACTCGAGTTATGGTGCTATGAACTCCAG (SEQ ID NO: 734) TRCN0000107501 CCCGCTGAACATTATCAGAAA (SEQ ID NO: 735) Direct: CCGGCCCGCTGAACATTATCAGAAACTCGAGTTTCTGATAATGTTCAGCGGGTTTTTG (SEQ ID NO: 736) Reverse: AATTCAAAACCCGCTGAACATTATCAGAAACTCGAGTTTCTGATAATGTTCAGCGGG (SEQ ID NO: 737) TRCN0000107503 AGCCACTAAGTCAAGCTGAAA (SEQ ID NO: 738) Direct: CCGGAGCCACTAAGTCAAGCTGAAACTCGAGTTTCAGCTTGACTTAGTGGCTTTTTTG (SEQ ID NO: 738) Reverse: AATTCAAAAAAGCCACTAAGTCAAGCTGAAACTCGAGTTTCAGCTTGACTTAGTGGCT (SEQ ID NO: 740) TRCN0000107500 CCAACTGGTCTAAGCAGCATT (SEQ ID NO: 741) Direct: CCGGCCAACTGGTCTAAGCAGCATTCTCGAGAATGCTGCTTAGACCAGTTGGTTTTTG (SEQ ID NO: 742) Reverse: AATTCAAAACCAACTGGTCTAAGCAGCATTCTCGAGAATGCTGCTTAGACCAGTTGG (SEQ ID NO:742) TRCN0000413154 GAAACGAGGACACAATGAAAT (SEQ ID NO: 743) Direct: CCGGGAAACGAGGACACAATGAAATCTCGAGATTTCATTGTGTCCTCGTTTCTTTTTG (SEQ ID NO: 744) Reverse: AATTCAAAAGAAACGAGGACACAATGAAATCTCGAGATTTCATTGTGTCCTCGTTTC (SEQ ID NO: 745) TRCN0000107502 GCCACTAAGTCAAGCTGAAAT (SEQ ID NO: 746) Direct: CCGGGCCACTAAGTCAAGCTGAAATCTCGAGATTTCAGCTTGACTTAGTGGCTTTTTG (SEQ ID NO: 747) Reverse: AATTCAAAAAGCCACTAAGTCAAGCTGAAATCTCGAGATTTCAGCTTGACTTAGTGGC (SEQ ID NO: 748) TRCN0000107504 CTGGAGTTCATAGCACCATAA (SEQ ID NO: 749) Direct: CCGGCTGGAGTTCATAGCACCATAACTCGAGTTATGGTGCTATGAACTCCAGTTTTTG (SEQ ID NO: 750) Reverse: AATTCAAAAACTGGAGTTCATAGCACCATAACTCGAGTTATGGTGCTATGAACTCCAG (SEQ ID NO: 751) TRCN0000107501 CCCGCTGAACATTATCAGAAA (SEQ ID NO: 752) Direct: CCGGCCCGCTGAACATTATCAGAAACTCGAGTTTCTGATAATGTTCAGCGGGTTTTTG (SEQ ID NO: 753) Reverse: AATTCAAAACCCGCTGAACATTATCAGAAACTCGAGTTTCTGATAATGTTCAGCGGG (SEQ ID NO: 754) TRCN0000185775 GATTGTTTCAACACAAGAGTA (SEQ ID NO: 755) Direct: CCGGGATTGTTTCAACACAAGAGTACTCGAGTACTCTTGTGTTGAAACAATCTTTTTG (SEQ ID NO: 756) Reverse: AATTCAAAAAGATTGTTTCAACACAAGAGTACTCGAGTACTCTTGTGTTGAAACAATC (SEQ ID NO: 757) TRCN0000107503 AGCCACTAAGTCAAGCTGAAA (SEQ ID NO: 758) Direct: CCGGAGCCACTAAGTCAAGCTGAAACTCGAGTTTCAGCTTGACTTAGTGGCTTTTTTG (SEQ ID NO: 759) Reverse: AATTCAAAAAAGCCACTAAGTCAAGCTGAAACTCGAGTTTCAGCTTGACTTAGTGGCT (SEQ ID NO: 760) TRCN0000107500 CCAACTGGTCTAAGCAGCATT (SEQ ID NO: 761) Direct: CCGGCCAACTGGTCTAAGCAGCATTCTCGAGAATGCTGCTTAGACCAGTTGGTTTTTG (SEQ ID NO: 762) Reverse: AATTCAAAACCAACTGGTCTAAGCAGCATTCTCGAGAATGCTGCTTAGACCAGTTGG (SEQ ID NO: 763) TRCN0000413154 GAAACGAGGACACAATGAAAT (SEQ ID NO: 764) Direct: CCGGGAAACGAGGACACAATGAAATCTCGAGATTTCATTGTGTCCTCGTTTCTTTTTG (SEQ ID NO: 765) Reverse: AATTCAAAAGAAACGAGGACACAATGAAATCTCGAGATTTCATTGTGTCCTCGTTTC (SEQ ID NO: 766) TRCN0000107502 GCCACTAAGTCAAGCTGAAAT (SEQ ID NO: 767) Direct: CCGGGCCACTAAGTCAAGCTGAAATCTCGAGATTTCAGCTTGACTTAGTGGCTTTTTG (SEQ ID NO: 768) Reverse: AATTCAAAAAGCCACTAAGTCAAGCTGAAATCTCGAGATTTCAGCTTGACTTAGTGGC (SEQ ID NO: 769) TRCN0000107504 CTGGAGTTCATAGCACCATAA (SEQ ID NO: 780) Direct: CCGGCTGGAGTTCATAGCACCATAACTCGAGTTATGGTGCTATGAACTCCAGTTTTTG (SEQ ID NO: 781) Reverse: AATTCAAAAACTGGAGTTCATAGCACCATAACTCGAGTTATGGTGCTATGAACTCCAG (SEQ ID NO: 782) TRCN0000107501 CCCGCTGAACATTATCAGAAA (SEQ ID NO: 783) Direct: CCGGCCCGCTGAACATTATCAGAAACTCGAGTTTCTGATAATGTTCAGCGGGTTTTTG (SEQ ID NO: 784) Reverse: AATTCAAAACCCGCTGAACATTATCAGAAACTCGAGTTTCTGATAATGTTCAGCGGG (SEQ ID NO: 785) TRCN0000107503 AGCCACTAAGTCAAGCTGAAA (SEQ ID NO: 786) Direct: CCGGAGCCACTAAGTCAAGCTGAAACTCGAGTTTCAGCTTGACTTAGTGGCTTTTTTG (SEQ ID NO: 787) Reverse: AATTCAAAAAAGCCACTAAGTCAAGCTGAAACTCGAGTTTCAGCTTGACTTAGTGGCT (SEQ ID NO: 788) TRCN0000107500 CCAACTGGTCTAAGCAGCATT (SEQ ID NO: 789) Direct: CCGGCCAACTGGTCTAAGCAGCATTCTCGAGAATGCTGCTTAGACCAGTTGGTTTTTG (SEQ ID NO: 790) Reverse: AATTCAAAACCAACTGGTCTAAGCAGCATTCTCGAGAATGCTGCTTAGACCAGTTGG (SEQ ID NO: 791) TRCN0000413154 GAAACGAGGACACAATGAAAT (SEQ ID NO: 792) Direct: CCGGGAAACGAGGACACAATGAAATCTCGAGATTTCATTGTGTCCTCGTTTCTTTTTG (SEQ ID NO: 793) Reverse: AATTCAAAAGAAACGAGGACACAATGAAATCTCGAGATTTCATTGTGTCCTCGTTTC (SEQ ID NO: 794) TRCN0000107502 GCCACTAAGTCAAGCTGAAAT (SEQ ID NO: 795) Direct: CCGGGCCACTAAGTCAAGCTGAAATCTCGAGATTTCAGCTTGACTTAGTGGCTTTTTG (SEQ ID NO: 796) Reverse: AATTCAAAAAGCCACTAAGTCAAGCTGAAATCTCGAGATTTCAGCTTGACTTAGTGGC (SEQ ID NO: 797) TRCN0000107504 CTGGAGTTCATAGCACCATAA (SEQ ID NO: 798) Direct: CCGGCTGGAGTTCATAGCACCATAACTCGAGTTATGGTGCTATGAACTCCAGTTTTTG (SEQ ID NO: 799) Reverse: AATTCAAAAACTGGAGTTCATAGCACCATAACTCGAGTTATGGTGCTATGAACTCCAG (SEQ ID NO: 800) TRCN0000107501 CCCGCTGAACATTATCAGAAA (SEQ ID NO: 801) Direct: CCGGCCCGCTGAACATTATCAGAAACTCGAGTTTCTGATAATGTTCAGCGGGTTTTTG (SEQ ID NO: 802) Reverse: AATTCAAAACCCGCTGAACATTATCAGAAACTCGAGTTTCTGATAATGTTCAGCGGG (SEQ ID NO: 803)

[00456] In some embodiments, the AIM2 inflammasome inhibitor is a miRNA (miR) that inhibits the expression of AIM2, or an agonist of a miR that inhibits the expression of AIM2. Exemplary miRs that inhibit AIM2 are miR-223 (Yang, Fan, et al. "MicroRNA-223 acts as an important regulator to Kupffer cells activation at the early stage of Con A-induced acute liver failure via AIM2 signaling pathway." Cellular Physiology and Biochemistry 34.6 (2014): 2137-2152). Accordingly, an AIM2 inhibitor for use in the present invention is miR-223 according to any one of SEQ ID NOs: 589-593.

[00457] The AIM2 inflammasome reconstituted in a cell-free in vitro system can be used as a tool for screening AIM2 inflammasome inhibitors according to the methods disclosed in Kaneko et al., 2015, or the methods disclosed in US application US2013/0158100A1, which is incorporated herein by reference in its entirety.

Caspase-1 inhibitors

[00458] In some embodiments, the inflammasome antagonist inhibits caspase-1. In some embodiments, the caspase-1 inhibitor for use in the methods and compositions is belnakazan (VX-765). VX-765 is an orally absorbable prodrug of VRT-043198, a potent and selective inhibitor of caspases belonging to the ICE/caspase-1 subfamily, and has the following formula:

(See Wannamaker W. et al., 2007. (S)-1-((S)-2-{[1-(4-amino-3-chloro-phenyl)-methanoyl]-amino}-3,3 -dimethyl-butanoyl)-pyrrolidine-2-carboxylic acid ((2R,3S)-2-ethoxy-5-oxo-tetrahydro-furan-3-yl)-amide (VX-765), an orally available selective interleukin (IL )-converting enzyme/caspase-1 inhibitor, exhibits potent anti-inflammatory activities by inhibiting the release of IL-1beta and IL-18.

[00459] In some embodiments, the caspase-1 inhibitor is Z-VAD-FMK, which has the following structure:

and is a cell-penetrating inhibitor of one or more caspases and a potent inhibitor of caspase-1 activation in NLRP3-induced cells (Dostert C. et al., 2009. Malarial hemozoin is a Nalp3 inflammasome activating danger signal. PLoS One. 4(8):e6510.). Z-VAD-FMK irreversibly binds to the catalytic site of caspase proteases (Slee EA. et al., 1996. Benzyloxycarbonyl-Val-Ala-Asp (OMe) fluoromethylketone (Z-VAD.FMK) inhibits apoptosis by blocking the processing of CPP32. Biochem J. 315 (Pt 1):21-4.).

[00460] In some embodiments, the caspase-1 inhibitor is Ac-YVAD-cmk, which has the following structure:

and is a caspase-1 inhibitor and a chloromethyl ketone tetrapeptide based on the target sequence of proIL-1β (YVHD). Ac-YVAD cmk has been reported to block inflammasome activation and therefore exhibit anti-inflammatory, anti-apoptotic and antipyroptotic effects.

[00461] In some embodiments, the caspase-1 inhibitor is Ac-YVAD-CHO, which has the following structure:

(Brenner, B., et al. 1998. Cell Death Differ. 5: 29-37. PMID: 10200443) Caspase-1 substrate (CAS 143305-11-7)

[00462] In some embodiments, the caspase-1 inhibitor is parthenolide, which has the following structure:

[00463] Parthenolide, a sesquiterpene lactone derived from pyrethrum, is a known inhibitor of NF-κB activation and a direct inhibitor of caspase-1 and multiple inflammasomes, including the NLRP3 and NLRP1 inflammasomes (Juliana C. et al., 2010. Anti-inflammatory compounds parthenolide and Bay 11-7082 are direct inhibitors of the inflammasome. J Biol Chem. 285(13):9792-802.). Parthenolide directly inhibits the NLRP3 inflammasome by disrupting the NLRP3 ATPase activity.

[00464] In some embodiments, the caspase-1 inhibitor is one or a combination of: pralnacasane (VX-740), which has the following structure:

[00465] Z-WEHD-FMK (also known as benzyloxycarbonyl-V-A-D-O-methyl fluoromethyl ketone).

[00466] In some embodiments, the caspase-1 inhibitor is shikonin or acetylshikonin, wherein shikonin is:

and acetylshikonin is:

Shikonin is a highly lipophilic naphthoquinone found in the roots of Lithospermum erythrorhizon, which is used in traditional Chinese medicine for its pleiotropic effects, and inhibits NLRP3 inflammasome activation (Zorman et al., PLOS One, 2016; 11(7); e0159826.).

[00467] In some embodiments, the caspase-1 inhibitor may be a small molecule inhibitor, as will be appreciated by one of skill in the art. Non-limiting examples include cyanopropanate-containing molecules such as (S)-3-((S)-1-((S)-2-(4-amino-3-chlorobenzamido)-3,3-dimethylbutanoyl)pyrrolidine-2-carboxamido)-3-cyanopropanoic acid, as well as other small molecule caspase-1 inhibitors such as ((2R,3S)-2-ethoxy-5-oxotetrahydrofuran-3-yl)amide (S)-1-((S)-2-{[1-(4-amino-3-chlorophenyl)methanoyl]amino}-3,3-dimethylbutanoyl)pyrrolidine-2-carboxylic acid. Such inhibitors can be synthesized chemically.

[00468] In some embodiments, the caspase-1 inhibitor may be a direct inhibitor of caspase-1 enzymatic activity or may be an indirect inhibitor that inhibits initiation of inflammasome assembly or inflammasome signal propagation. Caspase-1 inhibitors for use in the present invention may be antioxidants, including reactive oxygen species (ROS) inhibitors. Examples of such caspase-1 inhibitors include, but are not limited to, flavonoids, including flavones such as apigenin, luteolin, and diosmin; flavonols such as myricetin, fisetin, and quercetin; flavanols and polymers thereof such as catechin, gallocatechin, epicatechin, epigallocatechin, epigallocatechin-3-gallate, and theaflavin; isoflavone phytoestrogens; and stilbenoids such as resveratrol. Also included are phenolic acids and their esters such as gallic acid and salicylic acid; terpenoids or isoprenoids such as andrographolide and parthenolide; vitamins such as vitamins A, C and E; vitamin cofactors such as coenzyme Q10, manganese and iodide, other organic antioxidants such as citric acid, oxalic acid, phytic acid and alpha-lipoic acid, and Rhus verniciflua Stokes varnish tree extract. The caspase-1 inhibitor may be a combination of these compounds, for example a combination of alpha-lipoic acid, coenzyme Q10 and vitamin E, or a combination of a caspase 1 inhibitor(s) with another inflammasome inhibitor such as glyburide or a functionally equivalent precursor or derivative thereof.

[00469] Examples of dosages of some inflammasome inhibitors are listed below: apigenin (about 0.1-10 mg/kg), luteolin (about 1-100 mg), diosmin (about 100-900 mg), myricetin (about 10-300 mg), quercetin (about 10-1000 mg), fisetin (1-200 mg/kg), Rhus verniciflua Stokes extract (1-100 mg/kg), catechin (about 50-500 mg), gallocatechin (about 100-1000 mg), epicatechin (about 0.1-10 mg/kg), epigallocatechin (about 100-1000 mg), epigallocatechin-3-gallate (about 100-1,000 mg), theaflavin (approximately 75-750 mg), isoflavone phytoestrogens (approximately 25-250 mg), resveratrol (approximately 100-1,000 mg), andrographolide (approximately 100-500 mg), parthenolide (approximately 0.1-50 mg), vitamin A (approximately 5,000-20,000 IU), vitamin C (approximately 100-2,000 mg), coenzyme Q10 (approximately 30-500 mg), vitamin E (approximately 10-1,000 IU), alpha-lipoic acid (approximately 10-1,000 mg), coenzyme Q10 (30-500 mg), manganese (approximately 1-100 mg), alpha-lipoic acid, coenzyme Q10 and vitamin E (approximately 10-1000 mg, 30-500 mg, 10-1000 IU, respectively), glyburide (approximately 1-20 mg) and a glyburide derivative that lacks the cyclohexylurea group (approximately 1-200 mg).

[00470] In some embodiments, the caspase-1 inhibitor is any of the small molecule compounds described in U.S. Patents 6,355,618; 6,632,962, 5,756,466, or international applications WO2001/042216; WO2004/064713, WO98/16502, WO 97/24339, EP623592, and Dolle et al., J. Med. Chem. 39, 2438 (1996); Dolle et al., J. Med. Chem. 40, 1941 (1997), the contents of each of which are incorporated herein by reference in their entireties. In some embodiments, the caspase-1 inhibitor has been reported to be non-peptide caspase-1 inhibitors. US Patent No. (Bemis et al.);

[00471] In some embodiments, the caspase-1 inhibitor is an ICE (caspase-1) inhibitor having the structure:

wherein R ₁ is, but is not limited to, R ₃ CO-, R ₃ is, but is not limited to, C ₁ -C ₆ alkyl, aryl, heteroaryl, -(CHR) _n -aryl, and -(CHR) _n -heteroaryl, and R ₂ is selected from various groups. In some embodiments, the caspase-1 inhibitor is an ICE (caspase-1) inhibitor having the structure:

wherein R ₁ includes aryl and heteroaryl; A is an amino acid; n is 0-4; m is 0 or 1; and R ₂ is aryl. According to some embodiments, the caspase-1 inhibitor is an ICE (caspase-1) inhibitor having the structure:

wherein R ₁ includes aryl and heteroaryl; AA1 and AA2 are single bonds or amino acid residues; Tet is a tetrazole ring; Z is alkylene, alkenylene, O, S, etc.; and E is H, alkyl, etc.

[00472] In some embodiments of the compositions and methods described herein, the caspase-1 inhibitor is an RNA-based caspase-1 inhibitor, such as a siRNA specific for caspase-1. In some embodiments of the compositions and methods described herein, the RNA-based AIM2 inhibitor is encoded by ccDNA.

[00473] In some embodiments of the compositions and methods described herein, the caspase-1 inhibitor is an RNA-based caspase-1 inhibitor, such as a caspase-1-specific siRNA. In some embodiments of the compositions and methods described herein, the RNA-based caspase-1 inhibitor is encoded by ccDNA. Exemplary caspase-1 siRNA sequences contemplated for use in the kits and compositions described herein are disclosed in WO2008/033285; Keller, M., et al. Cell. 2008; 132(5): 818-831; Artlett, C. M., et al. Arthritis and Rheumatology. 2011 Jul; 63 (11): 3563-3574; Burdette, D., et al. J Gen Virology. 2012, 93: 235-246, which are incorporated herein by reference in their entirety. The siRNA sequences for caspase-1 are also commercially available and known to those of ordinary skill in the art.

[00474] The human caspase-1 protein is encoded by the CASP1 gene, which contains the nucleic acid sequence NM_033292.3 (SEQ ID NO: 611), and the human caspase-1 protein has the amino acid sequence NP_150634.1 (SEQ ID NO: 612). Caspase-1 inhibitors further include antisense polynucleotides that can be used to inhibit transcription of the caspase-1 gene and thereby inhibit caspase-1 and the downstream NLRP3 inflammasome and AIM2 inflammasome pathways. Polynucleotides that are complementary to a segment of a polynucleotide encoding caspase-1 (e.g., a polynucleotide as set forth in SEQ ID NO: 611) are designed to bind to mRNA encoding caspase-1 and to inhibit translation of such mRNA. Antisense polynucleotides may be encoded by a ccDNA vector disclosed herein and may optionally be operably linked to a tissue-specific or inducible promoter disclosed herein.

[00475] Inhibition of caspase-1 or procaspase-1 mRNA can be accomplished using RNAi gene silencing molecules according to methods commonly known to those skilled in the art. For example, oligonucleotide duplexes of siRNA gene silencing molecules specifically targeting human caspase-1 (NM_033292.3) can be easily used to suppress the expression of procaspase-1. Targeting caspase-1 mRNA can be successfully accomplished using siRNA; and other siRNA molecules can be easily prepared by those skilled in the art based on the known sequence of the target mRNA. Accordingly, to avoid any doubt, one of ordinary skill in the art can design nucleic acid inhibitors, such as RNAi agents (silencing RNAs), for the nucleic acid sequence of NM_033292.3, which is shown below:

1 atactttcag tttcagtcac acaagaaggg aggagagaaa agccatggcc gacaaggtcc

61 tgaaggagaa gagaaagctg tttatccgtt ccatgggtga aggtacaata aatggcttac

121 tggatgaatt attacagaca agggtgctga acaaggaaga gatggagaaa gtaaaacgtg

181 aaaatgctac agttatggat aagacccgag ctttgattga ctccgttatt ccgaaagggg

241 cacaggcatg ccaaatttgc atcacataca tttgtgaaga agacagttac ctggcaggga

301 cgctgggact ctcagcagat caaacatctg gaaattacct taatatgcaa gactctcaag

361 gagtactttc ttcctttcca gctcctcagg cagtgcagga caacccagct atgcccacat

421 cctcaggctc agaagggaat gtcaagcttt gctccctaga agaagctcaa aggatatgga

481 aacaaaagtc ggcagagatt tatccaataa tggacaagtc aagccgcaca cgtcttgctc

541 tcattatctg caatgaagaa tttgacagta ttcctagaag aactggagct gaggttgaca

601 tcacaggcat gacaatgctg ctacaaaatc tggggtacag cgtagatgtg aaaaaaaatc

661 tcactgcttc ggacatgact acagagctgg aggcatttgc acaccgccca gagcacaaga

721 cctctgacag cacgttcctg gtgttcatgt ctcatggtat tcgggaaggc atttgtggga

781 agaaacactc tgagcaagtc ccagatatac tacaactcaa tgcaatcttt aacatgttga

841 ataccaagaa ctgcccaagt ttgaaggaca aaccgaaggt gatcatcatc caggcctgcc

901 gtggtgacag ccctggtgtg gtgtggttta aagattcagt aggagtttct ggaaacctat

961 ctttaccaac tacagaagag tttgaggatg atgctattaa gaaagcccac atagagaagg

1021 attttatcgc tttctgctct tccacaccag ataatgtttc ttggagacat cccacaatgg

1081 gctctgtttt tattggaaga ctcattgaac atatgcaaga atatgcctgt tcctgtgatg

1141 tggaggaaat tttccgcaag gttcgatttt catttgagca gccagatggt agagcgcaga

1201 tgcccaccac tgaaagagtg actttgacaa gatgtttcta cctcttccca ggacattaaa

1261 ataaggaaac tgtatgaatg tctgtgggca ggaagtgaag agatccttct gtaaaggttt

1321 ttggaattat gtctgctgaa taataaactt ttttgaaata ataaatctgg tagaaaaatg

1381 aaaaaaaaa aaa (SEQ ID NO: 611)

[00476] In some embodiments, the caspase-1 inhibitor is an RNAi that is complementary to the RNAi target sequence in NM_033292.3 (SEQ ID NO: 611); also referred to as NCBI gene 834 (CASP1). Currently, wild-type caspase-1 transcripts include: NM_001223.4, NM_001257118.2, NM_001257119.2, NM_033292.3 (SEQ ID NO: 611), NM_033293.3, NM_033294.3, NM_033295.3, XM_017018393.1, XM_017018394.1, XM_017018395.1, XM_017018396.1. The RNAi agent that inhibits caspase-1 may be a nucleic acid complementary to the 17-21 consecutive bases of SEQ ID NOs: 613-619, which are shown in Table 5E.

[00477] Table 5E: RNAi target sequences for caspase-1 inhibition:

Target sequence SEQ ID NO: Clone ID CACACGTCTTGCTCTCATTAT 613 TRCN0000003504 CTACAACTCAATGCAATCTTT 614 TRCN0000003503 CCAGATATACTACAACTCAAT 615 TRCN0000003502 GAAGAGTTTGAGGATGATGCT 616 TRCN0000010796 CCATGGGTGAAGGTACAATAA 617 TRCN0000118461 GCTTTGATTGACTCCGTTATT 618 TRCN0000118459 GAAGGTACAATAAATGGCTTA 619 TRCN0000118460

[00478] In some embodiments, the caspase-1 inhibitor is an siRNA agent. Exemplary siRNA sequences that inhibit caspase-1 are shown in Table 5F.

[00479] Table 5F: Exemplary siRNAs that inhibit caspase-1

Transcript Clone ID Target sequence Forward and reverse oligonucleotide sequences NM_033294.3 TRCN0000003504 CACACGTCTTGCTCTCATTAT (SEQ ID NO: 620) Direct: CCGGCACACGTCTTGCTCTCATTATCTCGAGATAATGAGAGCAAGACGTGTGTTTTTG (SEQ ID NO: 621) Reverse: AATTCAAAACACACGTCTTGCTCTCATTATCTCGAGATAATGAGAGCAAGACGTGTG (SEQ ID NO: 622) NM_033294.3 TRCN0000003503 CTACAACTCAATGCAATCTTT (SEQ ID NO: 623) Direct: CCGGCTACAACTCAATGCAATCTTTCTCGAGAAAGATTGCATTGAGTTGTAGTTTTTG (SEQ ID NO: 624) Reverse: AATTCAAAACTACAACTCAATGCAATCTTTCTCGAGAAAGATTGCATTGAGTTGTAG (SEQ ID NO: 625) NM_033294.3 TRCN0000003502 CCAGATATACTACAACTCAAT (SEQ ID NO: 626) Direct: CCGGCCAGATATACTACAACTCAATCTCGAGATTGAGTTGTAGTATATCTGGTTTTTG (SEQ ID NO: 627) Reverse: AATTCAAAACCAGATATACTACAACTCAATCTCGAGATTGAGTTGTAGTATATCTGG (SEQ ID NO: 628) NM_033294.3 TRCN0000010795 TGTATGAATGTCTGCTGGGCA (SEQ ID NO: 629) Direct: CCGGTGTATGAATGTCTGCTGGGCACTCGAGTGCCCAGCAGACATTCATACATTTTTG (SEQ ID NO: 630) Reverse: AATTCAAAATGTATGAATGTCTGCTGGGCACTCGAGTGCCCAGCAGACATTCATACA (SEQ ID NO: 631) NM_033294.3 TRCN0000139687 CAAGGACCTGAAGGAGAAGAA (SEQ ID NO: 632) Direct: CCGGCAAGGACCTGAAGGAGAAACTCGAGTTCTTCTCCTTCAGGTCCTTGTTTTTG (SEQ ID NO: 633) Reverse: AATTCAAAACAAGGACCTGAAGGAGAAGAACTCGAGTTCTTCTCCTTCAGGTCCTTG (SEQ ID NO: 634) NM_033294.3 TRCN0000138367 CAATGTCTGTGGGAGGAAGAA (SEQ ID NO: 635) Direct: CCGGCAATGTCTGTGGGAGGAAACTCGAGTTCTTCCTCCCACAGACATTGTTTTTG (SEQ ID NO: 636) Reverse: AATTCAAAAACAATGTCTGTGGGAGGAAGAACTCGAGTTCTTCCTCCCACAGACATTG (SEQ ID NO: 637) NM_033294.3 TRCN0000072917 CAAGGTCCTGTAGGGAGAAGA (SEQ ID NO: 638) Direct: CCGGCAAGGTCCTGTAGGGAGAAGACTCGAGTCTTCTCCCTACAGGACCTTGTTTTTG (SEQ ID NO: 639) Reverse: AATTCAAAACAAGGTCCTGTAGGGAGAAGACTCGAGTCTTCTCCCTACAGGACCTTG (SEQ ID NO: 640) NM_033294.3 TRCN0000233250 CAAGGTCCTGTAGGGAGAAGA (SEQ ID NO: 641) Direct: CCGGCAAGGTCCTGTAGGGAGAAGACTCGAGTCTTCTCCCTACAGGACCTTGTTTTTG (SEQ ID NO: 642) Reverse: AATTCAAAACAAGGTCCTGTAGGGAGAAGACTCGAGTCTTCTCCCTACAGGACCTTG (SEQ ID NO: 643) NM_033294.3 TRCN0000321071 ACAAGCCCAAGGTGATCATTA (SEQ ID NO: 644) Direct: CCGGACAAGCCCAAGGTGATCATTACTCGAGTAATGATCACCTTGGGCTTGTTTTTTG (SEQ ID NO: 645) Reverse: AATTCAAAAAACAAGCCCAAGGTGATCATTACTCGAGTAATGATCACCTTGGGCTTGT (SEQ ID NO: 646) NM_033294.3 TRCN0000125361 CAAGGACTTGAAGGAGAAGAA (SEQ ID NO: 647) Direct: CCGGCAAGGACTTGAAGGAGAAGAACTCGAGTTCTTCTCCTTCAAGTCCTTGTTTTTG (SEQ ID NO: 648) Reverse: AATTCAAAACAAGGACTTGAAGGAGAAGAACTCGAGTTCTTCTCCTTCAAGTCCTTG (SEQ ID NO: 649) NM_033294.3 TRCN0000006653 CCCAAGTTTGAAGTACAAGTA (SEQ ID NO: 650) Direct: CCGGCCCAAGTTTGAAGTACAAGTACTCGAGTACTTGTACTTCAAACTTGGGTTTTTG (SEQ ID NO: 651) Reverse: AATTCAAAAACCCAAGTTTGAAGTACAAGTACTCGAGTACTTGTACTTCAAACTTGGG (SEQ ID NO: 652) NM_033294.3 TRCN0000058747 CCCAGGACATGATAATAAGAT (SEQ ID NO: 653) Direct: CCGGCCCAGGACATGATAATAAGATCTCGAGATCTTATTATCATGTCCTGGGTTTTTG (SEQ ID NO: 654) Reverse: AATTCAAAAACCCAGGACATGATAATAAGATCTCGAGATCTTATTATCATGTCCTGGG (SEQ ID NO: 655) NM_033294.3 TRCN0000153291 GAATTTGACAGTTTCCTGCCA (SEQ ID NO: 656) Direct: CCGGGAATTTGACAGTTTCCTGCCACTCGAGTGGCAGGAAACTGTCAAATTCTTTTTG (SEQ ID NO: 657) Reverse: AATTCAAAAAGAATTTGACAGTTTCCTGCCACTCGAGTGGCAGGAAACTGTCAAATTC (SEQ ID NO: 658) NM_033294.3 TRCN0000073644 CCCAAGTTTGAGGTCAAAGTT (SEQ ID NO: 659) Direct: CCGGCCCAAGTTTTGAGGTCAAAGTTCTCGAGAACTTTGACCTCAAACTTGGGTTTTTG (SEQ ID NO: 660) Reverse: AATTCAAAAACCCAAGTTTGAGGTCAAAGTTCTCGAGAACTTTGACCTCAAACTTGGG (SEQ ID NO: 661) NM_033294.3 TRCN0000038805 CGACAAGATGTTCTCCCTCAA (SEQ ID NO: 662) Direct: CCGGCGACAAGATGTTCTCCCTCAACTCGAGTTGAGGGAGAACATCTTGTCGTTTTTG (SEQ ID NO: 663) Reverse: AATTCAAAACGACAAGATGTTCTCCCTCAACTCGAGTTGAGGGAGAACATCTTGTCG (SEQ ID NO: 664)

[00480] In some embodiments, the caspase-1 inhibitor is an siRNA, thereby inhibiting caspase-1 mRNA (or procaspase-1 proprotein), which results in inhibition of the downstream NLRP3 inflammasome and/or AIM2 inflammasome pathways. In some embodiments, the caspase-1 inhibitor is GAA GGC CCA UAU AGA GAA A (SEQ ID NO: 904; sense strand sequence shown), which inhibits the expression of human caspase-1 or a fragment or homolog thereof that is at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90% identical thereto. Examples of caspase-1 siRNA sequences contemplated for use in the kits and compositions described herein are disclosed in WO 2008/033285 or US20090280058, Keller, M., et al. Cell. 2008; 132(5): 818-831; Artlett, C.M., et al. Arthritis and Rheumatology. 2011 Jul; 63 (11): 3563-3574; Burdette, D., et al. J Gen Virology. 2012, 93: 235-246; which are incorporated herein by reference in their entireties.

[00481] Custom siRNAs for NLRP3, AIM2, and caspase-1 can be made to order from Dharmacon Research, Inc., Lafayette, Colorado. Other sources for custom siRNAs include Xeragon Oligonucleotides, Huntsville, Alabama, and Ambion, Austin, Texas. Alternatively, siRNAs can be chemically synthesized using ribonucleoside phosphoamidites and a DNA/RNA synthesizer. In some embodiments, RNAi or siRNAs for NLRP3, AIM2, and caspase-1 can be encoded in the ccDNA disclosed herein.

[00482] In some embodiments, the caspase-1 inhibitor is a caspase-1 substrate (CAS 143305-11-7) having the structure:

and which has the following sequence: Asn-Glu-Ala-Tyr-Val-His-Asp-Ala-Pro-Val-Arg-Ser-Leu-Asn (SEQ ID NO: 538). In some embodiments of the compositions and methods described herein, the caspase-1 inhibitor is encoded by ccDNA administered to the subject (including, for example, subsequent delivery of ccDNA). In some embodiments of the compositions and methods described herein, the caspase-1 inhibitor encoded by ccDNA administered to the subject is a caspase-1 substrate (SEQ ID NO: 538).

[00483] RNAi can be designed to target a variety of mRNAs. A general strategy for designing RNAi, such as siRNA, involves starting at an AUG stop codon and then scanning the length of the target cDNA target for AA dinucleotide sequences. The 19 3'-terminal nucleotides adjacent to the AA sequences have been reported to be potential siRNA target sites. The potential target sites were then compared to a relevant genome database so that any target sequences that have significant homology to non-target genes could be discarded. Multiple target sequences were localized along the length of the gene, so that target sequences were derived from the 3', 5', and medial portions of the mRNA. Negative control siRNAs were prepared using the same nucleotide composition as the siRNAs of the present invention, but were scrambled and tested for lack of sequence homology to any genes in the transfected cells (Elbashir, S. M., et al., 2001, Nature, 411, 494-498; Ambion siRNA Design Protocol, at www.ambion.com).

[00484] Target sequences can be 17-25 bases in length, and optimally 21 bases in length, starting with AA. RNAi or siRNA that bind to target sequences have been modified with a thiol group at carbon 5 of C6 on one strand.

VII. Methods of application

[00485] The ccDNA vector for expressing, for example, an inhibitor of an immune response (e.g., an innate immune response) disclosed herein can also be used in a method of delivering a nucleotide sequence of interest (e.g., encoding an inhibitor of an innate immune response) to a target cell (e.g., a host cell). The method can be, in particular, a method of delivering an inhibitor of an immune response (e.g., an innate immune response) to a cell of a subject in need thereof and treating an immune disorder or weakening or suppressing the innate immune system. The present invention provides for in vivo expression of an inhibitor of an immune response (e.g., an innate immune response) encoded in a ccDNA vector in a cell in a subject such that a therapeutic effect of expressing an inflammasome antagonist occurs. These results are observed in both in vivo and in vitro modes of delivery of the ccDNA vector.

[00486] The present invention further provides a method for delivering an inhibitor of an immune response (e.g., an innate immune response), e.g., to a cell of a subject in need thereof, comprising multiple administrations of a ccDNA vector of the present invention encoding said inflammasome antagonist. Since the ccDNA vector of the present invention does not induce an immune response similar to that typically observed against encapsidated viral vectors, such a multiple administration strategy is likely to be more successful in a ccDNA-based system. The ccDNA vector is administered in amounts sufficient to transfect target tissue cells and provide sufficient levels of gene transfer and expression of the inhibitor of the immune response (e.g., an innate immune response), e.g., without undue adverse effects. Conventional and pharmaceutically acceptable routes of administration include, but are not limited to, intraretinal administration (e.g., subretinal injection, suprachoroidal injection, or intravitreal injection), intravenous administration (e.g., in a liposomal formulation), direct delivery to a selected organ (e.g., any one or more tissues selected from the liver, kidney, gallbladder, prostate, adrenal gland, heart, intestine, lung, and stomach), intramuscular, and other parenteral routes. Routes of administration may be combined if desired.

[00487] Delivery of a ccDNA vector for expression of, for example, an inhibitor of an immune response (e.g., an innate immune response) described herein is not limited to delivery of the expressed inhibitor. For example, ccDNA vectors produced by a conventional method (e.g., using a cell-based production method (e.g., insect cell production methods)) or a synthetic method as described herein can be used with other provided delivery systems to provide part of a gene therapy. One non-limiting example of a system that can be combined with ccDNA vectors in accordance with the present disclosure includes systems that separately deliver one or more cofactors or immune suppressors for efficient gene expression of a ccDNA vector expressing an inhibitor.

[00488] The present invention also provides a method of suppressing an immune response, such as an innate immune response, in a subject, comprising administering to a target cell in need thereof (in particular a muscle cell or tissue) of said subject a therapeutically effective amount of a ccDNA vector, optionally with a pharmaceutically acceptable carrier. Although said ccDNA vector may be administered in the presence of a carrier, such a carrier is not necessary. The selected ccDNA vector comprises a nucleotide sequence encoding an inhibitor of an immune response (e.g. an innate immune response), for example, which can be used to treat or suppress the immune system. In particular, said ccDNA vector may comprise an inflammasome antagonist target sequence operably linked to control elements capable of directing the transcription of an inflammasome antagonist target encoded by said exogenous DNA sequence upon administration to the subject. The said ccDNA vector may be administered by any suitable route as described above and elsewhere herein.

[00489] The compositions and vectors provided herein can be used to deliver an inhibitor of an immune response (e.g., an innate immune response), for example, for various purposes. In some embodiments, the transgene encodes an inhibitor of an immune response (e.g., an innate immune response) that is for use in research purposes, such as to create a somatic transgenic animal model carrying the transgene, such as to study the function of the inhibitor of the immune response (e.g., an innate immune response). In another example, the transgene encodes an inhibitor of an immune response (e.g., an innate immune response) that is for use in creating an animal model of a suppressed immune system or in an immunocompromised subject. In some embodiments, the encoded inhibitor of an immune response (e.g., an innate immune response) can be used to treat or prevent exaggerated immune responses or a state of innate immune hyperreactivity in a subject, such as in response to gene therapy or the like, in a mammalian subject. An inhibitor of an immune response (e.g., an innate immune response) can be transferred to a patient (e.g., expressed in the patient) in an amount sufficient to reduce or prevent exaggerated immune responses in the subject.

[00490] A ccDNA vector is not limited to a single ccDNA vector molecule. Thus, in another aspect, multiple ccDNA vectors expressing different proteins or the same immune response (e.g., innate immune response) inhibitors, for example, but operably linked to different promoters or cis-regulatory elements, can be delivered simultaneously or sequentially to a target cell, tissue, organ, or subject. Thus, this strategy can provide for simultaneous gene therapy or gene delivery of multiple inflammasome antagonists. It is also possible to place different portions of the inhibitor on separate ccDNA vectors (e.g., different domains and/or cofactors required for the functionality of the immune response (e.g., innate immune response) inhibitor), for example, which can be administered simultaneously or at different times and which can be separately regulated, thereby adding an additional level of control over the expression of one or more inhibitors. Delivery can also be performed multiple times and, importantly for gene therapy in clinical settings, with subsequent dose increases or decreases, given the lack of host immune response against the capsid due to the absence of the viral capsid. No anti-capsid response is expected due to the absence of the capsid.

[00491] The present invention also provides a method of suppressing an immune response, such as an innate immune response, in a subject, comprising administering to a target cell in need thereof (particularly a muscle cell or tissue) of said subject a therapeutically effective amount of a ccDNA vector disclosed herein, optionally with a pharmaceutically acceptable carrier. Although said ccDNA vector may be administered in the presence of a carrier, such a carrier is not required. An implemented ccDNA vector comprises a nucleotide sequence of interest, such as an immune response inhibitor, that can be used to suppress the innate immune system or alleviate a state of increased immunoreactivity in a subject. In particular, the ccDNA vector may comprise a target exogenous DNA sequence operably linked to control elements capable of directing the transcription of a target polypeptide, protein, or oligonucleotide encoded by said exogenous DNA sequence upon administration to the subject. The cDNA vector may be administered by any suitable route as described above and elsewhere in this document.

Ex vivo treatment

[00492] In some embodiments, cells are removed from a subject, introduced into the cells a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response), such as those disclosed herein, and then returned to the subject. Methods for removing cells from a subject for ex vivo treatment and then reintroducing them into the subject are known in the art (see, e.g., U.S. Patent No. 5,399,346; the disclosure of which is incorporated herein in its entirety). Alternatively, the ccDNA vector is introduced into cells obtained from another subject, into cultured cells, or into cells from any other suitable source, and the cells are administered to a subject in need thereof.

[00493] Cells transduced with a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response), such as those disclosed herein, are preferably administered to a subject in a "therapeutically effective amount" in combination with a pharmaceutical carrier. Those skilled in the art will appreciate that the therapeutic effects need not be complete or curative, so long as they are beneficial to the subject in some way.

[00494] In some embodiments, a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response), such as those disclosed herein, can encode an inflammasome antagonist described herein (sometimes referred to as a transgene or heterologous nucleotide sequence) that is to be produced in a cell in vitro, ex vivo, or in vivo. For example, in contrast to the use of ccDNA vectors described herein in the method of treatment discussed herein, in some embodiments, a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response) can be introduced into cultured cells and the expressed inflammasome antagonist can be isolated from the cells, such as to produce antibodies and fusion proteins. In some embodiments, cultured cells comprising a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response) disclosed herein can be used for the commercial production of antibodies or fusion proteins, such as as a source of cells for small-scale or large-scale biomanufacturing of antibodies or fusion proteins. In alternative embodiments, a ccDNA vector for expressing an inhibitor of an immune response (e.g., an innate immune response) disclosed herein is introduced into cells in a non-human host subject for the production of antibodies or fusion proteins in vivo, including small-scale production as well as commercial large-scale production of an inflammasome antagonist.

[00495] The cDNA vectors for expressing an inhibitor of an immune response (e.g., an innate immune response) disclosed herein can be used in both veterinary and medical applications. Suitable subjects for the ex vivo gene delivery methods described above include both birds (e.g., chickens, ducks, geese, quail, turkeys, and pheasants) and mammals (e.g., humans, cattle, ovine, caprine, equine, feline, canine, and lagomorph), with mammals being preferred. Human subjects are most preferred. Human subjects include neonates, infants, children and adolescents, and adult humans.

[00496] All patents and other publications; including literature references, issued patents, published patent applications, and patent applications co-pending with this application; mentioned in this specification are expressly incorporated herein by reference for the purpose of describing and disclosing, for example, methodologies described in such publications that may be used in connection with the technology described herein. These publications are provided solely by reason of their disclosure prior to the filing date of this application. Nothing in this regard should be construed as an admission that the inventors do not have rights to prior invention with respect to such disclosure, by virtue of prior invention or for any other reason. All statements regarding dates or representations regarding the contents of these documents are based on information available to applicants and do not constitute an admission in any way of the correctness of the dates or contents of these documents.

[00497] The description of embodiments of the present disclosure should not be considered exhaustive or to limit the present invention to the precise form disclosed. Although specific embodiments and examples of the disclosure have been described herein for illustrative purposes, various equivalent modifications are possible within the scope of the disclosure, as will be understood by those skilled in the art. For example, although the steps or functions of a method are presented in the order shown, in alternative embodiments the functions may be performed in a different order, or the functions may be performed substantially simultaneously. The principles of the present disclosure provided herein may be applied to other procedures or methods as appropriate. The various embodiments described herein may be combined to provide additional embodiments. Aspects of the present disclosure may be modified, as necessary, to employ the compositions, functions, and concepts of the above references and applied to provide additional embodiments of the present disclosure. In addition, certain changes in the protein structure may be made for reasons of biological functional equivalence without affecting the biological or chemical effect, either qualitatively or quantitatively. These and other changes may be made to the disclosure in light of the detailed description. All such modifications are intended to be included within the scope of the appended claims.

[00498] Specific elements of any of the above embodiments may be combined or replaced with elements of other embodiments. Furthermore, although advantages associated with some embodiments of the present disclosure have been described in the context of those embodiments, other embodiments may also exhibit such advantages, and not all embodiments need necessarily exhibit such advantages to fall within the scope of the disclosure.

[00499] The technology described herein is further illustrated by the following examples, which should in no way be construed as further limitations. It should be understood that the present invention is not limited to the particular methodology, protocols, and reagents, etc., described herein, and therefore admits variations. The terminology used herein is intended solely to describe particular embodiments, and is not intended to limit the scope of the present invention, which is defined solely by the claims.

EXAMPLES

EXAMPLE 1: Construction of TTX plasmids

[00500] TTX format plasmids were prepared, the structural diagram of which is shown in FIG. 4C (TTX-R) or FIG. 4D (TTX-L). Exemplary TTX-R and TTX-L plasmids are described in Table 6A below. TTX-R and TTX-L plasmids differ in the position of the mutated AAB2 ITR sequence, as shown in FIG. 4C and FIG. 4D, respectively. TTX-R plasmids (TTX plasmid 1, 3, 5, and 7) were prepared by the molecular cloning disclosed herein to produce TTX vectors. TTX-L plasmids (TTX plasmid 2, 4, 6, and 8) for use in producing TTX vectors (TTX vector 2, 4, 6, 8). Each of the TTX-R plasmids contains (a) a wild-type AAB2 inverted terminal repeat (ITR); (b) an expression cassette, and (c) a modified AAB2 inverted terminal repeat (ITR), as illustrated in FIG. 4D.

[00501] ccDNA plasmids (i.e., plasmids containing a ccDNA vector template used to subsequently produce a ccDNA vector) can be constructed using known techniques to preferably provide at least the following elements as operably linked components downstream of transcription: a 5'-ITR (mutant or wild-type AAB); control elements including a promoter, an exogenous DNA sequence of interest; a transcription termination region; and a 3'-ITR (mutant or wild-type of the corresponding AAB ITR). Notably, the nucleotide sequences in the ITRs substantially replace the regions encoding rep and cap. Although the rep sequences are ideally encoded by a helper plasmid or vector, they can alternatively be carried by the vector plasmid itself. In such cases, the rep sequences are preferably located outside the inter-ITR region, but can also be located within the inter-ITR region. The target exogenous DNA sequence is operably linked to control elements that direct transcription or expression of the encoded polypeptide, protein, or oligonucleotide thereof in a cell, tissue, organ, or in the subject's body (i.e., in vitro, ex vivo, or in vivo). Such control elements may comprise control sequences typically associated with the selected gene. Alternatively, heterologous control sequences may be used. Suitable heterologous control sequences typically include those derived from sequences encoding mammalian or viral genes.

[00502] The target exogenous DNA sequence in the ccDNA vector may be operably linked to control elements that direct the transcription or expression of the encoded polypeptide, protein, or oligonucleotide thereof in a cell, tissue, organ, or in the subject (i.e., in vitro, ex vivo, or in vivo). Such control elements may comprise control sequences typically associated with the gene of choice. Alternatively, heterologous control sequences may be used. Suitable heterologous control sequences typically include those derived from sequences encoding mammalian or viral genes. Examples include, but are not limited to, promoters such as the SV40 early promoter; the promoter from the mouse mammary tumor virus long terminal repeat (LTR); the adenovirus large late promoter (Ad MLP); herpes simplex virus (HSV) promoters; cytomegalovirus (CMV) promoter such as CMV immediate early promoter region (CMVIE); Rous sarcoma virus (RSV) promoter; synthetic promoters; hybrid promoters; and the like. In addition, sequences derived from non-viral genes such as the mouse metallothionein gene can also be used in the present invention. ITR sequences of many AAV serotypes are known.

[00503] The expression cassette of each TTX plasmid (both TTX-R and TTX-L) includes the following elements located between the ITR sequences: (i) an enhancer/promoter; (ii) a cloning site for the transgene; (iii) a WHP post-transcriptional response element (WPRE); and (iv) a polyadenylation signal from the bovine growth hormone gene (BGHpA). Unique restriction endonuclease recognition sites (R1-6) were also introduced between each component (e.g., see FIG. 4C and FIG. 4D) to facilitate the introduction of new genetic components at specific sites in the construct. R3 and R4 enzyme sites were engineered into the cloning site to introduce the open reading frame of the transgene. These sequences were cloned into pFastBac HT B plasmid obtained from ThermoFisher Scientific.

[00504] All TTX plasmids additionally contain an exogenous sequence, the open reading frame of which for the transgene (firefly luciferase or "Luc" or human factor IX or "FIX") was also obtained by inserting the exogenous sequence into the cloning site. Each of the structures of several exemplary TTX plasmids presented in Table 6A was constructed according to the scheme shown in FIG. 4D (right mutated ITR AAB) or FIG. 4C (left mutated ITR). Each TTX plasmid included an enhancer/promoter and a transgene (e.g. luciferase with various promoters or FIX with the CAG promoter), a post-translational regulatory element (WPRE), and a polyadenylation termination signal (poly(A) BGH), flanked by: (a) the mutated AAB2 inverted terminal repeat (ITR) polynucleotide sequence encoded in the plasmid on either the left (L) or right (R) side of the expression cassette, and (b) the wild-type (unmutated) AAB2 ITR sequence at the opposite end of the expression cassette.

[00505] The TTX plasmids in Table 6A were constructed with WPRE containing SEQ ID NO: 8 and BGHpA containing SEQ ID NO: 9 as components between the luciferase transgene and the right ITR. In addition, each of the TTX plasmids (TTX-1 through TTX-10) also contained an R3/R4 cloning site (SEQ ID NO: 7) on either side of the ORF of a luciferase or factor IX reporter sequence (Padua FIX of SEQ ID NO: 12 or FIX of SEQ ID NO: 11).

[00506] Regarding Table 6A:

• “wt-L” refers to the wild-type AAB2 ITR encoded in the plasmid on the left side of the expression cassette (containing the polynucleotide sequence SEQ ID NO: 51);

• “wt-R” refers to the wild-type AAB2 ITR encoded in the plasmid on the right side of the expression cassette (containing the polynucleotide sequence SEQ ID NO: 1);

• “mut-L” refers to the mutated AAB2 ITR sequence shown in SEQ ID NO: 52;

• “mut-R” refers to the mutated AAB2 ITR sequence shown in SEQ ID NO: 2;

• “CAG” refers to a synthetic promoter constructed from (C) the immediate early enhancer and promoter elements of cytomegalovirus, (A) the first exon and first intron of the chicken beta-actin gene, (G) the splice acceptor of the rabbit beta-globin gene, of SEQ ID NO: 3;

• “AAT w/SV40 intr” refers to AAT (human alpha-1 antitrypsin) with the SV40 large T antigen intron of SEQ ID NO: 4; and

• “hEF1-α” refers to the human elongation factor-1 alpha (EF-1 alpha) of SEQ ID NO: 6.

Table 6A

Plasmid ITR-L Promoter Transgene ITR-R TTX-1 wt-l CAG Luciferase mut-R TTX-2 mut-L CAG Luciferase wt-R TTX-3 wt-l AAT w/SV40 intr Luciferase mut-R TTX-4 mut-L AAT w/SV40 intr Luciferase wt-R TTX-5 wt-l LP1 w/SV40 intr Luciferase mut-R TTX-6 mut-L LP1 w/SV40 intr Luciferase wt-R TTX-7 wt-l hEF1-α Luciferase mut-R TTX-8 mut-L hEF1-α Luciferase wt-R TTX-9 wt-l CAG Padua FIX mut-R TTX-10 wt-l CAG FIX mut-R

Table 6B

Plasmid ITR-L Promoter Transgene ITR-R α (alpha) wt-l CAG Luc mut-R β (beta) wt-l LP-1β FIX mut-R

Each construct in Table 6B contains a modified SV40 poly(A) sequence (SEQ ID NO: 10) located in the 3´ untranslated region (UTR) between the transgene and the mut-R ITR.

"LP-1 β" refers to the LP-1 β promoter (SEQ ID NO: 16), which is similar to the LP-1 promoter (SEQ ID NO: 5) with 2 additional restriction enzyme sites.

[00507] According to one embodiment, the vector polynucleotide (ccDNA vector) comprises a pair of two different ITRs selected from the group consisting of: SEQ ID NO: 1 and SEQ ID NO: 52; and SEQ ID NO: 2 and SEQ ID NO: 51. According to one embodiment of each of these aspects, the vector polynucleotide or non-viral capsidless DNA vectors with covalently closed ends comprises (at) a pair of ITRs selected from the group consisting of: SEQ ID NO: 101 and SEQ ID NO: 102; SEQ ID NO: 103 and SEQ ID NO: 104, SEQ ID NO: 105 and SEQ ID NO: 106; SEQ ID NO: 107 and SEQ ID NO: 108; SEQ ID NO: 109 and SEQ ID NO: 110; SEQ ID NO: 111 and SEQ ID NO: 112; SEQ ID NO: 113 and SEQ ID NO: 114; and SEQ ID NO: 115 and SEQ ID NO: 116. In some embodiments, the ccDNA vectors do not have an ITR that comprises any sequence selected from SEQ ID NO: 500-529.

EXAMPLE 2: Bacmid and baculovirus for the production of linear, continuous and capsid-less DNA vectors

[00508] Competent DH10Bac cells (MAX Efficiency® DH10Bac™ Competent Cells, Thermo Fisher, catalog number 10361012) were transformed with either TTX or control plasmids according to the protocol provided by the supplier, available on their website (Thermo Fisher, can be found online at: https://www.thermofisher.com/order/catalog/product/10361012). To produce recombinant bacmids (“TTX bacmids”), recombination between the plasmid and the baculovirus shuttle vector was induced in DH10Bac cells. Recombinant bacmids were selected by positive selection based on screening for blue-white staining of E. coli (the Φ80dlacZΔM15 marker provides α-complementation of the β-galactosidase gene from the bacmid vector) on a bacterial agar plate containing X-gal and IPTG. White colonies were picked and cultured in 10 ml of media.

[00509] Recombinant bacmids ("TTX bacmids") were isolated from E. coli and transfected into Sf9 or Sf21 insect cells using FugeneHD™ to generate infectious baculovirus. Adherent Sf9 or Sf21 insect cells were cultured in 50 ml of media in T25 flasks at 25°C. After four days, the culture medium (containing P0 virus) was removed from the cells, filtered through a 0.45 μm filter, and infectious recombinant baculovirus particles ("TTX baculovirus" or "comparison baculovirus") were separated from the cells in culture.

[00510] Optionally, the first generation of baculovirus (P0) was amplified by infecting naive Sf9 or Sf21 insect cells in 50-500 ml of media. Cells were cultured at 130 rpm and 25°C, monitoring cell diameter and viability until cells reached a diameter of 18-19 nm (as opposed to the naive cell diameter of 14-15 nm) and a density of ~4.0 x 10 ⁶ cells/ml. At 3-8 days post infection, P1 baculovirus particles in the media were collected after centrifugation to remove cells and cell debris, followed by filtration through a 0.45 μm filter.

[00511] TTX baculovirus was collected and the infectivity of baculovirus was determined. Specifically, 4×20 ml of Sf9 cell cultures at 2.5×10 ⁶ cells/ml were treated with P1 baculovirus at the following dilutions of 1/1000, 1/10000, 1/50000, 1/100000 and incubated. Infectivity was determined daily for 4-5 days by the increase in cell diameter and cell cycle arrest, as well as changes in cell viability.

[00512] The Rep 78 sequence (SEQ ID NO: 13) was operably linked to a fragment of the IE1 promoter (SEQ ID NO: 15) and then inserted into the BamHI/KpnI restriction site of the pFASTBAC™ dual expression vector (ThermoFisher, catalog #10712024) such that the Rep 78 sequence is linked to the HSV TK poly(A) sequence at the 3' end. The Rep 52 sequence (SEQ ID NO: 14) was then cloned into the SalI-HindIII site of the vector to operably link the Rep52 sequence to the pPH promoter at the 5' end and the SV40 poly(A) sequence at the 3' end. The resulting construct is referred to herein as the "Rep plasmid."

[00513] The Rep plasmid was transformed into competent DH10Bac cells (MAX Efficiency® DH10Bac™ competent cells, Thermo Fisher, catalog number 10361012) according to the protocol provided by the supplier, available on their website (Thermo Fisher®, https://www.thermofisher.com/order/catalog/product/10361012). To produce recombinant bacmids (“Rep bacmid”), recombination was induced in DH10Bac cells between the Rep plasmid and the baculovirus shuttle vector. Recombinant bacmids were selected by positive selection based on blue-white staining screening of E. coli (Φ80dlacZΔM15 marker provides α-complementation of β-galactosidase gene from bacmid vector) on bacterial agar plate containing X-gal and IPTG. Single white colonies were picked and inoculated into 10 ml of selective media (kanamycin, gentamicin, tetracycline in LB broth). Recombinant bacmids (Rep bacmids) were isolated from E. coli, and Sf9 or Sf21 insect cells were transfected with the indicated Rep bacmids to generate infectious baculovirus.

[00514] Sf9 or Sf21 insect cells were cultured in 50 ml of media for 4 days and infectious recombinant baculovirus ("Rep baculovirus") was isolated from the culture. Optionally, first-generation Rep baculovirus (P0) was amplified by infecting naive Sf9 or Sf21 insect cells and cultured in 50-500 ml of media. Three to eight days after infection, P1 baculovirus particles in the media were collected by separating the cells by centrifugation or filtration, or using another fractionation method. Rep baculovirus was collected and the infectivity of the baculovirus was determined. Specifically, 4×20 ml of Sf9 cell cultures at 2.5×10 ⁶ cells/ml were treated with baculovirus P1 at the following dilutions: 1/1000, 1/10000, 1/50000, 1/100000 and incubated. Infectivity was determined daily for 4-5 days by the increase in cell diameter and cell cycle arrest, as well as changes in cell viability.

[00515] Sf cell culture media containing either (1) TTX or α (alpha) baculovirus or (2) the Rep baculovirus described above were then added to fresh Sf9 cell culture (2.5 x 10 ⁶ cells/mL, 20 mL) at a ratio of 1:1000 and 1:10,000, respectively. Cells were then cultured at 130 rpm at 25°C. Cell diameter and viability were detected 4-5 days after coinfection. When cell diameter reached 18-20 nm with ~70-80% viability, cell cultures were centrifuged, medium was removed, and cell pellets were collected. Cell pellets were first resuspended in a sufficient volume of aqueous medium, either water or buffer. TTX or α (alpha) vectors were isolated and purified from cells using the Qiagen Midi Plus purification protocol (Qiagen, catalog #12945, 0.2 mg cell pellet weight processed per column).

[00516] The yields of DNA vectors (e.g., TTX vectors) obtained and purified from Sf9 insect cells were initially determined based on UV absorbance at 260 nm. The yields of various TTX DNA vectors, based on UV absorbance, are presented in Table 7 below.

Table 7

Design Volume of culture Culture parameters
(Diameter in micrometers) Output (mg/l) Estimated yield (pg/cell) TTX-1 2×1 l Total: 6.02×10 ⁶
Viability: 53.3%
Diameter: 18.4 µm 15.8 5.23 TTX-9 1 l Total: 6.65×10 ⁶
Viability: 81.8%
Diameter: 18.7 µm 4.8 0.88 4 l Total: 2.45×10 ⁶
Viability: 74.5%
Diameter: 18.5 µm 5.9 3.5 1 l Total: 4.92×10 ⁶
Viability: 84.4%
Diameter: 19.7 µm 4.0 1.0 TTX-10 1 l Total: 5.55×10 ⁶
Viability: 77.4%
Diameter: 18.6 µm 6.5 1.3

EXAMPLE 3: Denaturing gel electrophoresis for identification of cDNA vector products

[00517] To qualitatively demonstrate that the isolated DNA vector material has covalently closed ends as required by their definition, the samples are digested with a restriction endonuclease identified based on the DNA vector sequence as a single restriction site endonuclease, which preferably results in two unequal sized digestion products (example: 1000 bp and 2000 bp). Following digestion and electrophoresis on a denaturing gel (which separates the two complementary strands of DNA), the linear, covalently open DNA will separate into 1000 bp fragments. and 2000 bp, while the covalently closed DNA will be separated into fragments of 2 times larger size (2000 bp and 4000 bp) because the two DNA strands are linked, already unwound, and are twice as long (but single-stranded). In addition, when cleaving the monomeric, dimeric, and n-mer forms of the vector DNA, they will all be separated into fragments of the same size due to the linked ends of the multimeric vector DNA (see FIG. 5B).

[00518] As used herein, the term "Analysis for identification of vector DNA by agarose gel electrophoresis under native and denaturing gel conditions" refers to the following assay. The restriction endonuclease is selected to be an enzyme that makes a single cut to produce products that are approximately 1/3× and 2/3× the length of the vector DNA. This ensures separation of bands on both native and denaturing gels. It is important to remove the buffer from the sample prior to denaturation. The Qiagen PCR Cleanup Kit (Qiagen catalog number 28104) or desalting "spin columns" such as GE HealthCare Ilustra™ MicroSpin™ G-25 columns (GE Healthcare catalog number 27532501) work well with endonuclease digestion.

1. Digest the DNA with the appropriate restriction endonuclease(s)

2. Add to Qiagen PCR Cleanup Kit, elute with _dH2O (30 µl)

3. Add 4 µl of 10x denaturing solution (10× = 0.5 M NaOH, 10 mM EDTA)

4. Add 6 µl of 10x gel loading solution (dye plus glycerol or Ficoll, NOT buffered)

5. DNA molecular weight markers can be prepared without the Qiagen kit by adding 10x denaturing solution to a final concentration of 4x.

6. Prepare 0.8-1.0% gel in _H2O in microwave until boiling, leave at ambient temperature for a few minutes.

7. Pour into a gel tray with a comb and place in a cold room to speed up the polymerization (2 hours)

8. Place the tray in an electrophoresis box and equilibrate with 1 mM EDTA and 200 mM NaOH for 2 hours, shaking occasionally to ensure equal NaOH concentration in the gel and in the gel box.

9. Prepare 1 L of 1x denaturing solution (50 mM NaOH, 1 mM EDTA)

10. Pour enough gel into the gel box to immerse the gel to a depth of more than 0.5 cm.

11. Run large gels (15-20 cm) overnight at 25 V. Run medium gels (8-11 cm) overnight at 20 V.

Follow-up gel analysis

12. Transfer the gel to a tray and rinse with _dH2O

13. Drain water and neutralize gel in 1x TBE or TAE (20 minutes with gentle agitation)

14. Transfer gel to _dH2O (or 1x TBE/TAE) with 1x SYBR Gold (20 min with gentle shaking)

Thermo Fisher, SYBR® Gold Nucleic Acid Gel Stain (10,000x Concentrate in DMSO), Cat. # S11494

15. Visualize the gel using epifluorescent light (blue) or UV (312 nm)

[00519] The isolated DNA vectors/vector are identified by agarose gel electrophoresis under native or denaturing conditions, as illustrated in FIG. 5 and FIG. 6. The DNA vector yields multiple bands on native gels, as shown in FIG. 5A. Each band may represent vectors having different conformations in the native state, such as continuous, discontinuous, monomeric, dimeric, etc.

[00520] The structures of the isolated ccDNA vector were further analyzed by digesting DNA obtained from coinfected Sf9 cells (as described herein) with restriction endonucleases selected based on a) the presence of only one cleavage site in the vector DNA, and b) the resulting fragments were large enough to be clearly visible when fractionated on a 0.8% denaturing agarose gel (>800 bp).

[00521] Specifically, equal amounts (2 μg based on OD260) of TTX plasmid and TTX vector were digested with restriction endonucleases at 37°C for 1 hour. After digestion, the vector DNA material was isolated using a QIAquick column and eluted with water. The samples were denatured in denaturing solution (0.05 M NaOH, 1 mM EDTA), while a 0.8% agarose gel prepared in water was pre-equilibrated for 2 hours in equilibration buffer (1 mM EDTA, 200 mM NaOH). Then, the samples immersed in 1x denaturing solution (50 mM NaOH, 1 mM EDTA) were analyzed on the gel overnight at 4°C. The next day, the gel was washed, neutralized in TBE for 20 min, soaked in 1x aqueous SYBR Gold for 1 h, and visualized under UV/blue light.

[00522] The presence of a DNA vector is initially identified based on the characteristic multi-band profiles (the primary and secondary bands are spaced apart to indicate that the secondary band represents material with a mass approximately twice that of the primary band) on a native gel, and then confirmed on a denatured gel based on the characteristic multi-band profile illustrated on the right side of FIG. 5A. As illustrated in FIG. 5B, the linear discontinuous DNA vectors and the linear and continuous TTX vector can be distinguished by the size of their reaction products - for example, a discontinuous DNA vector is expected to produce 1 kb and 2 kb fragments, while a continuous capsidless vector is expected to produce 2 kb and 4 kb fragments.

[00523] FIG. 6 is an illustrative image of an actual denaturing gel with TTX vectors 1 and 2, 3 and 4, 5 and 6, and 7 and 8 (all described in Table 1A above) with (+) or without (-) endonuclease digestion. After the endonuclease reaction, each TTX vector yielded two bands (*). The sizes of the two bands for these vectors, determined based on the size marker, are shown at the bottom of the image. The band sizes confirm that each of the TTX vectors has a continuous structure.

[00524] The contribution of the TTX plasmid to the UV absorption was estimated by comparing the fluorescence intensity of the TTX vector with a standard. For example, if it is determined from the UV absorption that 4 μg of TTX vector is loaded onto the gel, and the fluorescence intensity of the TTX vector is equivalent to the 2 kb band with a known mass of 1 μg, then the mass of the TTX vector is 1 μg. Thus, the TTX vector constitutes 25% of the total amount of UV absorbing material. The intensity of the band on the gel is then plotted against the calculated input amount that the band represents - for example, if the total amount of TTX vector represents 8 kb and the excised reference band represents 2 kb, then the intensity of that band on the plot would represent 25% of the total input, which in this case would be 0.25 μg for a 1.0 μg input. Using the TTX plasmid titration to construct a standard curve, the amount for the TTX vector band is calculated using the regression line equation, which can then be used to determine the percentage of the total input represented by TTX vector, or the percent purity (FIG. 7).

EXAMPLE 4: DNA vectors express transgene-encoded protein in vitro

[00525] The wild-type SA cDNA sequence for human factor IX mRNA ("wtFIX", SEQ ID NO: 11) or the Padua variant cDNA sequence ("PaduaFIX", SEQ ID NO: 12) was inserted into the cloning site of TTX plasmid 1 to generate TTX plasmid 1-wtFIX and TTX plasmid 1-PaduaFIX, respectively. These plasmids were introduced into Sf9 insect cells and used to generate TTX bacmid 1-wtFIX and TTX bacmid 1-PaduaFIX, as well as TTX baculovirus 1-wtFIX and TTX baculovirus 1-PaduaFIX, respectively, using the methods described herein. In vitro protein expression from TTX plasmids and TTX vectors was tested by transfecting HEK293 cells (2× ¹⁰⁵ cells/well, 96-well plate) with 250 ng/well of (1) TTX plasmid 1-wtFIX, (2) TTX plasmid 1-PaduaFIX, (3) TTX vector 1-wtFIX, (4) TTX vector 1-PaduaFIX, (5) β(beta) plasmid 1-wtFIX, or (6) β(beta) vector 1-wtFIX using Fugene6 transfection reagent (3:1 Fugene6:DNA). The result of Western blot analysis is shown in FIG. 8. The FIX antibody reaction revealed bands of 55 kDa, which correspond to the mass of the FIX proteins produced. Negative control lysates transfected with β(beta)-plasmid 1-wtFIX or β(beta)-vector 1-wtFIX did not produce detectable amounts of FIX protein. This result confirms that TTX-vector 1 can be used for efficient transfer and expression of a therapeutic gene, such as the gene encoding human factor IX.

[00526] ELISA: In general, culture media from transfected cells were added in duplicate to wells treated with anti-FIX antibody and incubated for 1 hour, followed by washing and incubation with detection antibody for 1 hour at room temperature. Samples were washed again, TMB substrate was added and staining was developed for 10 minutes, the reaction was stopped, and the absorbance of the samples at 450 nm was immediately read. An example of samples from reactions with TMB substrate is shown in FIG. 15A, and the concentration of FIX in each sample, determined based on the absorbance of the sample at 450 nm, is shown in FIG. 15A. High levels of FIX protein expression were detected from TTX plasmid 1 and TTX vector 1, while no significant FIX expression was detected from β (comparison) plasmid or β (comparison) vector.

[00527] This further confirms that TTX vector 1, derived from TTX plasmid 1, containing, in 5' to 3' direction, the WT replicative polynucleotide sequence (SEQ ID NO: 51), the CAG promoter (SEQ ID NO: 3), the R3/R4 cloning site (SEQ ID NO: 7), WPRE (SEQ ID NO: 8), BGHpA (SEQ ID NO: 9) and the modified replicative polynucleotide sequence (SEQ ID NO: 2), induces transgene expression significantly more efficiently compared to α (alpha) vector 1, derived from α (alpha) plasmid 1, which does not include WPRE (SEQ ID NO: 8) and BGHpA (SEQ ID NO: 9).

EXAMPLE 5: Preparation of ccDNA coexpressing factor IX and a cGAS inhibitor

[00528] The Kaposi's sarcoma-associated herpesvirus ORF52 protein (SEQ ID NO: 882), or a variant thereof that inhibits cGAS, or a truncated cytoplasmic isoform of LANA (LANAΔ161 or SEQ ID NO: 884) lacking amino acids 161-1162 of SEQ ID NO: 882), is operably linked to a promoter and inserted into the R5 restriction cloning site of a TTX 9 or TTX 10 plasmid encoding a factor IX transgene, as described in Example 1 and Example 4. Thus, a ccDNA is obtained that encodes both factor IX and a cGAS inhibitor, as described in Examples 2-3.

EXAMPLE 6: Confirmation of cGAS inhibitor expression expressed by ccDNA

[00529] Expression of a target cGAS inhibitor co-expressed by ccDNA, such as the Kaposi's sarcoma-associated herpesvirus ORF52 protein (SEQ ID NO: 882) or a variant thereof that inhibits cGAS, or the truncated cytoplasmic isoform LANA (SEQ ID NO: 884), can be confirmed using HeLa cells and antibodies specific for the cGAS inhibitor, such as the ORF52 antibody described in Li et al. ("Kaposi's sarcoma-associated herpesvirus inhibitor of cGAS (KicGAS) Encoded by ORF52, is an Abundant Tegument protein and Is Required for Production of Infectious Progeny Viruses," J. Virol. 2016, 90(11): 5329). For example, HeLa cells are cultured and transient transfections of constructs coexpressing factor IX and a target cGAS inhibitor are performed using, for example, Fusegene6 transfection reagent (3:1; fusgene6:DNA). Western blotting and/or flow cytometry techniques are used to detect expression of the cGAS inhibitor, as known to those skilled in the art. Expression of factor IX is confirmed as described in Example 4.

EXAMPLE 7: Preparation of cDNA coexpressing factor IX and a TLR-9 inhibitor

[00530] Oligonucleotides that can form a hairpin structure comprising the following sequences such as (TCCTGGCGGGGAAGT, SEQ ID NO: 889), ODN-2114 (TCCTGGAGGGGAAGT, SEQ ID NO: 890), poly(G) (GGGGGGGGGGGGGGGGGGGGGG, SEQ ID NO: 891), ODN-A151 (TTAGGGTTAGGGTTAGGGTTAGGG, SEQ ID NO: 892), G-ODN (CTCC-TATTGGGGGTTTCCTAT, SEQ ID NO: 893), IRS-869 (TCCTGGAGGGGTTGT, SEQ ID NO: 894), INH-1 (CCTGGATGGGAATTCCCATCCAGG, SEQ ID NO: 895), INH-4 (TTCCCATCCAGGCCTGGATGGGAA, SEQ ID NO: 896), (IRS-661 TGCTTGCAAGCTT-GCAAGCA, SEQ ID NO: 897), 4024 (TCCTGGATGGGAAGT, SEQ ID NO: 898), 4084F (CCTGGATGGGAA, SEQ ID NO: 899), INH-13 (CTTACCGCTGCACCTGGATGGGAA, SEQ ID NO: 900), INH-18 (CCTGGATGGGAACTTACCGCTGCA, SEQ ID NO: 901) and IRS-954 TGCTCCTGGAGGGTTGT, SEQ ID NO: 902) are designed to produce sticky ends after annealing of the complementary 5´-3´ and 3´-5´ strands so that they can be inserted by ligation into a pre-selected restriction cloning site, such as R5 or another site, of the TTX 9 or TTX 10 plasmid encoding the factor IX transgene, as described in Example 1 and Example 4.

[00531] For example, oligonucleotides with a suitable restriction site are annealed by mixing each strand in equal molar amounts in a suitable buffer: e.g., 100 mM potassium acetate; 30 mM HEPES, pH 7.5, heating to 94°C for 2 minutes, and gradually cooling. Thus, oligonucleotides with significant predicted secondary structure benefit from a more gradual cooling/annealing step. This can be accomplished by placing the oligonucleotide solution in a water bath or incubator and turning the device off/on. The annealed oligonucleotides can be diluted in a nuclease-free buffer and stored in double-stranded annealed form at 4°C. The ccDNA plasmid with the TLR-9 inhibitory oligonucleotide sequence is then purified (e.g., by gel electrophoresis or column) and used to produce a ccDNA vector. A ccDNA encoding factor IX and containing a TLR-9 antagonist can be produced.

EXAMPLE 8: Controlled expression of a transgene from a ccDNA: Expression of a transgene from a ccDNA vector in vivo can be maintained and/or increased by repeated dosing.

[00532] A ccDNA vector was prepared according to the methods described in Example 1 above using a ccDNA plasmid containing the CAG promoter (SEQ ID NO: 3), wherein the luciferase transgene (SEQ ID NO: 71) is used as a model inflammasome antagonist flanked by asymmetric ITRs (e.g., 5'-WT-ITR (SEQ ID NO: 1) and 3' mod-ITR (SEQ ID NO: 2)), and evaluated in various in vivo treatment programs. This ccDNA vector was used in all subsequent experiments described in Examples 6-10. In Example 6, the ccDNA vector was purified, formulated with a lipid nanoparticle (LNP-ccDNA), and injected into the tail vein of each CD-1® IGS mouse. Liposomes were prepared using a suitable lipid mixture containing four components for the formation of lipid nanoparticle (LNP) liposomes, including cationic lipids, auxiliary lipids, cholesterol, and PEG lipids.

[00533] To assess sustained in vivo transgene expression from the ccDNA vector over a long period of time, ccDNA LNPs were administered in sterile PBS by intravenous injection into the tail vein of approximately 5-7 week old CD-1® IGS mice. Three dose groups were evaluated: 0.1 mg/kg, 0.5 mg/kg, and 1.0 mg/kg, with ten mice per group (except for the 1.0 mg/kg group, which had 15 mice per group). Injections were given on day 0. Five mice from each group received an additional identical dose injection on day 28. Luciferase expression was measured by IVIS imaging after intravenous administration to CD-1® IGS mice (Charles River Laboratories; wild-type (WT) mice). Luciferase expression was assessed by IVIS imaging following intraperitoneal injection of 150 mg/kg luciferin substrate on days 3, 4, 7, 14, 21, 28, 31, 35, and 42 and regularly (e.g., weekly, twice weekly, or every 10 days or every 2 weeks) between days 42-110. Luciferase transgene expression, as a proxy for the inflammasome antagonist, was measured by IVIS imaging for at least 132 days after 3 different administration protocols (data not shown).

[00534] An extension study was performed to examine the effect of repeated dosing, such as repeated administration of luciferase-expressing LNP-cDNA, to subjects receiving LNP-cDNA. In particular, the ability to increase expression levels by one or more additional administrations of the cDNA vector was assessed.

[00535] In this study, the biodistribution of luciferase expression from the ccDNA vector was assessed by IVIS in CD-1® IGS mice following an initial intravenous administration of 1.0 mg/kg (i.e., the priming dose) on days 0 and 28 (Group A). The second administration of ccDNA vector was performed by tail vein injection at 3 mg/kg (Group B) or 10 mg/kg (Group C) in 1.2 ml into the tail vein on day 84. Five (5) CD-1® mice were used in each of Groups A, B, and C. IVIS visualization of luciferase expression in mice was performed prior to additional doses on days 49, 56, 63, and 70 as described above, and following booster doses on day 84 and on days 91, 98, 105, 112, and 132. Luciferase expression was assessed and detected in all three Groups A, B, and C until at least day 110 (the longest period assessed).

[00536] Luciferase expression levels were shown to be increased by repeated dosing (i.e., repeated administration of the ccDNA composition) of LNP-ccDNA-Luc, as determined by assessing luciferase activity in the presence of luciferin. Luciferase transgene expression, as a proxy for the inflammasome antagonist, was measured by IVIS imaging for at least 110 days following 3 different dosing protocols (Groups A, B, and C). Mice that did not receive any additional repeat dosing (priming dose of 1 mg/kg (i.e., Group A)) showed stable luciferase expression throughout the study. Mice in Group B that received a booster dose of 3 mg/kg ccDNA vector showed approximately a seven-fold increase in observed luciferase luminosity compared to mice in Group C. Surprisingly, mice that received a booster dose of 10 mg/kg ccDNA vector had a 17-fold increase in observed luciferase luminosity compared to mice that did not receive any booster dose (Group A).

[00537] Group A of CD-1® IGS mice show luciferase expression following intravenous administration of 1 mg/kg ccDNA vector via the tail vein on days 0 and 28. Groups B and C of CD-1® IGS mice show luciferase expression following administration of 1 mg/kg ccDNA vector at the first time point (day 0) and a booster dose of ccDNA vector at the second time point on day 84. The second administration (i.e., booster dose) of ccDNA vector increased expression by at least 7-fold and as much as 17-fold.

[00538] A 3-fold increase in the dose (i.e., amount) of ccDNA vector at the repeated dose in Group B (i.e., at the repeated dose of 3 mg/kg) resulted in a 7-fold increase in luciferase expression. Also unexpectedly, a 10-fold increase in the amount of ccDNA vector at the repeated dose (i.e., at the repeated dose of 10 mg/kg) in Group C resulted in a 17-fold increase in luciferase expression. Thus, the second administration (i.e., repeat dose) of ccDNA increased expression by at least 7-fold and even up to 17-fold. This shows that the increase in transgene expression following a repeat dose is greater than expected and is dependent on the dose or amount of ccDNA vector given at the repeat dose and appears to be synergistic with the initial transgene expression following the initial priming administration on day 0. Thus, the dose-dependent increase in transgene expression is not additive, rather the level of transgene expression is dose-dependent and super-proportional to the total amount of ccDNA vector given at each time point.

[00539] Both Groups B and C showed a significant dose-dependent increase in luciferase expression compared to control mice (Group A) that did not receive a second dose of the ccDNA vector at a second time point. Taken together, these data demonstrate that transgene expression from a ccDNA vector can be increased in a dose-dependent manner by a second dose (i.e., repeated administration) of the ccDNA vector at at least a second time point.

[00540] Taken together, these data demonstrate that the expression level of a transgene, such as an inflammasome antagonist, from ccDNA vectors can be maintained at a steady state for at least 84 days and can be increased in vivo following a second dose of the ccDNA vector administered at at least a second time point.

EXAMPLE 9: Synthetic nanocarriers with supersaturated amounts of rapamycin

[00541] Nanocarrier compositions comprising PLGA (lactide:glycolide 3:1, intrinsic viscosity 0.39 dl/g) and PLA-PEG (PEG block 5 kDa, intrinsic viscosity 0.36 dl/g) polymers and a rapamycin agent (RAPA) can be synthesized using an oil-in-water emulsion evaporation method. The organic phase is formed by dissolving the polymers and RAPA in dichloromethane. The emulsion is formed by homogenizing the organic phase in an aqueous phase containing a polyvinyl alcohol (PVA) surfactant. The emulsion is then combined with a large amount of aqueous buffer and stirred to ensure evaporation of the solvent. The RAPA content in the various compositions is varied such that as the RAPA content in the composition increases, the saturation limit of the RAPA system is exceeded. The saturation content of RAPA for the composition is calculated using the solubility of RAPA in the aqueous phase and in the dispersed phase of the nanocarrier. For compositions containing PVA as the main solute in the aqueous phase, the solubility of RAPA in the aqueous phase is found to be proportional to the concentration of PVA such that RAPA is soluble at a weight ratio of 1:125 to dissolved PVA. For compositions containing the described PLGA and PLA-PEG as nanocarrier polymers, the solubility of RAPA in the dispersed phase of the nanocarrier is found to be 7.2% w/w. The following formula can be used to calculate the saturation content of RAPA for the composition:

RAPA content = V(0.008c _PVA + 0.072c _pol )

[00542] where c _PVA is the mass concentration of PVA, c _pol is the total mass concentration of polymers, and V is the volume of the nanocarrier suspension after completion of evaporation.

Table 8

Sample ID Estimated supersaturation (%) RAPA load (%) Diameter (nm) 1 -50 2.5 143 2 -25 3.8 146 3 1 4.9 147 4 23 4.9 130 5 48 8.1 160 6 73 9.8 189 7 98 12.4 203

[00543] For 1, 2 and 3, a constant fraction of 60% RAPA cannot be isolated, indicating a sub-saturated equilibrium state between the aqueous and organic phases. For the remaining nanocarriers containing large amounts of RAPA, a constant mass of 6.8 mg RAPA cannot be isolated. This constant absolute mass loss indicates that the system is in a state of supersaturation (i.e., supersaturated in one or more phases).

EXAMPLE 10: Synthetic nanocarriers with supersaturated amounts of rapamycin eliminate or delay antibody production

[00544] Nanocarrier compositions comprising PLGA (lactide:glycolide 3:1, intrinsic viscosity 0.39 dl/g) and PLA-PEG (PEG block 5 kDa, intrinsic viscosity 0.36 dl/g) polymers and a RAPA agent can be synthesized using the oil-in-water emulsion evaporation method described in Example 5. The RAPA content in the various compositions is varied such that as the RAPA content in the composition increases, the saturation limit of the RAPA system is exceeded.

Table 9

Sample ID Estimated supersaturation (%) RAPA load (%) Diameter (nm) 1 -50 2.5 143 3 1 4.9 147 8 21 8.5 163 9 48 13.5 159

[00545] To evaluate the ability of the compositions to induce immune tolerance, mice were co-administered with the nanocarrier and keyhole limpet hemocyanin (KLH) by intravenous injection three times per week, and then stimulated weekly with KLH alone. Following KLH stimulation, mouse sera were then analyzed for the presence of KLH antibodies. Compositions prepared in a supersaturated state and having a final RAPA load of 8% or greater resulted in a greater absence or delay in the production of KLH antibodies than compositions made at or before saturation and having a final RAPA load of 5% or less.

EXAMPLE 11: Synthetic nanocarriers with supersaturated amounts of rapamycin

[00546] Nanocarrier compositions comprising PLA (intrinsic viscosity 0.41 dL/g) and PLA-PEG (PEG block 5 kDa, intrinsic viscosity 0.50 dL/g) polymers and a RAPA agent were synthesized using the oil-in-water emulsion evaporation method described in Example 9. The RAPA content of the various compositions was varied such that as the RAPA content in the compositions increased, the saturation limit of the RAPA system was exceeded. The saturation limit RAPA content of the composition was calculated using the method described in Example 9. For compositions comprising the described PLA and PLA-PEG as nanocarrier polymers, the solubility of RAPA in the continuous phase of the nanocarrier was found to be 8.4% w/w. The following formula can be used to calculate the saturation limit RAPA content of the composition:

RAPA content = V(0.008c _PVA + 0.084c _pol )

[00547] where c _PVA is the mass concentration of PVA, c _pol is the total mass concentration of polymers, and V is the volume of the nanocarrier suspension upon completion of evaporation. All batches of nanocarriers are filtered through 0.22 μm filters upon completion of fabrication.

Table 10

Sample ID Calculated supersaturation
(%) RAPA load (%) Without rinsing
Diameter (nm) Final diameter (nm) Filtered
Permeability (g/ ^m2 ) 10 -10 5.4 145 149 >171 11 0 6.2 150 155 >180 12 10 6.1 151 154 >170 13 20 6.1 148 148 80 14 30 6.2 171 151 28 15 40 5.8 202 154 16

Despite the addition of increasing amounts of RAPA to nanocarriers 12-15, the final RAPA content of the nanocarriers did not increase, while the filter throughput decreased. This indicates that the compositions were oversaturated with RAPA and the excess RAPA was removed during washing and/or filtration.

EXAMPLE 12: Factor IX or VIII for the treatment of hemophilia B using cctDNA encoding factor IX or factor VIII co-administered with rapamycin

[00548] The experiment is conducted in mice deficient in factor IX or factor VIII that contain a knock-in hFIX or hFVIII sequence with a deleterious mutation (e.g., R333Q for hFIX). Male mice with knock-out factor IX or FVIII receive single or repeated doses of ccDNA-LNPs (ccDNA-loaded lipid nanoparticles) co-administered with rapamycin or a rapamycin analog, wherein the ccDNA-LNPs and rapamycin or a rapamycin analog are contained in separate compositions. LNPs with ccDNA vectors are administered to appropriate mice at doses of 0.3 to 5 mg/kg in a volume of 1.2 ml, together with a nanocarrier with rapamycin (e.g., supersaturated rapamycin (e.g., SVP-rapamycin), as described in Examples 9-11), or an analog thereof, administered at, for example, 0.05 mg/kg, 0.1 mg/kg, and up to 5 mg/kg. Therapeutically effective doses are determined by monitoring the effectiveness of immune response inhibition (e.g., with single and repeated dosing) and measuring the target amount of transgene expression. Each dose can be administered by intravenous administration. SVP-Rap can be co-administered, for example, on day 0 and day 14.

[00549] Expression of factor IX or factor VIII in plasma is assessed by ELISA as described in Example 4 at various time points, such as 10, 20, 30, 40, 50, 1000, and 200 days or more, etc. Activated partial thromboplastin time and bleeding time can also be measured as a measure of the efficacy and effect of co-administration of rapamycin or an analog on expression of factor VIII or factor IX. Mice that receive a ccDNA vector co-administered with rapamycin are expected to have increased and/or sustained expression of factor IX or factor VIII over a longer period of time compared to mice that receive only the ccDNA vector but not rapamycin or an analog. In addition, upon repeated dosing, mice that receive a repeated dose of the ccDNA vector and rapamycin are expected to exhibit less activation of cytokine secretion and increased duration of transgene expression and therapeutic efficacy compared to mice that receive a repeated dose of the ccDNA vector but do not receive rapamycin. The co-administration times can be staggered to 0, 1, 2, 3, 4, 5, 6, 7, 8 hours.

EXAMPLE 13: Factor IX for the treatment of hemophilia B using cctDNA encoding factor IX and a cGAS antagonist

[00550] The experiment is carried out in factor IX-deficient mice that have an inserted hFIX sequence with a deleterious mutation (R333Q). Male mice with suppressed factor IX receive single or repeated doses of ccDNA-LNPs (lipid nanoparticles with ccDNA). Two variants of ccDNA-vector LNPs are used; 1) a ccDNA-vector LNP encoding human factor IX (either the native human sequence or Padua FIX variants) and encoding the ORF52 protein of Kaposi's sarcoma-associated herpesvirus; and a ccDNA-vector LNP encoding only factor IX but not the cGAS inhibitor as a comparative ccDNA vector. LNPs with ccDNA vectors are administered to the respective mice at doses of 0.3 to 5 mg/kg in a volume of 1.2 ml. Each dose should be administered by intravenous hydrodynamic injection. Expression of factor IX in plasma is assessed by ELISA as described herein at various time points, such as 10, 20, 30, 40, 50, 1000 and 200 days or more, etc. Activated partial thromboplastin time and bleeding time are also measured as a determination of efficacy. Mice that receive a ccDNA vector expressing both hFIX and ORF52 are expected to have increased and/or sustained expression of factor IX over a longer period of time compared to mice that receive a ccDNA vector expressing only factor IX but not ORF52 or another cGAS inhibitor. In addition, upon repeated dosing, mice that receive a repeated dose of a ccDNA vector containing both ORF52 and factor IX are expected to exhibit less activation of cytokine secretion and increased duration of transgene expression and therapeutic efficacy compared to mice that receive a repeated dose of a ccDNA vector encoding factor IX alone. The cGAS inhibitor and factor IX may be delivered on different ccDNA vectors, but are preferably encoded by the same vector and, accordingly, cGAS inhibition occurs in the same cell as the cell that receives the ccDNA vector encoding the transgene, such as factor IX.

EXAMPLE 14: Determination of the effects of co-administration of ccDNA and cGAS antagonists on innate immune responses and duration of factor IX expression

[00551] To examine the effects of co-administration of a cGAS inhibitor or antagonist of interest on an immune response (e.g., an innate immune response) in vitro, reporter cell lines can be used for functional assays that examine cGAS activation. A cGAS reporter cell line that can be used for such in vitro assays can be a stably co-transfected cell line that expresses full-length human cGAS and a reporter gene, such as a secreted alkaline phosphatase (SEAP) reporter gene, under the transcriptional control of a transcription factor response element, such as an NF-kB binding site, an AP-1 binding site, or a combination thereof. For example, reporter cells are seeded in 96-well plates. After a predetermined period of time, such as 16 hours, the cells are stimulated with varying amounts of compositions containing cGAS expressing factor IX, with or without a cGAS inhibitor. The activity of a reporter gene, such as SEAP, can be analyzed using any method or assay known to one skilled in the art to compare the level of cGAS activation in the presence of the ccDNA of interest with or without a cGAS inhibitor. It is expected that less activation of the reporter molecule will be observed in the presence of a cGAS inhibitor.

[00552] Additionally, cGAS-silenced reporter lines can be used, such as lines derived from human THP-1 monocytes, which is a cell line often used to study DNA sensing pathways because it expresses all of the cytosolic DNA sensors identified to date (except DAI). Such cGAS-silenced reporter lines can express one or more inducible secreted reporter genes, such as Lucia luciferase and SEAP (secreted embryonic alkaline phosphatase). The reporter gene can be under the control of a minimal ISG54 (interferon-stimulated gene) promoter in combination with one or more, such as five, IFN-stimulated response elements. The reporter gene can also be under the control of a minimal IFN-β promoter fused to one or more, such as five, copies of a response element, such as an NF-kB response element. cGAS activity in the presence of cGAS inhibitors in combination with the ccDNAs described herein can be compared in a gene-silenced cell line versus a parental cell line.

[00553] To examine the effects of co-administration of a cDNA of interest and a cGAS inhibitor or cGAS antagonist on activation of the cGAS and STING immune response (e.g., innate immune response) ex vivo, human monocytes can be isolated, for example, by density gradient centrifugation of peripheral blood and magnetic separation. These monocytes can be examined before and after exposure to and/or activation of the cDNA of interest with or without a cGAS inhibitor, with appropriate controls. After treatment, serum and cell supernatants are used to measure one or more cytokine pathways as a functional indicator of cGAS/STING pathway activation, such as interleukin (IL)-1β, IL-6, IL-8, interferon (IFN)-γ, monocyte chemoattractant protein (MCP)-1 and/or tumor necrosis factor (TNF)-α, using any assay or method known to those skilled in the art. In addition, nuclear extracts can be used to test for NF-κB activation using any assay or method known to those skilled in the art. It is expected that when ccDNA is administered in the presence of a cGAS inhibitor, there will be less activation of cytokine pathways and cytokine secretion, resulting in increased duration of transgene expression and therapeutic efficacy.

[00554] To examine the effects of co-administration of a ccDNA of interest and a cGAS inhibitor or cGAS antagonist on the activation of the cGAS and STING immune response (e.g., innate immune response) in vivo, a mouse model can be used. Mouse serum or lymphocyte samples are examined before and after exposure to and/or activation of a ccDNA expressing a transgene of interest, such as factor IX, with or without a cGAS inhibitor, with appropriate controls. Following treatment, serum and cell supernatants are used to measure one or more cytokine pathways as a functional indicator of cGAS/STING pathway activation, such as interleukin (IL)-1β, IL-6, IL-8, interferon (IFN)-γ, monocyte chemoattractant protein (MCP)-1, and/or tumor necrosis factor (TNF)-α, using any assay or method known to one of skill in the art. In addition, nuclear extracts can be used to test for NF-κB activation using any assay or method known to those skilled in the art. It is expected that when ccDNA is administered in the presence of a cGAS inhibitor, there will be less activation and cytokine secretion, resulting in increased duration of transgene expression and therapeutic efficacy.

EXAMPLE 15: Factor IX for the treatment of hemophilia B using cctDNA encoding factor IX and a TLR-9 antagonist

[00555] The experiment is carried out in factor IX-deficient mice that have an inserted hFIX sequence with a deleterious mutation (R333Q). Male mice with suppressed factor IX receive single or repeated doses of ccDNA-LNPs (lipid nanoparticles with ccDNA). Two variants of ccDNA-vectored LNPs are used; 1) ccDNA-LNP encoding human factor IX (either the native human sequence or Padua FIX variants) and encoding the ORF52 protein of Kaposi's sarcoma-associated herpesvirus; and ccDNA-LNP encoding only factor IX but not the cGAS inhibitor as a comparative ccDNA vector. The ccDNA-vectored LNPs are administered to the respective mice at doses of 0.3 to 5 mg/kg in a volume of 1.2 ml. Each dose should be administered by intravenous hydrodynamic injection. Expression of factor IX in plasma is assessed by ELISA as described in Example 4 at various time points, such as 10, 20, 30, 40, 50, 1000 and 200 days or more, etc. Activated partial thromboplastin time and bleeding time are also measured as a determination of efficacy. Mice that receive a ccDNA vector containing a TLR9 antagonist and expressing hFIX are expected to have increased and/or sustained expression of factor IX over a longer period of time compared to mice that receive a ccDNA vector expressing only factor IX but not a TLR-9 inhibitor. In addition, upon repeated dosing, mice that receive a repeated dose of a ccDNA vector containing a TLR9 inhibitor, such as a hairpin oligonucleotide sequence, and factor IX are expected to exhibit less activation of cytokine secretion and an increased duration of transgene expression and therapeutic efficacy compared to mice that receive a repeated dose of a ccDNA vector encoding factor IX alone. The TLR-9 inhibitor and factor IX may be delivered on different ccDNA vectors, in trans, but are preferably encoded by the same vector and, accordingly, TLR-9 inhibition occurs in the same cell as the cell that receives the ccDNA vector encoding the transgene, such as factor IX.

EXAMPLE 16: Determination of the effects of cDNA and TLR antagonists on innate immune responses and duration of transgene expression

[00556] To examine the effects of co-administration of a cDNA of interest and a TLR9 inhibitor or TLR9 antagonist on innate immune responses in vitro, reporter lines can be used for TLR9-dependent functional assays that examine the downstream effects of TLR9 activation. The TLR9 reporter cell line can be a stably co-transfected cell line that expresses full-length human Toll-like receptor 9 (TLR9) and a reporter gene, such as a secreted alkaline phosphatase (SEAP) reporter gene, under the transcriptional control of a transcription factor response element, such as an NF-kB binding site, an AP-1 binding site, or a combination thereof. For example, reporter cells are seeded in 96-well plates. After a predetermined period of time, such as 16 hours, the cells are stimulated with varying amounts of compositions containing cDNA expressing the transgene of interest, with or without a TLR9 antagonist. Such an antagonist may be an oligonucleotide that inhibits TLR. The activity of a reporter gene, such as SEAP, may be analyzed using any method or assay known to one skilled in the art to determine the level of TLR9 activation in the presence of the ccDNA of interest with or without a TLR9 antagonist. It is expected that in the presence of a TLR9 inhibitor, less activation of the reporter molecule will be observed.

[00557] To examine the effects of co-administration of a cDNA of interest and a TLR9 inhibitor or TLR9 antagonist on TLR9-mediated activation of innate immune responses ex vivo, human monocytes can be isolated, for example, by density gradient centrifugation of peripheral blood and magnetic separation. These monocytes can be examined before and after exposure to and/or activation of the cDNA of interest, with or without a TLR9 antagonist, with appropriate controls. Following treatment, serum and cell supernatants are used to measure one or more cytokine pathways as a functional indicator of TLR9 activation, such as interleukin (IL)-1β, IL-6, IL-8, interferon (IFN)-γ, monocyte chemoattractant protein (MCP)-1, and/or tumor necrosis factor (TNF)-α, using any assay or method known to one of skill in the art. In addition, 0914800, nuclear extracts can be used to test for NF-κB activation using any assay or method known to one skilled in the art. It is expected that when ccDNA is administered in the presence of a TLR9 inhibitor, there will be less activation of cytokine pathways and cytokine secretion, resulting in increased duration of transgene expression and therapeutic efficacy.

[00558] To examine the effects of co-administration of a cDNA of interest and a TLR9 inhibitor or TLR9 antagonist on TLR9-mediated activation of innate immune responses in vivo, a mouse model can be used. Mouse serum or lymphocyte samples are examined before and after exposure to and/or activation of cDNA expressing a transgene of interest, such as factor IX, with or without a TLR9 inhibitor, with appropriate controls. Following treatment, serum and cell supernatants are used to measure one or more cytokine pathways as a functional indicator of cGAS/STING pathway activation, such as interleukin (IL)-1β, IL-6, IL-8, interferon (IFN)-γ, monocyte chemoattractant protein (MCP)-1, and/or tumor necrosis factor (TNF)-α, using any assay or method known to one of skill in the art. In addition, nuclear extracts can be used to test for NF-κB activation using any assay or method known to those skilled in the art. It is expected that when ccDNA is administered in the presence of a TLR9 inhibitor, there will be less activation and cytokine secretion, resulting in increased duration of transgene expression and therapeutic efficacy.

EXAMPLE 17: Preparation of ccDNA composition with RAPA in the form of LNP vectors

In some embodiments, it may be desirable to package rapamycin directly into the ccDNA vector. One non-limiting example of such direct preparation of a ccDNA and RAPA formulation is described below.

Combinations of ccDNA with rapamycin in lipid nanoparticles (LNPs) can be prepared by mixing an alcoholic lipid solution containing rapamycin with an aqueous solution of ccDNA using a microfluidic device (e.g., NanoAssemblr™) in a ratio of 1:3 (v/v) at a total flow rate of 12 ml/min. The mass ratio of total lipids to ccDNA can be approximately 10:1 to 30:1. In general, an ionizable lipid (e.g., MC3), a non-cationic lipid (e.g., distearoylphosphatidylcholine (DSPC)), a membrane integrity component (such as a sterol, e.g., cholesterol), and a conjugated lipid molecule (e.g., a PEG lipid, e.g., 1-(monomethoxypolyethyleneglycol)-2,3-dimyristoylglycerol, with an average molecular weight of PEG 2000 (“PEG-DMG”)) are solubilized in an alcohol (e.g., ethanol) in a molar ratio of 50:10:38.5:1.5. Rapamycin is then dissolved in the lipid solution to a target concentration. The cDNA is diluted to 0.2 mg/mL in 25 mM sodium acetate buffer, pH 4. After LNP formation (using, for example, the NanoAssemblr™), the alcohol is removed and the sodium acetate buffer is replaced with PBS by dialysis. The alcohol removal and simultaneous buffer exchange can be accomplished, for example, by dialysis or tangential flow filtration. The resulting lipid nanoparticles are filtered through a sterile 0.2 μm filter and stored in the same manner as the cDNA vector-loaded LNPs described above.

EXAMPLE 18: Determining the effects of co-administration of a cDNA vector and rapamycin or a rapamycin analog on innate immune responses and duration of factor IX expression

[00559] To examine the effects of co-administration of a cDNA of interest and rapamycin or an analog on innate immune responses in vitro, reporter cell lines can be used for functional assays that examine the downstream effects of TLR and mTORC1 activation. The TLR9 reporter cell line can be a stably co-transfected cell line that expresses full-length human Toll-like receptor 9 (TLR9) and a reporter gene, such as a secreted alkaline phosphatase (SEAP) reporter gene, under the transcriptional control of a transcription factor response element, such as an NF-kB binding site, an AP-1 binding site, or a combination thereof. For example, reporter cells are seeded in 96-well plates. After a predetermined period of time, such as 16 hours, the cells are stimulated with varying amounts of compositions containing cDNA expressing the transgene of interest, with or without rapamycin or an analog thereof. The activity of a reporter gene, such as SEAP, can be analyzed using any method or assay known to one skilled in the art to determine the level of mTORC1 activation in the presence of the ccDNA of interest with or without rapamycin or an analog thereof. It is expected that in the presence of rapamycin, greater activation of the reporter molecule will be observed and that the induction of STAT3, IL-10, and other cytokines will be reduced.

[00560] To examine the effects of co-administration of a cDNA of interest and rapamycin on the activation of innate immune responses ex vivo, human monocytes can be isolated, for example, by density gradient centrifugation of peripheral blood and magnetic separation. These monocytes can be examined before and after exposure to and/or activation of the cDNA of interest with or without rapamycin or an analog thereof, with appropriate controls. Following treatment, serum and cell supernatants are used to measure one or more cytokine pathways as a functional index, such as mTORC1 and/or IL-10 activation, using any assay or method known to one of skill in the art. Additionally, nuclear extracts can be used to test for NF-κB activation using any assay or method known to one of skill in the art. It is expected that administration of ccDNA in the presence of rapamycin or its analogue will result in less activation of cytokine pathways and secretion of cytokines such as IL-10 and type I IFN, resulting in increased duration of transgene expression and therapeutic efficacy.

EXAMPLE 19: Production of a cDNA vector coexpressing factor IX and a TLR-9 inhibitor

[00561] Oligonucleotides that can form a hairpin structure comprising the following sequences such as (TCCTGGCGGGGAAGT, SEQ ID NO: 889), ODN-2114 (TCCTGGAGGGGAAGT, SEQ ID NO: 890), poly(G) (GGGGGGGGGGGGGGGGGGGGGG, SEQ ID NO: 891), ODN-A151 (TTAGGGTTAGGGTTAGGGTTAGGG, SEQ ID NO: 892), G-ODN (CTCC-TATTGGGGGTTTCCTAT, SEQ ID NO: 893), IRS-869 (TCCTGGAGGGGTTGT, SEQ ID NO: 894), INH-1 (CCTGGATGGGAATTCCCATCCAGG, SEQ ID NO: 895), INH-4 (TTCCCATCCAGGCCTGGATGGGAA, SEQ ID NO: 896), (IRS-661 TGCTTGCAAGCTT-GCAAGCA, SEQ ID NO: 897), 4024 (TCCTGGATGGGAAGT, SEQ ID NO: 898), 4084F (CCTGGATGGGAA, SEQ ID NO: 899), INH-13 (CTTACCGCTGCACCTGGATGGGAA, SEQ ID NO: 900), INH-18 (CCTGGATGGGAACTTACCGCTGCA, SEQ ID NO: 901) and IRS-954 TGCTCCTGGAGGGTTGT, SEQ ID NO: 902) are designed to produce sticky ends after annealing of the complementary 5´-3´ and 3´-5´ strands so that they can be inserted by ligation into a pre-selected restriction cloning site, such as R5 or another site, of the TTX 9 or TTX 10 plasmid encoding the factor IX transgene, as described in Example 1 and Example 4.

[00562] For example, oligonucleotides with a suitable restriction site are annealed by mixing each strand in equal molar amounts in a suitable buffer: e.g., 100 mM potassium acetate; 30 mM HEPES, pH 7.5, heating to 94°C for 2 minutes, and gradually cooling. Thus, oligonucleotides with predicted significant secondary structure benefit from a more gradual cooling/renaturation step. This can be accomplished by placing the oligonucleotide solution in a water bath or incubator and turning the device off/on. The annealed oligonucleotides can be diluted in nuclease-free buffer and stored in double-stranded annealed form at 4°C. The ccDNA vector with the TLR-9 inhibitory oligonucleotide sequence is then purified (e.g., by gel electrophoresis or a column) and used to produce a ccDNA vector. A ccDNA vector can be produced that encodes factor IX and contains a TLR-9 antagonist as described in Examples 2-3. Described herein are methods for determining the effects of co-administration of a ccDNA vector expressing a TLR-9 inhibitor and rapamycin or a rapamycin analog.

EXAMPLE 20: Production of a ccDNA vector coexpressing factor IX and a cGAS inhibitor

[00563] The Kaposi's sarcoma-associated herpesvirus ORF52 protein (SEQ ID NO: 882), or a variant thereof that inhibits cGAS, or a truncated cytoplasmic isoform of LANA (LANAΔ161 or SEQ ID NO: 884) lacking amino acids 161-1162 of SEQ ID NO: 883), is operably linked to a promoter and inserted into the R5 restriction cloning site of a TTX 9 or TTX 10 plasmid encoding a factor IX transgene, as described in Example 1 and Example 4. Thus, a ccDNA vector is obtained that encodes both factor IX and a cGAS inhibitor, as described in Examples 2-3. This document describes methods for determining the effects of co-administration of a ccDNA vector expressing a cGAS inhibitor and rapamycin or a rapamycin analog.

EXAMPLE 21: Stable in vivo transgene expression by ccDNA vectors formulated with LNPs

[00564] The reproducibility of the results obtained in Example 7 with a different lipid nanoparticle was assessed in vivo in mice. On day 0, mice were dosed with either a ccDNA vector containing a luciferase transgene driven by the CAG promoter that was encapsulated in LNPs different from those used in Example 6, or the same LNPs containing poly(C) but without the ccDNA or luciferase gene. Specifically, male CD-1® mice, approximately 4 weeks old, were dosed with a single injection of 0.5 mg/kg TTX-luciferase LNPs or control poly(C) LNPs administered intravenously via the lateral tail vein on day 0. On day 14, the animals were systemically dosed with 150 mg/kg luciferin by intraperitoneal injection at 2.5 ml/kg. Approximately 15 minutes after luciferin administration, each animal was imaged using the In Vivo Imaging System (IVIS).

[00565] Significant fluorescence was observed in the liver in all four mice receiving ccDNA, with very little fluorescence observed in animals except at the injection site, indicating that the LNPs mediated liver-specific delivery of the ccDNA construct and that the delivered ccDNA vector was capable of controlled, sustained expression of its transgene for at least two weeks after administration.

EXAMPLE 22: Stable transgene expression in the liver in vivo after administration of a ccDNA vector

[00566] In a separate experiment, the localization of ccDNA delivered by LNPs in the liver of treated animals was assessed. A ccDNA vector containing a functional transgene of interest was encapsulated in the same LNPs used in Example 17 and administered to mice in vivo at a dose level of 0.5 mg/kg by intravenous injection. After 6 hours, the mice were sacrificed and liver samples were collected, formalin fixed, and paraffin embedded using standard protocols. ^RNAscope® in situ hybridization assays were performed to visualize the ccDNA vectors in tissue using a probe specific for the ccDNA transgene and detect using a chromogenic reaction and hematoxylin staining (Advanced Cell Diagnostics). The visualization analysis confirmed that ccDNA was present in hepatocyte samples from treated mice. The skilled artisan will appreciate that luciferase in the ccDNA vector can be replaced by any nucleic acid sequence selected from Table 5.

EXAMPLE 23: Stable expression of a cfDNA transgene in the eye in vivo

[00567] The stability of expression of the ccDNA vector transgene in tissues other than the liver was assessed to determine the tolerability and expression of the ccDNA vector following ocular administration in vivo. Although luciferase was used as an exemplary transgene, one of ordinary skill in the art can easily replace the luciferase transgene with an inflammasome antagonist sequence from any of the sequences listed in Table 5A-5F.

[00568] On day 0, male Sprague Dawley rats, approximately 9 weeks of age, were injected subretinally with 5 μl of either a ccDNA vector containing the luciferase transgene formulated with jetPEI® Transfection Reagent (Polyplus) or plasmid DNA encoding luciferase formulated with jetPEI®, both at a concentration of 0.25 μg/μl. Four rats were tested per group. Animals were sedated and injected subretinally with study drug in the right eye using a 33-gauge needle. The left eye of each animal was left intact. Eyes were examined immediately after injection by optical coherence tomography or fundus imaging to confirm the presence of a subretinal bleb. Rats received buprenorphine and topical antibiotic ointment according to standard procedures.

[00569] On days 7, 14, 21, 28, and 35, animals in both groups were administered systemically freshly prepared luciferin at 150 mg/kg by intraperitoneal injection at 2.5 ml/kg. Five to 15 minutes after luciferin administration, all animals were imaged using IVIS under isoflurane anesthesia. Total flux [photons/s] and average flux (photons/s/sr/ ^cm2 ) in a region of interest including the eye were obtained with an exposure of 5 minutes. Results are presented graphically as the mean luminosity in the treated eye (“injected”) for each treatment group relative to the mean luminosity in the untreated eye (“uninjected”) for each treatment group. Significant fluorescence was readily detected in eyes injected with the ccDNA vector but was much weaker in eyes injected with the plasmid. After 35 days, plasmid-injected rats were sacrificed, and the study continued for ccDNA-treated rats with luciferin injection and IVIS imaging on days 42, 49, 56, 63, 70, and 99. The results demonstrate that the ccDNA vector, administered into the rat eye by a single injection, mediates transgene expression in vivo and that such expression was maintained at high levels for at least 99 days post-injection.

EXAMPLE 24: Hydrodynamic delivery of cDNA

[00570] Hydrodynamic tail vein injection is a well-established method for delivering nucleic acid into the liver of rodents. In this system, pressurized injection of a large volume of unencapsulated nucleic acid results in a transient increase in cellular permeability and delivery directly into tissues and cells. This provides an experimental mechanism for evading many host immune systems, such as macrophage recruitment. Accordingly, luciferase expression observed following hydrodynamic injection of a naked ccDNA vector was compared to that observed following a more traditional intravenous injection of LNP-encapsulated ccDNA. For this experiment, the ccDNA vectors used were the wild-type AAV2 left ITR and the mutated right ITR.

[00571] In general, a ccDNA vector encoding luciferase under the control of the CAG promoter was prepared and either encapsulated in LNPs or left unencapsulated. Adult male CD-1 mice were injected into the tail vein with either (i) the ccDNA vector encapsulated in LNPs at 0.5 mg/kg in a total volume of 5 ml/kg or (ii) the same vector, but unencapsulated, at 0.01 mg/kg in a total volume of 1.2 ml. There were three mice per treatment group. Body weights were recorded on days 1, 2, and 3. Intravital imaging was performed on days 1 and 3 using the in vivo imaging system (IVIS). For imaging, each mouse was injected intraperitoneally with 150 mg/kg luciferin at 2.5 ml/kg. After 15 minutes, each mouse was anesthetized and imaged.

[00572] Even when administered at a 50-fold lower dose, luciferase expression observed in mice that received hydrodynamic injection was much higher (~10 ¹⁰ maximum total flux) than in mice that did not receive hydrodynamic injection (~10 ⁷ maximum total flux) (FIG. 9). Previous studies have found that administration of LNPs alone without a ccDNA vector cargo did not elicit an immune response (data not shown), and thus the difference between the two dosing groups may be due to binding of the LNP-encapsulated ccDNA vector to one or more host immune systems and evasion of these systems by hydrodynamic injection.

EXAMPLE 25: Modulation of immune pathways in cultured cells and impact on expression of ccDNA vector

[00573] A cell-based assay was developed to facilitate the investigation of the contributions of various immune pathways to the host response to ccDNA administration. The assay utilizes THP-1 cells (an acute monocytic leukemia cell line) in several variants: THP-1 Dual™ cells (Invitrogen) with stable integration of reporter constructs to detect both NF-κB (TLR9 pathway, via SEAP detection using Quanti-Blue™) and IRF pathway activation (via secreted luciferase using Quanti-Luc™), THP-1 cells with constitutive suppression of the cGAS pathway, and THP-1 cells with constitutive suppression of the STING pathway. By using known inhibitors of specific pathways, the relative contributions of endogenous immune pathways to the observed immune response to a particular stimulus can be better understood.

[00574] In general, THP-1 cells in culture were diluted to 0.5 x 10 ⁶ /mL in Opti-MEM™ media (ThermoFisher) and 150 µL were added to each well of a 96-well plate. Cells were pretreated with inhibitors: target inhibitors were diluted in Opti-MEM™ and added to the designated sample wells. For this experiment, A151 (oligonucleotide TTAGGGTTAGGGTTAGGGTTAGGGTTAGGG (SEQ ID NO:892) and BX795 (N-[3-[[5-iodo-4-[[3-[(2-thienylcarbonyl)amino]propyl]amino]-2-pyrimidinyl]amino]phenyl]-1-pyrrolidinecarboxamide, CAS 702675-74-9) were used in each sample well at final concentrations of 0 μM, 0.625 μM, 1.25 μM, or 2.5 μM. Plates were incubated at 37°C for 2 hours. 200 ng of target ccDNA vector was diluted 1:3 in Lipofectamine™ 3000 and incubated for 5-10 min at room temperature. Then, the ccDNA vector-lipofectamine complex was added to the wells with samples. The plates were incubated for 24 hours at 37°C. The magnitude of NF-κB activation and IRF2 activation were quantified using the Quanti-Blue™ and Quanti-Luc™ kits, respectively, according to the manufacturer's instructions.

[00575] Administration of two different preparations of the ccDNA vector into THP-1 Dual™ reporter cells resulted in significant interferon induction for both preparations, indicating activation of at least one immune pathway (FIG. 10A). Notably, no interferon induction was observed when either of the two gene-silenced THP-1 lines was treated with the same concentration of ccDNA (FIG. 10A), indicating that the cGAS/STING pathway is involved in cytokine induction in response to ccDNA administration. A similar result was observed when THP-1 Dual™ reporter cells were treated with both ccDNA and BX795; BX795 is a specific inhibitor of the STING pathway, and its abolition of ccDNA-induced interferon induction again indicates involvement of the STING pathway (FIG. 10A). A151 is known to inhibit the cGAS/STING pathway, the TLR9 pathway, and inflammasome-mediated immune pathways. Its effect was similar to that observed with BX795 treatment (FIG. 10A).

[00576] In a second experiment, the concentrations of inhibitor required to observe a protective effect upon administration of ccDNA were analyzed (FIG. 10B). For both A151 and AS1411, the observed inhibition of interferon induction was concentration dependent, with maximal inhibition observed at a concentration of 2.5 μM (FIG. 10B).

EXAMPLE 26: Effect of modulation of unmethylated CpG content in ccDNA on the immune response

[00577] CpG motifs in the gene sequence are known to stimulate the TLR9 DNA-sensing pathway. Accordingly, the effect of reducing the number of CpG motifs in the ccDNA construct sequence on the activation of the innate immune pathway upon administration of this sequence in vivo was studied.

A. Cell-based assays to test the effects of minimizing unmethylated CpG content in ccDNA

[00578] Studies were performed to assess (i) the activation of the TLR9 pathway in response to ecDNA administration, and (ii) the effect of modulating the presence/methylation status of CpG on such activation. For this particular study, a ecDNA vector that expressed green fluorescent protein and contained a wild-type left ITR and a mutant right ITR was used.

[00579] HEK-293 cells expressing human TLR9 (HEK-BLUE.hTLR9 cells, InvivoGen) were seeded in a 96-well plate at 50,000 cells per well. The plates were incubated overnight at 37°C. For methylation-pretreated ccDNA samples, ccDNA vector, buffer, S-adenosylmethionine, CpG methyltransferase, and water were mixed to achieve a total reaction volume of 50 μl according to methods known in the art. The reaction mixture was incubated at 37°C for 1 hour, then stopped by heating to 65°C for 20 minutes. ccDNA was purified from the reaction mixture using a commercially available purification kit (PCR Purification Kit, Qiagen®), and the concentration of the resulting DNA was measured.

[00580] Cells were pretreated for 3 hours with any target inhibitors, in this experiment A151 was used at a final concentration of 10 μM per well. Following pretreatment, cells were transfected with 300 ng of ccDNA at a 1:3 ratio with Lipofectamine 3000 diluted in Opti-MEM™ or the positive control ODN2006, which is known to stimulate the TLR9 pathway. Cells were incubated for 24 hours at 37°C and 5% CO ₂ . SEAP expression (a component of the TLR9 pathway) was then measured using Quanti-BLUE™ (InvivoGen).

[00581] As shown in FIG. 11A, ODN2006 induced a robust NF-κB response; the cfDNA construct induced a less significant response, and when premethylated, the response was reduced to background levels. When combined with A151 (known to inhibit the TLR9 pathway), cfDNA-treated samples also showed minimal levels of NF-κB induction (FIG. 11B). First, this demonstrates that the TLR9 pathway contributes to the host immune response to cfDNA administration. Furthermore, minimization of CpG content by methylation abolished most of the TLR9 activation by cfDNA, and this effect could be mimicked by pretreating cells with A151 without altering CpG content or methylation status.

B. Mouse studies assessing the effects of minimizing unmethylated CpG content in ccDNA

[00582] The impact of minimizing the CpG content of ccDNA vectors was also assessed in mice. Cytokine responses and expression of ccDNA-encoded genes were measured upon administration of ccDNA vectors to mice.

[00583] Three different ccDNA vectors were used, each encoding luciferase as a transgene. The first ccDNA vector had a large amount of unmethylated CpG (~350) ("high-CpG ccDNA") and contained a constitutive CAG promoter; the second had a moderate amount of unmethylated CpG (~60) ("low-CpG ccDNA") and contained a liver-specific hAAT promoter; and the third was a methylated form of the second variant such that it contained no unmethylated CpG ("no-CpG ccDNA") and also contained the hAAT promoter. The ccDNA vectors were otherwise identical. Vectors were prepared as described above.

[00584] Four groups of four male CD-1 mice, approximately 4 weeks old, were treated with one of the LNP-encapsulated ccDNA vectors or a poly(C) control. On day 0, each mouse was given a single intravenous injection into the tail vein of 0.5 mg/kg ccDNA vector at a volume of 5 ml/kg. Body weights were recorded on days -1, -, 1, 2, 3, 7, and weekly thereafter until the mice were sacrificed. Whole blood and serum samples were collected on days 0, 1, and 35. Intravital imaging was performed on days 7, 14, 21, 28, and 35, and weekly thereafter using the in vivo imaging system (IVIS). For imaging, each mouse was given an intraperitoneal injection of 150 mg/kg luciferin at 2.5 ml/kg. After 15 min, each mouse was anesthetized and imaged. Mice were sacrificed on day 93 and terminal tissues, including liver and spleen, were collected. Cytokine measurements were performed 6 h post-dose on day 0.

[00585] Similar body weight loss was observed in each of the cfDNA-treated mouse groups (5-7%), followed by rapid recovery by day 7. Cytokine analyses from day 0 samples showed that while many of the cytokines assessed were similarly elevated in all treatment groups, interferon alpha, tumor necrosis factor alpha, and MIP-1 alpha were all decreased in the low-CpG or no-CpG samples compared to the high-CpG samples (FIG. 12A and FIG. 12B).

[00586] While mice treated with either low-CpG or high-CpG ccDNA showed significant fluorescence on days 7 and 14, mice treated with high-CpG ccDNA showed a rapid decline in fluorescence after day 14 and a continuous decline for the remainder of the study. In contrast, total flux for both low-CpG ccDNA and CpG-null ccDNA treated mice remained at a consistently high level (FIG. 12C), indicating that maintaining the presence of unmethylated CpG in the ccDNA vector below some threshold, and thus the absence of TLR9 pathway activation, helps avoid the more rapid decline in ccDNA-encoded protein expression (and thus fluorescence) seen in other cases in this study.

EXAMPLE 27: Expression and host response in neonatal mice

[00587] Previous experiments have shown that the cGAS/STING pathway is at least partially involved in the cytokine induction response observed upon introduction of the ccDNA vector into cells. This pathway is known to become active later in development, and neonatal mice with immature immune systems lack an active cGAS/STING pathway. Accordingly, an experiment was conducted in neonatal mice to examine the effect of the absence of this pathway on the expression and stability of the ccDNA vector.

[00588] A ccDNA vector encoding luciferase as a transgene with a wild-type AAB2 left ITR and a mutant right ITR and a CAG promoter was used. The ccDNA vector was prepared as described above. ccDNA vector samples or poly(C) control were administered by intravenous injection into the tail vein of neonatal (8-day-old) male CD-1 mice at a dose level of 0.1 or 0.5 mg/kg in a volume of up to 5 ml/kg. Five replicates were included in each sample group. Body weights were recorded on day 1 and for the next three days. Intravital imaging was performed on days 7, 14, and 21 using the in vivo imaging system (IVIS). For imaging, each mouse was given an intraperitoneal injection of 150 mg/kg luciferin at 2.5 ml/kg. After 15 minutes, each mouse was anesthetized and imaged.

[00589] Notably, no weight loss was observed in any treatment group following the day 0 injection. High levels of total flux (representative of luciferase expression from injected ccDNA vectors) were observed in all ccDNA-injected animals, with the 0.5 mg/kg dose resulting in expression levels approximately 1 log higher than the 0.1 mg/kg dose during the first 14 days (FIG. 13). Thereafter, expression levels stabilized and were maintained at the same level in both dosing groups. Compared to similar studies in adult CD-1 mice, ccDNA vector expression levels were at least two logs higher in neonatal mice even after 14 days (data not shown). This result suggests that avoidance of cGAS/STING pathway activation is beneficial for promoting ccDNA vector expression and stability.

EXAMPLE 28: Effect of modulation of multiple immune pathways on cDNA persistence and expression and cDNA cytokine induction

[00590] Previous studies have assessed the effects of TLR9 pathway modulation in both cultured cell systems and mouse systems. However, multiple molecular pathways are known to be involved in the host response to foreign DNA, and the effect of avoiding TLR9 pathway activation may not be easily detectable if one or more other pathways are still engaged by the introduction of ccDNA. To test this, CpG-minimized ccDNA vectors were tested in a goldenticket mouse strain that has a mutation that abolishes STING function. This experiment allowed the TLR9 pathway to be examined without the confounding effect of cGAS/STING pathway activity.

[00591] Three different ccDNA vectors were used, each encoding luciferase as a transgene. The first ccDNA vector had a large amount of unmethylated CpG (~350) (“high-CpG ccDNA”) and contained a constitutive promoter (cET), the second had a moderate amount of unmethylated CpG (~60) (“low-CpG ccDNA”), and the third had a small amount of CpG (~36) but was methylated such that it contained no unmethylated CpG (“no-CpG ccDNA”). The second and third constructs contained liver-specific hAAT promoters. The ccDNA vectors were otherwise identical. Vectors were prepared as described above.

[00592] Each of the ccDNA vector samples or the poly(C) control were administered by intravenous injection into the tail vein of adult male goldenticket mice (Tmem173 ^gt ) at a dose level of 0.5 mg/kg in a volume of 5 ml/kg. In some cases, mice were given a second dose of the ccDNA vector sample on day 22. Four replicates were included in each sample group. Body weights were recorded on the days of dosing and for the following three days. Whole blood and serum samples were collected on day 0 (6 hours post-dose) and day 22 (6 hours post-dose). Intravital imaging was performed on days 7, 14, 22, 29, 36, and 43 using the in vivo imaging system (IVIS). For imaging, each mouse was given an intraperitoneal injection of 150 mg/kg luciferin at 2.5 ml/kg. After 15 min, each mouse was anesthetized and imaged. Mice were sacrificed on day 43 and terminal tissues, including liver and spleen, were collected. Cytokine measurements were performed on blood collected on days 0 and 22.

[00593] Body weight loss following ccDNA administration was <5% and was essentially restored in all cases by day 3. Following rechallenge on day 22, treated mice again lost <5% of their body weight and rapidly regained it over the following days. Cytokine induction was assessed in blood samples from day 0 (FIG. 14A). With the exception of IL-18, levels of each cytokine analyzed correlated with the degree of CpG present in the ccDNA construct (FIG. 14A), with little or no induction of IFN-alpha, IFN-gamma, IL-6, IP-10, MCP-1, MIP-1 alpha, MIP-1 beta, and RANTES in mice treated with CpG-free ccDNA. In mice receiving the second dose, all cytokine levels were increased relative to day 0 levels in all samples (FIG. 14B), but again in all cases except IL-18, the degree of activation correlated with the amount of CpG present in the introduced ccDNA vector.

[00594] Luciferase expression was similar across treatment groups until day 22 (FIG. 14C). After this time point, samples from mice that received either a single dose or a repeat dose of high CpG ccDNA showed a dramatic decrease in total flux, while the low CpG ccDNA and no CpG ccDNA groups either maintained similar total flux measurements or showed a smaller loss of signal relative to the high CpG ccDNA group. Taken together, the results demonstrate that minimizing the CpG content of the administered ccDNA vectors and, as a result, avoiding engagement of the TLR9 innate immune pathway, contributed to the dramatic decrease in cytokine induction and more robust persistence of ccDNA gene expression in treated goldenticket mice.

[00595] EXAMPLE 29: Chronic Dosing and Repeated Dosing of ccDNA Vector in Rag2 Mice.

[00596] In situations where one or more transgenes encoded in the expression gene cassette of a ccDNA vector are expressed in a host environment (e.g., a cell or subject) where the expressed protein is recognized as foreign, there is a potential for the host to mount an adaptive immune response that may result in unwanted depletion of the expression product, which may be mistaken for a lack of expression. In some cases, this may occur with a reporter molecule that is heterologous to the normal host environment. Accordingly, ccDNA vector transgene expression was assessed in vivo in the Rag2 mouse model, which lacks B and T cells and therefore does not mount an adaptive immune response to non-native mouse proteins such as luciferase. In general, on day 0, gene-silenced c57bl/6 and Rag2 mice were injected intravenously into the tail vein with 0.5 mg/kg of the LNP-encapsulated ccDNA vector expressing luciferase or poly(C) control, and on day 21, some mice were given a booster dose of the same LNP-encapsulated ccDNA vector at the same dose level. All test groups consisted of 4 mice each. IVIS imaging was performed after luciferin injection at weekly intervals.

[00597] When comparing the total flux observed in the IVIS assays, the fluorescence observed in wild-type mice (an indirect measure of the presence of expressed luciferase) treated with the ccDNA vector-Luc LNPs gradually decreased after day 21, while Rag2 mice treated with the same drug exhibited relatively constant, robust luciferase expression throughout the 42 days of the experiment (FIG. 16A). The decrease observed around day 21 in wild-type mice corresponds to the time window in which an adaptive immune response might be expected to occur. Repeated administration of the ccDNA vector LNPs to Rag2 mice resulted in a marked increase in expression that was robust for at least the 21 days that it was monitored in this study (FIG. 16B). The results suggest that adaptive immunity may play a role when non-native protein is expressed from a ccDNA vector in the host, and that the observed decrease in expression at 20+ days after initial administration may indicate an interfering adaptive immune response to the expressed molecule rather than (or in addition to) a decrease in expression. It should be noted that this response is expected to be low in the case of native proteins expressed in the host, where the host would presumably correctly recognize the expressed molecules as self and would not mount such an immune response.

EXAMPLE 30: Effect of liver-specific expression and CpG modulation on robust expression

[00598] As described in Example 29, an unwanted host immune response may in some cases artificially attenuate what would otherwise be robust expression of one or more target transgenes from an introduced ccDNA vector. To assess the impact of avoiding and/or attenuating a potential host immune response on robust expression from a ccDNA vector, two approaches were used. First, because the ccDNA-Luc vector used in the preceding examples was under the control of a constitutive CAG promoter, a similar construct was made using a liver-specific promoter (hAAT) or another constitutive promoter (hEF-1) to determine whether avoiding prolonged exposure to myeloid cells or non-liver tissues would reduce any observed immune effects. Second, some ccDNA-luciferase constructs were modified to have reduced CpG content, a known trigger of the host immune response. Expression of the ccDNA-encoded luciferase gene was measured when these promoter-swapped ccDNA vectors were injected into mice.

[00599] Three different ccDNA vectors were used, each encoding luciferase as a transgene. The first ccDNA vector had a high amount of unmethylated CpG (~350) and contained a constitutive CAG promoter (“CAG ccDNA”); the second had a moderate amount of unmethylated CpG (~60) and contained a liver-specific hAAT promoter (“low CpG hAAT ccDNA”); and the third was a methylated form of the second that lacked the unmethylated CpG and also contained the hAAT promoter (“no CpG hAAT ccDNA”). The ccDNA vectors were otherwise identical. Vectors were prepared as described above.

[00600] Four groups of four male CD- ^1® mice, approximately 4 weeks old, were administered one of the LNP-encapsulated ccDNA vectors or a poly(C) control. On day 0, each mouse was given a single intravenous injection into the tail vein of 0.5 mg/kg ccDNA vector at a volume of 5 ml/kg. Body weights were recorded on days -1, -, 1, 2, 3, 7, and weekly thereafter until the mice were sacrificed. Whole blood and serum samples were collected on days 0, 1, and 35. Intravital imaging was performed on days 7, 14, 21, 28, and 35, and weekly thereafter using the in vivo imaging system (IVIS). For imaging, each mouse was given an intraperitoneal injection of 150 mg/kg luciferin at 2.5 ml/kg. After 15 min, each mouse was anesthetized and imaged. Mice were sacrificed on day 93 and terminal tissues, including liver and spleen, were collected. Cytokine measurements were performed 6 h post-dose on day 0.

[00601] Although all ccDNA-treated mice showed significant fluorescence on days 7 and 14, mice receiving CAG ccDNA showed rapid decline in fluorescence after day 14 and a more gradual decline throughout the remainder of the study. In contrast, mice receiving low-CpG and no-CpG hAAT ccDNA showed consistent high flux (FIG. 17). This suggests that targeting the ccDNA vector specifically to the liver resulted in robust and long-lasting transgene expression from the vector for at least 77 days following a single injection. Constructs with minimal or no CpG content had similar long-term persistent expression profiles, while the construct with a constitutive promoter and high CpG content showed a decrease in expression over time, suggesting that host immune activation upon administration of the ccDNA vector may play a role in any decrease in expression observed upon administration of such a vector to a subject. These results provide alternative methods to tailor the duration of the response to a desired level by selecting a tissue-restricted promoter and/or altering the CpG content of the ccDNA vector if a host immune response, potentially a transgene-specific response, is observed.

EXAMPLE 31: Expression of an inflammasome antagonist in vivo

[00602] Once proper protein expression and function in recipient cells is confirmed in vitro, a ccDNA vector with sequences encoding an inflammasome antagonist can be formulated with lipid nanoparticles and administered to mice deficient in functional expression of the corresponding protein product at various time points (in utero, neonatal, 4 weeks, and 8 weeks of age) to test protein expression and function in vivo.

[00603] LNPs with ccDNA vectors are administered to the appropriate mice at doses of 0.3 to 5 mg/kg in a volume of 1.2 ml. Each dose should be administered by intravenous hydrodynamic administration or will be administered, for example, by intraperitoneal injection. Administration to normal mice serves as a control and can also be used to detect the presence and amount of therapeutic protein.

[00604] Following a single administration, such as a single dose of LNP-ccDNA, expression in liver tissue in a recipient mouse will be determined at various time points, such as 10, 20, 30, 40, 50, 1000, and 200 days or more. etc. In particular, samples of the mouse liver and bile duct will be obtained and analyzed for the presence of protein using immunostaining of tissue sections. The presence of the protein will be assessed quantitatively, as well as with respect to proper localization in the tissue and the cells therein. Cells in the liver (e.g., hepatic and epithelial) and bile duct (e.g., cholangiocytes) will be assessed to determine protein expression.

EXAMPLE 32: Generation of ccDNA coexpressing a therapeutic gene (e.g., factor IX) and an inhibitor of the NLRP3 inflammasome pathway.

[00605] A151 (SEQ ID NO: 892) or a variant thereof that inhibits AIM2 is operably linked to a promoter and inserted into the R5 restriction cloning site of a ccDNA vector as described in Example 1. This produces a ccDNA that encodes both factor IX and an AIM2 inhibitor.

EXAMPLE 33: Confirmation of expression of NLRP3 inflammasome inhibitor expressed by ccDNA

[00606] Expression of a target NLRP3 or AIM2 inhibitor, or caspase-1 co-expressed by a ccDNA, such as A151 (SEQ ID NO: 892), can be confirmed using HeLa cells and antibodies specific for the inhibitor. For example, HeLa cells are cultured and transient transfections of constructs co-expressing factor IX and a target NLRP3 or AIM2 inhibitor, or caspase-1 are performed using, for example, Fusegene6 transfection reagent (3:1; fusgene6:DNA). Western blotting and/or flow cytometry techniques are used to detect expression of the NLRP3 or AIM2 inhibitor, or caspase-1, as known to those skilled in the art.

EXAMPLE 34: Factor IX for the treatment of hemophilia B using cctDNA encoding factor IX and an NLRP3 inflammasome pathway inhibitor.

[00607] The experiment is carried out in factor IX-deficient mice that have an inserted hFIX sequence with a deleterious mutation (R333Q). Male mice with suppressed factor IX receive single or repeated doses of ccDNA-LNPs (lipid nanoparticles with ccDNA). Two variants of ccDNA-vectored LNPs are used; 1) a ccDNA-LNP encoding human factor IX (either the native human sequence or Padua FIX variants) and encoding A151 (SEQ ID NO: 892); and a ccDNA-LNP encoding only factor IX but not the cGAS inhibitor as a comparative ccDNA vector. The ccDNA-vectored LNPs are administered to the respective mice at doses of 0.3 to 5 mg/kg in a volume of 1.2 ml. Each dose should be administered by intravenous hydrodynamic injection. Expression of factor IX in plasma is assessed by ELISA at various time points, such as 7, 14, 21 days or more, etc. Activated partial thromboplastin time and bleeding time are also measured as a determination of efficacy. Mice that receive a ccDNA vector expressing both hFIX and A151 are expected to have increased and/or sustained expression of factor IX over a longer period of time compared to mice that receive a ccDNA vector expressing only factor IX but not A151 or another NLRP3 inhibitor or AIM2 or caspase-1. In addition, following repeated dosing, mice that receive a repeated dose of a ccDNA vector containing both A151 and factor IX are expected to exhibit reduced activation of cytokine secretion and increased duration of transgene expression and therapeutic efficacy compared to mice that receive a repeated dose of a ccDNA vector encoding factor IX alone. The NLRP3 inflammasome pathway inhibitor and factor IX may be delivered on different ccDNA vectors, but are preferably encoded by the same vector and, accordingly, inhibition of the NLRP3 inflammasome pathway inhibitor occurs in the same cell as the cell that receives the ccDNA vector encoding a transgene such as factor IX.

EXAMPLE 35: Determining the effects of co-administration of cctDNA and an NLRP3 inflammasome inhibitor on innate immune responses and duration of factor IX expression

[00608] To examine the effects of co-administration of a ccDNA of interest and an inhibitor of the NLRP3 inflammasome pathway on innate immune responses in vitro, reporter cell lines can be used for functional assays that examine activation of the NLRP3 inflammasome or caspase-1. An NLRP3 inflammasome reporter cell line that can be used for such in vitro assays can be a stably co-transfected cell line that expresses full-length NLRP3 and a reporter gene, such as a secreted alkaline phosphatase (SEAP) reporter gene, under the transcriptional control of a transcription factor response element, such as an NF-kB binding site, an AP-1 binding site, or a combination thereof. For example, reporter cells are seeded in 96-well plates. After a predetermined period of time, such as 16 hours, the cells are stimulated with varying amounts of compositions containing factor IX-expressing cDNA with or without an NLRP3 inflammasome inhibitor. The activity of a reporter gene, such as SEAP, can be analyzed using any method or assay known to one skilled in the art to compare the level of caspase-1 activation or NLRP3 inflammasome activation in the presence of the cDNA of interest with or without an NLRP3 inflammasome pathway inhibitor. It is expected that less activation of the reporter molecule will be observed in the presence of an NLRP3 inflammasome inhibitor. A similar reporter assay can be used to evaluate caspase-1 inhibitors.

[00609] Similarly, to examine the effects of co-administration of a cDNA of interest and an inhibitor of the AIM2 inflammasome pathway on innate immune responses in vitro, reporter cell lines can be used for functional assays that examine activation of the AIM2 inflammasome or caspase-1. An AIM2 inflammasome reporter cell line that can be used for such in vitro assays can be a stably co-transfected cell line that expresses full-length human AIM2 and a reporter gene, such as the secreted alkaline phosphatase (SEAP) reporter gene, under the transcriptional control of a transcription factor response element, such as an NF-kB binding site, an AP-1 binding site, or a combination thereof. The assay can be performed in a similar manner to the NLRP3 inflammasome reporter assay, in which reporter cells, such as those plated in 96-well plates, are stimulated after a predetermined period of time with varying amounts of compositions containing factor IX-expressing cDNA, with or without an AIM2 inflammasome inhibitor. The activity of a reporter gene, such as SEAP, can be analyzed using any method or assay known to one skilled in the art to compare the level of caspase-1 activation or AIM2 inflammasome activation in the presence of the cDNA of interest, with or without an AIM2 inflammasome pathway inhibitor. It is expected that less activation of the reporter molecule will be observed in the presence of an AIM2 inflammasome inhibitor. A similar reporter assay can be used to evaluate caspase-1 inhibitors.

[00610] Additionally, reporter lines with suppression of the NLRP3 inflammasome or the AIM2 inflammasome can be used, such as THP1-defNLRP3 cells (InvivoGen) or THP-1 cells overexpressing TRIM11, which suppresses the AIM2 inflammasome (Liu et al., Cell Reports (2016) 16: 1988-2002), and other cell lines known in the art. Such reporter lines with suppression of AIM2 or NLRP3 can express one or more inducible genes of secreted reporters, such as Lucia luciferase and SEAP (secreted embryonic alkaline phosphatase). The reporter gene can be under the control of the minimal ISG54 (interferon-stimulated gene) promoter in combination with one or more, such as five, IFN-stimulated response elements. The reporter gene may also be under the control of a minimal IFN-β promoter fused to one or more, such as five, copies of a response element, such as an NF-kB response element. The activity of NLRP3 or AIM2 or caspase-1 in the presence of at least one NLRP3 or AIM2 or caspase-1 inhibitor in combination with the ccDNA described herein can be compared in a gene-silenced cell line versus a parent cell line.

[00611] To examine the effects of co-administration of a cDNA of interest and an NLRP3 inflammasome inhibitor and/or an AIM2 inflammasome inhibitor, or an NLRP3 antagonist, or an AIM2 antagonist on the NLRP3 inflammasome and/or AIM2 inflammasome activation pathway ex vivo, human monocytes can be isolated, for example, by density gradient centrifugation of peripheral blood and magnetic separation. These monocytes can be examined before and after exposure to and/or activation of the cDNA of interest with or without an NLRP3 inflammasome inhibitor and/or an AIM2 inflammasome inhibitor, or an NLRP3 antagonist, or an AIM2 antagonist, or a caspase-1 inhibitor, with appropriate controls. After processing, the serum and cell supernatants are used to measure one or more cytokine pathways as a functional indicator of NLRP3 inflammasome pathway activation and/or AIM2 inflammasome pathway inhibitor, such as interleukin (IL)-1β, IL-6, IL-8, IL-18, interferon (IFN)-γ, interferon (IFN)-α, monocyte chemoattractant protein (MCP)-1, IP-10 and/or tumor necrosis factor (TNF)-α, using any assay or method known to those skilled in the art. In addition, nuclear extracts can be used to test for NF-κB activation using any assay or method known to those skilled in the art. It is expected that administration of ccDNA in the presence of an NLRP3 and/or AIM2 inflammasome pathway inhibitor or a caspase 1 inhibitor will result in less cytokine pathway activation and cytokine secretion, thereby increasing the duration of transgene expression and therapeutic efficacy.

[00612] To examine the effects of co-administration of an ecDNA of interest and an NLRP3 and/or AIM2 inflammasome pathway inhibitor or a caspase 1 inhibitor on NLRP3 and/or AIM2 inflammasome pathway activation or caspase 1 activation in vivo, a mouse model can be used. Mouse serum or lymphocyte samples are examined before and after exposure to and/or activation of ecDNA expressing a transgene of interest, such as factor IX, with or without an NLRP3 and/or AIM2 inflammasome pathway inhibitor or a caspase-1 inhibitor, with appropriate controls. After processing, the serum and cell supernatants are used to measure one or more cytokine pathways as a functional indicator of NLRP3 and/or AIM2 inflammasome pathway activation or caspase 1 activation, such as interleukin (IL)-1β, IL-6, IL-8, IL-18, interferon (IFN)-γ, interferon (IFN)-α, monocyte chemoattractant protein (MCP)-1 and/or tumor necrosis factor (TNF)-α, using any assay or method known to those skilled in the art. In addition, nuclear extracts can be used to test for NF-κB activation using any assay or method known to those skilled in the art. It is expected that administration of ccDNA in the presence of an NLRP3 and/or AIM2 inflammasome pathway inhibitor or a caspase 1 inhibitor will result in less significant immune activation and cytokine secretion, thereby increasing the duration of transgene expression and therapeutic efficacy.

[00613] Co-administration of a cDNA of interest expressing human factor IX produced from the TTX-9 plasmid and an inhibitor of the NLRP3 inflammasome and caspase-1 was evaluated. Groups of C57bL mice (n = 8) were evaluated as shown in Table 11.

[00614] Table 11:

LNC Group cDNA dosage level Immunosuppression LNC 1 siRNA 0.5 mg/kg Absent 2 0.5 mg/kg TTX9 Absent 3 0.5 mg/kg TTX9 Cremophor (solvent control) via gastric tube 12-16 hours before TTX9, then one hour before TTX9. 4 0.5 mg/kg TTX9 VX765 100 mg/kg in cremophor 12-16 hours before TTX9 gavage and then one hour before dosing 5 0.5 mg/kg TTX9 MCC950 (NLRP3 inhibitor) 50 mg/kg ip 12-16 hr before TTX9 administration, then 1 hr before cDNA administration

[00615] In general, animals were pretreated with a macrophage activation inhibitor or control according to the groups as shown in Table 12. Animals were administered MCC950 (NLRP2 inhibitor) (Group 5) or VX765 (belnakazan; selective caspase-1 inhibitor) (Group 4) i.p. 12-16 hr and then also 1 hr prior to administration of 0.5 mg/kg ccDNA (TTX9-LNP) (Group 1) or LNP-siRNA (negative control) (Group 1) by i.v. administration via the lateral tail vein. The pretreatment control group received clondronate (vehicle control) only (Group 3). Whole blood was collected from each group on days 0, 1, 7, and 21 by blood sampling via the tail vein or facial vein, or from the orbital area.

Table 12: Introduction of pre-processing

Group No. Number of animals in the group Pre-processing Dose level Dose volume Treatment regimen, ROA 1 8 Not applicable Not applicable Not applicable not applicable 2 8 Not applicable Not applicable Not applicable 3 8 Solvent control 0.0 mg/kg 5 ml/kg Once on Days -1 ^a and 0 ^b
By 4 8 VX765 200 mg/kg 5 8 MCC950
(NLRP3 inhibitor) 50 mg/kg 5 ml/kg intravenously 1 hour before TA administration

[00616] ^a The first p/o administration will be performed 12-16 hours prior to cDNA processing on Day 0.

[00617] ^b The second oral administration will be performed 1 hour prior to cDNA treatment on Day 0.

[00618] # = number; ROA = route of administration; p.o. = gastric tube; i.p. = intraperitoneal; i.v. = intravenous; TA = study drug; NA = not applicable.

[00619] Cytokine levels were quantified and assessed using the ProcartaPlex multiplex immunoassay (Invitrogen) according to the manufacturer's instructions, which is a bead-based, quantitative, multiplex immunoassay for measuring levels of various cytokines and chemokines using the Luminex technology platform. Samples from mice were mixed with a premixed custom 8-plex mouse cytokine assay kit, magnetic beads, and assayed for levels of IFN-α, IFN-γ, IL-6, IP-10, IL-18, IL-1β, MCP-1, and TNF-alpha. FIGS. 18A-18H show assessment of cytokine levels following TTX-9 administration with pharmacological depletion of macrophages using an NLRP3 inhibitor (MCC950) or a caspase 1 inhibitor (VX765). IFNγ and IL-18 levels were significantly reduced by MCC950 (NLRP3 inhibitor) treatment (FIG. 18B and FIG. 18D), and IP-10 levels were reduced by MCC950 treatment (FIG. 18F). IL-18 levels were also reduced by VX765 (caspase-1 inhibitor) treatment (FIG. 18D).

List of sequences without formalities

Subsequence Description Subsequence SEQ ID NO:1 WT-ITR AAB2 (right) AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGctgcctgcagg SEQ ID NO:2 Modified ITR AAB2 (right) AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

SEQ ID NO:3 CAG promoter TCAATATTGGCCATTAGCCATATTATTCATTGGTTATATAGCATAAATCAATATTGGCTATTGGCCATTGCATACGTTGTATCTATATCATAATATGTACATTTATATTGGCTCATGTCCAATATGACCGCCATGTTGGCATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCC GCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTCCGCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTACGGGACTTTCCTACTTG GCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTCGAGGTGAGCCCCACGTTCTGCTTCACTCTCCCCCATCTCCCCCCCCTCCCCACCCCCAATTTTGTATTTATTTATTTTTTAATTATTTTGTGCAGCGATGGGGGCGGGGGGGGGGGGGGCGCGCCAGGCGGGGCGGGGCGGGGCGAGGGGCGGGGCGGGGCGAGGCGGAGAGGTGCGGCGGCAGCCAATCAGAGCGG CGCGCTCCGAAAGTTTCCTTTTATGGCGAGGCGGCGGCGGCGGGCGGCCCTATAAAAAGCGAAGCGCGCGGCGGGCGGGAGTCGCTGCGACGCTGCCTTCGCCCCGTGCCCCGCTCCGCCGCCGCCTCGCGCCGCCCGCCCCGGCTCTGACTGACCGCGTTACTCCCACAGGTGAGCGGGCGGGACGGCCCTTCTCCTCCGGGCTGTAATTAGCGCTTGGTTTAATGACGGCTTGTTTCTTTT TCTGTGGCTGCGTGAAAGCCTTGAGGGGCTCCGGGAGGGCCCTTTGTGCGGGGGGGAGCGGCTCGGGGGGTGCGTGCGTGTGTGTGTGCGTGGGGAGCGCCGCGTGCGGCCCGCGCTGCCCGGCGGCTGTGAGCGCTGCGGGCGCGGCGCGGGGCTTTGTGCGCTCCGCAGTGTGCGCGAGGGGAGCGCGGCCGGGGGCGGTGCCCCGCGGTGCGGGGGGGGCTGCGAGGGGAACAAAGG CTGCGTGCGGGGTGTGTGCGTGGGGGGGTGAGCAGGGGGTGTGGGCGCGGCGGTCGGGCTGTAACCCCCCCCTGCACCCCCCTCCCCGAGTTGCTGAGCACGGCCCGGCTTCGGGTGCGGGGCTCCGTACGGGGCGTGGCGCGGGGCTCGCCGTGCCGGGCGGGGGGTGGCGGCAGGTGGGGGTGCCGGGCGGGGCGGGGCCGCCTCGGGCCGGGGAGGGCTCGGGGGAGGGGCGCG GCGG CCCCCGGAGCGCCGGCGGCTGTCGAGGCGCGGCGAGCCGCAGCCATTGCCTTTTATGGTAATCGTGCGAGAGGGCGCAGGGACTTCCTTTGTCCCAAATCTGTGCGGAGCCGAAATCTGGGAGGCGCCGCCGCACCCCCTCTAGCGGGCGCGGGGCGAAGCGGTGCGGCGCCGGCAGGAAGGAAATGGGCGGGGAGGGCCTTCGTGCGTCGCCGCGCCGCCGTCCCCTTCTCCCTCCA GCCTCGGGGCTGTCCGCGGGGGGACGGCTGCCTTCGGGGGGGACGGGGCAGGGCGGGGTTCGGCTTCTGGCGTGTGACCGGCGGCTCTAGAGCCTCTGCTAACCATGTTTTAGCCTTCTTCTTTTTCCTACAGCTCCTGGGCAACGTGCTGGTTATTGTGCTGTCTCATCATTTGTCGACAGAATTCCTCGAAGATCCGAAGGGGTTCAAGCTTGGCATTCCGGTACTGTTGGTAAA GCCA

SEQ ID NO:4 AAT promoter AGGCTCAGAGGCACACAGGAGTTTCTGGGCTCACCCTGCCCCCTTCCAACCCCTCAGTTCCCATCCTCCAGCAGCTGTTTGTGTGCTGCCTCTGAAGTCCACACTGAACAAACTTCAGCCTACTCATGTCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTGGGCCCATGCCACCTCCAA CATCCACTCGACCCCTTGGAATTTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGA GGGTCCGGGTTCAAAACCACTTGCTGGGTGGGGAGTCGTCAGTAAGTGGCTATGCCCCGACCCCGAAGCCTGTTTCCCCATCTGTACAATGGAAATGATAAAGACGCCCATCTGATAGGGTTTTTGTGGCAAATAAACATTTGGTTTTTTTGTTTTGTTTTGTTTTGTTTTTTGAGATGGAGGTTTGCTCTGTCGCCCAGGCTGGAGTGCAGTGACACAATCTCATCTCACCACAACCTTC CCCTGCCTCAGCCTCCCAAGTAGCTGGGATTACAAGCATGTGCCACCACACCTGGCTAATTTTCTATTTTTAGTAGA GACGGGTTTCTCCATGTTGGTCAGCCTCAGCCTCCCAAGTAACTGGGATTACAGGCCTGTGCCACCACACCCGGCTAATTTTTTCTATTTTTGACAGGGACGGGGTTTCACCATGTTGGTCAGGCTGGTCTAGAGGTACCGGATCTTGCTACCAGTGGAACAGCCACTAAGGATCTGCAGTGAGAGCAGAGGGCCAGCTAAGTGGTACTCTCCCAGAGACTGTCTGACTCACGCCACCC CCTCCACCTGGACACAGGACGCTGTGGTTTCTGAGCCAGGTACAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGT ACACTGCCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGGCGACTCAGATCCCAGCCAGTGGACTTAGCCCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATCCGGACTCTAAGGTAAATATAAAATTTT TAAGTGTATAATGTGTTAAACTACTGATTCTAATTGTTTCTCTCTTTTAGATTCCAACCTTTGGAACTGA

SEQ ID NO:5 LP1 Promoter CCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTTTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGGAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGC GTCCGGGCAGCGTAGGCGGGCGACTC AGATCCCAGCCAGTGGACTTAGCCCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATCCGGACTCTAAGGTAAATATAAAATTTTTAAGTGTATAATGTGTTAAACTACTGATTCTAATTGTTTCTCTC TTTTAGATTCCAACCTTTGGAACTGA

SEQ ID NO:6 EF1-α promoter GGCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAAGTTGGGGGGAGGGGTCGGCAATTGAACCGGTGCCTAGAGAAGGTGGCGCGGGGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAACGGGTTTGCCGCCAGAACACAGG TAAGTGCCGTGTGTGGTTCCCGCGGGCCTGGCCTCTTTACGGGTTATGGCCCTTGCGTGCCT TGAATTACTTCCACCTGGCTGCAGTACGTGATTCTTGATCCCGAGCTTCGGGTTGGAAGTGGGTGGGAGAGTTCGAGGCCTTGCGCTTAAGGAGCCCCTTCGCCTCGTGCTTGAGTTGAGGC CTGGCCTGGGCGCTGGCCGCCGCGTGCGAATCTGGTGGCACCTTCGCGCCTGTCTCGCTGCTTTCGATAAGTCTCTAGCCATTTAAAATTTTTGATGACCTGCTGCGACGCTTTTTTTCT GGCAAGATAGTCTTGTAAATGCGGGCCAAGATCTGCACACTGGTATTTCGG TTTTTGGGGCCGCGGGCGGCGACGGGGCCCGTGCGTCCCAGCGCACATGTTCGGCGAGGCGGGGCCTGCGAGCGCGGCCACCGAGAATCGGACGGGGGTAGTCTCAAGCTGGCCGGCCTGCTCTGGTGCCTGGTCTCGCGCCGCCGTGTATCGCCCCGCCCTGGGCGGCAAGGCTGGCCCGGTCGGCACCAGTTGCGTGAGCGGAAAGATGGCCGCTTCCCGGCCCTGCTGCAGGGA GCTCAAAATGGAGGACGCGGCGCTCGGGAGAGCGGGCGGGTGAGTCACCCACACAAAG GAAAAGGGCCTTTCCGTCCTCAGCCGTCGCTTCATGTGACTCCACGGAGTACCGGGCGCCGTCCAGGCACCTCGATTAGTTCTCGAGCTTTTGGAGTACGTCGTCTTTAGGTTGGGGGGAGGGGTTTTATGCGATGGAGTTTCCCCACACTGAGTGGGTGGAGACTGAAGTTAGGCCAGCTTGGCACTTGATGTAATTCTCCTTGGAATTTGCCCTTTTTGAGTTTGGATCTTGGTTCATTC TCAAGCCTCAGACAGTGGTTCAAAGTTTTTTTCTTCCATTTCAGGTGTCGTGA SEQ ID NO:7 R3/R4 R3(PmeI) GTTTTAAAC; R4 (PacI) TTAATTAA

SEQ ID NO:8 WPR GAGCATCTTACCGCCATTTATTCCCATATTTGTTCTGTTTTTCTTGATTTGGGTATACATTTAAATGTTAATAAAACAAAATGGTGGGGCAATCATTTACATTTTTAGGGATATGTAATTACTAGTTCAGGTGTATTGCCACAAGACAAACATGTTAAGAAACTTTCCCGTTATTTACGCTCTGTTCCTGTTAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGATATTCTTAACTATGTTGCTC CTTTTACGCTGTGTGGATATGCTGCTTTATAGCCTCT GTATCTAGCTATTGCTTCCCGTACGGCTTTCGTTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTTAGAGGAGTTGTGGCCCGTTGTCCGTCAACGTGGCGTGGTGTGCTCTGTGTTTGCTGACGCAACCCCCACTGGCTGGGGCATTGCCACCACCTGTCAACTCCTTTCTGGGACTTTCGCTTTCCCCCTCCCGATCGCCACGGCAGAACTCATCGCCGCCTGCCTTGCCCG CTGCTGGACAGGGGCTAGGTTGCTGGGCACTGATAATTCCGTGGTGTTGTC SEQ ID NO:9 BGHpA TGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCGGTGGGCTCTATGGC SEQ ID NO:10 Modified SV40-pA TAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTTAACAACAACAATTGCATTCATTTTATGTTTCAGGTTCAGGGGGAGGTGTGGGAGGTTTTTTAAAGCAAGTAAAACCTCTACAAATGTGGTA

SEQ ID NO: 11 wtFIX
Note: The sequence was subsequently codon optimized by GenScript.

SEQ ID NO:12 PaduaFIX
Note: The sequence was subsequently codon optimized by GenScript.

SEQ ID NO:13 Rep 78 CGCAGCCACC ATGGCGGGGTTTTACGAGATTGTGATTAAGGTCCCCAGCGACCTTGACGGGCATCTGCCCGGCATTTCTGACAGCTTTGTGAACTGGGTGGCCGAGAAGGAATGGGAGTTGCCGCCAGATTCTGACATGGATCTGAATCTGATTGAGCAGGCACCCCTGACCGTGGCCGAGAAGCTGCAGCGCGACTTTCTGACGGAATGGCGCCGTGTGAGTAAGGCCCCGGAGGCCC TTTT CTTTGTGCAATTTGAGAAGGGAGAGAGCTACTTCCACATGCACGTGCTCGTGGAAACCACCGGGGTGAAATCCATGGTTTTGGGACGTTTCCTGAGTCAGATTCGCGAAAAACTGATTCAGAGAATTTACCGCGGGATCGAGCCGACTTTGCCAAACTGGTTCGCGGTCACAAAGACCAGAAATGGCGCGGAGGCGGGAACAAGGTGGTGGATGAGTGCTACATCCCCAATT ACTTGCTCCCCAAAACCCAGCCTGAGCTCCAGTGGGCGTGGACTAATATGGAACAGTATTTAAGCGCCTGTTTGAATCTCACGGAGCGTAAACGGTTGGTGGCGCAGCATCTGACGCACGTGTCGCAGACGCAGGAGCAGAACAAAGAGAATCAGAATCCCAATTCTGATGCGCCGGTGATCAGATCAAAAACTTCAGCCAGGTACATGGAGCTGGTCGGGTGGCTCGTGGAC AAGGGGATTACCTCGGAGAAGCAGTGGATCCAGGAGGACCAGGCCTCATACATCTCCTTCAATGCGGCCTCCAACTCGCGGTCCCAAATCAAGGCTGCCTTGGACAATGCGGGAAAGATTATGAGCCTGACTAAAACCGCCCCCGACTACCTGGTGGGCCAGCAGCCCGTGGAGGACATTTCCAGCAATCGGATTTATAAAATTTTGGAACTAAACGGGTACGATCCCCAATAT GCGGCTTCCGTCTTTCTGGGATGGGCCACGAAAAAGTTCGGCAAGAGGAACACCATCTGGCTGTTTGGGCCTGCAACTACCGGGAAGACCAACATCGCGGAGGCCATAGCCCACACTGTGCCCTTCTACGGGTGCGTAAACTGGACCAATGAGAACTTTCCCTTCAACGACTGTGTCGACAAGATGGTGATCTGGTGGGAGGAGGGGAAGATGACCGCCAAGGTCGTGGAGTC GGCCAAAGCCATTCTCGGAGGAAGCAAGGTGCGCGTGGACCAGAAATGCAAGTCCTCGGCCCAGATAGACCCGACTCCCGTGATCGTCACCTCCAACACCAACATGTGCGCCGTGATTGACGGGAACTCAACGACCTTCGAACACCAGCAGCCGTTGCAAGACCGGATGTTCAAATTTGAACTCACCCGCCGTCTGGATCATGACTTTGGGAAGGTCACCAAGCAGGAAGTCA AAGACTTTTTCCGGTGGGCAAAGGATCACGTGGTTGAGGTGGAGCATGAATTCTACGTCAAAAAGGGTGGAGCCAAGAAAAGACCCGCCCCCAGTGACGCAGATATAAGTGAGCCCAAACGGGTGCGCGAGTCAGTTGCGCAGCCATCGACGTCAGACGCGGAAGCTTCGATCAACTACGCAGACAGGTACCAAAACAAATGTTCTCGTCACGTGGGCATGAATCTGATGCTG TTTCCCTGCAGACAATGCGAGAGAATGAATCAGAATTCAAATATCTGCTTCACTCACGGACAGAAAGACTGTTTAGAGTGCTTTCCCGTGTCAGAATCTCAACCCGTTTCTGTCGTCAAAAGGCGTATCAGAAACTGTGCTACATTCATCATATCATGGGAAAGGTGCCAGACGCTTGCACTGCCTGCGATCTGGTCAATGTGGATTTGGATGACTGCATCTTTGAACAATAA

SEQ ID NO:14 Rep 52 ATGGAGCTGGTCGGGTGGCTCGTGGACAAGGGGATTACCTCGGAGAAGCAGTGGATCCAGGAGGACCAGCCTCATACATCTCCTTCAATGCGGCCTCCAACTCCGGGTCCCAAATCAAGGCTGCCTTGGACAATGCGGGAAAGATTATGAGCCTGACTAAAACCGCCCCCGACTACCTGGTGGGCCAGCAGCCCGTGGAGGACATTTCCAGCAATCGGATTTATAAAATTTTGGAACTAAACGGGTA CGATCCCCAATATGCGGCTTCCGTCTTTCTGGGATGGGGCCACGAAAAAGT TCGGCAAGAGGAACACCATCTGGCTGTTTGGGCCTGCAACTACCGGGAAGACCAACATCGCGGAGGCCATAGCCCACACTGTGCCCTTCTACGGGTGCGTAAACTGGACCAATGAGAACTTTCCCTTCAACGACTGTGTCGACAAGATGGTGATCTGGTGGGAGGAGGGGAAGATGACCGCCAAGGTCGTGGAGTCGGCCAAAGCCATTCTCGGAGGAAGCAAGGTGCGCGTGGACCAGAAATGCA AGTCCTCGGCCCAGATAGACCCGACTCCCGTGATCGTCACCTCCAACACCAAC ATGTGCGCCGTGATTGACGGGAACTCAACGACCTTCGAACACCAGCAGCCGTTGCAAGACCGGATGTTCAAATTTGAACTCACCCGCCGTCTGGATCATGACTTTGGGAAGGTCACCAAGCAGGAAGTCAAAGACTTTTTCCGGTGGGCAAAGGATCACGTGGTTGAGGTGGAGCATGAATTCTACGTCAAAAAGGTGGAGCCAAGAAAAGACCCGCCCCCAGTGACGCAGATATAAGT GAGCCCAAACGGGTGCGCGAGTCAGTTGCGCAGCCATCGACGTCAGACGCGGAAGCTT CGATCAACTACGCAGACCGCTACCAAAACAAATGTTCTCGTCACGTGGGCATGAATCTGATGCTGTTTCCCTGCAGACAATGCGAGAGAATGAATCAGAATTCAAATATCTGCTTCACTCACGGACAGAAAGACTGTTTAGAGTGCTTTCCCGTGTCAGAATCTCAACCCGTTTCTGTCGTCAAAAGGCGTATCAGAAACTGTGCTACATTCATCATATCATGGGAAAGGTGCCAGACGCTTGCACT GCCTGCGATCTGGTCAATGTGGATTTGGATGACTGCATCTTTGAACAATAA

SEQ ID NO:15 IE1 promoter fragment AATAAACGATAACGCCGTTGGTGGCGTGAGGCATGTAAAAGGTTACATCATTATCTTGTTCGCCATCCGGTTGGTATAAATAGACGTTCATGTTGGTTTTTGTTTCAGTTGCAAGTTGGCTGCGGCGCGCGCAGCACCTTT
SEQ ID NO:16 LP-1 β promoter CCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGGAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGC GTCCGGGCAGCGTAGGCGGGCGACTCAGATCCC AGCCAGTGGACTTAGCCCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACACTCGAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATCCGGACATCGATTCTAAGGTAAATATAAAATTTTTAAGTGTATAATTTGTTAAACTACTGATTCTAATTGTT TCTCTCTTTTAGATTCCAACCTTTGGAACTGA SEQ ID NO:17 Selected portion of SEQ ID NO: 2 containing RBE GCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO:18 RNA polymerase III promoter for human U6 snRNA
(human small nuclear RNA U6 promoter) GAGGGCCTATTTCCCATGATTCCTTCATATTTGCATATACGATACAAGGCTGTTAGAGAGATAATTGGAATTAATTTGACTGTAAACACAAAGATATTAGTACAAAATACGTGACGTAGAAAGTAATAATTTCTTGGGTAGTTTGCAGTTTTAAAATTATGTTTTAAAATGGACTATCATATGCTTACCGTAACTTGAAAGTATTTCGATTTCTTGGCTTTATATATCTTGTGGAAAGGAC

SEQ ID NO:19 human H1 RNA promoter
(Human H1 Promoter) GAACGCTGACGTCATCAACCCGCTCCAAGGAATCGCGGGCCCAGTGTCACTAGGCGGGAACACCCAGCGCGCGTGCGCCCTGGCAGGAAGATGGCTGTGAGGGACAGGGGAGTGGCGCCCTGCAATATTTGCATGTCGCTATGTGTTCTGGGAAATCACCATAAACGTGAAATGTCTTTGGATTTGGGAATCGTATAAGAACTGTATGAGACCAC SEQ ID NO:20 IE2 Promoter ATAAACGATAACGCCGTTGGTGGCGTGAGGCATGTAAAAGGTTACATCATTATCTTGTTCGCCATCCGGTTGGTATAAATAGACGTTCATGTTGGTTTTTGTTTCAGTTGCAAGTTGGCTGCGGCGCGCGCAGCACCTTTGCGGCCATCT SEQ ID NO:21-38 SEQ ID NO:39 Rep-binding site (RBS) for AAB2 GCGGCTCGCTCGCTC SEQ ID NO:40-50 SEQ ID NO:51 WT-ITR AAB2 (left) CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT SEQ ID NO:52 Modified ITR AAB2 (left) CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT

SEQ ID NO:53 Construction A CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGAAACCCGGGCGTGCCCGGGCGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGAACAGAGAAACAGGAGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGCCAAGAACAGTTGGAACAGCAGAATATGGGCCAAACAGGATATCTGTGG TAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCCCGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTCCAGGGTGCCCCAAGGACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAA TCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTCTGCTCCCCGAGCTCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTGGAGACGCCATCCACGCTGTTTTGACTTCCATAGAAGGCCGCCACCATGGAAGACGCCAAAACATAAAGAAAGGCCCGGCGCCATTCTATCCGCTGGAAGATGGAACCGCTGGAGAGCAACTGCATAAGGCTATGAAGAGATACGCCCTGGTTCCTG GAACAATTGCTTTACAGATGCACATATCGAGGTGGACATCACTTACGCTGAGTACTTCGAAATGTCCGTTCGGTTGGCAGAAGCTATGAAACGATATGGGCTGAATACAAATCACAGAATCGTCGTATGCAGTGAAAACTCTCT TCAATTCTTTATGCCGGTGTTGGGCGCGTTATTTATCGGAGTTGCAGTTGCGCCCGCGAACGACATTTATAATGAACGTGAATTGCTCAACAGTATGGGCATTTCGCAGCCTACCGTGGTGTTCGTTTCCAAAAAGGGGTTGCAAAAAATTTTGAACGTGCAAAAAAAGCTCCCAATCATCCAAAAAATTATTATCATGGATTCTAAAACGGATTACCAGGGATTTCAGTCGATGTACACGTTCGTC ACATCTCATCTACCTCCCGGTTTTAATGAATACGATTTTGTGCCAGAGTCCTTCGATAGGGACAAGACAATTGCACTGATCATGAACTCCTCTGGATCTACTGGTCTGCCTAAAGGTGTCGCTCTGCCTCATAGAACTGCCTG CGTGAGATTCTCGCATGCCAGAGATCCTATTTTTGGCAATCAAATCATTCCGGATACTGCGATTTTAAGTGTTGTTCCATTCCATCACGGTTTTGGAATGTTTACTACACTCGGATATTTGATATGTGGATTTCGAGTCGTCTTAATGTATAGATTTGAAGAAGAGCTGTTTCTGAGGAGCCTTCAGGATTACAAGATTCAAAGTGCGCTGCTGGTGCCAACCCTATTCTCCTTCTTC GCCAAAGCACTCTGATTGACAAATACGATTTATCTAATTTACACGAAATTGCTTCTGGTGGCGCTCCCCTCTCTAAGGAAGTCGGGGAAGCGGTTGCCAAGAGGTTCCATCTGCCAGGTATCAGGCAAGGATATGGGCTCACTGAGACTACA TCAGCTATTCTGATTACACCCGAGGGGGATGATAAACCGGGCGCGGTCGGTAAAGTTGTTCCATTTTTTGAAGCGAAGGTTGTGGATCTGGATACCGGGAAAACGCTGGGCGTTAATCAAAGAGGCGAACTGTGTGTGAGAGGTCCTATGATTATGTCCGGTTATGTAAACAATCCGGAAGCGACCAACGCCTTGATTGACAAGGATGGATGGCTACATTCTGGAGACATAGCTTACT GGGACGAAGACGAACACTTCTTCATCGTTGACCGCCTGAAGTCTCTGATTAAGTACAAAGGCTATCAGGTGGCTCCCGCTGAATTGGAATCCATCTTGCTCCAACACCCCAACATCTTCGACGCAGGTGTCGCAGGTCTTCCCGACGATGAC GCCGGTGAACTTCCCGCCGCCGTTGTTGTTTTGGAGCACGGAAAGACGATGACGGAAAAAGAGATCGTGGATTACGTCGCCAGTCAAGTAACAACCGCGAAAAAGTTGCGCGAGGAGTTGTGTTTGTGGACGAAGTACCGAAAGGTCTTACCGGAAAACTCGACGCAAGAAAAATCAGAGAGATCCTCATAAAGGCCAAGAAGGGCGGAAAGATCGCCGTGTAAGAGCATCTTACCGCCAT TTATTCCCATATTTGTTCTGTTTTTCTTGATTTGGGTATACATTTAAATGTTAATAAAACAAAATGGTGGGGCAATCATTTACATTTTTAGGGATATGTAATTACTAGTTCAGGTGTATTGCCACAAGACAAACATGTTAAGAAACTTT CCCGTTATTTACGCTCTGTTCCTGTTAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGATATTCTTAACTATGTTGCTCCTTTTACGCTGTGTGGATATGCTGCTTTATAGCCTCTGTATCTAGCTATTGCTTCCCGTACGGCTTTCGTTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTTAGAGGAGTTGTGGCCCGTTGTCCGTCAACGTGGCGTGGTG TGCTCTGTGTTTGCTGACGCAACCCCCACTGGCTGGGGCATTGCCACCACCTGTCAACTCCTTTCTGGGACTTTCGCTTTCCCCCTCCCGATCGCCACGGCAGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTAGGTTGCTGG GCACTGATAATTCCGTGGTGTTGTCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCGGTGGG CTCTATGGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

SEQ ID NO:54 Design B CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGAACAGAGAAACAGGAGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGTTGGAACAGCAGAATATGGGCC AAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCCCGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTCCAGGGTGCCCCAAGGACCTGAAATGACCCTGTGCCT TATTTGAACTAACCAATCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTCTGCTCCCCGAGCTCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTGGAGACGCCATCCACGCTGTTTTGACTTCCATAGAAGGCCGCCACCATGGAAGACGCCAAAACATAAAGAAAGGCCCGGCGCCATTCTATCCGCTGGAAGATGGAACCGCTGGAGAGCAACTGCATAAGGCTATGAAGAGATA CGCCCTGGTTCCTGGAACAATTGCTTTTACAGATGCACATATCGAGGTGGACATCACTTACGCTGAGTACTTCGAAATGTCCGTTCGGTTGGCAGAAGCTATGAAACGATATGGGCTGAATACAAATCACAGAATCGTCGTAT GCAGTGAAAACTCTCTTCAATTCTTTATTGCCGGTGTTGGGCGCGTTATTTATCGGAGTTGCAGTTGCGCCCGCGAACGACATTTATAATGAACGTGAATTGCTCAACAGTATGGGCATTTCGCAGCCTACCGTGGTGTTCGTTTCCAAAAGGGGTTGCAAAAAATTTTGAACGTGCAAAAAAAGCTCCCAATCATCCAAAAAATTATTATCATGGATTCTAAAACGGATTACCAGGGATTTCAGTCGA TGTACACGTTCGTCACATCTCATCTACCTCCCGGTTTTAATGAATACGATTTTGTGCCAGAGTCCTTCGATAGGGACAAGACAATTGCACTGATCATGAACTCCTCTGGATCTACTGGTCTGCCTAAAGGTGTCGCTCTG CCTCATAGAACTGCCTGCGTGAGATTCTCGCATGCCAGAGATCCTATTTTTGGCAATCAAATCATTCCGGATACTGCGATTTTAAGTGTTGTTCCATTCCATCACGGTTTTGGAATGTTTACTACACTCGGATATTTGATATGTGGATTTCGAGTCGTCTTAATGTATAGATTTGAAGAAGAGCTGTTTCTGAGGAGCCTTCAGGATTACAAGATTCAAAGTGCGCTGCTGGTGCCAACC CTATTCTCCTTCTTCGCCAAAAGCACTCTGATTGACAAATACGATTTATCTAATTTACACGAAATTGCTTCTGGTGGCGCTCCCCTCTCTAAGGAAGTCGGGGAAGCGGTTGCCAAGAGGTTCCATCTGCCAGGTATCAGGCAAGGATAT GGGCTCACTGAGACTACATCAGCTATTCTGATTACACCCGAGGGGGATGATAAACCGGGCGCGGTCGGTAAAGTTGTTCCATTTTTTGAAGCGAAGGTTGTGGATCTGGATACCGGGAAAACGCTGGGCGTTAATCAAAGAGGCGAACTGTGTGTGAGAGGTCCTATGATTATGTCCGGTTATGTAAACAATCCGGAAGCGACCAACGCCTTGATTGACAAGGATGGATGGCTACAT TCTGGAGACATAGCTTACTGGGACGAAGACGAACACTTCTTCATCGTTGACCGCCTGAAGTCTCTGATTAAGTACAAAGGCTATCAGGTGGCTCCCGCTGAATTGGAATCCATCTTGCTCCAACACCCCAACATCTTCGACGCAGGTGTCGC AGGTCTTCCCGACGATGACGCCGGTGAACTTCCCGCCGCCGTTGTTGTTTTGGAGCACGGAAAGACGATGACGGAAAAAGAGATCGTGGATTACGTCGCCAGTCAAGTAACAACCGCGAAAAAGTTGCGCGGAGGGAGTTGTGTTTGTGGACGAAGTACCGAAAGGTCTTACCGGAAAACTCGACGCAAGAAAAATCAGAGAGATCCTCATAAAGGCCAAGAAGGGCGGAAAGATCGCCGTG TAAGAGCATCTTACCGCCATTTATTCCCATATTTGTTCTGTTTTTCTTGATTTGGGTATACATTTAAATGTTAATAAAACAAAATGGTGGGGCAATCATTTACATTTTTAGGGATATGTAATTACTAGTTCAGGTGTATTGCCAAAGA CAAACATGTTAAGAAACTTTCCCGTTATTTACGCTCTGTTCCTGTTAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGATATTCTTAACTATGTTGCTCCTTTTACGCTGTGTGGATATGCTGCTTTATAGCCTCTGTATCTAGCTATTGCTTCCCGTACGGCTTTCGTTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTTAGAGGAGTTGTGGCCCGTTGTC CGTCAACGTGGCGTGGTGTGCTCTGTGTTTGCTGACGCAACCCCCACTGGCTGGGGCATTGCCACCACCTGTCAACTCCTTTCTGGGACTTTCGCTTTCCCCCTCCCGATCGCCACGGCAGAACTCATCGCCGCCTGCCTTGCCCGCTGCTG GACAGGGGCTAGGTTGCTGGGCACTGATAATTCCGTGGTGTTGTCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAACAATAGCAGG CATGCTGGGGATGCGGTGGGCTCTATGGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGAAACCCGGGCGTGCGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

SEQ ID NO:55 Construction C CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGAAACCCGGGCGTGCCCGGGCGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGAACAGAGAAACAGGAGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGCCAAGAACAGTTGGAACAGCAGAATATGGGCCAAACAGGATATCTGTGG TAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCCCGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTCCAGGGTGCCCCAAGGAC CTGAAATGACCCTGTGCCTTATTTGAACTAACCAATCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTCTGCTCCCCGAGCTCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTGGAGACGCCATCCACGCTGTTTTGACTTCCATAGAAGGCCGCCACCATGATCATGGCCGAGAGCCCTGGCCTGATCACCATCTGCCTGCTGGGCTACCTGCTGAGCGCCGAGTGCACCG TGTTCCTGGACCACGAGAACGCCAACAAGATCCTGAACCGGGCCCAAGAGATACAACAGCGGCAAGCTGGAGGAGTTCGTGCAGGGCAACCTGGAGAGGGAGTGCATGGAGGAGA AGTGCAGCTTCGAGGAGGCCAGGGAAGTGTTCGAGAACACCGAGCGGACCACCGAGTTCTGGAAGCAGTACGTGGACGGCGACCAGTGCGAGAGCAACCCTTGCCTGAACGGCGGCAGCTGCAAGGACGACATCAACAGCTACGAGTGCTGGTGCCCTTTCGGCTTCGAGGGCAAACTGCGAGCTGGACGTGACCTGCAACATCAAGAACGGCCGCTGCGAGCAGTTCTGCAAGAACAG CGCCGACAACAAAGTGGTGTGTAGCTGCACCGAGGGCTACAGACTGGCCGAGAACCAGAAGAGCTGCGAGCCCGCCGTGCCCTTCCCCTGCGGCAGAGTGAGCGTGTCCCAGAC CAGCAAGCTGACCAGAGCCGAGACCGTGTTCCCCGACGTGGACTACGTGAATAGCACCGAGGCCGAGACCATCCTGGACAACATCACCCAGAGCACCCAGTCCTTCAACGACTTCACCAGAGTTGTGGGCGGCGAGGACGCCAAGCCCGGCCAGTTCCCCCTGGCAGGTGGTGCTGAACGGCAAAGTGGATGCCTTCTGCGGCGGCAGCATCGTGAACGAGAAGTGGATCGTGACAGCC GCCCACTGCGTGGAGACCGGCGTGAAGATCACCGTGGTGGCCGGCGAACACAATATCGAGGAGACCGAGCACACCGAGCAGAAGCGGAACGTCATCCGGATTATCCCCCACCACAAC TACAACGCCGCCATCAACAAGTACAACCACGACATCGCCCTGCTGGAGCTGGACGAGCCTCTGGTGCTGAATAGCTACGTGACCCCCATCTGCATCGCCGACAAGGAGTACACCAACATCTTCCTGAAGTTCGGCAGCGGCTACGTGTCCGGCTGGGGCAGAGTGTTCCACAAGGGCAGAAGCGCCCTGGTGCTGCAGTACCTGAGAGTGCCCCTGGTGGACAGAGCCACCTGCCTGTTGAGC ACCAAGTTCACCATCTACAAACAACATGTTCTGCGCCGGCTTCCACGAGGGCGGCAGAGACAGCTGCCAGGGCGACAGCGGCGGACCCCACGTGACCGAAGTGGAGGGCACCA GCTTCCTGACCGGCATCATCAGCTGGGGCGAGGAGTGCGCCATGAAGGGCAAGTACGGCATCTACACCAAAGTGAGCCGGTACGTGAACTGGATCAAGGAGAAAACCAAGCTGACCTGAGAGCATCTTACCGCCATTTATTCCCATATTTGTTCTGTTTTTCTTGATTTGGGTATACATTTAAATGTTAATAAAACAAAATGGTGGGGCAATCATTTACATTTTTAGGGATATGTAATTACTAGTTCAGG TGTATTGCCACAAGACAAACATGTTAAGAAACTTTCCCGTTATTTACGCTCTGTTCCTGTTAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGATAT TCTTAACTATGTTGCTCCTTTTACGCTGTGTGGATATGCTGCTTTATAGCCTCTGTATCTAGCTATTGCTTCCCGTACGGCTTTCGTTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTTAGAGGAGTTGTGGCCCGTTGTCCGTCAACGTGGCGTGGTGTGCTCTGTGTTTGCTGACGCAACCCCCACTGGCTGGGGCATTGCCACCACCTGTCAACTCCTTTCTGGG ACTTTCGCTTTCCCCCTCCCGATCGCCACGGCAGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTAGGTTGCTGGGCACTGATAATTCCGTGGTGTTGTCTGTGCCTTCT AGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCGGTGGGCTCTATGGCAGGAACCCCTAGTGATGGAGTTGGCCACT CCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

SEQ ID NO:56 Construction D CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGAACAGAGAAACAGGAGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGTTGGAACAGCAGAATATGGG CCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCCCGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTCC AGGGTGCCCCCAAGGACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAATCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTCTGCTCCCCGAGCTCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTGGAGACGCCATCCACGCTGTTTTGACTTCCATAGAAGGCCGCCACCATGATCATGGCCGAGAGCCCTGGCCTGATCACCATCTGCCTGCTGGGCTACCTGCT GAGCGCCGAGTGCACCGTGTTCCTGGACCACGAGAACGCCAACAAGATCCTGAACCGGCCCAAGAGATACAACAGCGGCAAGCTGGAGGAGTTCGTGCAGGGCAACCTGGAGAG GGAGTGCATGGAGGAGAAGTGCAGCTTCGAGGAGGCCAGGGAAGTGTTCGAGAACACCGAGCGGACCACCGAGTTCTGGAAGCAGTACGTGGACGGCGACCAGTGCGAGAGCAACCCTTGCCTGAACGGCGGCAGCTGCAAGGACGACATCAACAGCTACGAGTGCTGGTGCCCTTTCGGCTTCGAGGGCAAACTGCGAGCTGGACGTGACCTGCAACATCAAGAACGGCCGCTGCGAGCAG TTCTGCAAGAACAGCGCCGACAACAAAGTGGTGTGTAGCTGCACCGAGGGCTACAGACTGGCCGAGAACCAGAAGAGCTGCGAGCCCGCCGTGCCCTTCCCCTGCGGCAG AGTGAGCGTGTCCCAGACCAGCAAGCTGACCAGAGCCGAGACCGTGTTCCCCGACGTGGACTACGTGAATAGCACCGAGGCCGAGACCATCCTGGACAACATCACCCAGAGCACCCAGTCCTTCAACGACTTCACCAGAGTTGTGGGCGGCGAGGACGCCAAGCCCGGCCAGTTCCCCCTGGCAGGTGGTGCTGAACGGCAAAGTGGATGCCTTCTGCGGCGGCAGCATCGTGAACGA GAAGTGGATCGTGACAGCCGCCCACTGCGTGGAGACCGGCGTGAAGATCACCGTGGTGGCCGGCGAACACAATATCGAGGAGACCGAGCACACCGAGCAGAAGCGGAACGTCATCCGG ATTATCCCCCACCACAACTACAACGCCGCCATCAACAAGTACAACCACGACATCGCCCTGCTGGAGCTGGACGAGCCTCTGGTGCTGAATAGCTACGTGACCCCCATCTGCATCGCCGACAAGGAGTACACCAACATCTTCCTGAAGTTCGGCAGCGGCTACGTGTCCGGCTGGGGCAGAGTGTTCCACAAGGGCAGAAGCGCCCTGGTGCTGCAGTACCTGAGAGTGCCCCTGGTGGACAGA GCCACCTGCCTGTTGAGCACCAAGTTCACCATCTACAAACAACATGTTCTGCGCCGGCTTCCACGAGGGCGGCAGAGACAGCTGCCAGGGCGACAGCGGCGGACCCCACGTG ACCGAAGTGGAGGGCACCAGCTTCCTGACCGGCATCATCAGCTGGGGCGAGGAGTGCGCCATGAAGGGCAAGTACGGCATCTACACCAAAGTGAGCCGGTACGTGAACTGGATCAAGGAGAAAACCAAGCTGACCTGAGAGCATCTTACCGCCATTTATTCCCATATTTGTTCTGTTTTTCTTGATTTGGGTATACATTTAAATGTTAATAAAACAAAATGGTGGGGCAATCATTTACATTTTTAGGGATA TGTAATTACTAGTTCAGGTGTATTGCCACAAGACAAACATGTTAAGAAACTTTCCCGTTATTTACGCTCTGTTCCTGTTAATCAACCTCTGGATTACAAAATTT GTGAAAGATTGACTGATATTCTTAACTATGTTGCTCCTTTTACGCTGTGTGGATATGCTGCTTTATAGCCTCTGTATCTAGCTATTGCTTCCCGTACGGCTTTCGTTTTCTCCTCCTTGTATAAAATCCTGGTTGCTGTCTCTTTTAGAGGAGTTGTGGCCCGTTGTCCGTCAACGTGGCGTGGTGTGCTCTGTGTTTGCTGACGCAACCCCCACTGGCTGGGGCATTGCCACCACCTG TCAACTCCTTTCTGGGACTTTCGCTTTCCCCCTCCCGATCGCCACGGCAGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTAGGTTGCTGGGCACTGATAATTCCG TGGTGTTGTCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCGGTGGGCTCTATGGCAGGAAC CCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGAAACCCGGGCGTGCGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

SEQ ID NO:57 Construction E CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGAAACCCGGGCGTGCCCGGGCGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGGCTCAGAGGCTCAGAGGCACACAGGAGTTTCTGGGCTCACCCTGCCCCCTTCCAACCCCTCAGTTCCCATCCTCCAGCAGCTGTTTGTGTGCTGCCTCTGAAGTCCACACTGAACAA ACTTCAGCCTACTCATGTCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGTCCGGGTTCAAAACCACTTGCTGGGTGGGGAGTCG TCAGTAAGTGGCTATGCCCCGACCCCGAAGCCTGTTTCCCCATCTGTACAATGGAAATGATAAAGACGCCCATCTGATAGGGTTTTTGTGGCAAATAAACATTTGGTTTTTTTGTTTTGTTTTGTTTTGTTTTTTGAGATGGAGGTTTGCTCTGTCGCCCAGGCTGGAGTGCAGTGACACAATCTCATCTCACCAACCTTCCCCTGCCTCAGCCTCCCAAGTAGCTGGGATTACAAG CATGT GCCACCACACCTGGCTAATTTTCTATTTTTAGTAGAGACGGGTTTCTCCATGTTGGTCAGCCTCAGCCTCCCAAGTAACTGGGATTACAGGCCTGTGCCACCACACCCGGCTAATTTTTTCTATTTTTGACAGGGACGGGGTTTCACCATGTTGGTCAGGCTGGTCTAGAGGTACCGGATCTTGCTACCAGTGGAACAGCCACTAAGGATTCTGCAGTGAGAGCAGAGGGCCAGCTAAG TGGTAC TCTCCCAGAGACTGTCTGACTCACGCCACCCCCTCCACCTTGGACACAGGACGCTGTGGTTTCTGAGCCAGGTACAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGGCGACTCAGATCCCAGCCAGTGGACTTAGCCCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCG TTGCCC CTCTGGATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGCCGCCACCATGGAAGACGCCAAAACATAAAGAAAGGCCCGGCGCCATTCTATCCGCTGGAAGATGGAACCGCTGGAGAGCAACTGCATAAGGCTATGAAGAGATACGCCCTGGTTCCTGGAACAATTGCTTTTACAGATGCACATATCGAGGTGGACATCAC TTACGCTGAGTACTTCGAAATGTCCGTTCGGTTGGCAGAAGCTATGAAACGATATGGGCTGAATACAAAATCACAGAATCGTCGTATGCAGTGAAAACTCTCTTCAATTCTTATGCCGGTGTTGGGCGCGTTATTTATCGGAGTTGCAGTTGCGCCCGCGAACGACATTTATAATGAACGTGAATTGCTCAACAGTATGGGCATTTCGCAGCCTACCGTGGTGTTCGTTTCCAAAAGG GGTTG CAAAAAATTTTGAACGTGCAAAAAAAGCTCCCAATCATCCAAAAAATTATTATCATGGATTCTAAAACGGATTACCAGGGATTTCAGTCGATGTACACGTTCGTCACATCTCATCTACCTCCCGGTTTTTAATGAATACGATTTTGTGCCAGAGTCCTTCGATAGGGACAAGACAATTGCACTGATCATGAACTCCTCTGGATCTACTGGTCTGCCTAAAGGTGTCGCTCTGCCTCATAGAACTGC CTGCGTGAGATTCTCGCATGCCAGAGATCCTATTTTTGGCAATCAAATCATTCCGGATACTGCGATTTTAAGTGTTGTTCCATTCCATCACGGTTTTGGAATGTTTACTACACTCGGATATTTGATATGTGGATTTCGAGTCGTCTTAATGTATAGATTTGAAGAAGAGCTGTTTCTGAGGAGCCTTCAGGATTACAAGATTCAAAGTGCGCTGCTGGTGCCAACCCTATTCTCCTTC TTCGCC AAAAGCACTCTGATTGACAAATACGATTTATCTAATTTACACGAAATTGCTTCTGGTGGCGCTCCCCTCTCTAAGGAAGTCGGGGAAGCGGTTGCCAAGAGGTTCCATCTGCCAGGTATCAGGCAAGGATATGGGCTCACTGAGACTACATCAGCTATTCTGATTACACCCGAGGGGGATGATAAACCGGGCGCGGTCGGTAAAGTTGTTCCATTTTTTGAAGCGAAGGTTGTGGATCTGG ATAC CGGGAAAACGCTGGGCGTTAATCAAAGAGGCGAACTGTGTGTGAGAGGTCCTATGATTATGTCCGGTTATGTAAACAATCCGGAAGCGACCAACGCCTTGATTGACAAGGATGGATGGCTACATTCTGGAGACATAGCTTACTGGGACGAAGACGAACACTTCTTCATCGTTGACCGCCTGAAGTCTCTGATTAAGTACAAAGGCTATCAGGTGGCTCCCGCTGAATTGGAATCCATCTTGCTC CAACACCCCAACATCTTCGACGCAGGTGTCGCAGGTCTTCCCGACGATGACGCCGGTGAACTTCCCGCCGCCGTTGTTGTTTTGGAGCACGGAAAGACGATGACGGAAAAAGAGATCGTGGATTACGTCGCCAGTCAAGTAACAACCGCGAAAAAGTTGCGCGGAGGAGTTGTGTTTGTGGACGAAGTACCGAAAGGTCTTACCGGAAAACTCGACGCAAGAAAAATCAGAGAGATCCT CATAAA GGCCAAGAAGGGCGGAAAGATCGCCGTGTAAGAGCATCTTACCGCCATTTATTCCCATATTTGTTCTGTTTTTCTTGATTTGGGTATACATTTAAATGTTAATAAAACAAAATGGTGGGGCAATCATTTACATTTTTAGGGATATGTAATTACTAGTTCAGGTGTATTGCCACAAGACAAACATGTTAAGAAACTTTCCCGTTATTTACGCTCTGTTCCTGTTAATCAACCTCTGGATTACAAA ATTTGTGAAAGATTGACTGATATTCTTAACTATGTTGCTCCTTTTACGCTGTGTGGATATGCTGCTTTATAGCCTCTGTATCTAGCTATTGCTTCCCGTACGGCTTTCGTTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTTAGAGGAGTTGTGGCCCGTTGTCCGTCAACGTGGCGTGGTGTGCTCTGTGTTTGCTGACGCAACCCCCACTGGCTGGGGCATTGCCACCA CCTGTCA ACTCCTTTCTGGGACTTTCGCTTTCCCCCTCCCGATCGCCACGGCAGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTAGGTTGCTGGGCACTGATAATTCCGTGGTGTTGTCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTG AGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCGGTGGGCTCTATGGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

SEQ ID NO:58 Construction F CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGGCTCAGAGGCTCAGAGGCACACAGGAGTTTCTGGGCTCACCCTGCCCCCTTCCAACCCCTCAGTTCCCATCCTCCAGCAGCTGTTTGTGTGCTGCCTCT GAA GTCCACACTGAACAAACTTCAGCCTACTCATGTCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGTCCGGGTTCAAAACCACTT GCTGGGTGGGGAGTCGTCAGTAAGTGGCTATGCCCCGACCCCGAAGCCTGTTTCCCCATCTGTACAATGGAAATGATAAAGACGCCCATCTGATAGGGTTTTTGTGGCAAATAAACATTTGGTTTTTTTGTTTTGTTTTGTTTTGTTTTTTGAGATGGAGGTTTGCTCTGTCGCCCAGGCTGGAGTGCAGTGACACAATCTCATCTCACCAACACCTTCCCCTGCCTCAGCCTCCCA AGTAGCT GGGATTACAAGCATGTGCCACCACACCTGGCTAATTTTCTATTTTTAGTAGAGACGGGTTTCTCCATGTTGGTCAGCCTCAGCCTCCCAAGTAACTGGGATTACAGGCCTGTGCCACCACACCCGGCTAATTTTTTCTATTTTTGACAGGGACGGGGTTTCACCATGTTGGTCAGGCTGGTCTAGAGGTACCGGATCTTGCTACCAGTGGAACAGCCACTAAGGATTCTGCAGTGAGAG CAGAG GGCCAGCTAAGTGGTACTCTCCCAGAGACTGTCTGACTCACGCCACCCCCTCCACCTTGGACACAGGACGCTGTGGTTTCTGAGCCAGGTACAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGGCGACTCAGATCCCAGCCAGTGGACTTAGCCCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCA CCAGC AGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGCCGCCACCATGGAAGACGCCAAAACATAAAGAAAGGCCCGGCGCCATTCTATCCGCTGGAAGATGGAACCGCTGGAGAGCAACTGCATAAGGCTATGAAGAGATACGCCCTGGTTCCTGGAACAATTGCTTTTACAGATGCACA TATCGAGGTGGACATCACTTACGCTGAGTACTTCGAAATGTCCGTTCGGTTGGCAGAAGCTATGAAACGATATGGGCTGAATACAAAATCACAGAATCGTCGTATGCAGTGAAAACTCTCTTCAATTCTTATGCCGGTGTTGGGCGCGTTATTTATCGGAGTTGCAGTTGCGCCCGCGAACGACATTTATAATGAACGTGAATTGCTCAACAGTATGGGCATTTCGCAGCCTACCGTGGTG TTC GTTTCCAAAAAGGGGTTGCAAAAAATTTTGAACGTGCAAAAAAAGCTCCCAATCATCCAAAAAATTATTATCATGGATTCTAAAACGGATTACCAGGGATTTCAGTCGATGTACACGTTCGTCACATCTCATCTACCTCCCGGTTTTTAATGAATACGATTTTGTGCCAGAGTCCTTCGATAGGGACAAGACAATTGCACTGATCATGAACTCCTCTGGATCTACTGGTCTGCCTAAAGGTGTCGC TCTGCCTCATAGAACTGCCTGCGTGAGATTCTCGCATGCCAGAGATCCTATTTTTGGCAATCAAATCATTCCGGATACTGCGATTTTAAGTGTTGTTCCATTCCATCACGGTTTTGGAATGTTTACTACACTCGGATATTTGATATGTGGATTTCGAGTCGTCTTAATGTATAGATTTGAAGAAGAGCTGTTTCTGAGGAGCCTTCAGGATTACAAGATTCAAAGTGCGCTGCTGGTG CCAACC CTATTCTCCTTCTTCGCCAAAAGCACTCTGATTGACAAATACGATTTATCTAATTTACACGAAATTGCTTCTGGTGGCGCTCCCCTCTCTAAGGAAGTCGGGGAAGCGGTTGCCAAGAGGTTCCATCTGCCAGGTATCAGGCAAGGATATGGGCTCACTGAGACTACATCAGCTATTCTGATTACACCCGAGGGGGATGATAAACCGGGCGCGGTCGGTAAAGTTGTTCCATTTTTTGAAGCGA AGGTTGTGGATCTGGATACCGGGAAAACGCTGGGCGTTAATCAAAGAGGCGAACTGTGTGTGAGAGGTCCTATGATTATGTCCGGTTATGTAAACAATCCGGAAGCGACCAACGCCTTGATTGACAAGGATGGATGGCTACATTCTGGAGACATAGCTTACTGGGACGAAGACGAACACTTCTTCATCGTTGACCGCCTGAAGTCTCTGATTAAGTACAAAGGCTATCAGGTGGCTCCCG CTGA ATTGGAATCCATCTTGCTCCAACACCCCAACATCTTCGACGCAGGTGTCGCAGGTCTTCCCGACGATGACGCCGGTGAACTTCCCGCCGCCGTTGTTGTTTTGGAGCACGGAAAGACGATGACGGAAAAAGAGATCGTGGATTACGTCGCCAGTCAAGTAACAACCGCGAAAAAGTTGCGCGGAGGAGTTGTGTTTGTGGACGAAGTACCGAAAGGTCTTACCGGAAAACTCGACGC AAGAAAA ATCAGAGAGATCCTCATAAAGGCCAAGAAGGGCGGAAAGATCGCCGTGTAAGAGCATCTTACCGCCATTTATTCCCATATTTGTTCTGTTTTTCTTGATTTGGGTATACATTTAAATGTTAATAAAACAAAATGGTGGGGCAATCATTTACATTTTTAGGGATATGTAATTACTAGTTCAGGTGTATTGCCACAAGACAAACATGTTAAGAAACTTTCCCGTTATTTACGCTCTGTTCCTGTTA ATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGATATTCTTAACTATGTTGCTCCTTTTACGCTGTGTGGATATGCTGCTTTATAGCCTCTGTATCTAGCTATTGCTTCCCGTACGGCTTTCGTTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTTAGAGGAGTTGTGGCCCGTTGTCCGTCAACGTGGCGTGGTGTGCTCTGTGTTTGCTGACGCAACCCCCACT GGCTG GGGCATTGCCACCACCTGTCAACTCCTTTCTGGGACTTTCGCTTTCCCCCTCCCGATCGCCACGGCAGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTAGGTTGCTGGGCACTGATAATTCCGTGGTGTTGTCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAG GAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCGGTGGGCTCTATGGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGAAACCCGGGCGTGCGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

SEQ ID NO:59 G design CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGAAACCCGGGCGTGCCCGGGCGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGGCTCAGAGGCTCAGAGGCACACAGGAGTTTCTGGGCTCACCCTGCCCCCTTCCAACCCCTCAGTTCCCATCCTCCAGCAGCTGTTTGTGTGCTGCCTCTGAAGTCCACACTGAAC AAACTTCAGCCTACTCATGTCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGG GTCCGGGTTCAAAACCACTTGCTGGGTGGGGAGTCGTCAGTAAGTGGCTATGCCCCGACCCCGAAGCCTGTTTCCCCATCTGTACAATGGAAATGATAAAGACGCCCATCTGATAGGGTTTTTGTGGCAAATAAACATTTGGTTTTTTTGTTTTGTTTTGTTTTGTTTTTTGAGATGGAGGTTTGCTCTGTCGCCCAGGCTGGAGTGCAGTGACACAATCTCATCTC ACCACAACCTTCCCCTGCCTCAGCCTCCCAAGTAGCTGGGATTACAAGCATGTGCCACCACACCTGGCTAATTTTCTATTTTTAGTAGAGACGGGTTTCTCCATGTTGGTCAGCCTCAGCCTCCCAAGTAACTGGGATTACAGGCCTGTGCCACCACACCCGGCTAATTTTTTCTATTTTTGACAGGGACGGGGTTTCACCATGTTGGTCAGGCTGGTCTAGAGGTA CCGGATCTTGCTACCAGTGGAACAGCCACTAAGGATCTGCAGTGAGAGCAGAGGGCCAGCTAAGTGGTACTCTCCCAGAGACTGTCTGACTCACGCCACCCCCTCCACCTTGGACACAGGACGCTGTGGTTTCTGAGCCAGGTACAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGGCGACTCAGATCCCAGCCAG TGGACTTAGCCCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGCCGCCACCATGATCATGGCCGAGAGCCCTGGCCTGATCACCATCTGCCTGCTGGGCT ACCTGCTGAGCGCCGAGTGCACCGTGTTCCTGGACCACGAGAACGCCAACAAGATCCTGAACCGGCCCAAGAGATACAACAGCGGCAAGCTGGAGGAGTTCGTGCAGGGCAACCTGGAGAGGGAGTGCATGGAGGAGAAGTGCAGCTTCGAGGAGGCCAGGGAAGTGTTCGAGAACACCGAGCGGACCACCGAGTTCTGGAAGCAGTACGTGGACGGCGACCAGTGCGAGAGCAACCCTTGC CTGAACGGCGGCAGCTGCAAGGACGACATCAACAGCTACGAGTGCTGGTGCCCTTTCGGCTTCGAGGGCAAGAACTGCGAGCTGGACGTGACCTGCAACATCAAGAACGGCCGCTGCGAGCAGTTCTGCAAGAACAGCGCCGACAACAAAGTGGTGTGTAGCTGCACCGAGGGCTACAGACTGGCCGAGAACCAGAAGAGCTGCGAGCCCGC CGTGCCCTTCCCCTGCGGCAGAGTGAGCGTGTCCCAGACCAGCAAGCTGACCAGAGCCGAGACCGTGTTCCCCGACGTGGACTACGTGAATAGCACCGAGGCCGAGACCATCCTGGACAACATCACCCAGAGCACCCAGTCCTTCAACGACTTCACCAGAGTTGTGGGCGGCGAGGACGCCAAGCCCGGCCAGTTCCCCCTGGCAGGTGGTGCTGAACGGCAAAGTGGATGCCTTCT GCGGCGGCAGCATCGTGAACGAGAAGTGGATCGTGACAGCCGCCCACTGCGTGGAGACCGGCGTGAAGATCACCGTGGTGGCCGGCGAACACAATATCGAGGAGACCGAGCACACCGAGCAGAAGCGGAACGTCATCCGGATTATCCCCCACCACAACTACAACGCCGCCATCAACAAGTACAACCACGACATCGCCCTGCTGGAGCTGGACGAGCC TCTGGTGCTGAATAGCTACGTGACCCCCATCTGCATCGCCGACAAGGAGTACACCAACATCTTCCTGAAGTTCGGCAGCGGCTACGTGTCCGGCTGGGGCAGAGTGTTCCACAAGGGCAGAAGCGCCCTGGTGCTGCAGTACCTGAGAGTGCCCCTGGTGGACAGAGCCACCTGCCTGTTGAGCACCAAGTTCACCATCTACAACAACATGTTCTGCGCCGGCTTCCACGAGGGCGGCAGAGA CAGCTGCCAGGGCGACAGCGGCGGACCCCACGTGACCGAAGTGGAGGGCACCAGCTTCCTGACCGGCATCATCAGCTGGGGCGAGGAGTGCGCCATGAAGGGCAAGTACGGCATCTACACCAAAGTGAGCCGGTACGTGAACTGGATCAAGGAGAAAACCAAGCTGACCTGAGAGCATCTTACCGCCATTTATTCCCATATTTGTTCTGTT TTTCTTGATTTGGGTATACATTTAAATGTTAATAAAACAAAATGGTGGGGCAATCATTTACATTTTTAGGGATATGTAATTACTAGTTCAGGTGTATTGCCACAAGACAAACATGTTAAGAAACTTTCCCGTTATTTACGCTCTGTTCCTGTTAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGATATTCTTAACTATGTTGCTCCTTTTACGCTGTGTGGATATGCTGCTTTATAGCCT CTGTATCTAGCTATTGCTTCCCGTACGGCTTTCGTTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTTAGAGGAGTTGTGGCCCGTTGTCCGTCAACGTGGCGTGGTGTGCTCTGTGTTTGCTGACGCAACCCCCACTGGCTGGGGCATTGCCACCACCTGTCAACTCCTTTCTGGGACTTTCGCTTTCCCCCTCCCGA TCGCCACGGCAGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTAGGTTGCTGGGCACTGATAATTCCGTGGTGTTGTCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATT CTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCGGTGGGCTCTATGGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

SEQ ID NO:60 H design CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGGCTCAGAGGCTCAGAGGCACACAGGAGTTTCTGGGCTCACCCTGCCCCCTTCCAACCCCTCAGTTCCCATCCTCCAGCAGCTGTTTGTGTGCTGCCTCT GAAGTCCACACTGAACAAACTTCAGCCTACTCATGTCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTT AGGTAGTGTGAGAGGGTCCGGGTTCAAAACCACTTGCTGGGTGGGGAGTCGTCAGTAAGTGGCTATGCCCCGACCCCGAAGCCTGTTTCCCCATCTGTACAATGGAAATGATAAAGACGCCCATCTGATAGGGTTTTTGTGGCAAATAAACATTTGGTTTTTTTGTTTTGTTTTGTTTTGTTTTTTGAGATGGAGGTTTGCTCTGTCGCCCAGGCTGGAGTGCAGT GACACAATCTCATCTCACCACAACCTTCCCCTGCCTCAGCCTCCCAAGTAGCTGGGATTACAAGCATGTGCCACCACACCTGGCTAATTTTCTATTTTTAGTAGAGACGGGTTTCTCCATGTTGGTCAGCCTCAGCCTCCCAAGTAACTGGGATTACAGGCCTGTGCCACCACACCCGGCTAATTTTTTCTATTTTTGACAGGGACGGGGTTTCACCATGTTGGTCA GGCTGGTCTAGAGGTACCGGATCTTGCTACCAGTGGAACAGCCACTAAGGATTCTGCAGTGAGAGCAGAGGGCCAGCTAAGTGGTACTCTCCCAGAGACTGTCTGACTCACGCCACCCCCTCCACCTTGGACACAGGACGCTGTGGTTTCTGAGCCAGGTACAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGG CGACTCAGATCCCAGCCAGTGGACTTAGCCCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGCCGCCACCATGATCATGGCCGAGAGCCCTGGCCTGATCAC CATCTGCCTGCTGGGCTACCTGCTGAGCGCCGAGTGCACCGTGTTCCTGGACCACGAGAACGCCAACAAGATCCTGAACCGGCCCAAGAGATACAACAGCGGCAAGCTGGAGGAGTTCGTGCAGGCAACCTGGAGAGGGAGTGCATGGAGGAGAAGTGCAGCTTCGAGGAGGCCAGGGAAGTGTTCGAGAACACCGAGCGGACCACCGAGTTCTGGAAGCAGTACGTGGACGGCGACCAG TGCGAGAGCAACCCTTGCCTGAACGGCGGCAGCTGCAAGGACGACATCAACAGCTACGAGTGCTGGTGCCCTTTCGGCTTCGAGGGCAAGAACTGCGAGCTGGACGTGACCTGCAACATCAAGAACGGCCGCTGCGAGCAGTTCTGCAAGAACAGCGCCGACAACAAAGTGGTGTGTAGCTGCACCGAGGGCTACAGACTGGCCGAGAACCA GAAGAGCTGCGAGCCCGCCGTGCCCTTCCCCTGCGGCAGAGTGAGCGTGTCCCAGACCAGCAAGCTGACCAGAGCCGAGACCGTGTTCCCCGACGTGGACTACGTGAATAGCACCGAGGC CGAGACCATCCTGGACAACATCACCCAGAGCACCCAGTCCTTCAACGACTTCACCAGAGTTGTGGGCGGCGAGGACGCCAAGCCCGGCCAGTTCCCCCTGGCAGGTGGTGCTGAACGGCAAA GTGGATGCCTTCTGCGGCGGCAGCATCGTGAACGAGAAGTGGATCGTGACAGCCGCCCACTGCGTGGAGACCGGCGTGAAGATCACCGTGGTGGCCGGCGAACACAATATCGAGGAGACCGAGCACACCGAGCAGAAGCGGAACGTCATCCGGATTATCCCCCACCACAACTACAACGCCGCCATCAACAAGTACAACCACGACATCGCCCT GCTGGAGCTGGACGAGCCTCTGGTGCTGAATAGCTACGTGACCCCCATCTGCATCGCCGACAAGGAGTACACCAACATCTTCCTGAAGTTCGGCAGCGGCTACGTGTCCGGCTGGGGCAGAGTGTTCCACAAGGGCAGAAGCGCCCTGGTGCTGCAGTACCTGAGAGTGCCCCTGGTGGACAGAGCCACCTGCCTGTTGAGCACCAAGTTCACCATCTTACAACAACATGTTCTGCGCCGGCTT CCACGAGGGCGGCAGAGACAGCTGCCAGGGCGACAGCGGCGGACCCCACGTGACCGAAGTGGAGGGCACCAGCTTCCTGACCGGCATCATCAGCTGGGGCGAGGAGTGCGCCATGAAGGGCAAGTACGGCATCTACACCAAAGTGAGCCGGTACGTGAACTGGATCAAGGAGAAAACCAAGCTGACCTGAGAGCATCTTACCGCCATTTA TTCCCATATTTGTTCTGTTTTTCTTGATTTGGGTATACATTTAAATGTTAATAAAACAAAATGGTGGGGCAATCATTTACATTTTTAGGGATATGTAATTACTAGTTCAGGTGTATTGCCACAAGA CAAACATGTTAAGAAACTTTCCCGTTATTTACGCTCTGTTCCTGTTAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGATATTCTTAACTATGTTGCTCCTTTACGCTGTGTGGATA TGCTGCTTTATAGCCTCTGTATCTAGCTATTGCTTCCCGTACGGCTTTCGTTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTTAGAGGAGTTGTGGCCCGTTGTCCGTCAACGTGGCGTGGTGTGCTCTGTGTTTGCTGACGCAACCCCCACTGGCTGGGGCATTGCCACCACCTGTCAACTCCTTTCTGGGAC TTTCGCTTTCCCCCTCCCGATCGCCACGGCAGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTAGGTTGCTGGGCACTGATAATTCCGTGGTGTTGTCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGT CTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCGGTGGGCTCTATGGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGAAACCCGGGCGTGCGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

SEQ ID NO:61 Construction α CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTTGTAGTTAATGATTAACCCGCCATGCTACTTATCTACGTAGCCATGCTCTAGAGCGGCCGCACGCGTAGATCTTCAATATTGGCCATTAGCCATATTATTCATTG GTTATATAGCATAAATCAAT ATTGGCTATTGGCCATTGCATACGTTGTATCTATATCATAATATGTACATTTATATTGGCTCATGTCCAATATGACCGCCATGTTGGCATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAG TAACGCCAATAGGGAC TTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTCCGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTACGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTCGAGGTGAGCCCCACGTTCTGCTTCACTCTCCCCATCTCC CCCCCCTCCCCACCCCCAATTTTGT ATTTATTTATTTTTTAATTATTTTGTGCAGCGATGGGGGCGGGGGGGGGGGGGGCGCGCGCCAGGCGGGGCGGGGCGGGGCGAGGGGGCGGGGCGGGGCGAGGCGGAGAGGTGCGGCGGCAGCCAATCAGAGCGGCGCGCTCCGAAAGTTTCCTTTTATGGCGAGGGCGGCGGCGGCGGCGGCCCTATAAAAAGCGAAGCGCGCGGCGGGCGGGAGTCGCTGCGACGCTGCCTTCGCCCCG TGCCCCGCTCCGCCGCCGCCTCGCG CCGCCCGCCCCGGCTCTGACTGACCGCGTTACTCCCACAGGTGAGCGGGCGGGACGGCCCTTCTCCTCCGGGCTGTAATTAGCGCTTGGTTTAATGACGGCTTGTTTCTTTTCTGTGGCTGCGTGAAAGCCTTGAGGGGCTCCGGGAGGGCCCTTTGTGCGGGGGGGAGCGGCTCGGGGGGTGCGTGCGTGTGTGTGTGCGTGGGGAGCGCCGCGTGCGGCCCGCGCTGCCCGGCGGCTGTG AGCGCTGCGGGCGCGGCGCGGGGC TTTGTGCGCTCCGCAGTGTGCGCGAGGGGAGCGCGGCCGGGGGCGGTGCCCCGCGGTGCGGGGGGGGCTGCGAGGGGAACAAAGGCTGCGTGCGGGGTGTGTGCGTGGGGGGGTGAGCAGGGGGTGTGGGCGCGGCGGTCGGGCTGTAACCCCCCCCTGCACCCCCCTCCCCGAGTTGCTGAGCACGGCCCGGCTTCGGGTGCGGGGCTCCGTACGGGGCGTGGCGCGGGGCTCGCC GTGCGGGCGGGGGGTGGCGGCAGGTGGG GGTGCCGGGCGGGGCGGGGCCGCCTCGGGCCGGGGAGGGCTCGGGGGAGGGGCGGCGGCCCCCGGAGCGCCGGCGGCTGTCGAGGCGCGGCGAGCCGCAGCCATTGCCTTTTATGGTAATCGTGCGAGAGGGCGCAGGGACTTCCTTTGTCCCAAATCTGTGCGGAGCCGAAATCTGGGAGGCGCCGCCGCACCCCCTCTAGCGGGCGCGGGGCGAAGCGGTGCGGCGCCGGCAGGAAGG AAATGGGCGGGGAGGGCCTTCGTG CGTCGCCGCGCCGCCGTCCCCTTCTCCCTCTCCAGCCTCGGGGCTGTCCGCGGGGGGACGGCTGCCTTCGGGGGGGACGGGGCAGGGCGGGGTTCGGCTTCTGGCGTGTGACCGGCGGCTCTAGAGCCTCTGCTAACCATGTTTTAGCCTTCTTCTTTTTCCTACAGCTCCTGGGCAACGTGCTGGTTATTGTGCTGTCTCATCATTTGTCGACAGAATTCCTCGAAGATCCGAAGGGG TTCAAGCTTGGCATTCCGGTACTGTTG GTAAAGCCACCATGGAAGACGCCAAAACATAAAGAAAGGCCCGGCGCCATTCTATCCGCTGGAAGATGGAACCGCTGGAGAGCAACTGCATAAGGCTATGAAGAGATACGCCCTGGTTCCTGGAACAATTGCTTTTACAGATGCACATATCGAGGTGGACATCACTTACGCTGAGTACTTCGAAATGTCCGTTCGGTTGGCAGAAGCTATGAAACGATATGGGCTGAATACAAATCACAGAATCGTCGTA TGCAGTGAAAACTCT CTTCAATTCTTTATGCCGGTGTTGGGCGCGTTATTTATCGGAGTTGCAGTTGCGCCCGCGAACGACATTTATAATGAACGTGAATTGCTCAACAGTATGGGCATTTCGCAGCCTACCGTGGTGTTCGTTTCCAAAAAGGGGTTGCAAAAAATTTTGAACGTGCAAAAAAAGCTCCCAATCATCCAAAAAATTATTATCATGGATTCTAAAACGGATTACCAGGGATTTCAGTCGATGTACACGTTCGTC ACATCTCATCTACCTCC CGGTTTTTAATGAATACGATTTTGTGCCAGAGTCCTTCGATAGGGACAAGACAATTGCACTGATCATGAACTCCTCTGGATCTACTGGTCTGCCTAAAGGTGTCGCTCTGCCTCATAGAACTGCCTGCGTGAGATTCTCGCATGCCAGAGATCCTATTTTTGGCAATCAAATCATTCCGGATACTGCGATTTTAAGTGTTGTTCCATTCCATCACGGTTTTGGAATGTTTACTACACTCGGATATT TGATATGTGGATTTCGAGTCG TCTTAATGTATAGATTTGAAGAAGAGCTGTTTCTGAGGAGCCTTCAGGATTACAAGATTCAAAGTGCGCTGCTGGTGCCAACCCTATTCTCCTTCTTCGCCAAAAGCACTCTGATTGACAAATACGATTTATCTAATTTACACGAAATTGCTTCTGGTGGCGCTCCCCTCTCTAAGGAAGTCGGGGAAGCGGTTGCCAAGAGGTTCCATCTGCCAGGTATCAGGCAAGGATATGGGCTC ACTGAGACTACATCAGCTATTCTGATT ACACCCGAGGGGGATGATAAACCGGGCGCGGTCGGTAAAGTTGTTCCATTTTTTGAAGCGAAGGTTGTGGATCTGGATACCGGGAAAACGCTGGGCGTTAATCAAAGAGGCGAACTGTGTGTGAGAGGTCCTATGATTATGTCCGGTTATGTAAACAATCCGGAAGCGACCAACGCCTTGATTGACAAGGATGGATGGCTACATTCTGGAGACATAGCTTACTGGGACGAAGACGAAC ACTTCTTCATCGTTGACCGCCTGAAGTC TCTGATTAAGTACAAAGGCTATCAGGTGGCTCCCGCTGAATTGGAATCCATCTTGCTCCAACACCCCAACATCTTCGACGCAGGTGTCGCAGGTCTTCCCGACGATGACGCCGGTGAACT TCCCGCCGCCGTTGTTGTTTTGGAGCACGGAAAGACGATGACGGAAAAAGAGATCGTGGATTACGTCGCCAGTCAAGTAACAACCGCGAAAAAGTTGCGCGGAGGAGTTGTGTTTGTGGA CGAAGTACCGAAAGGTCTTACCGGAA AACTCGACGCAAGAAAAATCAGAGAGATCCTCATAAAGGCCAAGAAGGGCGGAAAGATCGCCGTGTAATTCTAGAGTCGGGGCGGCCGGCCGCTTCGAGCAGACATGATAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTTAACAACAACAATTGCATTCATTT TATGTTTCAGGTT CAGGGGGAGGTGTGGGAGGTTTTTTAAAGCAAGTAAAACCTCTACAAATGTGGTAAAATCGATAAGTGCGGACCGAGCGGCCGCTCTAGAGCATGGCTACGTAGATAAGTAGCATGGCGGGTTAATCATTAACTACACCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTG GCCAACTCCATCACTAGGGGTTCCT

SEQ ID NO:62 Construction β CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTTGTAGTTAATGATTAACCCGCCATGCTACTTATCTACGTAGCCATGCTCTAGAGCGGCCGCGCTAGCCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACA CACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTCTGGGCCCAT GCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGGAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGGCGACTCAGATCCCAGCCAGTGGACTTAGCCCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAG CAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGACGAGGACACTCGAGGGCCCTGTCTCCTCAGC TTCAGGCACCACCACTGACCTGGGACAGTGAATCCGGACATCGATTCTAAGGTAAATATAAAATTTTTAAGTGTATAATTTGTTAAACTACTGATTCTAATTGTTTCTCTCTTTTAGATTCCAACCTTTGGAACTGAATTCTAGACCACCATGCAGAGGGTGAACATGATCATGGCTGAGAGCCCTGGCCTGATCACCATCTGCCTGCTGGGCTACCTGCTGTCTGCTGAGTGCACTGTGTTCCT GGACCATGAGAATGCCAACAAGATCCTGAACAGGCCCAAGAGATACAACTCTGGCAAGCTGGAGGAGTT TGTGCAGGGCAACCTGGAGAGGGAGTGCATGGAGGAGAAGTGCAGCTTTGAGGAGGCCAGGGAGGTGTTTGAGAACACTGAGAGGACCACTGAGTTCTGGAAGCAGTATGTGGATGGGGACCAGTGTGAGAGCAACCCCTGCCTGAATGGGGGCAGCTGCAAGGATGACATCAACAGCTATGAGTGCTGGTGCCCCTTTGGCTTTGAGGGCAAGAACTGTGAGCTGGATGTGACCTGC AACATCAAGAATGGCAGATGTGAGCAGTTCTGCAAGAACTCTGCTGACAACAAGGTGGTGTGCAGCTGCACTGAGG GCTACAGGCTGGCTGAGAACCAGAAGAGCTGTGAGCCTGCTGTGCCATTCCCATGTGGCAGAGTGTCTGTGAGCCAGACCAGCAAGCTGACCAGGGCTGAGGCTGTGTTCCCTGATGTGGACTATGTGAACAGCACTGAGGCTGAAACCATCCTGGACAACATCACCCAGAGCACCCAGAGCTTCAATGACTTCACCAGGGTGGTGGGGGGGGAGGATGCCAAGCCTGGCCAGTTCCC CTGGCAAGTGGTGCTGAATGGCAAGGTGGATGCCTTCTGTGGGGGCAGCATTGTGAATGAGAAGTGGATTGTGAC TGCTGCCCACTGTGTGGAGACTGGGGTGAAGATCACTGTGGTGGCTGGGGAGCACAACATTGAGGAGACTGAGCACACTGAGCAGAAGAGGAATGTGATCAGGATCCCCCACCACAACTACAATGCTGCCATCAACAAGTACAACCATGACATTGCCCTGCTGGAGCTGGATGAGCCCCTGGTGCTGAACAGCTATGTGACCCCCATCTGCATTGCTGACAAGGAGTACACCAACATCTTC CTGAAGTTTGGCTCTGGCTATGTGTCTGGCTGGGGCAGGGTGTTCCACAAGGGCAGGTCTGCCCTGGTGC TGCAGTACCTGAGGGTGCCCCTGGTGGACAGGGCCACCTGCCTGAGGAGCACCAAGTTCACCATCTACAACAACATGTTCTGTGCTGGCTTCCATGAGGGGGGCAGGGACAGCTGCCAGGGGGACTCTGGGGGCCCCCATGTGACTGAGGTGGAGGGCACCAGCTTCCTGACTGGCATCATCAGCTGGGGGGAGGAGTGTGCCATGAAGGGCAAGTATGGCATCTACACCAAAGTCTCCAGATA TGTGAACTGGATCAAGGAGAAGACCAAGCTGACCTAATGACTCCATGGTTCGAATGCTTTATTTGTGAAA TTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTTAACAACAACAATTGCATTCATTTTATGTTTCAGGTTCAGGGGGAGGTGTGGGAGGTTTTTTAAAACTAGTGCGGCCGCTCTAGAGCATGGCTACGTAGATAAGTAGCATGGCGGGTTAATCATTAACTACAAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCC GGGCGACCAAAGGTCGCCCGACGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

SEQ ID NO:63 Mut2-L CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGGAAACCCGGGCGTGCCCGGGCGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT SEQ ID NO:64 Mut3-R AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGAAACCCGGGCGTGCGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG SEQ ID NO:65 Selected portion of SEQ ID NO: 52 containing RBE GCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO:66 IE1 promoter fragment AATAAACGATAACGCCGTTGGTGGCGTGAGGCATGTAAAAGGTTACATCATTATCTTGTTCGCCATCCGGTTGGTATAAATAGACGTTCATGTTGGTTTTTGTTTCAGTTGCAAGTTGGCTGCGGCGCGCGCAGCACCTTT

SEQ ID NO:67 Nucleotide sequence of Rep 78 (including the underlined Kozak sequence) CGCAGCCACC-ATGGCGGGGTTTTACGAGATTGTGATTAAGGTCCCCAGCGACCTTGACGAGCATCTGCCCGGCATTTCTGACAGCTTTGTGAACTGGGTGGCCGAGAAGGAATGGGAGTTGCCGCCAGATTCTGACATGGATCTGAATCTGATTGAGCAGGCACCCCTGACCGTGGCCGAGAAGCTGCAGCGCGACTTTCTGACGGAATGGCGCCGTGTGAGTAAGGCCCCGG AGGCCCTTTTCTTTGTGCAATTTGAGAAGGGAGAGAGCTACTTCCACATGCACGTGCTCGTGGAAACCACCGGGGTGAAATCCATGGTTTTGGGACGTTTCCTGAGTCAGATTCGCGAAAAACTGATTCAGAGAATTTACCGCGGGATCGAGCCGACTTTGCCAAACTGGTTCGCGGTCACAAAGACCAGAAATGGCGCGGAGGCGGGAACAAGGTGGTGGATGAGTGCTACAT CCCCAATTACTTGCTCCCCAAAACCCAGCCTGAGCTCCAGTGGGCGTGGACTAATATGGAACAGTATTTAAGCGCCTGTTTGAATCTCACGGAGCGTAAACGGTTGGTGGCGCAGCATCTGACGCACGTGTCGCAGACGCAGGAGCAGAACAAAGAGAATCAGAATCCCAATTCTGATGCGCCGGTGATCAGATCAAAAACTTCAGCCAGGTACATGGAGCTGGTCGGGTGGCT CGTGGACAAGGGGATTACCTCGGAGAAGCAGTGGATCCAGGAGGACCAGCCTCATACATCTCCTTCAATGCGGCCTCCAACTCCGCGTCCCAAATCAAGGCTGCCTTGGACAATGCGGGAAAGATTATGAGCCTGACTAAAACCGCCCCCGACTACCTGGTGGGCCAGCAGCCCGTGGAGGACATTTCCAGCAATCGGATTTATAAAATTTTGGAACTAAACGGGTACGATCCC CAATATGCGGCTTCCGTCTTTCTGGGATGGGCCACGAAAAAGTTCGGCAAGAGGAACACCATCTGGCTGTTTGGGCCTGCAACTACCGGGAAGACCAACATCGCGGAGGCCATAGCCCACACTGTGCCCTTCTACGGGTGCGTAAACTGGACCAATGAGAACTTTCCCTTCAACGACTGTGTCGACAAGATGGTGATCTGGTGGGAGGAGGGGAAGATGACCGCCAAGGTCGTG GAGTCGGCCAAAGCCATTCTCGGAGGAAGCAAGGTGCGCGTGGACCAGAAATGCAAGTCCTCGGCCCAGATAGACCCGACTCCCGTGATCGTCACCTCCAACACCAACATGTGCGCCGTGATTGACGGGAACTCAACGACCTTCGAACACCAGCAGCCGTTGCAAGACCGGATGTTCAAATTTGAACTCACCCGCCGTCTGGATCATGACTTTGGGAAGGTCACCAAGCAGGAAG TCAAAGACTTTTTCCGGTGGGCAAAGGATCACGTGGTTGAGGTGGAGCATGAATTCTACGTCAAAAAGGGTGGAGCCAAGAAAAGACCCGCCCCCAGTGACGCAGATATAAGTGAGCCCAAACGGGTGCGCGAGTCAGTTGCGCAGCCATCGACGTCAGACGCGGAAGCTTCGATCAACTACGCAGACAGGTACCAAAACAAATGTTCTCGTCACGTGGGCATGAATCTGATGCT GTTTCCCTGCAGACAATGCGAGAGAATGAATCAGAATTCAAATATCTGCTTCACTCACGGACAGAAAGACTGTTTAGAGTGCTTTCCCGTGTCAGAATCTCAACCCGTTTCTGTCGTCAAAAGGCGTATCAGAAACTGTGCTACATTCATCATATCATGGGAAAGGTGCCAGACGCTTGCACTGCCTGCGATCTGGTCAATGTGGATTTGGATGACTGCATCTTTGAACAATAA

SEQ ID NO:68 Polyhedrin promoter sequence ATCATGGAGATAATTAAAATGATAACCATCTCGCAAATAAATAAGTATTTTACTGTTTTCGTAACAGTTTTGTAATAAAAAAACCTATAAATATTCCGGATTATTCATACCGTCCCACCATCGGGCGCG

SEQ ID NO:69 Rep58 DNA sequence GCCGCCACC-

SEQ ID NO:70 MND Promoter GAACAGAGAAACAGGAGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGTTGGAACAGCAGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGCCAAGAACAGATGGTCCCCAGATGCGGTCCCGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTCCAGGTGCCCCAAGGACCTGAAATGAC CCTGTGCCTTATTTGAACTAACCAATCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTCTGCTCCCCGAGCTCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTGGAGACGCCATCCACGCTGTTTTGACTTCCATAGAAG

SEQ ID NO:71 Luciferase (with Kozak sequence underlined) CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGAAACCCGGGCGTGCCCGGGCGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGGCTCAGAGGCTCAGAGGCACACAGGAGTTTCTGGGCTCACCCTGCCCCCTTCCAACCCCTCAGTTCCCATCCTCCAGCAGCTGTTTGTGTGCTGCCTCTGAAGTCCACACTGAAC AAACTTCAGCCTACTCATGTCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGG GTCCGGGTTCAAAACCACTTGCTGGGTGGGGAGTCGTCAGTAAGTGGCTATGCCCCGACCCCGAAGCCTGTTTCCCCATCTGTACAATGGAAATGATAAAGACGCCCATCTGATAGGGTTTTTGTGGCAAATAAACATTTGGTTTTTTTGTTTTGTTTTGTTTTGTTTTTTGAGATGGAGGTTTGCTCTGTCGCCCAGGCTGGAGTGCAGTGACACAATCTCATCTC ACCACAACCTTCCCCTGCCTCAGCCTCCCAAGTAGCTGGGATTACAAGCATGTGCCACCACACCTGGCTAATTTTCTATTTTTAGTAGAGACGGGTTTCTCCATGTTGGTCAGCCTCAGCCTCCCAAGTAACTGGGATTACAGGCCTGTGCCACCACACCCGGCTAATTTTTTCTATTTTTGACAGGGACGGGGTTTCACCATGTTGGTCAGGCTGGTCTAGAGGTA CCGGATCTTGCTACCAGTGGAACAGCCACTAAGGATCTGCAGTGAGAGCAGAGGGCCAGCTAAGTGGTACTCTCCCAGAGACTGTCTGACTCACGCCACCCCCTCCACCTTGGACACAGGACGCTGTGGTTTCTGAGCCAGGTACAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGGCGACTCAGATCCCAGCCAG TGGACTTAGCCCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGCCGCCACCATGATCATGGCCGAGAGCCCTGGCCTGATCACCATCTGCCTGCTGGGCT ACCTGCTGAGCGCCGAGTGCACCGTGTTCCTGGACCACGAGAACGCCAACAAGATCCTGAACCGGCCCAAGAGATACAACAGCGGCAAGCTGGAGGAGTTCGTGCAGGGCAACCTGGAGAGGGAGTGCATGGAGGAGAAGTGCAGCTTCGAGGAGGCCAGGGAAGTGTTCGAGAACACCGAGCGGACCACCGAGTTCTGGAAGCAGTACGTGGACGGCGACCAGTGCGAGAGCAACCCTTGC CTGAACGGCGGCAGCTGCAAGGACGACATCAACAGCTACGAGTGCTGGTGCCCTTTCGGCTTCGAGGGCAAGAACTGCGAGCTGGACGTGACCTGCAACATCAAGAACGGCCGCTGCGAGCAGTTCTGCAAGAACAGCGCCGACAACAAAGTGGTGTGTAGCTGCACCGAGGGCTACAGACTGGCCGAGAACCAGAAGAGCTGCGAGCCCGC CGTGCCCTTCCCCTGCGGCAGAGTGAGCGTGTCCCAGACCAGCAAGCTGACCAGAGCCGAGACCGTGTTCCCCGACGTGGACTACGTGAATAGCACCGAGGCCGAGACCATCCTGGACAACATCACCCAGAGCACCCAGTCCTTCAACGACTTCACCAGAGTTGTGGGCGGCGAGGACGCCAAGCCCGGCCAGTTCCCCCTGGCAGGTGGTGCTGAACGGCAAAGTGGATGCCTTCT GCGGCGGCAGCATCGTGAACGAGAAGTGGATCGTGACAGCCGCCCACTGCGTGGAGACCGGCGTGAAGATCACCGTGGTGGCCGGCGAACACAATATCGAGGAGACCGAGCACACCGAGCAGAAGCGGAACGTCATCCGGATTATCCCCCACCACAACTACAACGCCGCCATCAACAAGTACAACCACGACATCGCCCTGCTGGAGCTGGACGAGCC TCTGGTGCTGAATAGCTACGTGACCCCCATCTGCATCGCCGACAAGGAGTACACCAACATCTTCCTGAAGTTCGGCAGCGGCTACGTGTCCGGCTGGGGCAGAGTGTTCCACAAGGGCAGAAGCGCCCTGGTGCTGCAGTACCTGAGAGTGCCCCTGGTGGACAGAGCCACCTGCCTGTTGAGCACCAAGTTCACCATCTACAACAACATGTTCTGCGCCGGCTTCCACGAGGGCGGCAGAGA CAGCTGCCAGGGCGACAGCGGCGGACCCCACGTGACCGAAGTGGAGGGCACCAGCTTCCTGACCGGCATCATCAGCTGGGGCGAGGAGTGCGCCATGAAGGGCAAGTACGGCATCTACACCAAAGTGAGCCGGTACGTGAACTGGATCAAGGAGAAAACCAAGCTGACCTGAGAGCATCTTACCGCCATTTATTCCCATATTTGTTCTGTT TTTCTTGATTTGGGTATACATTTAAATGTTAATAAAACAAAATGGTGGGGCAATCATTTACATTTTTAGGGATATGTAATTACTAGTTCAGGTGTATTGCCACAAGACAAACATGTTAAGAAACTTTCCCGTTATTTACGCTCTGTTCCTGTTAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGATATTCTTAACTATGTTGCTCCTTTTACGCTGTGTGGATATGCTGCTTTATAGCCT CTGTATCTAGCTATTGCTTCCCGTACGGCTTTCGTTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTTAGAGGAGTTGTGGCCCGTTGTCCGTCAACGTGGCGTGGTGTGCTCTGTGTTTGCTGACGCAACCCCCACTGGCTGGGGCATTGCCACCACCTGTCAACTCCTTTCTGGGACTTTCGCTTTCCCCCTCCCGA TCGCCACGGCAGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTAGGTTGCTGGGCACTGATAATTCCGTGGTGTTGTCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATT CTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCGGTGGGCTCTATGGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

SEQ ID NO:72 WPR GAGCATCTTACCGCCATTTATTCCCATATTTGTTCTGTTTTTCTTGATTTGGGTATACATTTAAATGTTAATAAAACAAAATGGTGGGGCAATCATTTACATTTTTAGGGATATGTAATTACTAGTTCAGGTGTATTGCCACAAGACAAACATGTTAAGAAACTTTCCCGTTATTTACGCTCTGTTCCTGTTAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGATATTCTTAACTATGTTGCTC CTTTTACGCTGTGTGGATATGCTGCTTTATAGCCTCT GTATCTAGCTATTGCTTCCCGTACGGCTTTCGTTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTTAGAGGAGTTGTGGCCCGTTGTCCGTCAACGTGGCGTGGTGTGCTCTGTGTTTGCTGACGCAACCCCCACTGGCTGGGGCATTGCCACCACCTGTCAACTCCTTTCTGGGACTTTCGCTTTCCCCCTCCCGATCGCCACGGCAGAACTCATCGCCGCCTGCCTTGCCCG CTGCTGGACAGGGGCTAGGTTGCTGGGCACTGATAATTCCGTGGTGTTGTC SEQ ID NO:73 BGH-poly(A) TGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCGGTGGGCTCTATGGC

SEQ ID NO:74 HLCR-AAT promoter GGCTCAGAGGCTCAGAGGCACACAGGAGTTTCTGGGCTCACCCTGCCCCCTTCCAACCCCTCAGTTCCCATCCTCCAGCAGCTGTTTGTGTGCTGCCTCTGAAGTCCACACTGAACAAACTTCAGCCTACTCATGTCCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTTCTGGGCCCATGCCACC TCCAACATCCACTCGACCCCTTGGAATTTTCGGTGGAGAGGAGCAGAG GTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGTCCGGGTTCAAAACCACTTGCTGGGTGGGGAGTCGTCAGTAAGTGGCTATGCCCCGACCCCGAAGCCTGTTTCCCCATCTGTACAATGGAAATGATAAAGACGCCCATCTGATAGGGTTTTTGTGGCAAATAAACATTTGGTTTTTTTGTTTTGTTTTTGTTTTGTTTTTTGAGATGGAGGTTTGCTCTGTCGCCCAGGCTGGAGTG CAGTGACACAATCTCATCTCACCACAACCTTCCCCTGCCTCAGCCTCCCAAGTA GCTGGGATTACAAGCATGTGCCACCACACCTGGCTAATTTTCTATTTTTAGTAGAGACGGGTTTCTCCATGTTGGTCAGCCTCAGCCTCCCAAGTAACTGGGATTACAGGCCTGTGCCACCACACCCGGCTAATTTTTTCTATTTTTGACAGGGACGGGGTTTCACCATGTTGGTCAGGCTGGTCTAGAGGTACCGGATCTTGCTACCAGTGGAACAGCCACTAAGGATCTGCAGTG AGAGCAGAGGGCCAGCTAAGTGGTACTCTCCCAGAGACTGTCTGACTCACGCCACC CCCTCCACCTTGGACACAGGACGCTGTGGTTTCTGAGCCAGGTACAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGGCGACTCAGATCCCAGCCAGTGGACTTAGCCCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAG GACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGT SEQ ID NO:75-100

SEQ ID NO: 101 Left ITR-2 GCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGGAAACCCGGGCGTGCGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 102 Right ITR-2 GCGCGCTCGCTCGCTCACTGAGGCGCACGCCCGGGTTTCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 103 Left ITR-3 GCGCGCTCGCTCGCTCACTGAGGCCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 104 Right ITR-3 GCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 105 Left ITR-4 GCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 106 Right ITR-4 GCGCGCTCGCTCGCTCACTGAGGCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 107 Left ITR-10 GCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCTTTGCCCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 108 Right ITR-10 GCGCGCTCGCTCGCTCACTGAGGCCGGGCAAAGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 109 Left ITR-17 GCGCGCTCGCTCGCTCACTGAGGCCGAAACGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 110 Right ITR-17 GCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGTTTCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 111 Left ITR-6 GCGCGCTCGCTCGCTCACTGAGGCAAAGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 112 Right ITR-6 GCGCGCTCGCTCGCTCACTGAGGCTTTGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 113 Left ITR-1 GCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 114 Right ITR-1 GCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGC

SEQ ID NO: 115 Left ITR-5 GCGCGCTCGCTCGCTCACTGAGGCGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 116 Right ITR-5 GCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 117 Left ITR-7 GCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACTTTGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 118 Right ITR-7 GCGCGCTCGCTCGCTCACTGAGGCCGGGCGACAAAGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 119 Left ITR-8 GCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGATTTTTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 120 Right ITR-8 GCGCGCTCGCTCGCTCACTGAGGCCGGGCGAAAATCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 121 Left ITR-9 GCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGTTTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 122 Right ITR-9 GCGCGCTCGCTCGCTCACTGAGGCCGGGCGAAACGCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 123 Left ITR-11 GCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGGAAACCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 124 Right ITR-11 GCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGTTTCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 125 Left ITR-12 GCGCGCTCGCTCGCTCACTGAGGCCGCCCGGAAACCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 126 Right ITR-12 GCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGTTTCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGC

SEQ ID NO: 127 Left ITR-13 GCGCGCTCGCTCGCTCACTGAGGCCGCCCGAAACGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 128 Right ITR-13 GCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGTTTCGGGCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 129 Left ITR-14 GCGCGCTCGCTCGCTCACTGAGGCCGCCCAAAGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 130 Right ITR-14 GCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCTTTGGGCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 131 Left ITR-15 GCGCGCTCGCTCGCTCACTGAGGCCGCCAAAGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 132 Right ITR-15 GCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCTTTGGCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 133 Left ITR-16 GCGCGCTCGCTCGCTCACTGAGGCCGCAAAGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGC SEQ ID NO: 134 Right ITR-16 GCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCTTTGCGGCCTCAGTGAGCGAGCGAGCGCGC

SEQ ID NO: 135 HAAT promoter CCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTTTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGGAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGC GTCCGGGCAGCGTAGGCGGGCGACTC AGATCCCAGCCAGTGGACTTAGCCCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATCCGGACTCTAAGGTAAATATAAAATTTTTAAGTGTATAATGTGTTAAACTACTGATTCTAATTGTTTCTCTC TTTTAGATTCCAACCTTTGGAACTG SEQ ID NO: 136-150

SEQ ID NO: 151 ARSA
(CR456383.1 Homo sapiens ARSA full-length open reading frame (ORF) cDNA clone (pGEM.ARSA.V2 C22ORF cDNA clone)) ATGTCCATGGGGGCACCGCGGTCCCTCCTCCTGGCCCTGGCTGCTGGCCTGGCCGTTGCCCGTCCGCCCAACATCGTGCTGATCTTTGCCGACGACCTCGGCTATGGGGACCTGGGCTGCTATGGGCACCCCAGCTCTACCACTCCCAACCTGGACCAGCTGGCGGCGGGAGGGCTGCGGTTCACAGACTTCTACGTGCCTGTGTCTCTGTGCACACCCTCTAGGGCCGCCCTCCTGACCGG CCGGCTCCCGGTTCGGATGGGCATGTACCCTGGCGTCCTGGTGCCCAGCTCCCGGGGGGGCCTGCCCCTGGAGGAGGTGACCGTGGCCGAAGTCCTGGCTGCCCGAGGCTACCTCACAGGAATGGCCGGCAAGTGGCACC TTGGGGTGGGGCCTGAGGGGGCCTTCCTGCCCCCCCATCAGGGCTTCCATCGATTTCTAGGCATCCCGTACTCCCACGACCAGGGCCCCTGCCAGAACCTGACCTGCTTCCCGCCGGCCACTCCTTGCGACGGTGGCTGTGACCAGGGCCTGGTCCCCATCCCACTGTTGGCCAACCTGTCCGTGGAGGCGCAGCCCCCCTGGCTGCCCGGACTAGAGGCCCGCTACATGGCTTTCGCCCATGAC CTCATGGCCGACGCCCAGCGCCAGGATCGCCCCTTCTTCCTGTACTATGCCTCTCACCACACCCACTACCCTCAGTTCAGTGGGCAGAGCTTTGCAGAGCGTTCAGGCCGCGGGCCATTTGGGGACTCCCTGATGGAG CTGGATGCAGCTGTGGGGACCCTGATGACAGCCATAGGGGACCTGGGGCTGCTTGAAGAGACGCTGGTCATCTTCACTGCAGACAATGGACCTGAGACCATGCGTATGTCCCGAGGCGGCTGCTCCGGTCTCTTGCGGTGTGGAAAGGGAACGACCTACGAGGGCGGTGTCCGAGAGCCTGCCTTGGCCTTCTGGCCAGGTCATATCGCTCCCGGCGTGACCCACGAGCTGGCCAGCTCCCTGGA CCTGCTGCCTACCCTGGCAGCCCTGGCTGGGGCCCCACTGCCCAATGTCACCTTGGATGGCTTTGACCTCAGCCCCCTGCTGCTGGGCACAGGCAAGAGCCCTCGGCAGTCTCTCTTCTTCTACCCGTCCTACCCAG ACGAGGTCCGTGGGGTTTTTGCTGTGCGGACTGGAAAGTACAAGGCTCACTTCTTCACCCAGGGCTCTGCCCACAGTGATACCACTGCAGACCCTGCCTGCCACGCCTCCAGCTCTCTGACTGCTCATGAGCCCCCGCTGCTCTATGACCTGTCCAAGGACCCTGGTGAGAACTACAACCTGCTGGGGGGTGTGGCCGGGGCCACCCCAGAGGTGCTGCAAGCCCTGAAACAGCTTCAGCTGCTCA AGGCCCAGTTAGACGCAGCTGTGACCTTCGGCCCCAGCCAGGTGGCCCGGGGCGAGGACCCCGCCCTGCAGATCTGCTGTCATCCTGGCTGCACCCCCCGCCCAGCTTGCTGCCATTGCCCAGATCCCCATGCCTGA

SEQ ID NO: 152 I2S
(Iduronate-2-sulfatase (IDS) Genbank Homo sapiens, RefSeqGene on chromosome X) ATGCCGCCACCCCGGACCGGCCGAGGCCTTCTCTGGCTGGGTCTGGTTCTGAGCTCCGTCTGCGTCGCCCTCGGATCCGAAACGCAGGCCAACTCGACCACAGATGCTCTGAACGTTCTTCTCATCATCGTGGATGACCTGCGCCCCTCCCTGGGCTGTTATGGGGATAAGCTGGTGAGGTCCCCAAATATTGACCAACTGGCATCCCACAGCCTCCTCTTCCAGAATGCCTTTGCGCAGCAAG CAGTGTGCGCCCCGAGCCGCGTTTCTTTCCTCACTGGCAGGAGACCTGACACCACCCGCCTGTACGACTTCAACTCCTACTGGAGGGTGCACGCTGGAAACTTCTCCACCATCCCCCAGTACTTCAAGGAGAATGGCTATGTGACCATGTCGGTGGGAAAAGTCTTTCA CCCTGGGATATCTTCTAACCATACCGATGATTCTCCGTATAGCTGGTCTTTTTCCACCTTATCATCCTTTCCTCTGAGAAGTATGAAAACACTAAGACATGTCGAGGGCCAGATGGAGAACTCCATGCCAACCTGCTTTGCCCTGTGGATGTGCTGGATGTTCCCGAGGGCACCTTGCCTGACAAACAGAGCACTGAGCAAGCCATACAGTTGTTGGAAAAGATGAAAACGTCAGCCAGTCCTTTCTTC CTGGCCGTTGGGTATCATAAGCCACACATCCCCTTCAGATACCCCAAGGAATTTCAGAAGTTGTATCCCTTGGAGAACATCACCCTGGCCCCCGATCCCGAGGTCCCTGATGGCCTACCCCCTGTGGCCTACAACCCCTGGATGGACATCAGGCAACGGGAAGACG TCCAAGCCTTAAACATCAGTGTGCCGTATGGTCCAATTCCTGTGGACTTTCAGCGGAAAATCCGCCAGAGCTACTTTGCCTCTGTGTCATATTTGGATACACAGGTCGGCCGCCTCTTGAGTGCTTTGGACGATCTTCAGCTGGCCAACAGCACCATCATTGCATTTACCTCGGATCATGGGTGGGCTCTAGGTGAACATGGAGAATGGGCCAAATACAGCAATTTTGATGTTGCTACCCATG TTCCCCTGATATTCTATGTTCCTGGAAGGACGGCTTCACTTCCGGAGGCAGGCGAGAAGCTTTTCCCTTACCTCGACCCTTTTGATTCCGCCTCACAGTTGATGGAGCCAGGCAGGCAATCCATGGACCTTGTGGAACTTGTGTCTCTTTTTCCCACGCTGGCTGGACTT GCAGGACTGCAGGTTCCACCTCGCTGCCCCGTTCCTTCATTTCACGTTGAGCTGTGCAGAGAAGGCAAGAACCTTCTGAAGCATTTTCGATTCCGTGACTTGGAAGAGGATCCGTACCTCCCTGGTAATCCCCGTGAACTGATTGCCTATAGCCAGTATCCCCGGCCTTCAGACATCCCTCAGTGGAATTCTGACAAGCCGAGTTTAAAAGATAAAAGATCATGGGCTATTCCATACGCAC CATAGACTATAGGTATACTGTGTGGGTTGGCTTCAATCCTGATGAATTTCTAGCTAACTTTTCTGACATCCATGCAGGGGAACTGTATTTTGTGGATTCTGACCCATTGCAGGATCACAATATGTATAATGATTCCCAAGGTGGAGATCTTTTCCAGTTGTTGATGCCTTGA

SEQ ID NO:301 RBE-1 GCGGCTCGCTCGCTC SEQ ID NO:302 Spacer ACTGAGGC SEQ ID NO:303 Loop shoulder CGGGCGACCAAAGGTCGCCCGA SEQ ID NO:304 Truncated shoulder CGCCCGGGCG SEQ ID NO:305 Complementary sequence to spacer GCCTCAGT SEQ ID NO:306 RBE-2 GAGCGAGCGAGCGCGC SEQ ID NO:307 SEQ ID NO:308 Enhancer SV40 GGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCAGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCATAGTCCCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCGCCCATGGCTGACTAATTTTTTTTATTTATGCAGAG GCCGAGGCCGCCTCGGCCTCTGAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGCTTTGCAAA

SEQ ID NO:309 CMV Enhancer TAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCT ATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAG TACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTA CGGTGGGAGGTCTATATAAGCAGAGCTGGTTTAGTGAACCGTCAG

SEQ ID NO:310 Rat EF1-α promoter
(Rattus norvegicus BAC CH230-35L12 (Children's Hospital Oakland Research Institute), complete sequence. Sequence ID: gi|49615137|AC097023.6) GGAGCCGAGAGTAATTCATACAAAAGGAGGGATCGCCTTCGCAAGGGGAGAGCCCAGGGACCGTCCCTAAATTCTCACAGACCCAAATCCCTGTAGCCGCCCCACGACAGCGCGAGGAGCATGCGCCCAGGGCTGAGCGCGGGTAGATCAGAGCACACAAGCTCACAGTCCCCGGCGGTGGGGGGAGGGGCGCGCTGAGCGGGGGCCAGGGAGCTGGCGCGGGGCAAACTGGGAAAGTGGTGTC GTGTGCTGGCTCCGCCCTCTTCCCGAGGGTGGGGGGAACGGTATATAAGTGCGGTAGTCGCCTTGGACGTTCTTTTTCGCAAC GGGTTTGCCGTCAGAACGCAGGTGAGTGGCGGGTGTGGCTTCCGCGGGCCCCGGAGCTGGAGCCCTGCTCTGAGCGGGCCGGGCTGATATGCGAGTGTCGTCCGCAGGGTTTAGCTGTGAGCATTCCCACTTCGAGTGGCGGGCGGTGCGGGGGTGAGAGTGCGAGGCCTAGCGGCAACCCCGTAGCCTCGCCTCGTGTCCGGCTTGAGGCCTAGCGTGGTGTCCGCCGCCGCGTGCC ACTCCGGCCGCACTATGCGTTTTTTGTCCTTGCTGCCCTCGATTGCCTTCCAGCAGCATGGGCTAACAAAGGGAGGGTGTGGGGCTCACT CTTAAGGAGCCCATGAAGCTTACGTTGGATAGGAATGGAAGGGCAGGAGGGGCGACTGGGGCCCGCCCGCCTTCGGAGCACATGTCCGACGCCACCTGGATGGGGCGAGGCCTGTGGCTTTCCGAAGCAATCGGGCGTGAGTTTAGCCTACCTGGGCCATGTGGCCCTAGCACTGGGCACGGTCTGGCCTGGCGGTGCCGCGTTCCCTTGCCTCCCAACAAGGGTGAGGCCGTCCCGCCCGG CACCAGTTGCTTGCGCGGAAAGATGGCCGCTCCCGGGGCCCTGTTGCAAGGAGCTCAAAATGGAGGACGCGGCAGCCCGGTGGAGC GGGCGGGTGAGTCACCCACACAAAGGAAGAGGGCCTTGCCCCTCGCCGGCCGCTGCTTCCTGTGACCCCGTGGTCTATCGGCCGCATAGTCACCTCGGGCTTCTCTTGAGCACCGCTCGTCGCGGCGGGGGGAGGGGATCTAATGGCGTTGGAGTTTGTTCACATTTGGTGGGTGGAGACTAGTCAGGCCAGCCTGGCGCTGGAAGTCATTCTTGGAATTTGCCCCTTTGAGTTTGGAGCGA GGCTAATTCTCAAGCCTCTTAGCGGTTCAAAGGTATTTTCTAAACCCGTTTCCAGGTGTTGTGAAAGCCACCGCTAATTCAAAGCAA

SEQ ID NO:313 Secretory leader sequence VH1-02 MDWTWRILFLVAAATGAHS SEQ ID NO:314 Secretory leader sequence VK A26 MLPSQLIGFLLLWVPASRG SEQ ID NO:315 Large T-antigen of SV40 virus PKKKRKV SEQ ID NO:316 Nucleoplasmin KRPAATKKAGQAKKKK SEQ ID NO:317 c-myc PAAKRVKLD SEQ ID NO:318 c-myc RQRRNELKRSP SEQ ID NO:319 hRNPA1 M9 NQSSNFGPMKGGNFGGRSSGPYGGGGQYFAKPRNQGGY SEQ ID NO:320 IBB domain importin-alpha RMRIZFKNKGKDTAELRRRRVEVSVELRKAKKDEQILKRRNV SEQ ID NO:321 Myoma T protein VSRKRPRP SEQ ID NO:323 p53 human PQPKKKPL SEQ ID NO:324 c-abl IV mouse SALIKKKKKMAP SEQ ID NO:325 NS1 influenza virus DRLRR SEQ ID NO:326 NS1 influenza virus PKQKKRK SEQ ID NO:327 Hepatitis delta virus antigen RKLKKKIKKL

SEQ ID NO:328 Mouse Mx1 protein REKKKFLKRR SEQ ID NO:329 Human poly(ADP-ribose) polymerase KRKGDEVDGVDEVAKKKSKK SEQ ID NO:330 Glucocorticoid steroid hormone receptor (human) RKCLQAGMNLEARKTKK SEQ ID NO:331-499 SEQ ID: 500 GCCCGCTGGTTTCCAGCGGGCTGCGGGCCCGAAACGGGCCCGC SEQ ID: 501 CGGGCCCGTGCGGGCCCAAAGGGCCCGC SEQ ID: 502 GCCCGGGCACGCCCGGGTTTCCCGGGCG SEQ ID: 503 CGTGCGGGCCCAAAGGGCCCGC Selected polynucleotide sequences of the RIGHT side ITR: C-C' B-B´ The whole sequence WT-ITR-R CGGGCGACCAAAGGTCGCCG (SEQ ID: 504) A CGCCCGGGCTTTGCCCGGGC (SEQ ID: 505) CGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGC (SEQ ID: 506) TTX1-R (ITR folding prediction) CGGGCGACCAAAGGTCGCCG (SEQ ID: 507) A CGCCCGGGC(TTTGCCCG)GGC (SEQ ID: 508) CGGGCGACCAAAGGTCGCCCGACGCCCGGGCGGC
(SEQ ID: 509)

Kotin 2 (p.11) CGGG(CGACCAAAGGTC)GCCCG (SEQ ID: 510) A CGCCCGGGCTTTGCCCGGGC (SEQ ID: 511) CGGGGCCCGACGCCCGGGCTTTGCCCGGGC (SEQ ID: 512) Kotin 2 (p.11) CGGG(CGACCAAAGGTCG)CCCG (SEQ ID: 513) A CGCCCGGGCTTTGCCCGGGC (SEQ ID: 514) CGGGCCCGACGCCCGGGCTTTGCCCGGGC (SEQ ID: 515) Kotin 2 (p.11) [CGGGCGACCAAAGGTCGCCCG] all or part of the deletions in the square brackets can be used to create asymmetric interrupted self-complementary sequences; (SEQ ID: 516) A CGCCCGGGCTTTGCCCGGGC (SEQ ID: 517) Selected polynucleotide sequences of the LEFT side ITR: C-C' B-B´ The whole sequence WT-ITR-L GCCCGGGCAAAGCCCGGGCG (SEQ ID: 518) T CGGGCGACCTTTGGTCGCCG (SEQ ID: 519) GCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCG (SEQ ID: 520)

SEQ ID: 22 [GCCCGGGCAAA]GCCCGGGCG (SEQ ID: 521) T CGGGCGACCTTTGGTCGCCG (SEQ ID: 522) GCCCGGGCGTCGGGCGACCTTTGGTCGCCG (SEQ ID: 522) TTX1-L (ITR stacking prediction) GCC(CGGGCAAA)GCCCGGGCG (SEQ ID: 523) T CGGGCGACCTTTGGTCGCCCG (SEQ ID: 524) GCCGCCCGGGCGACGGGCGACCTTTGGTCGCCG (SEQ ID: 525) Kotin 2 (p.11) GCCC(GGGCAAAGCCC)GGGCG (SEQ ID: 526) T CGGGCGACCTTTGGTCGCCG (SEQ ID: 527) GCCCGGGCGTCGGGCGACCTTTGGTCGCCG (SEQ ID: 528) Kotin 2 (p.11) [GCCCGGGCAAAGCCCGGGCG] all or part of the deletions in the square brackets can be used to create asymmetric discontinuous self-complementary sequences; (SEQ ID: 528) T CGGGCGACCTTTGGTCGCCG (SEQ ID: 529)

SEQ ID NO: 530 _{NM_004895.1} CAGGGCAGCCTTCAGTCTGATTCAGGAGAACGAGGTCCTCTTCACCATGTGCTTCATCCCCCTGGTCTGCTGGATCGTGTGCACTGGACTGAAACAGCAGATGGAGAGTGGCAAGAGCCTTGCCCAGACATCCAAGACCTCCACCGCGGTGTACGTCTTCTTCCTTTCCAGTTTGCTGCAGCCCCGGGGAGGGAGCCAGGAGCACGGCCTCTGCGCCCACCTCTGGGGGCTCTGCTCTTTGGCTG CAGATGGAATCTGGAACCAGAAAATCCTGTTT GAAGAGTCCGACCTCAGGAATCATGGACTGCAGAAGGCGGATGTGTCTGCTTTCCTGAGGATGAACCTGTTCCAAAAGGAAGTGGACTGCGAGAAGTTCTACAGCTTCATCCACATGACTTTCCAGGAGTTCTTTGCCGCCATGTACTACCTGCTGGAAGAGGAAAAGGAAGGAAGGACGAACGTTCCAGGGAGTCGTTTTGAAGCTTCCCAGCCGAGACGTGACAGTCCTTCTGGAAAACT ATGGCAAATTCGAAAAGGGGTATTTGATTTTTTGTTG TACGTTTCCTCTTTGGCCTGGTAAACCAGGAGAGGACCTCCTACTTGGAGAAGAAATTAAGTTGCATGATCTCTCAGCAAATCAGGCTGGAGCTGCTGAAATGGATTGAAGTGAAAGCCAAAGCTAAAAAGCTGCATGATCAGCCCAGCCAGCTGGAATTGTTCTACTGTTTGTACGAGATGCAGGAGGAGGACTTCGTGCAAAGGGCCATGGACTATTTCCCCAAGATTGAGATCAATCTCT CCACCAGAATGGACCACATGGTTTCCTCCTTTTG CATTGAGAACTGTCATCGGGTGGAGTCACTGTCCCTGGGGTTTCTCCATAACATGCCCAAGGAGGAAGAGGAGGAGGAAAAGGAAGGCCGACACCTTGATATGGTGCAGTGTGTCCTCCCAAGCTCCCTCATGCTGCCTGTTCTCATGGGTTGGGGCGCTGTGGCCTCTCCCATGAGTGCTGCTTCGACATCTCCTTGGTCCTCAGCAGCAACCAGAAGCTGGTGGAGCTGGACCTGAGT GACAACGCCCTCGGTGACTTCGGAATCAGACTTCTGT GTGTGGGACTGAAGCACCTGTTGTGCAATCTGAAGAAGCTCTGGTTGGTGAATTCTGCCTTACGTCAGTCTGTTGTTCAGCTTTGTCCTCGGTACTCAGCACTAATCAGAATCTCACGCACCTTTACTGCGAGGCAACACTCTCGGAGACAAGGGATCAAACTACTCTGTGAGGGACTCTTGCACCCCGACTGCAAGCTTCAGGTGTTGGAATTAGACAACTGCAACCTCACGTCACACTGCTGCT GGGATCTTTCCACACTTCTGACCTCCAGCCA GAGCCTGCGAAAGCTGAGCCTGGGCAACAATGACCTGGGCGACCTGGGGGTCATGATGTTCTGTGAAGTGCTGAAACAGCAGAGCTGCCTCCTGCAGAACCTGGGGTTGTCTGAAATGTATTTCAATTATGAGACAAAAAGTGCGTTAGAAACACTTCAAGAAGAAAAGCCTGAGCTGACCGTCGTCTTTTGAGCCTTCTTGGTAGGAGTGGAAACGGGGCTGCCAGACGCCAGTGTTCTCC GGTCCCTCCAGCTGGGGGCCCTCAGGTGGAGAGAGC TGCGATCCATCCAGGCCAAGACCACAGCTCTGTGATCCTTCCGGTGGAGTGTCGGAGAAGAGAGCTTGCCGACGATGCCTTCCTGTGCAGAGCTTGGGCATCTCCTTTACGCCAGGGTGAGGAAGACACCAGGACAATGACAGCATCGGGTGTTGTTGTCATCACAGCGCCTCAGTTAGAGGATGTTCCTCTGGTGACCTCATGTAATTAGCTCATTCAATAAAGCACTTTCTTTATTTTT CTCTTCTCTGTCTAACCTTCTTTTTCCTATCTTTTT TTCTTCTTTGTTCTGTTTACTTTTGCTCATATCATCATTCCCGCTATCTTTCTATTAACTGACCATAACACAGAACTAGTTGACTATATATTATGTTGAAATTTTATGGCAGCTATTTATTTATTTAAATTTTTTGTAATAGTTTTGTTTTTCTAATAAGAAAAATCCATGCTTTTTGTAGCTGGTTGAAAATTCAGGAATATGTAAAACTTTTTGGTATTTAATTAAATTGATTCCTTTTCTTAAT TTTAAAAAAAAAAAAAAAAAAAAAAAAAAA

SEQ ID NO: 531 _{NM_183395} GTTCCTGAGGCTGGCATCTGGATGAGGAAACTGAAGTTGAGGAATAGTGAAGAGTTTGTCCAATGTCATAGCCCCGTAATCAACGGGACAAAAATTTTCTTGCTGATGGGTCAAGATGGCATCGTGAAGTGGTTGTTCACCGTAAACTGTAATACAATCCTGTTTATGGATTTGTTTGCATATTTTTCCCTCCATAGGGAAACCTTTTCTTCCATGGCTCAGGACACACTCCTGGATCGAG CCAACAGGAGAACTTTCTGGTAAGCATTTGGCTAACTTTTTTTTTTTTGAGATGGAGTCTTGCTGTGTCGCCTAGGCTGGAGTGCAGTGGCGTGATCTTGGCTCACTGCAGCTCCACTTCCCGGGTTCAATCAATTCTCCTACCTCAACTTCCTGAGTAGCTGGGATTACAGGCGCCCGCCACCACACCCGGCTCATTTTTGTACTTTTAGTAGAGACACAGTTTTGCCATGTTGGCCA GGCTGGTCTTGAATTCCTCAGCTCAGGTGATCTGCCTGCCTTGGCCTCTCAAAGTGCTGGGATTACAGGCGTGAGCCACTGTGCCCGGCCTTGGCTAACTTTTCAAAATTAAAGATTTTGACTTGTTACAGTCATGTGACATTTTTTTCTTTCTGTTTGCTGAGTTTTTGATAATTTATATCTCTCAAAGTGGAGACTTTAAAAAAGACTCATCCGTGTGCCGTGTTCACTGCCTGGTAT CTTAGTGTGGACCGAAGCCTAAGGACCCTGAAAACAGCTGCAGATGAAGATGGCAAGCACCCGCTGCAAGCTGGCCAGGTACCTGGAGGACCTGGAGGATGTGGACTTGAAGAAATTTAAGATGCACTTAGAGGACTATCCTCCCCAGAAGGGCTGCATCCCCCCTCCCGAGGGGTCAGACAGAAGGCAGACCATGTGGATCTAGCCACGCTAATGATCGACTTCAATGGGGAGGAGAAG GCGTGGGCCATGGCCGTGTGGATCTTCGCTGCGATCAACAGGAGAGACCTTTATGAGAAAGCAAAAAGAGATGAGCCGAAGTGGGGTTCAGATAATGCACGTGTTTCGAATCCCACTGTGATATGCCAGGAAGACAGCATTGAAGAGGAGTGGATGGGTTTACTGGAGTACCTTTCGAGAATCTCTATTTGTAAAATGAAGAAAGATTACCGTAAGAAGTACAGAAAGTACGTGAGAAGC AGATTCCAGTGCATTGAAGACAGGAATGCCCGTCTGGGTGAGAGTGTGAGCCTCAACAAACGCTACACACGACTGCGTCTCATCAAGGAGCACCGGAGCCAGCAGGAGAGGGAGCAGGAGCTTCTGGCCATCGGCAAGACCAAGACGTGTGAGAGCCCCGTGAGTCCCATTAAGATGGAGTTGCTGTTTGACCCCGATGATGAGCATTCTGAGCCTGTGCACACCGTGGTGTTCCAGGGGG GCGGCAGGGATTGGGAAAACAATCCTGGCCAGGAAGATGATGTTGGACTGGGCGTCGGGGACACTCTACCAAGACAGGTTTGACTATCTGTTCTATATCCACTGTCGAGAGGTGAGCCTTGTGACACAGAGGAGCCTGGGGGACCTGATCATGAGCTGCTGCCCCGACCCAAACCCACCCATCCACAAGATCGTGAGAAAACCCTCCAGAATCCTTCCTCATGGACGGCTTCGATGAG CTGCAAGGTGCCTTTGACGAGCACATAGGACCGCTCTGCACTGACTGGCAGAAGGCCGAGCGGGGAGACATTCCTGAGCAGCCTCATCAGAAAGAAGCTGCTTCCCGAGGCCTCTCTGCTCATCACCACGAGACCTGTGGCCCTGGAGAAACTGCAGCACTTGCTGGACCATCCTCGGCATGTGGAGATCCTGGGTTTCTCCGAGGCCAAAAGGAAAGAGTACTTCTTCAAGTACTTCT CTGATGAGGCCCAAGCCAGGGCAGCCTTCAGTCTGATTCAGGAGAACGAGGTCCTCTTCACCATGTGCTTCATCCCCCTGGTCTGCTGGATCGTGTGCACTGGACTGAAACAGCAGATGGAGAGTGGCAAGAGCCTTGCCCAGACATCCAAGACCACCACCGCGGTGTACGTCTTCTTCCTTTCCAGTTTGCTGCAGCCCCGGGGAGGGAGCCAGGAGCACGGCCTCTGCGCCCACCTCT GGGGGCTCTGCTCTTTGGCTGCAGATGGAATCTGGAACCAGAAAATCCTGTTTGAGGAGTCCGACCTCAGGAATCATGGACTGCAGAAGGCGGATGTGTCTGCTTTCCTGAGGATGAACCTGTTCCAAAAGGAAGTGGACTGCGAGAAGTTCTACAGCTTCATCCACATGACTTTCCAGGAGTTCTTTGCCGCCATGTACTACCTGCTGGAAGAGGAAAAGGAAGGAAGGACGAACGTTC CAGGGAGTCGTTTGAAGCTTCCCAGCCGAGACGTGACAGTCCTTCTGGAAAACTATGGCAAATTCGAAAAGGGGTATTTGATTTTTGTTGTACGTTTCCTCTTTGGCCTGGTAAACCAGGAGAGGACCTCCTACTTGGAGAAGAAATTAAGTTGCAAGATCTCTCAGCAAATCAGGCTGGAGCTGCTGAAATGGATTGAAGTGAAAGCCAAAGCTAAAAAGCTGCAGATCCAGCCCAGCC AGCTGGAATTGTTCTACTGTTTGTACGAGATGCAGGAGGAGGACTTCGTGCAAAGGGCCATGGACTATTTCCCCAAGATTGAGATCAATCTCTCCACCAGAATGGACCACATGGTTTCTTCCTTTTGCATTGAGAACTGTCATCGGGTGGAGTCACTGTCCCTGGGGTTTCTCCATAACATGCCCAAGGAGGAAGAGGAGGAGGAAAAGGAAGGCCGACACCTTGATATGGTGCAGTGTGT CCTCCCAAGCTCCTCTCATGCTGCCTGTTCTCATGGGTTGGGGCGCTGTGGCCTCTCGCATGAGTGCTGCTTCGACATCTCCTTGGTCCTCAGCAGCAACCAGAAGCTGGTGGAGCTGGACCTGAGTGACAACGCCCTCGGTGACTTCGGAATCAGACTTCTGTGTGTGGGACTGAAGCACCTGTTGTGCAATCTGAAGAAGCTCTGGTTGGTGAATTCTGGCCTTACGTCAGTCTGTTG TTCAGCTTTGTCCTCGGTACTCAGCACTAATCAGAATCTCACGCACCTTTACCTGCGAGGCAACACTCTCGGAGACAAGGGGATCAAACTACTCTGTGAGGGACTCTTGCACCCCGACTGCAAGCTTCAGGTGTTGGAATTAGACAACTGCAACCTCACGTCACACTGCTGCTGGGATCTTTCCACACTTCTGACCTCCAGCCAGAGCCTGCGAAAGCTGAGCCTGGGCAACAATGACCTG GGCGACCTGGGGGTCATGATGTTCTGTGAAGTGCTGAAACAGCAGAGCTGCCTCCTGCAGAACCTGGGGTTGTCTGAAATGTATTTCAATTATGAGACAAAAAGTGCGTTAGAAACACTTCAAGAAGAAAAGCCTGAGCTGACCGTCGTCTTTTGAGCCTTCTTGGTAGGAGTGGAAACGGGGCTGCCAGACGCCAGTGTTCTCCGGTCCCTCCAGCTGGGGGCCCTCAGGTGGAGAGAGC TGCGATCCATCCAGGCCAAGACCACAGCTCTGTGATCCTTCCGGTGGAGTGTCGGAGAAGAGAGCTTGCCGACGATGCCTTCCTGTGCAGAGCTTGGGCATCTCCTTTACGCCAGGGTGAGGAAGACACCAGGACAATGACAGCATCGGGTGTTGTTGTCATCACAGCGCCTCAGTTAGAGGATGTTCCTCTTGGTGACCTCATGTAATTAGCTCATTCAATAAAGCACTTTCTTTTATTTT SEQ ID NO:532 _{NM_001079821} GTTCCTGAGGCTGGCATCTGGGGAAACCTTTTCTTCCATGGCTCAGGACACACTCCTGGATCGAGCCAACAGGAGAACTTTCTGTGTGGACCGAAGCCTAAGGACCCTGAAAACAGCTGCAGATGAAGATGGCAAGCACCCGCTGCAAGCTGGCCAGGTACCTGGAGGACCTGGAGGATGTGGACTTGAAGAAATTTAAGATGCACTTAGAGGACTATCCTCCCCAGAAGGGCTGCATCCCCCCTCCCGAGG GGTCAGACAGAGAAGGCAGACCATGTGGATCTAGCCACGCTAATGATCGACTTCAATGGGGAGGAGAAGGCGTGGGCCATGGCCGTGTGGATCTTCGCTGCGATCAACAGGAGAGACCTTTATGAGAAAGCAAAAAGAGATGAGCCGAAGTGGGGTTCAGATAATGCACGTGTTTCGAATCCCACTGTGATAT GCCAGGAAGACAGCATTGAAGAGGAGTGGATGGGTTTACTGGAGTACCTTTCGAGAATCTCTATTTGTAAAATGAAGAAAGATTACCGTAAGAAGTACAGAAAGTACGTGAGAAGCAGATTCCAGTGCATTGAAGACAGGAATGCCCGTCTGGGTGAGAGTGTGAGCCTCAACAAACGCTACACACGACTGCGTCTCATCAAGGAGCACCGGAGCCAGCAGGAGAGGGAGCAGGAGCTTCTG GCCATCGGCAAGACCAAGACGTGTGAGAGCCCCGTGAGTCCCATTAAGATGGAGTTGCTGTTTGACCCCGATGATGAGCATTCTGAGCCTGTGCACACCGTGGTGTTCCAGGGGGCGGCAGGGATTGGGAAAACAATCCTGGCCAGGAAGATGATGTTGGACTGGGCGTCGGGGACACTCTACCAAGACAGGTTTGACTAT CTGTTCTATATCCACTGTCGAGAGGTGAGCCTTGTGACACAGAGGAGCCTGGGGGACCTGATCATGAGCTGCTGCCCCGACCCAAACCCACCCATCCACAAGATCGTGAGAAAACCCTCCAGAATCCTTCCTCATGGACGGCTTCGATGAGCTGCAAGGTGCCTTTGACGAGCACATAGGACCGCTCTGCACTGACTGGCAGAAGGCCGAGCGGGGAGACATTCTCCTGAGCAGCCTCATCAGAAAGA AGCTGCTTCCCGAGGCCTCTCTGCTCATCACCACGAGACCTGTGGCCCTGGAGAAACTGCAGCACTTGCTGGACCATCCTCGGCATGTGGAGATCCTGGGTTTCTCCGAGGCCAAAAGGAAAGAGTACTTCTTCAAGTACTTCTCTGATGAGGCCCAAGCCAGGGCAGCCTTCAGTCTGATTCAGGAGAACGA GGTCCTCTTCACCATGTGCTTCATCCCCCTGGTCTGCTGGATCGTGTGCACTGGACTGAAACAGCAGATGGAGAGTGGCAAGAGCCTTGCCCAGACATCCAAGACCACCACCGCGGTGTACGTCTTCTTCCTTTCCAGTTTGCTGCAGCCCCGGGGAGGGAGCCAGGAGCACGGCCTCTGCGCCCACCTCTGGGGGCTCTGCTCTTTGGCTGCAGATGGAATCTGGAACCAGAAAATCCTGTTTG AGGAGTCCGACCTCAGGAATCATGGACTGCAGAAGGCGGATGTGTCTGCTTTCCTGAGGATGAACCTGTTCCAAAAGGAAGTGGACTGCGAGAAGTTCTACAGCTTCATCCACATGACTTTCCAGGAGTTCTTTGCCGCCATGTACTACCTGCTGGAAGAGGAAAAGGAAGGAAGGACGAACGTTCCAGGGAGTCGTT TGAAGCTTCCCAGCCGAGACGTGACAGTCCTTCTGGAAAACTATGGCAAATTCGAAAAGGGGTATTTGATTTTTGTTGTACGTTTCCTCTTTGGCCTGGTAAACCAGGAGAGGACCTCCTACTTGGAGAAGAAATTAAGTTGCAAGATCTCTCAGCAAATCAGGCTGGAGCTGCTGAAATGGATTGAAGTGAAAGCCAAAGCTAAAAAGCTGCAGATCCAGCCCAGCCAGCTGGAATTGTTCTACT GTTTGTACGAGATGCAGGAGGAGGACTTCGTGCAAAGGGCCATGGACTATTTCCCCAAGATTGAGATCAATCTCTCCACCAGAATGGACCACATGGTTTCTTCCTTTTGCATTGAGAACTGTCATCGGGTGGAGTCACTGTCCCTGGGGTTTCTCCATAACATGCCCAAGGAGGAAGAGGAGGAGGAAAAGGAAGGC CGACACCTTGATATGGTGCAGTGTGTCCTCCCAAGCTCCTCTCATGCTGCCTGTTCTCATGGATTGGTGAACAGCCACCTCACTTCCAGTTTTTGCCGGGGCCTCTTTTCAGTTCTGAGCACCAGCCAGAGTCTAACTGAATTGGACCTCAGTGACAATTCTCTGGGGGACCCAGGGATGAGAGTGTTGTGTGAAACGCTCCAGCATCCTGGCTGTAACATTCGGAGATTGTGGTTGGGG CGCTGTGGCCTCTCGCATGAGTGCTGCTTCGACATCTCCTTGGTCCTCAGCAGCAACCAGAAGCTGGTGGAGCTGGACCTGAGTGACAACGCCCTCGGTGACTTCGGAATCAGACTTCTGTGTGTGGGACTGAAGCACCTGTTGTGCAATCTGAAGAAGCTCTGGTTGGTCAGCTGCTGCCTCACATCAGCATGTTGTCAGGA TCTTGCATCAGTATTGAGCACCAGCCATTCCCTGACCAGACTCTATGTGGGGGAGAATGCCTTGGGAGACTCAGGAGTCGCAATTTTATGTGAAAAAGCCAAGAATCCACAGTGTAACCTGCAGAAACTGGGGTTGGTGAATTCTGGCCTTACGTCAGTCTGTTGTTCAGCTTTGTCCTCGTACTCAGCACTAATCAGAATCTCACGCACCTTTACCTGCGAGGCAACACTCTCGGAGACA AGGGGATCAAACTACTCTGTGAGGGACTCTTGCACCCCGACTGCAAGCTTCAGGTGTTGGAATTAGACAACTGCAACCTCACGTCACACTGCTGCTGGGATCTTTCCACACTTCTGACCTCCAGCCAGAGCCTGCGAAAGCTGAGCCTGGGCAACAATGACCTGGGCGACCTGGGGGTCATGATGTTCTGTGAAGTGCTGA AACAGCAGAGCTGCCTCCTGCAGAACCTGGGGTTGTCTGAAATGTATTTCAATTATGAGACAAAAAGTGCGTTAGAAACACTTCAAGAAGAAAAGCCTGAGCTGACCGTCGTCTTTTGAGCCTTCTTGGTAGGAGTGGAAACGGGGCTGCCAGACGCCAGTGTTCTCCGGTCCCTCCAGCTGGGGGCCCTCAGGTGGAGAGCTGCGATCCATCCAGGCCAAGACCACAGCTCTGTGATCC TTCCGGTGGAGTGTCGGAGAAGAGAGCTTGCCGACGATGCCTTCCTGTGCAGAGCTTGGGCATCTCCTTTACGCCAGGGTGAGGAAGACACCAGGACAATGACAGCATCGGGTGTTGTTGTCATCACAGCGCCTCAGTTAGAGGATGTTCCTCTTGGTGACCTCATGTAATTAGCTCATTCAATAAAGCACTTTCTTTTATTTT SEQ ID NO: 533 _{NM_001127461} GTTCCTGAGGCTGGCATCTGGATGAGGAAACTGAAGTTGAGGAATAGTGAAGAGTTTGTCCAATGTCATAGCCCCGTAATCAACGGGACAAAATTTTCTTGCTGATGGGTCAAGATGGCATCGTGAAGTGGTTGTTCACCGTAAACTGTAATACAATCCTGTTTATGGATTTGTTTGCATATTTTTCCCTCCATAGGGAAACCTTTTCTTCCATGGCTCAGGACACACTCCTGGATCGAGCCAACAG GAGA ACTTTCTGGTAAGCATTTGGCTAACTTTTTTTTTTTTGAGATGGAGTCTTGCTGTGTCGCCTAGGCTGGAGTGCAGTGGCGTGATCTTGGCTCACTGCAGCCTCCACTTCCCGGGTTCAATCAATTCTCCTACCTCAACTTCCTGAGTAGCTGGGATTACAGGCGCCCGCCACCACACCCGGCTCATTTTTGTACTTTTAGTAGAGACACAGTTTTGCCATGTTGGCCAGGCTGGTC TTGAATTCCTCAGC TCAGGTGATCTGCCTGCTTGGCCTCTCAAAGTGCTGGGATTACAGGCGTGAGCCACTGTGCCCGGCCTTGGCTAACTTTTCAAAATTAAAGATTTTGACTTGTTACAGTCATGTGACATTTTTTTCTTTCTGTTTGCTGAGTTTTTGATAATTTATATCTCTCAAAGTGGAGACTTTAAAAAAGACTCATCCGTGTGCCGTGTTCACTGCCTGGTATCTTAGTGTGGACCGAAGCCTAAGGA CCCTGAAA ACAGCTGCAGATGAAGATGGCAAGCACCCGCTGCAAGCTGGCCAGGTACCTGGAGGACCTGGAGGATGTGGACTTGAAGAAATTTAAGATGCACTTAGAGGACTATCCTCCCCAGAAGGGCTGCATCCCCCCTCCCGAGGGGTCAGACAGAAGGCAGACCATGTGGATCTAGCCACGCTAATGATCGACTTCAATGGGGAGGAGAAGGCGTGGGCCATGGCCGTGTGGATCTTCGCTGCGAT CAACAGGA GAGACCTTTATGAGAAAGCAAAAAGAGATGAGCCGAAGTGGGGTTCAGATAATGCACGTGTTTCGAATCCCACTGTGATATGCCAGGAAGACAGCATTGAAGAGGAGTGGATGGGTTTACTGGAGTACCTTTCGAGAATCTCTATTTGTAAAATGAAGAAAGATTACCGTAAGAAGTACAGAAAGTACGTGAGAAGCAGATTCCAGTGCATTGAAGACAGGAATGCCCGTCTGGGTGAGAGTG TGAGCCTC AACAAACGCTACACACGACTGCGTCTCATCAAGGAGCACCGGAGCCAGCAGGAGAGGGAGCAGGAGCTTCTGGCCATCGGCAAGACCAAGACGTGTGAGAGCCCCGTGAGTCCCATTAAGATGGAGTTGCTGTTTGACCCCGATGATGAGCATTCTGAGCCTGTGCACACCGTGGTGTTCCAGGGGGCGGCAGGGATTGGGAAAACAATCCTGGCCAGGAAGATGATGTTGGACTGGGCGTC GGGGACACT CTACCAAGACAGGTTTGACTATCTGTTCTATATCCACTGTCGAGAGGTGAGCCTTGTGACACAGAGGAGCCTGGGGGACCTGATCATGAGCTGCTGCCCCGACCCAAACCCACCCATCCACAAGATCGTGAGAAAACCCTCCAGAATCCTCTTCCTCATGGACGGCTTCGATGAGCTGCAAGGTGCCTTTGACGAGCACATAGGACCGCTCTGCACTGACTGGCAGAAGGCCGAGCGGGGAGACATTCTC C TGAGCAGCCTCATCAGAAAGAAGCTGCTTCCCGAGGCCTCTCTGCTCATCACCACGAGACCTGTGGCCCTGGAGAAACTGCAGCACTTGCTGGACCATCCTCGGCATGTGGAGATCCTGGGTTTCTCCGAGGCCAAAAGGAAAGAGTACTTCTTCAAGTACTTCTCTGATGAGGCCCAAGCCAGGGCAGCCTTCAGTCTGATTCAGGAGAACGAGGTCCTCTTCACCATGTGCTTCATCCCCCT GGTCTGC TGGATCGTGTGCACTGGACTGAAACAGCAGATGGAGAGTGGCAAGAGCCTTGCCCAGACATCCAAGACCACCACCGCGGTGTACGTCTTCTTCCTTTCCAGTTTGCTGCAGCCCCGGGGAGGGAGCCAGGAGCACGGCCTCTGCGCCCACCTCTGGGGGCTCTGCTCTTTGGCTGCAGATGGAATCTGGAACCAGAAATCCTGTTTGAGGAGTCCGACCTCAGGAATCATGGACTGCAGAAGGCGGA TGT GTCTGCTTTCCTGAGGATGAACCTGTTCCAAAAGGAAGTGGACTGCGAGAAGTTCTACAGCTTCATCCACATGACTTTCCAGGAGTTCTTTGCCGCCATGTACTACCTGCTGGAAGAGGAAAAGGAAGGAAGGACGAACGTTCCAGGGAGTCGTTTGAAGCTTCCCAGCCGAGACGTGACAGTCCTTCTGGAAAACTATGGCAAATTCGAAAAGGGGTATTTGATTTTTGTTGTACGTTTC CTCTTTGGCC TGGTAAACCAGGAGAGGACCTCCTACTTGGAGAAGAAATTAAGTTGCAAGATCTCTCAGCAAATCAGGCTGGAGCTGCTGAAATGGATTGAAGTGAAAGCCAAAGCTAAAAAGCTGCAGATCCAGCCCAGCCAGCTGGAATTGTTCTACTGTTTGTACGAGATGCAGGAGGAGGACTTCGTGCAAAGGGCCATGGACTATTTCCCCAAGATTGAGATCAATCTCTCCACCAGAATGGACCACATGGTT TCT TCCTTTTGCATTGAGAACTGTCATCGGGTGGAGTCACTGTCCCTGGGGTTTCTCCATAACATGCCCAAGGAGGAAGAGGAGGAGGAAAAGGAAGGCCGACACCTTGATATGGTGCAGTGTGTCCTCCCAAGCTCCCTCTCATGCTGCCTGTTCTCATGGATTGGTGAACAGCCACCTCACTTCCAGTTTTTGCCGGGGCCTCTTTTCAGTTCTGAGCACCAGCCAGAGTCTAACTGAATTGGA CCTCAGTGA CAATTCTCTGGGGGACCCAGGGATGAGAGTGTTGTGTGAAACGCTCCAGCATCCTGGCTGTAACATTCGGAGATTGTGGTTGGGGCGCTGTGGCCTCTCGCATGAGTGCTGCTTCGACATCTCCTTGGTCCTCAGCAGCAACCAGAAGCTGGTGGAGCTGGACCTGAGTGACAACGCCCTCGGTGACTTCGGAATCAGACTTCTGTGTGTGGGACTGAAGCACCTGTTGTGCAATCTGAAGA AGCTCTGGT TGGTGAATTCTGGCCTTACGTCAGTCTGTTGTTCAGCTTTGTCCTCGGTACTCAGCACTAATCAGAATCTCACGCACCTTTACCTGCGAGGCAACACTCTCGGAGACAAGGGGATCAAACTACTCTGTGAGGGACTCTTGCACCCCGACTGCAAGCTTCAGGTGTTGGAATTAGACAACTGCAACCTCACGTCACACTGCTGCTGGGATCTTTCCACACTTCTGACCTCCAGCCAGAGCCTGCG AAAGCTG AGCCTGGGCAACAATGACCTGGGCGACCTGGGGGTCATGATGTTCTGTGAAGTGCTGAAACAGCAGAGCTGCCTCCTGCAGAACCTGGGGTTGTCTGAAATGTATTTCAATTATGAGACAAAAAGTGCGTTAGAAACACTTCAAGAAGAAAAGCCTGAGCTGACCGTCGTCTTTTGAGCCTTCTTGGTAGGAGTGGAAACGGGGCTGCCAGACGCCAGTGTTCTCCGGTCCCTCCAGCTG GGGGCCCTCAGG TGGAGAGAGCTGCGATCCATCCAGGCCAAGACCACAGCTCTGTGATCCTTCCGGTGGAGTGTCGGAGAAGAGCTTGCCGACGATGCCTTCCTGTGCAGAGCTTGGGCATCTCCTTTACGCCAGGGTGAGGAAGACACCAGGACAATGACAGCATCGGGTGTTGTTGTCATCACAGCGCCTCAGTTAGAGGATGTTCCTCTTGGTGACCTCATGTAATTAGCTCATTCAATAAAGCACTT TCTTTATTTT SEQ ID NO: 534 _{NM_001127462} GTTCCTGAGGCTGGCATCTGGATGAGGAAACTGAAGTTGAGGAATAGTGAAGAGTTTGTCCAATGTCATAGCCCCGTAATCAACGGGACAAAAATTTTCTTGCTGATGGGTCAAGATGGCATCGTGAAGTGGTTGTTCACCGTAAACTGTAATACAATCCTGTTTATGGATTTGTTTGCATATTTTTCCCTCCATAGGGAAACCTTTTCTTCCATGGCTCAGGACACACTCCTGGATCGAGCCA ACAGGAGA ACTTTCTGGTAAGCATTTGGCTAACTTTTTTTTTTTTGAGATGGAGTCTTGCTGTGTCGCCTAGGCTGGAGTGCAGTGGCGTGATCTTGGCTCACTGCAGCCTCCACTTCCCGGGTTCAATCAATTCTCCTACCTCAACTTCCTGAGTAGCTGGGATTACAGGCGCCCGCCACCACACCCGGCTCATTTTTGTACTTTTAGTAGAGACACAGTTTTGCCATGTTGGCCAGGCTGGTC TTGAATTCCTCAGC TCAGGTGATCTGCCTGCTTGGCCTCTCAAAGTGCTGGGATTACAGGCGTGAGCCACTGTGCCCGGCCTTGGCTAACTTTTCAAAATTAAAGATTTTGACTTGTTACAGTCATGTGACATTTTTTTCTTTCTGTTTGCTGAGTTTTTGATAATTTATATCTCTCAAAGTGGAGACTTTAAAAAAGACTCATCCGTGTGCCGTGTTCACTGCCTGGTATCTTAGTGTGGACCGAAGCCTAAGGA CCCTGAAA ACAGCTGCAGATGAAGATGGCAAGCACCCGCTGCAAGCTGGCCAGGTACCTGGAGGACCTGGAGGATGTGGACTTGAAGAAATTTAAGATGCACTTAGAGGACTATCCTCCCCAGAAGGGCTGCATCCCCCCTCCCGAGGGGTCAGACAGAAGGCAGACCATGTGGATCTAGCCACGCTAATGATCGACTTCAATGGGGAGGAGAAGGCGTGGGCCATGGCCGTGTGGATCTTCGCTGCGAT CAACAGGA GAGACCTTTATGAGAAAGCAAAAAGAGATGAGCCGAAGTGGGGTTCAGATAATGCACGTGTTTCGAATCCCACTGTGATATGCCAGGAAGACAGCATTGAAGAGGAGTGGATGGGTTTACTGGAGTACCTTTCGAGAATCTCTATTTGTAAAATGAAGAAAGATTACCGTAAGAAGTACAGAAAGTACGTGAGAAGCAGATTCCAGTGCATTGAAGACAGGAATGCCCGTCTGGGTGAGAGTG TGAGCCTC AACAAACGCTACACACGACTGCGTCTCATCAAGGAGCACCGGAGCCAGCAGGAGAGGGAGCAGGAGCTTCTGGCCATCGGCAAGACCAAGACGTGTGAGAGCCCCGTGAGTCCCATTAAGATGGAGTTGCTGTTTGACCCCGATGATGAGCATTCTGAGCCTGTGCACACCGTGGTGTTCCAGGGGGCGGCAGGGATTGGGAAAACAATCCTGGCCAGGAAGATGATGTTGGACTGGGCGTC GGGGACACT CTACCAAGACAGGTTTGACTATCTGTTCTATATCCACTGTCGAGAGGTGAGCCTTGTGACACAGAGGAGCCTGGGGGACCTGATCATGAGCTGCTGCCCCGACCCAAACCCACCCATCCACAAGATCGTGAGAAAACCCTCCAGAATCCTCTTCCTCATGGACGGCTTCGATGAGCTGCAAGGTGCCTTTGACGAGCACATAGGACCGCTCTGCACTGACTGGCAGAAGGCCGAGCGGGGAGACATTCTC C TGAGCAGCCTCATCAGAAAGAAGCTGCTTCCCGAGGCCTCTCTGCTCATCACCACGAGACCTGTGGCCCTGGAGAAACTGCAGCACTTGCTGGACCATCCTCGGCATGTGGAGATCCTGGGTTTCTCCGAGGCCAAAAGGAAAGAGTACTTCTTCAAGTACTTCTCTGATGAGGCCCAAGCCAGGGCAGCCTTCAGTCTGATTCAGGAGAACGAGGTCCTCTTCACCATGTGCTTCATCCCCCT GGTCTGC TGGATCGTGTGCACTGGACTGAAACAGCAGATGGAGAGTGGCAAGAGCCTTGCCCAGACATCCAAGACCACCACCGCGGTGTACGTCTTCTTCCTTTCCAGTTTGCTGCAGCCCCGGGGAGGGAGCCAGGAGCACGGCCTCTGCGCCCACCTCTGGGGGCTCTGCTCTTTGGCTGCAGATGGAATCTGGAACCAGAAATCCTGTTTGAGGAGTCCGACCTCAGGAATCATGGACTGCAGAAGGCGGA TGT GTCTGCTTTCCTGAGGATGAACCTGTTCCAAAAGGAAGTGGACTGCGAGAAGTTCTACAGCTTCATCCACATGACTTTCCAGGAGTTCTTTGCCGCCATGTACTACCTGCTGGAAGAGGAAAAGGAAGGAAGGACGAACGTTCCAGGGAGTCGTTTGAAGCTTCCCAGCCGAGACGTGACAGTCCTTCTGGAAAACTATGGCAAATTCGAAAAGGGGTATTTGATTTTTGTTGTACGTTTC CTCTTTGGCC TGGTAAACCAGGAGAGGACCTCCTACTTGGAGAAGAAATTAAGTTGCAAGATCTCTCAGCAAATCAGGCTGGAGCTGCTGAAATGGATTGAAGTGAAAGCCAAAGCTAAAAAGCTGCAGATCCAGCCCAGCCAGCTGGAATTGTTCTACTGTTTGTACGAGATGCAGGAGGAGGACTTCGTGCAAAGGGCCATGGACTATTTCCCCAAGATTGAGATCAATCTCTCCACCAGAATGGACCACATGGTT TCT TCCTTTTGCATTGAGAACTGTCATCGGGTGGAGTCACTGTCCCTGGGGTTTCTCCATAACATGCCCAAGGAGGAAGAGGAGGAGGAAAAGGAAGGCCGACACCTTGATATGGTGCAGTGTGTCCTCCCAAGCTTCCTCTCATGCTGCCTGTTCTCATGGGTTGGGGCGCTGTGGCCTCTCGCATGAGTGCTGCTTCGACATCTCCTTGGTCCTCAGCAGCAACCAGAAGCTGGTGGAGCTGGA CCTGAGTGA CAACGCCCTCGGTGACTTCGGAATCAGACTTCTGTGTGTGGGACTGAAGCACCTGTTGTGCAATCTGAAGAAGCTCTGGTTGGTCAGCTGCTGCCTCACATCAGCATGTTGTCAGGATCTTGCATCAGTATTGAGCACCAGCCATTCCCTGACCAGACTCTATGTGGGGGAGAATGCCTTGGGAGACTCAGGAGTCGCAATTTTATGTGAAAAAGCCAAGAATCCACAGTGTAACCTGCAGAA ACTGGGGT TGGTGAATTCTGGCCTTACGTCAGTCTGTTGTTCAGCTTTGTCCTCGGTACTCAGCACTAATCAGAATCTCACGCACCTTTACCTGCGAGGCAACACTCTCGGAGACAAGGGGATCAAACTACTCTGTGAGGGACTCTTGCACCCCGACTGCAAGCTTCAGGTGTTGGAATTAGACAACTGCAACCTCACGTCACACTGCTGCTGGGATCTTTCCACACTTCTGACCTCCAGCCAGAGCCTGCG AAAGCTG AGCCTGGGCAACAATGACCTGGGCGACCTGGGGGTCATGATGTTCTGTGAAGTGCTGAAACAGCAGAGCTGCCTCCTGCAGAACCTGGGGTTGTCTGAAATGTATTTCAATTATGAGACAAAAAGTGCGTTAGAAACACTTCAAGAAGAAAAGCCTGAGCTGACCGTCGTCTTTTGAGCCTTCTTGGTAGGAGTGGAAACGGGGCTGCCAGACGCCAGTGTTCTCCGGTCCCTCCAGCTG GGGGCCCTCAGG TGGAGAGAGCTGCGATCCATCCAGGCCAAGACCACAGCTCTGTGATCCTTCCGGTGGAGTGTCGGAGAAGAGCTTGCCGACGATGCCTTCCTGTGCAGAGCTTGGGCATCTCCTTTACGCCAGGGTGAGGAAGACACCAGGACAATGACAGCATCGGGTGTTGTTGTCATCACAGCGCCTCAGTTAGAGGATGTTCCTCTTGGTGACCTCATGTAATTAGCTCATTCAATAAAGCACTT TCTTTATTTT SEQ ID NO: 535-537 SEQ ID NO: 538 Caspase-1 substrate Asn-Glu-Ala-Tyr-Val-His-Asp-Ala-Pro-Val-Arg-Ser-Leu-Asn SEQ ID NO: 539 NLRP3 protein corresponding to NM-004895.1 MKMASTRCKLARYLEDVDLKKFKMHLEDYPPQKGCIPLPRGQTEKADHVDLATLMIDFNGEEKAWAMAVWIFAAINRRDLYEKAKRDEPKWGSDNARVSNPTVICQEDSIEEEWMGLLEYLSRISICKMKKDYRKKYRKYVRSRFQCIEDRNARLGESVSLNKRYTRLRLIKEHRSQQEREQELLAIGKTKTCESPVSPI KMELLFDPDDEHSEPVHTVVFQGAAGIGKTILARKMMLDWASGTLYQDRFDYLFYI HCREVSLVTQRSLGDLIMSCCPDPNPPIHKIVRKPSRILFLMDGFDELQGAFDEHIGPLCTDWQKAERGDILLSSLIRKKLLPEASLLITTRPVALEKLQHLLDHPRHVEILGFSEAKRKEYFFKYFSDEAQARAAFSLIQENEVLFTMCFIPLVCWIVCTGLKQQMESGKSLAQTSKTTTAVYVFFLSSLLQPRGGS QEHGLCAHLWGLCSLAADGIWNQKILFEESDLRNHGLQKADVSAFLRMNLFQKEVDCEKFY SFIHMTFQEFFAAMYYLLEEEKEGRTNVPGSRLKLPSRDVTVLLENYGKFEKGYLIFVVRFLFGLVNQERTSYLEKKLSCKISQQIRLELLKWIEVKAKAKKLQIQPSQLELFYCLYEMQEEDFVQRAMDYFPKIEINLSTRMDHMVSSFCIENCHRVESLSLGFLHNMPKEEEEEEKEGRHLDMVQCVLPSSSHA ACSHGLVNSHLTSSFFCRGLFSVLSTSQSLTELDLSDNSLGDPGMRVLCETLQHPGCNIRRLWL GRCGLSHECCFDISLVLSSNQKLVELDLSDNALGDFGIRLLCVGLKHLLCNLKKLWLVSCCLTSACCQDLASVLSTSHSLTRLYVGENALGDSGVAILCEKAKNPQCNLQKLGLVNSGLTSVCCSALSSVLSTNQNLTHLYLRGNTLGDKGIKLLCEGLLHPDCKLQVLELDNCNLTSHCCWDLSTLLTSSQSLRKLSLG NNDLGDLGVMMFCEVLKQQSCLLQNLGLSEMYFNYETKSALETLQEEKPELTVVFEPSW SEQ ID NO: 540 NLRP3 inflammasome inhibitor GUGCAUUGAAGACAGGAAUTT SEQ ID NO:541 GGCTGTAACATTCGGAGATTG SEQ ID NO:542 TCATCATTCCCGCTATCTTTC SEQ ID NO: 543 CCGTAAGAAGTACAGAAAGTA SEQ ID NO:544 GAGACTCAGGAGTCGCAATTT SEQ ID NO: 545 CCTCATGTAATTAGCTCATTC SEQ ID NO:546 GTGGATCTAGCCACGCTAATG SEQ ID NO: 547 CCACAGTGTAACCTGCAGAAA SEQ ID NO: 548 CCAGCCAGAGTCTAACTGAAT SEQ ID NO: 549 GCGTTAGAAACACTTCAAGAA SEQ ID NO: 550 GCTGGAATTGTTCTACTGTTT SEQ ID NO: 551 CCACATGACTTTCCAGGAGTT SEQ ID NOs: 552-586 See Table 5B in the description SEQ ID NO:587 Mature miR-9 (MIMAT0000441) UCU UUG GUU AUC U AG CUG UAU GA SEQ ID NO:588 hsa-miR-9-5p UCUUUGGUUAUCUAGCUGUAUGA SEQ ID NO: 589 miR-223 TGGGGTATTTGACAAACTGACA SEQ ID NO: 590 cbn-mir-233 MI0024890 UCGCCCAUCCCGUUGUUCCAAUAUUCCAACAACAAGUGAUUAUUGAGCAAUGCGCAUGUGCGG SEQ ID NO:591 cbr-mir-233 MI0000530 AAGCAUUUUUCUGUCCCGCGCAUCCCUUUGUUCCAAUAUUCAAACCAGUAGAAAGAUUAUUGAGCAAUGCGCAUGUGCGGGACAGAUUGAAUAGCUG SEQ ID NO: 592 cel-mir-233 MI0000308 AUAUAGCAUCUUUCUGUCUCGCCCAUCCCGUUGCUCCAAUAUUCUAACAACAAGUGAUUAUUGAGCAAUGCGCAUGUGCGGGAUAGACUGAUGGCUGC SEQ ID NO: 593 crm-mir-233 MI0011059 UGAAGCGUCUCUCUGUCCCGCUCAUCCUGUUGUUCCAAUAUUCCAACAGCCCAGUGAUUAUUGAGCAAUGCGCAUGUGCGGGACAGAUUGUAUGCUGCCAU SEQ ID NO: 594 hsa-miR-22-5p MIMAT000449 AGUUCUUCAGUGGCAAGCUUUA SEQ ID NO: 595 hsa-mir-22 MI0000078 GGCUGAGCCGCAGUAGUUCUUCAGUGGCAAGCUUUAUGUCCUGACCCAGCUAAAGCUGCCAGUUGAAGAACUGUUGCCCUCUGCC SEQ ID NO: 596 mmu-miR-33-5p or MIMAT0000667 GUGCAUUGUAGUUGCAUUGCA SEQ ID NO: 597 mmu-mir-33 MI0000707 CUGUGGUGCAUUGUAGUUGCAUUGCAUGUUCUGGCAAUACCUGUGCAAUGUUUCCACAGUGCAUCACGG SEQ ID NO: 598 AIM2 (NP_004824.1) MESKYKEILLLTGLDNITDEELDRFKFFLSDEFNIATGKLHTANRIQVATLMIQNAGAVSAVMKTIRIFQKLNYMLLAKRLQEEKEKVDKQYKSVTKPKPLSQAEMSPAASAAIRNDVAKQRAAPKVSPHVKPEQKQMVAQQESIREGFQKRCLPVMVLKAKKPFTFETQEGKQEMFHATVATEKEFFF VKVFNTLLKDKFIPKRIIIIARYYRHSGFLEVNSASRVLDAESDQKVNVPLNIIRKAGETPKINTLQTQPLGTIVNGLFVVQKVTEKKKNILFDLSDNTGKMEVLGVRNEDTMKCKEGDKVRLTFFTLSKNGEKLQLTSGVHSTIKVIKAKKKT SEQ ID NO: 599 SEQ ID NO:600 Aim 2 people (NM_004833.2) ATAGACATTTTCTTCTGTGGCTGCTAGTGAGAACCCAAACCAGCTCAGCCAATTAGAGCTCCAGTTGTCACTCCTACCCACACTGGGCCTGGGGGTGAAGGGAAGTGTTATTAGGGGTACATGTGAAGCCGTCCAGAAGTGTCAGAGTCTTTGTAGCTTTGAAAGTCACCTAGGTTATTTGGGCATGCTCTCCTGAGTCCTCTGCTAGTTAAGCTCTCTGAAAGAAGGTGGCAGAC CCGGTTTGCTGATCGCCCCAGGGATCAGGAGGCTGATCCCAAAGTTGTCAGATGGAGAGTAAATACAAGGAGATACTCTTGCTAACAGGCCTGGATAACATCACTGATGAGGAACTGGATAGGTTTAAGTTCTTTCTTTCAGACGAGTTTA ATATTGCCACAGGCAAACTACATACTGCAAACAGAATACAAGTAGCTACCTTGATGATTCAAAATGCTGGGGCGGTGTCTGCAGTGATGAAGACCATTCGTATTTTTCAGAAGTTGAATTATATGCTTTTGGCAAAACGTCTTCAGGAGGAGAAGGAGAAAGTTGATAAGCAATACAAATCGGTAACAAAACCAAAGCCACTAAGTCAAGCTGAAATGAGTCCTGCTGCATCTGCAGCCATCAGAAATG ATGTCGCAAAGCAACGTGCTGCACCAAAAGTCTCTCCTCATGTTAAGCCTGAACAGAAACAGATGGTGGCCCAGCAGGAATCTATCAGAGAAGGGTTTCAGAAGCGCTGTTTGCCAGTTATGGTACTGAAAGCAAAGAAGC CCTTCACGTTTGAGACCCAAGAAGGCAAGCAGGAGATGTTTCATGCTACAGTGGCTACAGAAAAGGAATTCTTCTTTGTAAAAGTTTTTAATACACTGCTGAAAGATAAATTCATTCCAAAGAGAATAATTATAATAGCAAGATATTATCGGCACAGTGGTTTCTTAGAGGTAAATAGCGCCTCACGTGTGTTAGATGCTGAATCTGACCAAAAGGTTAATGTCCCGCTGAACATTATCAGAAAAGCTGG TGAAACCCCGAAGATCAACACGCTTCAAACTCAGCCCCTTGGAACAATTGTGAATGGTTTGTTTGTAGTCCAGAAGGTAACAGAAAAGAAGAAAAACATATTATTTGACCTAAGTGACAACACTGGGAAAATGGAAGTA CTGGGGGTTAGAAACGAGGACACAATGAAATGTAAGGAAGGAGATAAGGTTCGACTTACATTCTTCACACTGTCAAAAAATGGAGAAAAACTACAGCTGACATCTGGAGTTCATAGCACCATAAAGGTTATTAAGGCCAAAAAAAAAACATAGAGAAGTAAAAAGGACCAATTCAAGCCAACTGGTCTAAGCAGCATTTAATTGAAGAATATGTGATACAGCCTCTTCAATCAGATTGTAAGTTACCTGAAAGCTGC AGTTCACAGGCTCCTCTCTCCACCAAATTAGGATAGAATAATTGCTGGATAAACAAATTCAGAATATCAACAGATGATCACAATAAACATCTGTTTCTCATTCAAAAAAAAAAAAAAAAAAAAAAAAAAA SEQ ID NO:601 AIM2 inflammasome inhibitor CCCGAAGATCAACACGCTTCA SEQ ID NO:602 A151 TTAGGGTTAGGGTTAGGGTTAGGG SEQ ID NO:603 C151 TTCAAATTCAAATTCAAATTCAAA SEQ ID NO:604 TTATGGG SEQ ID NO: 605-610 See Table 5C in the description SEQ ID NO:611 NM_033292.3 ATACTTTCAGTTTCAGTCACACAAGAAGGGAGGAGAGAAAAGCCATGGCCGACAAGGTCCTGAAGGAGAAGAGAAAGCTGTTTATCCGTTCCATGGGTGAAGGTACAATAAATGGCTTACTGGATGAATTATTACAGACAAGGGTGCTGAACAAGGAAGAGATGGAGAAAGTAAAACGTGAAAATGCTACAGTTATGGATAAGACCCGAGCTTTGATTGACTCCGTTATTCCGAAAGGGGCACAGG CATGCCAAATTTGCATCACATACATTTGTGAAGAAGACAGTTACCTGGCAGGGACGCTGGGACTCTCAGCAGATCAAACATCTGGAAATTACCTTAATATGC AAGACTCTCAAGGAGTACTTTCTTCCTTTCCAGCTCCTCAGGCAGTGCAGGACAACCCAGCTATGCCCACATCCTCAGGCTCAGAAGGGAATGTCAAGCTTTGCTCCCTAGAAGAAGCTCAAAGGATATGGAAACAAAAGTCGGCAGAGATTTATCCAATAATGGACAAGTCAAGCCGCACACGTCTTGCTCTCATTATCTGCAATGAAGAATTTGACAGTATTCCTAGAAGAACTGGAGCTGA GGTTGACATCACAGGCATGACAATGCTGCTACAAAATCTGGGGTACAGCGTAGATGTGAAAAAAAATCTCACTGCTTCGGACATGACTACAGAGCTGGAGGCAT TTGCACACCGCCCAGAGCACAAGACCTCTGACAGCACGTTCCTGGTGTTCATGTCTCATGGTATTCGGGAAGGCATTTGTGGGAAGAAACACTCTGAGCAAGTCCCAGATATACTACAACTCAATGCAATCTTTAACATGTTGAATACCAAGAACTGCCCAAGTTTGAAGGACAAACCGAAGGTGATCATCATCCAGGCCTGCCGTGGTGACAGCCCTGGTGTGGTGTGGTTTTAAAGAT TCAGTAGGAGTTTCTGGAAACCTATCTTTACCAACTACAGAAGAGTTTTGAGGATGATGCTATTAAGAAAGCCCACATAGAGAAGGATTTTATCGCTTTCTGCTCTTCCA CACCAGATAATGTTTCTTGGAGACATCCCACAATGGGCTCTGTTTTTATTGGAAGACTCATTGAACATATGCAAGAATATGCCTGTTCCTGTGATGTGGAGGAAATTTTCCGCAAGGTTCGATTTTCATTTGAGCAGCCAGATGGTAGAGCGCAGATGCCCACCACTGAAAGAGTGACTTTGACAAGATGTTTCTACCTCTTCCAGGACATTAAAATAAGGAAACTGTATGAATGTCTGTGGGCA GGAAGTGAAGAGATCCTTCTGTAAAGGTTTTTGGAATTATGTCTGCTGAATAATAAACTTTTTTGAAATAATAAATCTGGTAGAAAAATGAAAAAAAAAAAAA SEQ ID NO:612 NP_150634.1 human caspase-1 protein MADKVLKEKRKLFIRSMGEGTINGLLDELLQTRVLNKEEMEKVKRENATVMDKTRALIDSVIPKGAQACQICITYICEEDSYLAGTLGLSADQTSGNYLNMQDSQGVLSSFPAPQAVQDNPAMPTSSGSEGNVKLCSLEEAQRIWKQKSAEIYPIMDKSSRTRLALIICNEEFDSIPRRTGAEVDITGMTMLLQNLGYSV DVKKNLTASDMTTELEAFAHRPEHKTSDSTFLVFMSHGIREGICGKKHSEQVPDILQLNAIFNMLNTKNCPSLKDKPKVIIIQACRGDSPGVVWFKDSVG VSGNLSLPTTEEFEDDAIKKAHIEKDFIAFCSSTPDNVSWRHPTMGSVFIGRLIEHMQEYACSCDVEEIFRKVRFSFEQPDGRAQMPTTERVTLTRCFYL FPGH SEQ ID NO: 613-619 See Table 5E in the description SEQ ID NO: 620-664 See Table 5F in the description SEQ ID NO:665 AIM2 inflammasome inhibitor SEQ ID NO: 666-803 See Table 5D in the description SEQ ID NO:803 RBS sequence GCGCGCTCGCTCGCTC° SEQ ID NO:804 TRS Sequence GGTTGA SEQ ID NO: 805 TRS Sequence AGTT SEQ ID NO:806 TRS Sequence GGTTGG SEQ ID NO:807 TRS Sequence AGTTTGG SEQ ID NO: 808 TRS Sequence AGTTGA SEQ ID NO: 809 Another motive RRR ... SEQ ID NO: 882 ORF52 protein of Kaposi's sarcoma-associated herpesvirus MAAPRGRPKKDLTMEDLTAKISQLTVENRELRKALGSTADPRDRPLTATEKEAQLTATVGALSAAAAKKIEARVRTIFSKVVTQKQVDDALKGLSLRIDVCMSDGGTAKPPPGANNRRRRGASTTRAGVDD SEQ ID NO:883 Cytoplasmic isoform of LANA (ORF73) MAPPGMRLRSGRSTGAPLTRGSCRKRNRSPERCDLGDDLHLQPRRKHVADSVDGRECGPHTLPIPGSPTVFTSGLPAFVSSPTLPVAPIPSPAPATPLPPPALLPPVTTSSSPIPPSHPVSPGTTDTHSPSPALPPTQSPESSQRPPLSSPTGRPDSSTPMRPPPSQQTTPPHSPTTPPPEPPSKSSPDSLAPSTLRSLRKRRLSSPQGPSTLNPICQSPPVSP PRCDFANRSVYPPWATESPIYVGSSSDGDTPPRQPPTSPISIGSSSPSEGSWGDDTAM LVLLAEIAEEASKNEKECSENNQAGEDNGDNEISKESQVDKDDNDNKDDEEEQETDEEDEEEDDEEDDEEDDEEDDEEDDEEDDEEDDEEEDEEEDEEEDEEEDEEEEEEDDDDEDNEDEEDDEEEDKKEDEEDGGDGNKTLSIQSSQQQQEPQQQEPQQQEPQQQEPQQQEPQQQEPQQQEPQQQEPQQREP QQREPQQREPQQREPQQREPQQREPQQREPQQREPQQREPQQREPQQREPQQREPQQQEPQQQEPQQQEPQQQEPQQQEPQQQEPQQQ EPQQQEPQQQEPQQQEPQQQEPQQQEPQQQDEQQQDEQQQDEQQQDEQQQDEQQQDEQQQDEQQQDEQEQQDEQQQDEQQQQDEQEQQEEQEQQEEQQQDEQQQDEQQQDEQQQDEQEQQDEQQQDEQQQQDEQEQQEEQEQQEEQEQQEE QEQQEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQELEEQEQEQELEEVEEQEQEQEEQELEEVEE QEQEQEEQEEQELEEVEEQEEQELEEVEEQEEQELEEVEEQEQQGVEQQEQETVEEPIILHGSSSEDEMEVDYPVVSTHEQIASSPPGDNTPDDDPQPGPSREYRYVLRTSPPHRPGVRMRRVPVTHPKKPHPRYQQPPVPYRQIDDCPAKARPQHIFYRRFLGKDGRRDPKCQWKFAVIFWGNDPY GLKKLSQAFQFGGVKAGPVSCLPHPGPDQSPITYCVYVYCQNKDTSKKVQMARLAWEASHPLAGNLQSSIVKFKKPLPLTQPGENQGPGDSPQEMT SEQ ID NO: 884 Truncated cytoplasmic isoform of LANA (ORF73) MAPPGMRLRSGRSTGAPLTRGSCRKRNRSPERCDLGDDLHLQPRRKHVADSVDGRECGPHTLPIPGSPTVFTSGLPAFVSSPTLPVAPIPSPAPATPLPPPALLPPVTTSSSPIPPSHPVSPGTTDTHSPSPALPPTQSPESSQRPPLSSPTGRPDSSTP SEQ ID NO: 885 miR-25 GGCCAGTGTTGAGAGGCGGAGACTTGGGCAATTGCTGGACGCTGCCCTGGGCATTGCACTTGTCTCGGTCTGACAGTGCCGGCC SEQ ID NO: 886 miR-93 CTGGGGGCTCCAAAGTGCTGTTCGTGCAGGTAGTGTGATTACCCAACCTACTGCTGAGCTAGCACTTCCCGAGCCCCCGG SEQ ID NO: 887 TLR9 inhibitory oligonucleotide 5'-CCTN(3-5)G(3-5)RR-3' SEQ ID NO: 888 TLR9 inhibitory oligonucleotide TTATGGGn SEQ ID NO: 889 ODN-2088 TCCTGGCGGGGAAGT SEQ ID NO: 890 ODN-2114 TCCTGGAGGGGAAGT SEQ ID NO:891 poly(G) GGGGGGGGGGGGGGGGGGGG SEQ ID NO:892 ODN-A151 TTAGGGTTAGGGTTAGGGTTAGGG SEQ ID NO:893 G-ODN CTCCTATTGGGGGTTTCCTAT SEQ ID NO:894 IRS-869 TCCTGGAGGGGTTGT SEQ ID NO:895 INH-1 CCTGGATGGGAATTCCCATCCAGG SEQ ID NO:896 INH-4 TTCCCATCCAGGCCTGGATGGGAA SEQ ID NO:897 IRS-661 TGCTTGCAAGCTTGCAAGCA SEQ ID NO:898 4024 TCCTGGATGGGAAGT SEQ ID NO:899 4084F CCTGGATGGGAA SEQ ID NO:900 INH-13 CTTACCGCTGCACCTGGATGGGAA SEQ ID NO:901 INH-18 CCTGGATGGGAACTTACCGCTGCA SEQ ID NO:902 IRS-954 TGCTCCTGGAGGGGTTGT SEQ ID NO:903 AS1411 GGTGGTGGTGGTTGTGGTGGTGGTGG SEQ ID NO:904 Caspase-1 inhibitor GAA GGC CCA UAU AGA GAA A SEQ ID NO:905 AAB1 5' WT-ITR (LEFT) TTGCCCACTCCCTCTCTGCGCGCTCGCTCGCTCGGTGGGGCCTGCGGACCAAAGGTCCGCAGACGGCAGAGGTCTCCTCTGCCGGCCCCACCGAGCGAGCGACGCGCGCAGAGAGGGAGTGGGCAACTCCATCACTAGGGTAA SEQ ID NO:906 AAB1 3' WT-ITR (RIGHT) TTACCCTAGTGATGGAGTTGCCCACTCCCTCTCTGCGCGCGTCGCTCGCTCGGTGGGGCCGGCAGAGGAGACCTCTGCCGTCTGCGGACCTTTGGTCCGCAGGCCCCACCGAGCGAGCGAGCGCGCAGAGAGGGAGTGGGCAA SEQ ID NO:907 AAB2 5' WT-ITR (LEFT) CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT SEQ ID NO:908 AAB2 3' WT-ITR (RIGHT) AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGctgcctgcagg SEQ ID NO: 909 AAB3 5' WT-ITR (LEFT) TTGGCCACTCCCTCTATGCGCACTCGCTCGCTCGGTGGGGCCTGGCGACCAAAGGTCGCCAGACGGACGTGGGTTTCCACGTCCGGCCCCACCGAGCGAGCGAGTGCGCATAGAGGGAGTGGCCAACTCCATCACTAGAGGTAT SEQ ID NO:910 AAB3 3' WT-ITR (RIGHT) ATACCTCTAGTGATGGAGTTGGCCACTCCCTCTATGCGCACTCGCTCGCTCGGTGGGGCCGGACGTGGAAACCCACGTCCGTCTGGCGACCTTTGGTCGCCAGGCCCCACCGAGCGAGCGAGTGCGCATAGAGGGAGTGGCCAA SEQ ID NO:911 AAB4 5' WT-ITR (LEFT) TTGGCCACTCCCTCTATGCGCGCTCGCTCACTCACTCGGCCCTGGAGACCAAAGGTCTCCAGACTGCCGGCCTCTGGCCGGCAGGGCCGAGTGAGTGAGCGAGCGCGCATAGAGGGAGTGGCCAACT SEQ ID NO:912 AAB4 3' WT-ITR (RIGHT) AGTTGGCCACATTAGCTATGCGCGCTCGCTCACTCACTCGGCCCTGGAGACCAAAGGTCTCCAGACTGCCGGCCTCTGGCCGGCAGGGCCGAGTGAGTGAGCGAGCGCGCATAGAGGGAGTGGCCAA SEQ ID NO:913 AAB5 5' WT-ITR (LEFT) TCCCCCCTGTCGCGTTCGCTCGCTCGCTGGCTCGTTTGGGGGGGCGACGGCCAGAGGGCCGTCGTCTGGCAGCTCTTTGAGCTGCCACCCCCCCAAACGAGCCAGCGAGCGAGCGAACGCGACAGGGGGGAGAGTGCCACACTCTCAAGCAAGGGGGTTTTGTAAG SEQ ID NO:914 AAB5 3' WT-ITR (RIGHT) CTTACAAAACCCCCTTGCTTGAGAGTGTGGCACTCTCCCCCCTGTCGCGTTCGCTCGCTCGCTGGCTCGTTTGGGGGGGTGGCAGCTCAAAGAGCTGCCAGACGACGGCCCTCTGGCCGTCGCCCCCCCAAACGAGCCAGCGAGCGAGCGAACGCGACAGGGGGGA SEQ ID NO:915 AAB6 5' WT-ITR (LEFT) TTGCCCACTCCCTTCTAATGCGCGCTCGCTCGCTCGGTGGGGCCTGCGGACCAAAGGTCCGCAGACGCAGAGGTCTCCTCTGCCGGCCCCACCGAGCGAGCGAGCGCGCATAGAGGGAGTGGGCAACTCCATCACTAGGGGTAT SEQ ID NO:916 AAB6 3' WT-ITR (RIGHT) ATACCCCTAGTGATGGAGTTGCCCACTCCCTCTATGCGCGCTCGCTCGCTCGGTGGGGCCGGCAGAGGAGACCTCTGCCGTCTGCGGACCTTTGGTCCGCAGGCCCCACCGAGCGAGCGAGCGCGCATTAGAGGGAGTGGGCAA

Name Subsequence SEQ ID NO: ITR-18 right AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCGCACGCCCGGGTTTCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG 917 ITR-19 right AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG 918 ITR-20 right AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG 919 ITR-21 right AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCTTTGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG 920 ITR-22 right AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACAAAGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG 921 ITR-23 right AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGAAAATCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG 922 ITR-24 right AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGAAACGCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG 923 ITR-25 right AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCAAAGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG 924 ITR-26 right AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGTTTCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG 925 ITR-27 right AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGTTTCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG 926 ITR-28 right AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGTTTCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG 927 ITR-29 right AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCTTTGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG 928 ITR-30 right AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCTTTGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG 929 ITR-31 right AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCTTTGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG 930 ITR-32 right AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGTTTCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG 931 ITR-49 right AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGGCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG 932 ITR-50 right AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG 933 ITR-33 left CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGGAAACCCGGGCGTGCGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT 934 ITR-34 left CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT 935 ITR-35 left CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT 936 ITR-36 left CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT 937 ITR-37 left CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCAAAGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT 938 ITR-38 left CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACTTTGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT 939 ITR-39 left CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGATTTTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT 940 ITR-40 left CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGTTTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT 941 ITR-41 left CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCTTTGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT 942 ITR-42 left CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGGAAACCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT 943 ITR-43 left CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGAAACCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT 944 ITR-44 left CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGAAACGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT 945 ITR-45 left CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCAAAGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT 946 ITR-46 left CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCAAAGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT 947 ITR-47 left CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCAAAGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT 948 ITR-48 left CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGAAACGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT 949 VH-02 MDWTWRILFLVAAATGAHS 950 VK-A26 MLPSQLIGFLLLWVPASRG 951

Sources

1. Intracellular Nucleic Acid Detection in Autoimmunity. J.T. Crowl et al., Annu. Rev. Immunol. 2017, 35: 313-336

2. Innate Immune Recognition of DNA: a recent history. A. Dempsey and A.G. Bowie, Virology, May 2015, 0: 146-152

3. Cutting Edge: Antimalarial Drugs Inhibit IFN-β Production through Blockade of Cyclic GMP-AMP Synthase-DNA Interaction. J. An et al., J. Immunol. March 27, 2015

4. Small molecule inhibition of cGAS reduces interferon expression in primary macrophages from autoimmune mice. J. Vincent et al., Nature Communications, 8:750

5. Discovery of PF-06928215 as a high affinity inhibitor of cGAS enabled by a novel fluorescence polarization assay. PLOS ONE. September 21, 2017

6. The functional interactome of PYHIN immune regulators reveals IFIX is a sensor of viral DNA. B.A. Diner et al., Molecular Systems Biology, 2015, 11:787

7. Activation and Regulation of DNA-Driven Immune Responses. S.R. Paluden. Microbiology and Molecular Biology Reviews. 2015. 79(2): 225

8. Viral Evasion of DNA-stimulated innate immune responses. M.H. Christnesen and S.R. Paluden. Cellular and Molecular Immunology. 2017. 14:4-13

9. Sequence-specific activation of the DNA sensor cGAS by Y-form DNA structures as found in primary HIV-1 cDNA. A-M Herzner et al., 2015. Nature Immunology

10. RNA Polymerase III Detects Cytosolic DNA and Induces Type-I Interferons Through the RIG-I Pathway. Y-H Chiu et al. 2009. Cell. 138(3): 576-591

11. Mi-R-25/93 mediates hypoxia-induced immunosuppression by repressing cGAS. M.Z. Wu et al. 2017. Nat. Cell Biol. 19(10):1286-1296

12. Cytoplasmic isoforms of Kaposi sarcoma herpesvirus LANA recruit and antagonize the innate immune DNA sensor cGAS. Zhang G. et al., Proc Natl Acad Sci U S A., 23 Feb 2016;113(8):E1034-43.

13. Kaposi´s sarcoma-associated herpesvirus inhibitor of cGAS (KicGAS) Encoded by ORF52, is an Abundant Tegument protein and Is Required for Production of Infectious Progeny Viruses. W. Li, et al., J. Virol. 2016, 90(11): 5329.

--->

SEQUENCE LIST

<110> GENERATION BIO CO.

<120> Closed-end DNA (ccDNA) and its application in methods for reducing

immune response associated with gene therapy or therapy

nucleic acid

<130> 131698-03320

<140> PCT/US2020/015026

<141> 2020-01-24

<150> 62/857,542

<151> 2019-06-05

<150> 62/814,424

<151> 2019-03-06

<150> 62/814,414

<151> 2019-03-06

<150> 62/800,303

<151> 2019-02-01

<150> 62/800,285

<151> 2019-02-01

<150> 62/796,450

<151> 2019-01-24

<150> 62/796,417

<151> 2019-01-24

<160> 1220

<170> PatentIn version 3.5

<210> 1

<211> 141

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc 120

gagcgcgcag ctgcctgcag g 141

<210> 2

<211> 130

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 2

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc 120

tgcctgcagg 130

<210> 3

<211> 1923

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 3

tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta 60

ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc 120

aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg 180

gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 240

gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 300

agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 360

ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga 420

cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg 480

gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg 540

cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta 600

attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgcgccag gcggggcggg 660

gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg 720

cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg 780

aagcgcgcgg cgggcgggag tcgctgcgac gctgccttcg ccccgtgccc cgctccgccg 840

ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc 900

gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt 960

ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc gggggggagc 1020

ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg cccgcgctgc 1080

ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg 1140

aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag 1200

gctgcgtgcg gggtgtgtgc gtggggggggt gagcaggggg tgtgggcgcg gcggtcgggc 1260

tgtaaccccc ccctgcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg 1320

gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg 1380

ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga ggggcgcggc 1440

ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg 1500

taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct 1560

gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg cgccggcagg 1620

aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc 1680

cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg 1740

ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tttagccttc 1800

ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca tcatttgtcg 1860

acagaattcc tcgaagatcc gaaggggttc aagcttggca ttccggtact gttggtaaag 1920

cca 1923

<210> 4

<211> 1272

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 4

aggctcagag gcacacagga gtttctgggc tcaccctgcc cccttccaac ccctcagttc 60

ccatcctcca gcagctgttt gtgtgctgcc tctgaagtcc acactgaaca aacttcagcc 120

tactcatgtc cctaaaatgg gcaaacattg caagcagcaa acagcaaaca cacagccctc 180

cctgcctgct gaccttggag ctggggcaga ggtcagagac ctctctgggc ccatgccacc 240

tccaacatcc actcgacccc ttggaatttc ggtggagagg agcagaggtt gtcctggcgt 300

ggtttaggta gtgtgagagg gtccgggttc aaaaccactt gctgggtggg gagtcgtcag 360

taagtggcta tgccccgacc ccgaagcctg tttccccatc tgtacaatgg aaatgataaa 420

gacgcccatc tgatagggtt tttgtggcaa ataaacattt ggtttttttg ttttgttttg 480

ttttgttttt tgagatggag gtttgctctg tcgcccaggc tggagtgcag tgacacaatc 540

tcatctcacc acaaccttcc cctgcctcag cctcccaagt agctgggatt acaagcatgt 600

gccaccacac ctggctaatt ttctattttt agtagagacg ggtttctcca tgttggtcag 660

cctcagcctc ccaagtaact gggattacag gcctgtgcca ccacacccgg ctaatttttt 720

ctatttttga cagggacggg gtttcaccat gttggtcagg ctggtctaga ggtaccggat 780

cttgctacca gtggaacagc cactaaggat tctgcagtga gagcagaggg ccagctaagt 840

ggtactctcc cagagactgt ctgactcacg ccaccccctc caccttggac acaggacgct 900

gtggtttctg agccaggtac aatgactcct ttcggtaagt gcagtggaag ctgtacactg 960

cccaggcaaa gcgtccgggc agcgtaggcg ggcgactcag atcccagcca gtggacttag 1020

cccctgtttg ctcctccgat aactggggtg accttggtta atattcacca gcagcctccc 1080

ccgttgcccc tctggatcca ctgcttaaat acggacgagg acagggccct gtctcctcag 1140

cttcaggcac caccactgac ctgggacagt gaatccggac tctaaggtaa atataaaatt 1200

tttaagtgta taatgtgtta aactactgat tctaattgtt tctctctttt agattccaac 1260

ctttggaact ga 1272

<210> 5

<211> 547

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 5

ccctaaaatg ggcaaacatt gcaagcagca aacagcaaac acacagccct ccctgcctgc 60

tgaccttgga gctggggcag aggtcagaga cctctctggg cccatgccac ctccaacatc 120

cactcgaccc cttggaattt ttcggtggag aggagcagag gttgtcctgg cgtggtttag 180

gtagtgtgag aggggaatga ctcctttcgg taagtgcagt ggaagctgta cactgcccag 240

gcaaagcgtc cgggcagcgt aggcgggcga ctcagatccc agccagtgga cttagcccct 300

gtttgctcct ccgataactg gggtgacctt ggttaatatt caccagcagc ctcccccgtt 360

gcccctctgg atccactgct taaatacgga cgaggacagg gccctgtctc ctcagcttca 420

ggcaccacca ctgacctggg acagtgaatc cggactctaa ggtaaatata aaatttttaa 480

gtgtataatg tgttaaacta ctgattctaa ttgtttctct cttttagatt ccaacctttg 540

gaactga 547

<210> 6

<211> 1179

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 6

ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 60

ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 120

gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 180

gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc 240

gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt 300

acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 360

gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 420

cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 480

ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 540

caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 600

gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 660

gcgcggccac cgagaatcgg acggggggtag tctcaagctg gccggcctgc tctggtgcct 720

ggtctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 780

gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 840

acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 900

tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 960

tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 1020

gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 1080

ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 1140

gtggttcaaa gtttttttct tccatttcag gtgtcgtga 1179

<210> 7

<211> 8

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 7

gtttaaac 8

<210> 8

<211> 581

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 8

gagcatctta ccgccattta ttcccatatt tgttctgttt ttcttgattt gggtatacat 60

ttaaatgtta ataaaacaaa atggtggggc aatcatttac atttttaggg atatgtaatt 120

actagttcag gtgtattgcc acaagacaaa catgttaaga aactttcccg ttatttacgc 180

tctgttcctg ttaatcaacc tctggattac aaaatttgtg aaagattgac tgatattctt 240

aactatgttg ctccttttac gctgtgtgga tatgctgctt tatagcctct gtatctagct 300

attgcttccc gtacggcttt cgttttctcc tccttgtata aatcctggtt gctgtctctt 360

ttagaggagt tgtggcccgt tgtccgtcaa cgtggcgtgg tgtgctctgt gtttgctgac 420

gcaaccccca ctggctgggg cattgccacc acctgtcaac tcctttctgg gactttcgct 480

ttccccctcc cgatcgccac ggcagaactc atcgccgcct gccttgcccg ctgctggaca 540

ggggctaggt tgctgggcac tgataattcc gtggtgttgt c 581

<210> 9

<211> 225

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 9

tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct 60

ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct 120

gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg 180

ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggc 225

<210> 10

<211> 213

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 10

taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta 60

tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag 120

ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt 180

tttaaagcaa gtaaaacctc tacaaatgtg gta 213

<210> 11

<211> 1386

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 11

atgcagcgcg tgaacatgat catggccgag agccccggcc tgatcaccat ctgcctgctg 60

ggctacctgc tgagcgccga gtgcaccgtg ttcctggacc acgagaacgc caacaagatc 120

ctgaaccgcc ccaagcgcta caacagcggc aagctggagg agttcgtgca gggcaacctg 180

gagcgcgagt gcatggagga gaagtgcagc ttcgaggagg cccgcgaggt gttcgagaac 240

accgagcgca ccaccgagtt ctggaagcag tacgtggacg gcgaccagtg cgagagcaac 300

ccctgcctga acggcggcag ctgcaaggac gacatcaaca gctacgagtg ctggtgcccc 360

ttcggcttcg agggcaagaa ctgcgagctg gacgtgacct gcaacatcaa gaacggccgc 420

tgcgagcagt tctgcaagaa cagcgccgac aacaaggtgg tgtgcagctg caccgaggc 480

taccgcctgg ccgagaacca gaagagctgc gagcccgccg tgcccttccc ctgcggcagg 540

gtgagcgtga gccagaccag caagctgacc cgcgccgagg ccgtgttccc cgacgtggac 600

tacgtgaaca gcaccgaggc cgagaccatc ctggacaaca tcacccagag cacccagagc 660

ttcaacgact tcacccgcgt ggtgggcggc gaggacgcca agcccggcca gttcccctgg 720

caggtggtgc tgaacggcaa ggtggacgcc ttctgcggcg gcagcatcgt gaacgagaag 780

tggatcgtga ccgccgccca ctgcgtggag accggcgtga agatcaccgt ggtggccggc 840

gagcacaaca tcgaggagac cgagcacacc gagcagaagc gcaacgtgat ccgcatcatc 900

ccccaccaca actacaacgc cgccatcaac aagtacaacc acgacatcgc cctgctggag 960

ctggacgagc ccctggtgct gaacagctac gtgaccccca tctgcatcgc cgacaaggag 1020

tacaccaaca tcttcctgaa gttcggcagc ggctacgtga gcggctgggg ccgcgtgttc 1080

cacaagggcc gcagcgccct ggtgctgcag tacctgcgcg tgcccctggt ggaccgcgcc 1140

acctgcctgc gcagcaccaa gttcaccatc tacaacaaca tgttctgcgc cggcttccac 1200

gagggcggca gggacagctg ccagggcgac agcggcggcc cccacgtgac cgaggtggag 1260

ggcaccagct tcctgaccgg catcatcagc tggggcgagg agtgcgccat gaagggcaag 1320

tacggcatct acaccaaggt gagccgctac gtgaactgga tcaaggagaa gaccaagctg 1380

actaa 1386

<210> 12

<211> 1386

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 12

atgcagcgcg tgaacatgat catggccgag agccccggcc tgatcaccat ctgcctgctg 60

ggctacctgc tgagcgccga gtgcaccgtg ttcctggacc acgagaacgc caacaagatc 120

ctgaaccgcc ccaagcgcta caacagcggc aagctggagg agttcgtgca gggcaacctg 180

gagcgcgagt gcatggagga gaagtgcagc ttcgaggagg cccgcgaggt gttcgagaac 240

accgagcgca ccaccgagtt ctggaagcag tacgtggacg gcgaccagtg cgagagcaac 300

ccctgcctga acggcggcag ctgcaaggac gacatcaaca gctacgagtg ctggtgcccc 360

ttcggcttcg agggcaagaa ctgcgagctg gacgtgacct gcaacatcaa gaacggccgc 420

tgcgagcagt tctgcaagaa cagcgccgac aacaaggtgg tgtgcagctg caccgaggc 480

taccgcctgg ccgagaacca gaagagctgc gagcccgccg tgcccttccc ctgcggcagg 540

gtgagcgtga gccagaccag caagctgacc cgcgccgagg ccgtgttccc cgacgtggac 600

tacgtgaaca gcaccgaggc cgagaccatc ctggacaaca tcacccagag cacccagagc 660

ttcaacgact tcacccgcgt ggtgggcggc gaggacgcca agcccggcca gttcccctgg 720

caggtggtgc tgaacggcaa ggtggacgcc ttctgcggcg gcagcatcgt gaacgagaag 780

tggatcgtga ccgccgccca ctgcgtggag accggcgtga agatcaccgt ggtggccggc 840

gagcacaaca tcgaggagac cgagcacacc gagcagaagc gcaacgtgat ccgcatcatc 900

ccccaccaca actacaacgc cgccatcaac aagtacaacc acgacatcgc cctgctggag 960

ctggacgagc ccctggtgct gaacagctac gtgaccccca tctgcatcgc cgacaaggag 1020

tacaccaaca tcttcctgaa gttcggcagc ggctacgtga gcggctgggg ccgcgtgttc 1080

cacaagggcc gcagcgccct ggtgctgcag tacctgcgcg tgcccctggt ggaccgcgcc 1140

acctgcctgc tgagcaccaa gttcaccatc tacaacaaca tgttctgcgc cggcttccac 1200

gagggcggca gggacagctg ccagggcgac agcggcggcc cccacgtgac cgaggtggag 1260

ggcaccagct tcctgaccgg catcatcagc tggggcgagg agtgcgccat gaagggcaag 1320

tacggcatct acaccaaggt gagccgctac gtgaactgga tcaaggagaa gaccaagctg 1380

actaa 1386

<210> 13

<211> 1876

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 13

cgcagccacc atggcggggt tttacgagat tgtgattaag gtccccagcg accttgacgg 60

gcatctgccc ggcatttctg acagctttgt gaactgggtg gccgagaagg aatgggagtt 120

gccgccagat tctgacatgg atctgaatct gattgagcag gcacccctga ccgtggccga 180

gaagctgcag cgcgactttc tgacggaatg gcgccgtgtg agtaaggccc cggaggccct 240

tttctttgtg caatttgaga agggagagag ctacttccac atgcacgtgc tcgtggaaac 300

caccggggtg aaatccatgg ttttgggacg tttcctgagt cagattcgcg aaaaactgat 360

tcagagaatt taccgcggga tcgagccgac tttgccaaac tggttcgcgg tcacaaagac 420

cagaaatggc gccggaggcg ggaacaaggt ggtggatgag tgctacatcc ccaattactt 480

gctccccaaa acccagcctg agctccagtg ggcgtggact aatatggaac agtatttaag 540

cgcctgtttg aatctcacgg agcgtaaacg gttggtggcg cagcatctga cgcacgtgtc 600

gcagacgcag gagcagaaca aagagaatca gaatcccaat tctgatgcgc cggtgatcag 660

atcaaaaact tcagccaggt acatggagct ggtcgggtgg ctcgtggaca aggggattac 720

ctcggagaag cagtggatcc aggaggacca ggcctcatac atctccttca atgcggcctc 780

caactcgcgg tcccaaatca aggctgcctt ggacaatgcg ggaaagatta tgagcctgac 840

taaaaccgcc cccgactacc tggtgggcca gcagcccgtg gaggacatt ccagcaatcg 900

gatttataaa attttggaac taaacgggta cgatccccaa tatgcggctt ccgtctttct 960

gggatgggcc acgaaaaagt tcggcaagag gaacaccatc tggctgtttg ggcctgcaac 1020

taccgggaag accaacatcg cggaggccat agcccacact gtgcccttct acgggtgcgt 1080

aaactggacc aatgagaact ttcccttcaa cgactgtgtc gacaagatgg tgatctggtg 1140

ggaggagggg aagatgaccg ccaaggtcgt ggagtcggcc aaagccattc tcggaggaag 1200

caaggtgcgc gtggaccaga aatgcaagtc ctcggcccag atagacccga ctcccgtgat 1260

cgtcacctcc aacaccaaca tgtgcgccgt gattgacggg aactcaacga ccttcgaaca 1320

ccagcagccg ttgcaagacc ggatgttcaa atttgaactc acccgccgtc tggatcatga 1380

ctttgggaag gtcaccaagc aggaagtcaa agactttttc cggtgggcaa aggatcacgt 1440

ggttgaggtg gagcatgaat tctacgtcaa aaagggtgga gccaagaaaa gacccgcccc 1500

cagtgacgca gatataagtg agcccaaacg ggtgcgcgag tcagttgcgc agccatcgac 1560

gtcagacgcg gaagcttcga tcaactacgc agacaggtac caaaacaaat gttctcgtca 1620

cgtgggcatg aatctgatgc tgtttccctg cagacaatgc gagagaatga atcagaattc 1680

aaatatctgc ttcactcacg gacagaaaga ctgtttagag tgctttcccg tgtcagaatc 1740

tcaacccgtt tctgtcgtca aaaaggcgta tcagaaactg tgctacattc atcatatcat 1800

gggaaaggtg cgacgctt gcactgcctg cgatctggtc aatgtggatt tggatgactg 1860

catctttgaa caataa 1876

<210> 14

<211> 1194

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 14

atggagctgg tcgggtggct cgtggacaag gggattacct cggagaagca gtggatccag 60

gaggaccagg cctcatacat ctccttcaat gcggcctcca actcgcggtc ccaaatcaag 120

gctgccttgg acaatgcggg aaagattatg agcctgacta aaaccgcccc cgactacctg 180

gtgggccagc agcccgtgga ggacatttcc agcaatcgga tttataaaat tttggaacta 240

aacgggtacg atccccaata tgcggcttcc gtctttctgg gatgggccac gaaaaagttc 300

ggcaagagga acaccatctg gctgtttggg cctgcaacta ccgggaagac caacatcgcg 360

gaggccatag cccacactgt gcccttctac gggtgcgtaa actggaccaa tgagaacttt 420

cccttcaacg actgtgtcga caagatggtg atctggtggg aggaggggaa gatgaccgcc 480

aaggtcgtgg agtcggccaa agccattctc ggaggaagca aggtgcgcgt ggaccagaaa 540

tgcaagtcct cggcccagat agacccgact cccgtgatcg tcacctccaa caccaacatg 600

tgcgccgtga ttgacgggaa ctcaacgacc ttcgaacacc agcagccgtt gcaagaccgg 660

atgttcaaat ttgaactcac ccgccgtctg gatcatgact ttgggaaggt caccaagcag 720

gaagtcaaag actttttccg gtgggcaaag gatcacgtgg ttgaggtgga gcatgaattc 780

tacgtcaaaa agggtggagc caagaaaaga cccgccccca gtgacgcaga tataagtgag 840

cccaaacggg tgcgcgagtc agttgcgcag ccatcgacgt cagacgcgga agcttcgatc 900

aactacgcag accgctacca aaacaaatgt tctcgtcacg tgggcatgaa tctgatgctg 960

tttccctgca gacaatgcga gagaatgaat cagaattcaa atatctgctt cactcacgga 1020

cagaaagact gtttagagtg ctttcccgtg tcagaatctc aacccgtttc tgtcgtcaaa 1080

aaggcgtatc agaaactgtg ctacattcat catatcatgg gaaaggtgcc agacgcttgc 1140

actgcctgcg atctggtcaa tgtggatttg gatgactgca tctttgaaca ataa 1194

<210> 15

<211> 141

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 15

aataaacgat aacgccgttg gtggcgtgag gcatgtaaaa ggttacatca ttatcttgtt 60

cgccatccgg ttggtataaa tagacgttca tgttggtttt tgtttcagtt gcaagttggc 120

tgcggcgcgc gcagcacctt t 141

<210> 16

<211> 556

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 16

ccctaaaatg ggcaaacatt gcaagcagca aacagcaaac acacagccct ccctgcctgc 60

tgaccttgga gctggggcag aggtcagaga cctctctggg cccatgccac ctccaacatc 120

cactcgaccc cttggaattt cggtggagag gagcagaggt tgtcctggcg tggtttaggt 180

agtgtgagag gggaatgact cctttcggta agtgcagtgg aagctgtaca ctgcccaggc 240

aaagcgtccg ggcagcgtag gcgggcgact cagatcccag ccagtggact tagcccctgt 300

ttgctcctcc gataactggg gtgaccttgg ttaatattca ccagcagcct cccccgttgc 360

ccctctggat ccactgctta aatacggacg aggacactcg agggccctgt ctcctcagct 420

tcaggcacca ccactgacct gggacagtga atccggacat cgattctaag gtaaatataa 480

aatttttaag tgtataattt gttaaactac tgattctaat tgtttctctc ttttagattc 540

caacctttgg aactga 556

<210> 17

<211> 80

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 17

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggcggcct 60

cagtgagcga gcgagcgcgc 80

<210> 18

<211> 241

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 18

gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag 60

ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga 120

aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat 180

atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga 240

c 241

<210> 19

<211> 215

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 19

gaacgctgac gtcatcaacc cgctccaagg aatcgcgggc ccagtgtcac taggcgggaa 60

cacccagcgc gcgtgcgccc tggcaggaag atggctgtga gggacagggg agtggcgccc 120

tgcaatattt gcatgtcgct atgtgttctg ggaaatcacc ataaacgtga aatgtctttg 180

gatttgggaa tcgtataaga actgtatgag accac 215

<210> 20

<211> 150

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 20

ataaacgata acgccgttgg tggcgtgagg catgtaaaag gttacatcat tatcttgttc 60

gccatccggt tggtataaat agacgttcat gttggttttt gtttcagttg caagttggct 120

gcggcgcgcg cagcaccttt gcggccatct 150

<210> 21

<400> 21

000

<210> 22

<400> 22

000

<210> 23

<400> 23

000

<210> 24

<400> 24

000

<210> 25

<400> 25

000

<210> 26

<400> 26

000

<210> 27

<400> 27

000

<210> 28

<400> 28

000

<210> 29

<400> 29

000

<210> 30

<400> 30

000

<210> 31

<400> 31

000

<210> 32

<400> 32

000

<210> 33

<400> 33

000

<210> 34

<400> 34

000

<210> 35

<400> 35

000

<210> 36

<400> 36

000

<210> 37

<400> 37

000

<210> 38

<400> 38

000

<210> 39

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 39

gcgcgctcgc tcgctc 16

<210> 40

<400> 40

000

<210> 41

<400> 41

000

<210> 42

<400> 42

000

<210> 43

<400> 43

000

<210> 44

<400> 44

000

<210> 45

<400> 45

000

<210> 46

<400> 46

000

<210> 47

<400> 47

000

<210> 48

<400> 48

000

<210> 49

<400> 49

000

<210> 50

<400> 50

000

<210> 51

<211> 141

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 51

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc t 141

<210> 52

<211> 130

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 52

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 60

ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 120

aggggttcct 130

<210> 53

<211> 3123

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 53

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggaaacc cgggcgtgcc 60

cgggcgcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg 120

gttcctgaac agagaaacag gagaatatgg gccaaacagg atatctgtgg taagcagttc 180

ctgccccggc tcagggccaa gaacagttgg aacagcagaa tatgggccaa acaggatatc 240

tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc 300

ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa 360

atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc 420

ttctgctccc cgagctctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga 480

gacgccatcc acgctgtttt gacttccata gaaggccgcc accatggaag acgccaaaaa 540

cataaagaaa ggcccggcgc cattctatcc gctggaagat ggaaccgctg gagagcaact 600

gcataaggct atgaagagat acgccctggt tcctggaaca attgctttta cagatgcaca 660

tatcgaggtg gacatcactt acgctgagta cttcgaaatg tccgttcggt tggcagaagc 720

tatgaaacga tatgggctga atacaaatca cagaatcgtc gtatgcagtg aaaactctct 780

tcaattcttt atgccggtgt tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa 840

cgacatttat aatgaacgtg aattgctcaa cagtatgggc atttcgcagc ctaccgtggt 900

gttcgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaagc tcccaatcat 960

ccaaaaaatt attatcatgg attctaaaac ggattaccag ggatttcagt cgatgtacac 1020

gttcgtcaca tctcatctac ctcccggttt taatgaatac gattttgtgc cagagtcctt 1080

cgatagggac aagacaattg cactgatcat gaactcctct ggatctactg gtctgcctaa 1140

aggtgtcgct ctgcctcata gaactgcctg cgtgagattc tcgcatgcca gagatcctat 1200

ttttggcaat caaatcattc cggatactgc gattttaagt gttgttccat tccatcacgg 1260

ttttggaatg tttactacac tcggatattt gatatgtgga tttcgagtcg tcttaatgta 1320

tagatttgaa gaagagctgt ttctgaggag ccttcaggat tacaagattc aaagtgcgct 1380

gctggtgcca accctattct ccttcttcgc caaaagcact ctgattgaca aatacgattt 1440

atctaattta cacgaaattg cttctggtgg cgctcccctc tctaaggaag tcggggaagc 1500

ggttgccaag aggttccatc tgccaggtat caggcaagga tatgggctca ctgagactac 1560

atcagctatt ctgattacac ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt 1620

tccatttttt gaagcgaagg ttgtggatct ggataccggg aaaacgctgg gcgttaatca 1680

aagaggcgaa ctgtgtgtga gaggtcctat gattatgtcc ggttatgtaa acaatccgga 1740

agcgaccaac gccttgattg acaaggatgg atggctacat tctggagaca tagcttactg 1800

ggacgaagac gaacacttct tcatcgttga ccgcctgaag tctctgatta agtacaaagg 1860

ctatcaggtg gctcccgctg aattggaatc catcttgctc caacacccca acatcttcga 1920

cgcaggtgtc gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt 1980

tttggagcac ggaaagacga tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt 2040

aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct 2100

taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca agaagggcgg 2160

aaagatcgcc gtgtaagagc atcttaccgc catttattcc catatttgtt ctgtttttct 2220

tgatttgggt atacatttaa atgttaataa aacaaaatgg tggggcaatc atttacattt 2280

ttagggatat gtaattacta gttcaggtgt attgccacaa gacaaacatg ttaagaaact 2340

ttcccgttat ttacgctctg ttcctgttaa tcaacctctg gattacaaaa tttgtgaaag 2400

attgactgat attcttaact atgttgctcc ttttacgctg tgtggatatg ctgcttttata 2460

gcctctgtat ctagctattg cttcccgtac ggctttcgtt ttctcctcct tgtataaatc 2520

ctggttgctg tctcttttag aggagttgtg gcccgttgtc cgtcaacgtg gcgtggtgtg 2580

ctctgtgttt gctgacgcaa cccccactgg ctggggcatt gccaccacct gtcaactcct 2640

ttctgggact ttcgctttcc ccctcccgat cgccacggca gaactcatcg ccgcctgcct 2700

tgcccgctgc tggacagggg ctaggttgct gggcactgat aattccgtgg tgttgtctgt 2760

gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 2820

aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 2880

taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 2940

agacaatagc aggcatgctg gggatgcggt gggctctatg gcaggaaccc ctagtgatgg 3000

agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg 3060

cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc 3120

agg 3123

<210> 54

<211> 3117

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 54

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc tgaacagaga aacaggaa tatgggccaa acaggatatc 180

tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gttggaacag cagaatatgg 240

gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg 300

tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg 360

ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg 420

cttctgttcg cgcgcttctg ctccccgagc tctatataag cagagctcgt ttagtgaacc 480

gtcagatcgc ctggagacgc catccacgct gttttgactt ccatagaagg ccgccaccat 540

ggaagacgcc aaaaacataa agaaaggccc ggcgccattc tatccgctgg aagatggaac 600

cgctggagag caactgcata aggctatgaa gagatacgcc ctggttcctg gaacaattgc 660

ttttacagat gcacatatcg aggtggacat cacttacgct gagtacttcg aaatgtccgt 720

tcggttggca gaagctatga aacgatatgg gctgaataca aatcacagaa tcgtcgtatg 780

cagtgaaaac tctcttcaat tctttatgcc ggtgttgggc gcgttattta tcggagttgc 840

agttgcgccc gcgaacgaca tttataatga acgtgaattg ctcaacagta tgggcatttc 900

gcagcctacc gtggtgttcg tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa 960

aaagctccca atcatccaaa aaattattat catggattct aaaacggatt accagggatt 1020

tcagtcgatg tacacgttcg tcacatctca tctacctccc ggttttaatg aatacgattt 1080

tgtgccagag tccttcgata gggacaagac aattgcactg atcatgaact cctctggatc 1140

tactggtctg cctaaaggtg tcgctctgcc tcatagaact gcctgcgtga gattctcgca 1200

tgccagagat cctatttttg gcaatcaaat cattccggat actgcgattt taagtgttgt 1260

tccattccat cacggttttg gaatgtttac tacactcgga tatttgatat gtggatttcg 1320

agtcgtctta atgtatagat ttgaagaaga gctgtttctg aggagccttc aggattacaa 1380

gattcaaagt gcgctgctgg tgccaaccct attctccttc ttcgccaaaa gcactctgat 1440

tgacaaatac gatttatcta atttacacga aattgcttct ggtggcgctc ccctctctaa 1500

ggaagtcggg gaagcggttg ccaagaggtt ccatctgcca ggtatcaggc aaggatatgg 1560

gctcactgag actacatcag ctattctgat tacacccgag ggggatgata aaccgggcgc 1620

ggtcggtaaa gttgttccat tttttgaagc gaaggttgtg gatctggata ccgggaaaac 1680

gctgggcgtt aatcaaagag gcgaactgtg tgtgagaggt cctatgatta tgtccggtta 1740

tgtaaacaat ccggaagcga ccaacgcctt gattgacaag gatggatggc tacattctgg 1800

agacatagct tactgggacg aagacgaaca cttcttcatc gttgaccgcc tgaagtctct 1860

gattaagtac aaaggctatc aggtggctcc cgctgaattg gaatccatct tgctccaaca 1920

ccccaacatc ttcgacgcag gtgtcgcagg tcttcccgac gatgacgccg gtgaacttcc 1980

cgccgccgtt gttgttttgg agcacggaaa gacgatgacg gaaaaagaga tcgtggatta 2040

cgtcgccagt caagtaacaa ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga 2100

agtaccgaaa ggtcttaccg gaaaactcga cgcaagaaaa atcagagaga tcctcataaa 2160

ggccaagaag ggcggaaaga tcgccgtgta agagcatctt accgccattt attcccatat 2220

ttgttctgtt tttcttgatt tgggtataca tttaaatgtt aataaaacaa aatggtgggg 2280

caatcattta catttttagg gatatgtaat tactagttca ggtgtattgc cacaagacaa 2340

acatgttaag aaactttccc gttatttacg ctctgttcct gttaatcaac ctctggatta 2400

caaaatttgt gaaagattga ctgatattct taactatgtt gctcctttta cgctgtgtgg 2460

atatgctgct ttatagcctc tgtatctagc tattgcttcc cgtacggctt tcgttttctc 2520

ctccttgtat aaatcctggt tgctgtctct tttagaggag ttgtggcccg ttgtccgtca 2580

acgtggcgtg gtgtgctctg tgtttgctga cgcaaccccc actggctggg gcattgccac 2640

cacctgtcaa ctcctttctg ggactttcgc tttccccctc ccgatcgcca cggcagaact 2700

catcgccgcc tgccttgccc gctgctggac aggggctagg ttgctgggca ctgataattc 2760

cgtggtgttg tctgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc 2820

ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc 2880

atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa 2940

gggggaggat tgggaagaca atagcaggca tgctggggat gcggtggggct ctatggcagg 3000

aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg 3060

cccgggaaac ccgggcgtgc gcctcagtga gcgagcgagc gcgcagctgc ctgcagg 3117

<210> 55

<211> 2841

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 55

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggaaacc cgggcgtgcc 60

cgggcgcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg 120

gttcctgaac agagaaacag gagaatatgg gccaaacagg atatctgtgg taagcagttc 180

ctgccccggc tcagggccaa gaacagttgg aacagcagaa tatgggccaa acaggatatc 240

tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc 300

ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa 360

atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc 420

ttctgctccc cgagctctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga 480

gacgccatcc acgctgtttt gacttccata gaaggccgcc accatgatca tggccgagag 540

ccctggcctg atcaccatct gcctgctggg ctacctgctg agcgccgagt gcaccgtgtt 600

cctggaccac gagaacgcca acaagatcct gaaccggccc aagagataca acagcggcaa 660

gctggaggag ttcgtgcagg gcaacctgga gagggagtgc atggaggaga agtgcagctt 720

cgaggaggcc agggaagtgt tcgagaacac cgagcggacc accgagttct ggaagcagta 780

cgtggacggc gaccagtgcg agagcaaccc ttgcctgaac ggcggcagct gcaaggacga 840

catcaacagc tacgagtgct ggtgcccttt cggcttcgag ggcaagaact gcgagctgga 900

cgtgacctgc aacatcaaga acggccgctg cgagcagttc tgcaagaaca gcgccgacaa 960

caaagtggtg tgtagctgca ccgagggcta cagactggcc gagaaccaga agagctgcga 1020

gcccgccgtg cccttcccct gcggcagagt gagcgtgtcc cagaccagca agctgaccag 1080

agccgagacc gtgttccccg acgtggacta cgtgaatagc accgaggccg agaccatcct 1140

ggacaacatc acccagagca cccagtcctt caacgacttc accaagttg tgggcggcga 1200

ggacgccaag cccggccagt tcccctggca ggtggtgctg aacggcaaag tggatgcctt 1260

ctgcggcggc agcatcgtga acgagaagtg gatcgtgaca gccgcccact gcgtggagac 1320

cggcgtgaag atcaccgtgg tggccggcga acacaatatc gaggagaccg agcacaccga 1380

gcagaagcgg aacgtcatcc ggattatccc ccaccacaac tacaacgccg ccatcaacaa 1440

gtacaaccac gacatcgccc tgctggagct ggacgagcct ctggtgctga atagctacgt 1500

gacccccatc tgcatcgccg acaaggagta caccaacatc ttcctgaagt tcggcagcgg 1560

ctacgtgtcc ggctggggca gagtgttcca caagggcaga agcgccctgg tgctgcagta 1620

cctgagagtg cccctggtgg acagagccac ctgcctgttg agcaccaagt tcaccatcta 1680

caacaacatg ttctgcgccg gcttccacga gggcggcaga gacagctgcc agggcgacag 1740

cggcggaccc cacgtgaccg aagtggaggg caccagcttc ctgaccggca tcatcagctg 1800

gggcgaggag tgcgccatga agggcaagta cggcatctac accaaagtga gccggtacgt 1860

gaactggatc aaggagaaaa ccaagctgac ctgagagcat cttaccgcca tttattccca 1920

tatttgttct gtttttcttg atttgggtat acatttaaat gttaataaaa caaaatggtg 1980

gggcaatcat ttacattttt agggatatgt aattactagt tcaggtgtat tgccacaaga 2040

caaacatgtt aagaaacttt cccgttattt acgctctgtt cctgttaatc aacctctgga 2100

ttacaaaatt tgtgaaagat tgactgatat tcttaactat gttgctcctt ttacgctgtg 2160

tggatatgct gctttatagc ctctgtatct agctattgct tcccgtacgg ctttcgtttt 2220

ctcctccttg tataaatcct ggttgctgtc tcttttagag gagttgtggc ccgttgtccg 2280

tcaacgtggc gtggtgtgct ctgtgtttgc tgacgcaacc cccactggct ggggcattgc 2340

caccacctgt caactccttt ctgggacttt cgctttcccc ctcccgatcg ccacggcaga 2400

actcatcgcc gcctgccttg cccgctgctg gacaggggct aggttgctgg gcactgataa 2460

ttccgtggtg ttgtctgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt 2520

gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat 2580

tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag 2640

caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc 2700

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 2760

ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc 2820

gagcgcgcag ctgcctgcag g 2841

<210> 56

<211> 2835

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 56

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc tgaacagaga aacaggaa tatgggccaa acaggatatc 180

tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gttggaacag cagaatatgg 240

gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg 300

tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg 360

ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg 420

cttctgttcg cgcgcttctg ctccccgagc tctatataag cagagctcgt ttagtgaacc 480

gtcagatcgc ctggagacgc catccacgct gttttgactt ccatagaagg ccgccaccat 540

gatcatggcc gagagccctg gcctgatcac catctgcctg ctgggctacc tgctgagcgc 600

cgagtgcacc gtgttcctgg accacgagaa cgccaacaag atcctgaacc ggcccaagag 660

atacaacagc ggcaagctgg aggagttcgt gcagggcaac ctggagaggg agtgcatgga 720

ggagaagtgc agcttcgagg aggccaggga agtgttcgag aacaccgagc ggaccaccga 780

gttctggaag cagtacgtgg acggcgacca gtgcgagagc aacccttgcc tgaacggcgg 840

cagctgcaag gacgacatca acagctacga gtgctggtgc cctttcggct tcgagggcaa 900

gaactgcgag ctggacgtga cctgcaacat caagaacggc cgctgcgagc agttctgcaa 960

gaacagcgcc gacaacaaag tggtgtgtag ctgcaccgag ggctacagac tggccgagaa 1020

ccagaagagc tgcgagcccg ccgtgccctt cccctgcggc agagtgagcg tgtcccagac 1080

cagcaagctg accagagccg agaccgtgtt ccccgacgtg gactacgtga atagcaccga 1140

ggccgagacc atcctggaca acatcaccca gagcaccag tccttcaacg acttcaccag 1200

agttgtgggc ggcgaggacg ccaagcccgg ccagttcccc tggcaggtgg tgctgaacgg 1260

caaagtggat gccttctgcg gcggcagcat cgtgaacgag aagtggatcg tgacagccgc 1320

ccactgcgtg gagaccggcg tgaagatcac cgtggtggcc ggcgaacaca atatcgagga 1380

gaccgagcac accgagcaga agcggaacgt catccggatt atcccccacc acaactacaa 1440

cgccgccatc aacaagtaca accacgacat cgccctgctg gagctggacg agcctctggt 1500

gctgaatagc tacgtgaccc ccatctgcat cgccgacaag gagtacacca acatcttcct 1560

gaagttcggc agcggctacg tgtccggctg gggcagagtg ttccacaagg gcagaagcgc 1620

cctggtgctg cagtacctga gagtgcccct ggtggacaga gccacctgcc tgttgagcac 1680

caagttcacc atctacaaca acatgttctg cgccggcttc cacgagggcg gcagagacag 1740

ctgccagggc gacagcggcg gaccccacgt gaccgaagtg gagggcacca gcttcctgac 1800

cggcatcatc agctggggcg aggagtgcgc catgaagggc aagtacggca tctacaccaa 1860

agtgagccgg tacgtgaact ggatcaagga gaaaaccaag ctgacctgag agcatcttac 1920

cgccatttat tcccatattt gttctgtttt tcttgatttg ggtatacatt taaatgttaa 1980

taaaacaaaa tggtggggca atcatttaca tttttaggga tatgtaatta ctagttcagg 2040

tgtattgcca caagacaaac atgttaagaa actttcccgt tatttacgct ctgttcctgt 2100

taatcaacct ctggattaca aaatttgtga aagattgact gatattctta actatgttgc 2160

tccttttacg ctgtgtggat atgctgcttt atagcctctg tatctagcta ttgcttcccg 2220

tacggctttc gttttctcct ccttgtataa atcctggttg ctgtctcttt tagaggagtt 2280

gtggcccgtt gtccgtcaac gtggcgtggt gtgctctgtg tttgctgacg caacccccac 2340

tggctggggc attgccacca cctgtcaact cctttctggg actttcgctt tccccctccc 2400

gatcgccacg gcagaactca tcgccgcctg ccttgcccgc tgctggacag gggctaggtt 2460

gctgggcact gataattccg tggtgttgtc tgtgccttct agttgccagc catctgttgt 2520

ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta 2580

ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg 2640

ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc 2700

ggtgggctct atggcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc 2760

gctcgctcac tgaggccgcc cgggaaaccc gggcgtgcgc ctcagtgagc gagcgagcgc 2820

gcagctgcct gcagg 2835

<210> 57

<211> 3912

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 57

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggaaacc cgggcgtgcc 60

cgggcgcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg 120

gttcctggct cagaggctca gaggcacaca ggagtttctg ggctcaccct gcccccttcc 180

aacccctcag ttcccatcct ccagcagctg tttgtgtgct gcctctgaag tccacactga 240

acaaacttca gcctactcat gtccctaaaa tgggcaaaca ttgcaagcag caaacagcaa 300

acacacagcc ctccctgcct gctgaccttg gagctggggc agaggtcaga gacctctctg 360

ggcccatgcc acctccaaca tccactcgac cccttggaat ttcggtggag aggagcagag 420

gttgtcctgg cgtggtttag gtagtgtgag agggtccggg ttcaaaacca cttgctgggt 480

ggggagtcgt cagtaagtgg ctatgccccg accccgaagc ctgtttcccc atctgtacaa 540

tggaaatgat aaagacgccc atctgatagg gtttttgtgg caaataaaca tttggttttt 600

ttgttttgtt ttgttttgtt ttttgagatg gaggtttgct ctgtcgccca ggctggagtg 660

cagtgacaca atctcatctc accacaacct tcccctgcct cagcctccca agtagctggg 720

attacaagca tgtgccacca cacctggcta attttctatt tttagtagag acgggtttct 780

ccatgttggt cagcctcagc ctcccaagta actgggatta caggcctgtg ccaccacacc 840

cggctaattt tttctatttt tgacagggac ggggtttcac catgttggtc aggctggtct 900

agaggtaccg gatcttgcta ccagtggaac agccactaag gattctgcag tgagagcaga 960

gggccagcta agtggtactc tcccagagac tgtctgactc acgccacccc ctccaccttg 1020

gacacaggac gctgtggttt ctgagccagg tacaatgact cctttcggta agtgcagtgg 1080

aagctgtaca ctgcccaggc aaagcgtccg ggcagcgtag gcgggcgact cagatcccag 1140

ccagtggact tagcccctgt ttgctcctcc gataactggg gtgaccttgg ttaatattca 1200

ccagcagcct cccccgttgc ccctctggat ccactgctta aatacggacg aggacagggc 1260

cctgtctcct cagcttcagg caccaccact gacctgggac agtgccgcca ccatggaaga 1320

cgccaaaaac ataaagaaag gcccggcgcc attctatccg ctggaagatg gaaccgctgg 1380

agagcaactg cataaggcta tgaagata cgccctggtt cctggaacaa ttgcttttac 1440

agatgcacat atcgaggtgg acatcactta cgctgagtac ttcgaaatgt ccgttcggtt 1500

ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg tatgcagtga 1560

aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag ttgcagttgc 1620

gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgggca tttcgcagcc 1680

taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc aaaaaaagct 1740

cccaatcatc caaaaaatta ttatcatgga ttctaaaacg gattaccagg gatttcagtc 1800

gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg attttgtgcc 1860

agagtccttc gatagggaca agacaattgc actgatcatg aactcctctg gatctactgg 1920

tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc gtgagattct cgcatgccag 1980

agatcctatt tttggcaatc aaatcattcc ggatactgcg attttaagtg ttgttccatt 2040

ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat ttcgagtcgt 2100

cttaatgtat agatttgaag aagagctgtt tctgaggagc cttcaggatt acaagattca 2160

aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc aaaagcactc tgattgacaa 2220

atacgattta tctaatttac acgaaattgc ttctggtggc gctcccctct ctaaggaagt 2280

cggggaagcg gttgccaaga ggttccatct gccaggtatc aggcaaggat atgggctcac 2340

tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg gcgcggtcgg 2400

taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga aaacgctggg 2460

cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg attatgtccg gttatgtaaa 2520

caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt ctggagacat 2580

agcttactgg gacgaagacg aacacttctt catcgttgac cgcctgaagt ctctgattaa 2640

gtacaaaggc tatcaggtgg ctcccgctga attggaatcc atcttgctcc aacaccccaa 2700

catcttcgac gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac ttcccgccgc 2760

cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc 2820

cagtcaagta acaaccgcga aaaagttgcg cggagggagtt gtgtttgtgg acgaagtacc 2880

gaaaggtctt acggaaaac tcgacgcaag aaaaatcaga gagatcctca taaaggccaa 2940

gaagggcgga aagatcgccg tgtaagagca tcttaccgcc atttattccc atatttgttc 3000

tgtttttctt gatttgggta tacatttaaa tgttaataaa acaaaatggt ggggcaatca 3060

tttacatttt tagggatatg taattactag ttcaggtgta ttgccacaag acaaacatgt 3120

taagaaactt tcccgttatt tacgctctgt tcctgttaat caacctctgg attacaaaat 3180

ttgtgaaaga ttgactgata ttcttaacta tgttgctcct tttacgctgt gtggatatgc 3240

tgctttatag cctctgtatc tagctattgc ttcccgtacg gctttcgttt tctcctcctt 3300

gtataaatcc tggttgctgt ctcttttaga ggagttgtgg cccgttgtcc gtcaacgtgg 3360

cgtggtgtgc tctgtgtttg ctgacgcaac ccccactggc tggggcattg ccaccacctg 3420

tcaactcctt tctgggactt tcgctttccc cctcccgatc gccacggcag aactcatcgc 3480

cgcctgcctt gcccgctgct ggacaggggc taggttgctg ggcactgata attccgtggt 3540

gttgtctgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 3600

gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 3660

ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 3720

ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg caggaacccc 3780

tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac 3840

caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca 3900

gctgcctgca gg 3912

<210> 58

<211> 3906

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 58

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc tggctcagag gctcagaggc acacaggagt ttctgggctc 180

accctgcccc cttccaaccc ctcagttccc atcctccagc agctgtttgt gtgctgcctc 240

tgaagtccac actgaacaaa cttcagccta ctcatgtccc taaaatgggc aaacattgca 300

agcagcaaac agcaaacaca cagccctccc tgcctgctga ccttggagct ggggcagagg 360

tcagagacct ctctgggccc atgccacctc caacatccac tcgacccctt ggaatttcgg 420

tggagaggag cagaggttgt cctggcgtgg tttaggtagt gtgagagggt ccgggttcaa 480

aaccacttgc tgggtgggga gtcgtcagta agtggctatg ccccgacccc gaagcctgtt 540

tccccatctg tacaatggaa atgataaaga cgcccatctg atagggtttt tgtggcaaat 600

aaacatttgg tttttttgtt ttgttttgtt ttgttttttg agatggaggt ttgctctgtc 660

gcccaggctg gagtgcagtg acacaatctc atctcaccac aaccttcccc tgcctcagcc 720

tcccaagtag ctgggattac aagcatgtgc caccacacct ggctaatttt ctatttttag 780

tagagacggg tttctccatg ttggtcagcc tcagcctccc aagtaactgg gattacaggc 840

ctgtgccacc acacccggct aattttttct atttttgaca gggacggggt ttcaccatgt 900

tggtcaggct ggtctagagg taccggatct tgctaccagt ggaacagcca ctaaggattc 960

tgcagtgaga gcagagggcc agctaagtgg tactctccca gagactgtct gactcacgcc 1020

accccctcca ccttggacac aggacgctgt ggtttctgag ccaggtacaa tgactccttt 1080

cggtaagtgc agtggaagct gtacactgcc caggcaaagc gtccgggcag cgtaggcggg 1140

cgactcagat cccagccagt ggacttagcc cctgtttgct cctccgataa ctggggtgac 1200

cttggttaat attcaccagc agcctccccc gttgcccctc tggatccact gcttaaatac 1260

ggacgaggac agggccctgt ctcctcagct tcaggcacca ccactgacct gggacagtgc 1320

cgccaccatg gaagacgcca aaaacataaa gaaaggcccg gcgccattct atccgctgga 1380

agatggaacc gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg 1440

aacaattgct tttacagatg cacatatcga ggtggacatc acttacgctg agtacttcga 1500

aatgtccgtt cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat 1560

cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat 1620

cggagttgca gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat 1680

gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa 1740

cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc atggattcta aaacggatta 1800

ccagggattt cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga 1860

atacgatttt gtgccagagt ccttcgatag ggacaagaca attgcactga tcatgaactc 1920

ctctggatct actggtctgc ctaaaggtgt cgctctgcct catagaactg cctgcgtgag 1980

attctcgcat gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt 2040

aagtgttgtt ccattccatc acggttttgg aatgtttact acactcggat atttgatatg 2100

tggatttcga gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga ggagccttca 2160

ggattacaag attcaaagtg cgctgctggt gccaacccta ttctccttct tcgccaaaag 2220

cactctgatt gacaaatacg atttatctaa tttacacgaa attgcttctg gtggcgctcc 2280

cctctctaag gaagtcgggg aagcggttgc caagaggttc catctgccag gtatcaggca 2340

aggatatggg ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa 2400

accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac 2460

cgggaaaacg ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc ctatgattat 2520

gtccggttat gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct 2580

acattctgga gacatagctt actgggacga agacgaacac ttcttcatcg ttgaccgcct 2640

gaagtctctg attaagtaca aaggctatca ggtggctccc gctgaattgg aatccatctt 2700

gctccaacac cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg 2760

tgaacttccc gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat 2820

cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt 2880

tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat 2940

cctcataaag gccaagaagg gcggaaagat cgccgtgtaa gagcatctta ccgccattta 3000

ttcccatatt tgttctgttt ttcttgattt gggtatacat ttaaatgtta ataaaacaaa 3060

atggtggggc aatcatttac atttttaggg atatgtaatt actagttcag gtgtattgcc 3120

acaagacaaa catgttaaga aactttcccg ttatttacgc tctgttcctg ttaatcaacc 3180

tctggattac aaaatttgtg aaagattgac tgatattctt aactatgttg ctccttttac 3240

gctgtgtgga tatgctgctt tatagcctct gtatctagct attgcttccc gtacggcttt 3300

cgttttctcc tccttgtata aatcctggtt gctgtctctt ttagaggagt tgtggcccgt 3360

tgtccgtcaa cgtggcgtgg tgtgctctgt gtttgctgac gcaaccccca ctggctgggg 3420

cattgccacc acctgtcaac tcctttctgg gactttcgct ttccccctcc cgatcgccac 3480

ggcagaactc atcgccgcct gccttgcccg ctgctggaca ggggctaggt tgctgggcac 3540

tgataattcc gtggtgttgt ctgtgccttc tagttgccag ccatctgttg tttgcccctc 3600

ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga 3660

ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca 3720

ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc 3780

tatggcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca 3840

ctgaggccgc ccgggaaacc cgggcgtgcg cctcagtgag cgagcgagcg cgcagctgcc 3900

tgcagg 3906

<210> 59

<211> 3630

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 59

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggaaacc cgggcgtgcc 60

cgggcgcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg 120

gttcctggct cagaggctca gaggcacaca ggagtttctg ggctcaccct gcccccttcc 180

aacccctcag ttcccatcct ccagcagctg tttgtgtgct gcctctgaag tccacactga 240

acaaacttca gcctactcat gtccctaaaa tgggcaaaca ttgcaagcag caaacagcaa 300

acacacagcc ctccctgcct gctgaccttg gagctggggc agaggtcaga gacctctctg 360

ggcccatgcc acctccaaca tccactcgac cccttggaat ttcggtggag aggagcagag 420

gttgtcctgg cgtggtttag gtagtgtgag agggtccggg ttcaaaacca cttgctgggt 480

ggggagtcgt cagtaagtgg ctatgccccg accccgaagc ctgtttcccc atctgtacaa 540

tggaaatgat aaagacgccc atctgatagg gtttttgtgg caaataaaca tttggttttt 600

ttgttttgtt ttgttttgtt ttttgagatg gaggtttgct ctgtcgccca ggctggagtg 660

cagtgacaca atctcatctc accacaacct tcccctgcct cagcctccca agtagctggg 720

attacaagca tgtgccacca cacctggcta attttctatt tttagtagag acgggtttct 780

ccatgttggt cagcctcagc ctcccaagta actgggatta caggcctgtg ccaccacacc 840

cggctaattt tttctatttt tgacagggac ggggtttcac catgttggtc aggctggtct 900

agaggtaccg gatcttgcta ccagtggaac agccactaag gattctgcag tgagagcaga 960

gggccagcta agtggtactc tcccagagac tgtctgactc acgccacccc ctccaccttg 1020

gacacaggac gctgtggttt ctgagccagg tacaatgact cctttcggta agtgcagtgg 1080

aagctgtaca ctgcccaggc aaagcgtccg ggcagcgtag gcgggcgact cagatcccag 1140

ccagtggact tagcccctgt ttgctcctcc gataactggg gtgaccttgg ttaatattca 1200

ccagcagcct cccccgttgc ccctctggat ccactgctta aatacggacg aggacagggc 1260

cctgtctcct cagcttcagg caccaccact gacctgggac agtgccgcca ccatgatcat 1320

ggccgagagc cctggcctga tcaccatctg cctgctgggc tacctgctga gcgccgagtg 1380

caccgtgttc ctggaccacg agaacgccaa caagatcctg aaccggccca agagatacaa 1440

cagcggcaag ctggagagt tcgtgcaggg caacctggag agggagtgca tggaggagaa 1500

gtgcagcttc gaggaggcca gggaagtgtt cgagaacacc gagcggacca ccgagttctg 1560

gaagcagtac gtggacggcg accagtgcga gagcaaccct tgcctgaacg gcggcagctg 1620

caaggacgac atcaacagct acgagtgctg gtgccctttc ggcttcgagg gcaagaactg 1680

cgagctggac gtgacctgca acatcaagaa cggccgctgc gagcagttct gcaagaacag 1740

cgccgacaac aaagtggtgt gtagctgcac cgagggctac agactggccg agaaccagaa 1800

gagctgcgag cccgccgtgc ccttcccctg cggcagagtg agcgtgtccc agaccagcaa 1860

gctgaccaga gccgagaccg tgttccccga cgtggactac gtgaatagca ccgaggccga 1920

gaccatcctg gacaacatca cccagagcac ccagtccttc aacgacttca cccagttgt 1980

gggcggcgag gacgccaagc ccggccagtt cccctggcag gtggtgctga acggcaaagt 2040

ggatgccttc tgcggcggca gcatcgtgaa cgagaagtgg atcgtgacag ccgcccactg 2100

cgtggagacc ggcgtgaaga tcaccgtggt ggccggcgaa cacaatatcg aggagaccga 2160

gcacaccgag cagaagcgga acgtcatccg gattatcccc caccacaact acaacgccgc 2220

catcaacaag tacaaccacg acatcgccct gctggagctg gacgagcctc tggtgctgaa 2280

tagctacgtg acccccatct gcatcgccga caaggagtac accaacatct tcctgaagtt 2340

cggcagcggc tacgtgtccg gctggggcag agtgttccac aagggcagaa gcgccctggt 2400

gctgcagtac ctgagagtgc ccctggtgga cagagccacc tgcctgttga gcaccaagtt 2460

caccatctac aacaacatgt tctgcgccgg cttccacgag ggcggcagag acagctgcca 2520

gggcgacagc ggcggacccc acgtgaccga agtggagggc accagcttcc tgaccggcat 2580

catcagctgg ggcgaggagt gcgccatgaa gggcaagtac ggcatctaca ccaaagtgag 2640

ccggtacgtg aactggatca aggagaaaac caagctgacc tgagagcatc ttaccgccat 2700

ttattcccat atttgttctg tttttcttga tttgggtata catttaaatg ttaataaaac 2760

aaaatggtgg ggcaatcatt tacattttta gggatatgta attactagtt caggtgtatt 2820

gccacaagac aaacatgtta agaaactttc ccgttattta cgctctgttc ctgttaatca 2880

acctctggat tacaaaattt gtgaaagatt gactgatatt cttaactatg ttgctccttt 2940

tacgctgtgt ggatatgctg ctttatagcc tctgtatcta gctattgctt cccgtacggc 3000

tttcgttttc tcctccttgt ataaatcctg gttgctgtct cttttagagg agttgtggcc 3060

cgttgtccgt caacgtggcg tggtgtgctc tgtgtttgct gacgcaaccc ccactggctg 3120

gggcattgcc accacctgtc aactcctttc tgggactttc gctttccccc tcccgatcgc 3180

cacggcagaa ctcatcgccg cctgccttgc ccgctgctgg acaggggcta ggttgctggg 3240

cactgataat tccgtggtgt tgtctgtgcc ttctagttgc cagccatctg ttgtttgccc 3300

ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa 3360

tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg 3420

gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg 3480

ctctatggca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc 3540

tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag 3600

tgagcgagcg agcgcgcagc tgcctgcagg 3630

<210> 60

<211> 3624

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 60

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc tggctcagag gctcagaggc acacaggagt ttctgggctc 180

accctgcccc cttccaaccc ctcagttccc atcctccagc agctgtttgt gtgctgcctc 240

tgaagtccac actgaacaaa cttcagccta ctcatgtccc taaaatgggc aaacattgca 300

agcagcaaac agcaaacaca cagccctccc tgcctgctga ccttggagct ggggcagagg 360

tcagagacct ctctgggccc atgccacctc caacatccac tcgacccctt ggaatttcgg 420

tggagaggag cagaggttgt cctggcgtgg tttaggtagt gtgagagggt ccgggttcaa 480

aaccacttgc tgggtgggga gtcgtcagta agtggctatg ccccgacccc gaagcctgtt 540

tccccatctg tacaatggaa atgataaaga cgcccatctg atagggtttt tgtggcaaat 600

aaacatttgg tttttttgtt ttgttttgtt ttgttttttg agatggaggt ttgctctgtc 660

gcccaggctg gagtgcagtg acacaatctc atctcaccac aaccttcccc tgcctcagcc 720

tcccaagtag ctgggattac aagcatgtgc caccacacct ggctaatttt ctatttttag 780

tagagacggg tttctccatg ttggtcagcc tcagcctccc aagtaactgg gattacaggc 840

ctgtgccacc acacccggct aattttttct atttttgaca gggacggggt ttcaccatgt 900

tggtcaggct ggtctagagg taccggatct tgctaccagt ggaacagcca ctaaggattc 960

tgcagtgaga gcagagggcc agctaagtgg tactctccca gagactgtct gactcacgcc 1020

accccctcca ccttggacac aggacgctgt ggtttctgag ccaggtacaa tgactccttt 1080

cggtaagtgc agtggaagct gtacactgcc caggcaaagc gtccgggcag cgtaggcggg 1140

cgactcagat cccagccagt ggacttagcc cctgtttgct cctccgataa ctggggtgac 1200

cttggttaat attcaccagc agcctccccc gttgcccctc tggatccact gcttaaatac 1260

ggacgaggac agggccctgt ctcctcagct tcaggcacca ccactgacct gggacagtgc 1320

cgccaccatg atcatggccg agagccctgg cctgatcacc atctgcctgc tgggctacct 1380

gctgagcgcc gagtgcaccg tgttcctgga ccacgagaac gccaacaaga tcctgaaccg 1440

gcccaagaga tacaacagcg gcaagctgga ggagttcgtg cagggcaacc tggagaggga 1500

gtgcatggag gagaagtgca gcttcgagga ggccagggaa gtgttcgaga acaccgagcg 1560

gaccaccgag ttctggaagc agtacgtgga cggcgaccag tgcgagagca acccttgcct 1620

gaacggcggc agctgcaagg acgacatcaa cagctacgag tgctggtgcc ctttcggctt 1680

cgagggcaag aactgcgagc tggacgtgac ctgcaacatc aagaacggcc gctgcgagca 1740

gttctgcaag aacagcgccg acaacaaagt ggtgtgtagc tgcaccgagg gctacagact 1800

ggccgagaac cagaagagct gcgagcccgc cgtgcccttc ccctgcggca gagtgagcgt 1860

gtcccagacc agcaagctga cgagccga gaccgtgttc cccgacgtgg actacgtgaa 1920

tagcaccgag gccgagacca tcctggacaa catcacccag agcacccagt ccttcaacga 1980

cttcaccaga gttgtgggcg gcgaggacgc caagcccggc cagttcccct ggcaggtggt 2040

gctgaacggc aaagtggatg ccttctgcgg cggcagcatc gtgaacgaga agtggatcgt 2100

gacagccgcc cactgcgtgg agaccggcgt gaagatcacc gtggtggccg gcgaacacaa 2160

tatcgaggag accgagcaca ccgagcagaa gcggaacgtc atccggatta tcccccacca 2220

caactacaac gccgccatca acaagtacaa ccacgacatc gccctgctgg agctggacga 2280

gcctctggtg ctgaatagct acgtgacccc catctgcatc gccgacaagg agtacaccaa 2340

catcttcctg aagttcggca gcggctacgt gtccggctgg ggcagagtgt tccacaaggg 2400

cagaagcgcc ctggtgctgc agtacctgag agtgcccctg gtggacagag ccacctgcct 2460

gttgagcacc aagttcacca tctacaacaa catgttctgc gccggcttcc acgagggcgg 2520

cagagacagc tgccagggcg acagcggcgg accccacgtg accgaagtgg agggcaccag 2580

cttcctgacc ggcatcatca gctggggcga ggagtgcgcc atgaagggca agtacggcat 2640

ctacaccaaa gtgagccggt acgtgaactg gatcaaggag aaaaccaagc tgacctgaga 2700

gcatcttacc gccatttatt cccatatttg ttctgttttt cttgatttgg gtatacattt 2760

aaatgttaat aaaacaaaat ggtggggcaa tcatttacat ttttagggat atgtaattac 2820

tagttcaggt gtattgccac aagacaaaca tgttaagaaa ctttcccgtt atttacgctc 2880

tgttcctgtt aatcaacctc tggattacaa aatttgtgaa agattgactg atattcttaa 2940

ctatgttgct ccttttacgc tgtgtggata tgctgcttta tagcctctgt atctagctat 3000

tgcttcccgt acggctttcg ttttctcctc cttgtataaa tcctggttgc tgtctctttt 3060

agaggagttg tggcccgttg tccgtcaacg tggcgtggtg tgctctgtgt ttgctgacgc 3120

aacccccact ggctggggca ttgccaccac ctgtcaactc ctttctggga ctttcgcttt 3180

ccccctcccg atcgccacgg cagaactcat cgccgcctgc cttgcccgct gctggacagg 3240

ggctaggttg ctgggcactg ataattccgt ggtgttgtct gtgccttcta gttgccagcc 3300

atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt 3360

cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct 3420

ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc 3480

tggggatgcg gtggggctcta tggcaggaac ccctagtgat ggagttggcc actccctctc 3540

tgcgcgctcg ctcgctcact gaggccgccc gggaaacccg ggcgtgcgcc tcagtgagcg 3600

agcgagcgcg cagctgcctg cagg 3624

<210> 61

<211> 4257

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 61

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt 180

agccatgctc tagagcggcc gcacgcgtag atcttcaata ttggccatta gccatattat 240

tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat 300

atcataatat gtacatttat attggctcat gtccaatatg accgccatgt tggcattgat 360

tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg 420

agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc 480

gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt 540

gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc 600

atatgccaag tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg 660

cccagtacat gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg 720

ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct 780

ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg 840

gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga 900

ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg 960

cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg 1020

cgacgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc 1080

tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt 1140

aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag 1200

gggctccggg agggcccttt gtgcgggggg gagcggctcg gggggtgcgt gcgtgtgtgt 1260

gtgcgtgggg agcgccgcgt gcggcccgcg ctgcccggcg gctgtgagcg ctgcgggcgc 1320

ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc 1380

ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg 1440

gggtgagcag ggggtgtggg cgcggcggtc gggctgtaac ccccccctgc acccccctcc 1500

ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc gtggcgcggg 1560

gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc 1620

tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg gcggctgtcg 1680

aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg cgcagggact 1740

tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca ccccctctag 1800

cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg agggccttcg 1860

tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc cgcgggggga 1920

cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg 1980

ctctagagcc tctgctaacc atgttttagc cttcttcttt ttcctacagc tcctgggcaa 2040

cgtgctggtt attgtgctgt ctcatcattt gtcgacagaa ttcctcgaag atccgaaggg 2100

gttcaagctt ggcattccgg tactgttggt aaagccacca tggaagacgc caaaaacata 2160

aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga gcaactgcat 2220

aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc 2280

gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc agaagctatg 2340

aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa 2400

ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac 2460

atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac cgtggtgttc 2520

gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa 2580

aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc 2640

gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga gtccttcgat 2700

agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct gcctaaaggt 2760

gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga tcctattttt 2820

ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt 2880

ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga 2940

tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag tgcgctgctg 3000

gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata cgatttatct 3060

aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg ggaagcggtt 3120

gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga gactacatca 3180

gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca 3240

ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcaaaga 3300

ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa tccggaagcg 3360

accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac 3420

gaagacgaac acttcttcat cgttgaccgc ctgaagtctc tgattaagta caaaggctat 3480

caggtggctc ccgctgaatt ggaatccatc ttgctccaac accccaacat cttcgacgca 3540

ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg 3600

gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca 3660

accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc 3720

ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag 3780

atcgccgtgt aattctagag tcggggcggc cggccgcttc gagcagacat gataagatac 3840

attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa 3900

atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac 3960

aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc 4020

aagtaaaacc tctacaaatg tggtaaaatc gataagtgcg gaccgagcgg ccgctctaga 4080

gcatggctac gtagataagt agcatggcgg gttaatcatt aactacacct gcaggcagct 4140

gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct 4200

cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttcct 4257

<210> 62

<211> 2510

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 62

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt 180

agccatgctc tagagcggcc gcgctagccc ctaaaatggg caaacattgc aagcagcaaa 240

cagcaaacac acagccctcc ctgcctgctg accttggagc tggggcagag gtcagagacc 300

tctctgggcc catgccacct ccaacatcca ctcgacccct tggaatttcg gtggagagga 360

gcagaggttg tcctggcgtg gtttaggtag tgtgagaggg gaatgactcc tttcggtaag 420

tgcagtggaa gctgtacact gcccaggcaa agcgtccggg cagcgtaggc gggcgactca 480

gatcccagcc agtggactta gcccctgttt gctcctccga taactggggt gaccttggtt 540

aatattcacc agcagcctcc cccgttgccc ctctggatcc actgcttaaa tacggacgag 600

gacactcgag ggccctgtct cctcagcttc aggcaccacc actgacctgg gacagtgaat 660

ccggacatcg attctaaggt aaatataaaa tttttaagtg tataatttgt taaactactg 720

attctaattg tttctctctt ttagattcca acctttggaa ctgaattcta gaccaccatg 780

cagagggtga acatgatcat ggctgagagc cctggcctga tcaccatctg cctgctgggc 840

tacctgctgt ctgctgagtg cactgtgttc ctggaccatg agaatgccaa caagatcctg 900

aacaggccca agagatacaa ctctggcaag ctggagggagt ttgtgcaggg caacctggag 960

agggagtgca tggaggagaa gtgcagcttt gaggaggcca gggaggtgtt tgagaacact 1020

gagaggacca ctgagttctg gaagcagtat gtggatgggg accagtgtga gagcaacccc 1080

tgcctgaatg ggggcagctg caaggatgac atcaacagct atgagtgctg gtgccccttt 1140

ggctttgagg gcaagaactg tgagctggat gtgacctgca acatcaagaa tggcagatgt 1200

gagcagttct gcaagaactc tgctgacaac aaggtggtgt gcagctgcac tgaggctac 1260

aggctggctg agaaccagaa gagctgtgag cctgctgtgc cattcccatg tggcagagtg 1320

tctgtgagcc agaccagcaa gctgaccagg gctgaggctg tgttccctga tgtggactat 1380

gtgaacagca ctgaggctga aaccatcctg gacaacatca cccagagcac cccagcttc 1440

aatgacttca cccaggtggt ggggggggag gatgccaagc ctggccagtt cccctggcaa 1500

gtggtgctga atggcaaggt ggatgccttc tgtggggggca gcattgtgaa tgagaagtgg 1560

attgtgactg ctgcccactg tgtggagact ggggtgaaga tcactgtggt ggctggggag 1620

cacaacattg aggagactga gcacactgag cagaagagga atgtgatcag gatcatcccc 1680

caccacaact acaatgctgc catcaacaag tacaaccatg acattgccct gctggagctg 1740

gatgagcccc tggtgctgaa cagctatgtg acccccatct gcattgctga caaggagtac 1800

accaacatct tcctgaagtt tggctctggc tatgtgtctg gctggggcag ggtgttccac 1860

aagggcaggt ctgccctggt gctgcagtac ctgagggtgc ccctggtgga cagggccacc 1920

tgcctgagga gcaccaagtt caccatctac aacaacatgt tctgtgctgg cttccatgag 1980

gggggcaggg acagctgcca gggggactct gggggccccc atgtgactga ggtggagggc 2040

accagcttcc tgactggcat catcagctgg ggggaggagt gtgccatgaa gggcaagtat 2100

ggcatctaca ccaaagtctc cagatatgtg aactggatca aggagaagac caagctgacc 2160

taatgactcc atggttcgaa tgctttattt gtgaaatttg tgatgctatt gctttatttg 2220

taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc 2280

aggttcaggg ggaggtgtgg gaggtttttt aaaactagtg cggccgctct agagcatggc 2340

tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag 2400

ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc 2460

cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg 2510

<210> 63

<211> 126

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 63

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggaaacc cgggcgtgcc 60

cgggcgcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg 120

gttcct 126

<210> 64

<211> 120

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 64

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgcccggga aacccgggcg tgcgcctcag tgagcgagcg agcgcgcagc tgcctgcagg 120

<210> 65

<211> 80

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 65

gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct 60

cagtgagcga gcgagcgcgc 80

<210> 66

<211> 141

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 66

aataaacgat aacgccgttg gtggcgtgag gcatgtaaaa ggttacatca ttatcttgtt 60

cgccatccgg ttggtataaa tagacgttca tgttggtttt tgtttcagtt gcaagttggc 120

tgcggcgcgc gcagcacctt t 141

<210> 67

<211> 1876

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 67

cgcagccacc atggcggggt tttacgagat tgtgattaag gtccccagcg accttgacga 60

gcatctgccc ggcatttctg acagctttgt gaactgggtg gccgagaagg aatgggagtt 120

gccgccagat tctgacatgg atctgaatct gattgagcag gcacccctga ccgtggccga 180

gaagctgcag cgcgactttc tgacggaatg gcgccgtgtg agtaaggccc cggaggccct 240

tttctttgtg caatttgaga agggagagag ctacttccac atgcacgtgc tcgtggaaac 300

caccggggtg aaatccatgg ttttgggacg tttcctgagt cagattcgcg aaaaactgat 360

tcagagaatt taccgcggga tcgagccgac tttgccaaac tggttcgcgg tcacaaagac 420

cagaaatggc gccggaggcg ggaacaaggt ggtggatgag tgctacatcc ccaattactt 480

gctccccaaa acccagcctg agctccagtg ggcgtggact aatatggaac agtatttaag 540

cgcctgtttg aatctcacgg agcgtaaacg gttggtggcg cagcatctga cgcacgtgtc 600

gcagacgcag gagcagaaca aagagaatca gaatcccaat tctgatgcgc cggtgatcag 660

atcaaaaact tcagccaggt acatggagct ggtcgggtgg ctcgtggaca aggggattac 720

ctcggagaag cagtggatcc aggaggacca ggcctcatac atctccttca atgcggcctc 780

caactcgcgg tcccaaatca aggctgcctt ggacaatgcg ggaaagatta tgagcctgac 840

taaaaccgcc cccgactacc tggtgggcca gcagcccgtg gaggacatt ccagcaatcg 900

gatttataaa attttggaac taaacgggta cgatccccaa tatgcggctt ccgtctttct 960

gggatgggcc acgaaaaagt tcggcaagag gaacaccatc tggctgtttg ggcctgcaac 1020

taccgggaag accaacatcg cggaggccat agcccacact gtgcccttct acgggtgcgt 1080

aaactggacc aatgagaact ttcccttcaa cgactgtgtc gacaagatgg tgatctggtg 1140

ggaggagggg aagatgaccg ccaaggtcgt ggagtcggcc aaagccattc tcggaggaag 1200

caaggtgcgc gtggaccaga aatgcaagtc ctcggcccag atagacccga ctcccgtgat 1260

cgtcacctcc aacaccaaca tgtgcgccgt gattgacggg aactcaacga ccttcgaaca 1320

ccagcagccg ttgcaagacc ggatgttcaa atttgaactc acccgccgtc tggatcatga 1380

ctttgggaag gtcaccaagc aggaagtcaa agactttttc cggtgggcaa aggatcacgt 1440

ggttgaggtg gagcatgaat tctacgtcaa aaagggtgga gccaagaaaa gacccgcccc 1500

cagtgacgca gatataagtg agcccaaacg ggtgcgcgag tcagttgcgc agccatcgac 1560

gtcagacgcg gaagcttcga tcaactacgc agacaggtac caaaacaaat gttctcgtca 1620

cgtgggcatg aatctgatgc tgtttccctg cagacaatgc gagagaatga atcagaattc 1680

aaatatctgc ttcactcacg gacagaaaga ctgtttagag tgctttcccg tgtcagaatc 1740

tcaacccgtt tctgtcgtca aaaaggcgta tcagaaactg tgctacattc atcatatcat 1800

gggaaaggtg cgacgctt gcactgcctg cgatctggtc aatgtggatt tggatgactg 1860

catctttgaa caataa 1876

<210> 68

<211> 129

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 68

atcatggaga taattaaaat gataaccatc tcgcaaataa ataagtattt tactgttttc 60

gtaacagttt tgtaataaaa aaacctataa atattccgga ttattcatac cgtcccacca 120

tcgggcgcg 129

<210> 69

<211> 1203

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 69

gccgccacca tggagttggt gggctggctc gtggacaaag gcattacttc ggaaaagcag 60

tggattcagg aggatcaggc atcttacatc tcattcaacg ctgccagtaa ctcgaggtcc 120

cagatcaagg cagcgctgga caacgcggga aagattatga gtctgaccaa aactgctcca 180

gactacctcg ttggtcagca accggtggaa gatatctcca gcaacaggat ctacaagatt 240

ctggagctca acggctacga ccctcaatac gctgcctcag tgttcttggg ttgggccacc 300

aagaaattcg gcaagagaaa cactatctgg ctgttcggcc ccgctaccac tggaaagaca 360

aacatcgcag aagcgattgc tcacacggtg ccattctacg gctgcgtcaa ctggacaaac 420

gagaacttcc cgttcaacga ctgtgtcgat aagatggtta tctggtggga ggaaggaaag 480

atgacggcca aagtggtcga aagcgccaag gcaattctgg gtggctctaa agtgcgcgtc 540

gaccagaagt gcaaatcttc agctcaaatc gatcctaccc ccgttattgt gacatcaaac 600

acgaacatgt gtgccgtgat cgacggaaac agtacaacgt tcgaacacca gcaacctctc 660

caggatcgta tgttcaagtt cgagctcacc cgccgtttgg accatgattt cggcaaggtc 720

actaaacaag aggttaagga cttcttccgc tgggctaaag atcacgttgt ggaggttgaa 780

catgagttct acgtcaagaa aggaggtgct aagaaacgtc cagccccgtc ggacgcagat 840

atctccgaac ctaagagggt gagagagtcg gtcgcacagc caagcacttc tgacgcagaa 900

gcttccatta actacgcaga taggtaccaa aacaagtgca gcagacacgt gggtatgaac 960

ttgatgctgt tcccatgccg ccagtgtgag cgtatgaacc aaaactctaa catctgtttc 1020

acacatggcc agaaggactg cctcgaatgt ttccctgtgt cagagagtca gcccgtctca 1080

gtcgttaaga aagcttacca aaagttgtgc tacatccacc atattatggg taaagtccct 1140

gatgcctgta ccgcttgtga tctggtcaac gtggatttgg acgactgtat tttcgagcaa 1200

taa 1203

<210> 70

<211> 388

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 70

gaacagagaa acaggagaat atgggccaaa caggatatct gtggtaagca gttcctgccc 60

cggctcaggg ccaagaacag ttggaacagc agaatatggg ccaaacagga tatctgtggt 120

aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtcccgccc 180

tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc 240

ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc 300

tccccgagct ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc 360

atccacgctg ttttgacttc catagaag 388

<210> 71

<211> 1662

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 71

gccgccacca tggaagacgc caaaaacata aagaaaggcc cggcgccatt ctatccgctg 60

gaagatggaa ccgctggaga gcaactgcat aaggctatga agagatacgc cctggttcct 120

ggaacaattg cttttacaga tgcacatatc gaggtggaca tcacttacgc tgagtacttc 180

gaaatgtccg ttcggttggc agaagctatg aaacgatatg ggctgaatac aaatcacaga 240

atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg cgcgttattt 300

atcggagttg cagttgcgcc cgcgaacgac atttataatg aacgtgaatt gctcaacagt 360

atgggcattt cgcagcctac cgtggtgttc gtttccaaaa aggggttgca aaaaattttg 420

aacgtgcaaa aaaagctccc aatcatccaa aaaattatta tcatggattc taaaacggat 480

taccagggat ttcagtcgat gtacacgttc gtcacatctc atctacctcc cggttttaat 540

gaatacgatt ttgtgccaga gtccttcgat agggacaaga caattgcact gatcatgaac 600

tcctctggat ctactggtct gcctaaaggt gtcgctctgc ctcatagaac tgcctgcgtg 660

agattctcgc atgccagaga tcctattttt ggcaatcaaa tcattccgga tactgcgatt 720

ttaagtgttg ttccattcca tcacggtttt ggaatgttta ctacactcgg atatttgata 780

tgtggatttc gagtcgtctt aatgtataga tttgaagaag agctgtttct gaggagcctt 840

caggattaca agattcaaag tgcgctgctg gtgccaaccc tattctcctt cttcgccaaa 900

agcactctga ttgacaaata cgatttatct aatttacacg aaattgcttc tggtggcgct 960

cccctctcta aggaagtcgg ggaagcggtt gccaagaggt tccatctgcc aggtatcagg 1020

caaggatatg ggctcactga gactacatca gctattctga ttacacccga gggggatgat 1080

aaaccgggcg cggtcggtaa agttgttcca ttttttgaag cgaaggttgt ggatctggat 1140

accgggaaaa cgctgggcgt taatcaaaga ggcgaactgt gtgtgagagg tcctatgatt 1200

atgtccggtt atgtaaacaa tccggaagcg accaacgcct tgattgacaa ggatggatgg 1260

ctacattctg gagacatagc ttactgggac gaagacgaac acttcttcat cgttgaccgc 1320

ctgaagtctc tgattaagta caaaggctat caggtggctc ccgctgaatt ggaatccatc 1380

ttgctccaac accccaacat cttcgacgca ggtgtcgcag gtcttcccga cgatgacgcc 1440

ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa agacgatgac ggaaaaagag 1500

atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg aggagttgtg 1560

tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa aatcagagag 1620

atcctcataa aggccaagaa gggcggaaag atcgccgtgt aa 1662

<210> 72

<211> 581

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 72

gagcatctta ccgccattta ttcccatatt tgttctgttt ttcttgattt gggtatacat 60

ttaaatgtta ataaaacaaa atggtggggc aatcatttac atttttaggg atatgtaatt 120

actagttcag gtgtattgcc acaagacaaa catgttaaga aactttcccg ttatttacgc 180

tctgttcctg ttaatcaacc tctggattac aaaatttgtg aaagattgac tgatattctt 240

aactatgttg ctccttttac gctgtgtgga tatgctgctt tatagcctct gtatctagct 300

attgcttccc gtacggcttt cgttttctcc tccttgtata aatcctggtt gctgtctctt 360

ttagaggagt tgtggcccgt tgtccgtcaa cgtggcgtgg tgtgctctgt gtttgctgac 420

gcaaccccca ctggctgggg cattgccacc acctgtcaac tcctttctgg gactttcgct 480

ttccccctcc cgatcgccac ggcagaactc atcgccgcct gccttgcccg ctgctggaca 540

ggggctaggt tgctgggcac tgataattcc gtggtgttgt c 581

<210> 73

<211> 225

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 73

tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct 60

ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct 120

gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg 180

ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggc 225

<210> 74

<211> 1177

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 74

ggctcagagg ctcagaggca cacaggagtt tctgggctca ccctgccccc ttccaacccc 60

tcagttccca tcctccagca gctgtttgtg tgctgcctct gaagtccaca ctgaacaaac 120

ttcagcctac tcatgtccct aaaatgggca aacattgcaa gcagcaaaca gcaaacacac 180

agccctccct gcctgctgac cttggagctg gggcagaggt cagagacctc tctgggccca 240

tgccacctcc aacatccact cgaccccttg gaatttcggt ggagaggagc agaggttgtc 300

ctggcgtggt ttaggtagtg tgagagggtc cgggttcaaa accacttgct gggtggggag 360

tcgtcagtaa gtggctatgc cccgaccccg aagcctgttt ccccatctgt acaatggaaa 420

tgataaagac gcccatctga tagggttttt gtggcaaata aacatttggt ttttttgttt 480

tgttttgttt tgttttttga gatggaggtt tgctctgtcg cccaggctgg agtgcagtga 540

cacaatctca tctcaccaca accttcccct gcctcagcct cccaagtagc tgggattaca 600

agcatgtgcc accacacctg gctaattttc tatttttagt agagacgggt ttctccatgt 660

tggtcagcct cagcctccca agtaactggg attacaggcc tgtgccacca cacccggcta 720

attttttcta tttttgacag ggacggggtt tcaccatgtt ggtcaggctg gtctagaggt 780

accggatctt gctaccagtg gaacagccac taaggattct gcagtgagag cagaggcca 840

gctaagtggt actctcccag agactgtctg actcacgcca ccccctccac cttggacaca 900

ggacgctgtg gtttctgagc caggtacaat gactcctttc ggtaagtgca gtggaagctg 960

tacactgccc aggcaaagcg tccgggcagc gtaggcgggc gactcagatc ccagccagtg 1020

gacttagccc ctgtttgctc ctccgataac tggggtgacc ttggttaata ttcaccagca 1080

gcctcccccg ttgcccctct ggatccactg cttaaatacg gacgaggaca gggccctgtc 1140

tcctcagctt caggcaccac cactgacctg ggacagt 1177

<210> 75

<400> 75

000

<210> 76

<400> 76

000

<210> 77

<400> 77

000

<210> 78

<400> 78

000

<210> 79

<400> 79

000

<210> 80

<400> 80

000

<210> 81

<400> 81

000

<210> 82

<400> 82

000

<210> 83

<400> 83

000

<210> 84

<400> 84

000

<210> 85

<400> 85

000

<210> 86

<400> 86

000

<210> 87

<400> 87

000

<210> 88

<400> 88

000

<210> 89

<400> 89

000

<210> 90

<400> 90

000

<210> 91

<400> 91

000

<210> 92

<400> 92

000

<210> 93

<400> 93

000

<210> 94

<400> 94

000

<210> 95

<400> 95

000

<210> 96

<400> 96

000

<210> 97

<400> 97

000

<210> 98

<400> 98

000

<210> 99

<400> 99

000

<210> 100

<400> 100

000

<210> 101

<211> 70

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 101

gcgcgctcgc tcgctcactg aggccgcccg ggaaacccgg gcgtgcgcct cagtgagcga 60

gcgagcgcgc 70

<210> 102

<211> 70

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 102

gcgcgctcgc tcgctcactg aggcgcacgc ccgggtttcc cgggcggcct cagtgagcga 60

gcgagcgcgc 70

<210> 103

<211> 72

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 103

gcgcgctcgc tcgctcactg aggccgtcgg gcgacctttg gtcgcccggc ctcagtgagc 60

gagcgagcgc gc 72

<210> 104

<211> 72

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 104

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacggc ctcagtgagc 60

gagcgagcgc gc 72

<210> 105

<211> 72

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 105

gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggc ctcagtgagc 60

gagcgagcgc gc 72

<210> 106

<211> 72

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 106

gcgcgctcgc tcgctcactg aggccgacgc ccgggctttg cccgggcggc ctcagtgagc 60

gagcgagcgc gc 72

<210> 107

<211> 83

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 107

gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg ctttgcccgg 60

cctcagtgag cgagcgagcg cgc 83

<210> 108

<211> 83

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 108

gcgcgctcgc tcgctcactg aggccgggca aagcccgacg cccgggcttt gcccgggcgg 60

cctcagtgag cgagcgagcg cgc 83

<210> 109

<211> 77

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 109

gcgcgctcgc tcgctcactg aggccgaaac gtcgggcgac ctttggtcgc ccggcctcag 60

tgagcgagcg agcgcgc 77

<210> 110

<211> 77

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 110

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgtt tcggcctcag 60

tgagcgagcg agcgcgc 77

<210> 111

<211> 51

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 111

gcgcgctcgc tcgctcactg aggcaaagcc tcagtgagcg agcgagcgcg c 51

<210> 112

<211> 51

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 112

gcgcgctcgc tcgctcactg aggctttgcc tcagtgagcg agcgagcgcg c 51

<210> 113

<211> 80

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 113

gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct 60

cagtgagcga gcgagcgcgc 80

<210> 114

<211> 80

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 114

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggcggcct 60

cagtgagcga gcgagcgcgc 80

<210> 115

<211> 79

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 115

gcgcgctcgc tcgctcactg aggcgcccgg gcgtcgggcg acctttggtc gcccggcctc 60

agtgagcgag cgagcgcgc 79

<210> 116

<211> 79

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 116

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggcgcctc 60

agtgagcgag cgagcgcgc 79

<210> 117

<211> 89

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 117

gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgactttgtc 60

gcccggcctc agtgagcgag cgagcgcgc 89

<210> 118

<211> 89

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 118

gcgcgctcgc tcgctcactg aggccgggcg acaaagtcgc ccgacgcccg ggctttgccc 60

gggcggcctc agtgagcgag cgagcgcgc 89

<210> 119

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 119

gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgattttcgc 60

ccggcctcag tgagcgagcg agcgcgc 87

<210> 120

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 120

gcgcgctcgc tcgctcactg aggccgggcg aaaatcgccc gacgcccggg ctttgcccgg 60

gcggcctcag tgagcgagcg agcgcgc 87

<210> 121

<211> 85

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 121

gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgtttcgccc 60

ggcctcagtg agcgagcgag cgcgc 85

<210> 122

<211> 85

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 122

gcgcgctcgc tcgctcactg aggccgggcg aaacgcccga cgcccgggct ttgcccgggc 60

ggcctcagtg agcgagcgag cgcgc 85

<210> 123

<211> 89

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 123

gcgcgctcgc tcgctcactg aggccgcccg ggaaacccgg gcgtcgggcg acctttggtc 60

gcccggcctc agtgagcgag cgagcgcgc 89

<210> 124

<211> 89

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 124

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggtttccc 60

gggcggcctc agtgagcgag cgagcgcgc 89

<210> 125

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 125

gcgcgctcgc tcgctcactg aggccgcccg gaaaccgggc gtcgggcgac ctttggtcgc 60

ccggcctcag tgagcgagcg agcgcgc 87

<210> 126

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 126

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cggtttccgg 60

gcggcctcag tgagcgagcg agcgcgc 87

<210> 127

<211> 85

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 127

gcgcgctcgc tcgctcactg aggccgcccg aaacgggcgt cgggcgacct ttggtcgccc 60

ggcctcagtg agcgagcgag cgcgc 85

<210> 128

<211> 85

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 128

gcgcgctcgc tcgctcactg aggccggggcg accaaaggtc gcccgacgcc cgtttcgggc 60

ggcctcagtg agcgagcgag cgcgc 85

<210> 129

<211> 83

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 129

gcgcgctcgc tcgctcactg aggccgccca aagggcgtcg ggcgaccttt ggtcgcccgg 60

cctcagtgag cgagcgagcg cgc 83

<210> 130

<211> 83

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 130

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc ctttgggcgg 60

cctcagtgag cgagcgagcg cgc 83

<210> 131

<211> 81

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 131

gcgcgctcgc tcgctcactg aggccgccaa aggcgtcggg cgacctttgg tcgcccggcc 60

tcagtgagcg agcgagcgcg c 81

<210> 132

<211> 81

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 132

gcgcgctcgc tcgctcactg aggccggggcg accaaaggtc gcccgacgcc tttggcggcc 60

tcagtgagcg agcgagcgcg c 81

<210> 133

<211> 79

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 133

gcgcgctcgc tcgctcactg aggccgcaaa gcgtcgggcg acctttggtc gcccggcctc 60

agtgagcgag cgagcgcgc 79

<210> 134

<211> 79

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 134

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgct ttgcggcctc 60

agtgagcgag cgagcgcgc 79

<210> 135

<211> 546

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 135

ccctaaaatg ggcaaacatt gcaagcagca aacagcaaac acacagccct ccctgcctgc 60

tgaccttgga gctggggcag aggtcagaga cctctctggg cccatgccac ctccaacatc 120

cactcgaccc cttggaattt ttcggtggag aggagcagag gttgtcctgg cgtggtttag 180

gtagtgtgag aggggaatga ctcctttcgg taagtgcagt ggaagctgta cactgcccag 240

gcaaagcgtc cgggcagcgt aggcgggcga ctcagatccc agccagtgga cttagcccct 300

gtttgctcct ccgataactg gggtgacctt ggttaatatt caccagcagc ctcccccgtt 360

gcccctctgg atccactgct taaatacgga cgaggacagg gccctgtctc ctcagcttca 420

ggcaccacca ctgacctggg acagtgaatc cggactctaa ggtaaatata aaatttttaa 480

gtgtataatg tgttaaacta ctgattctaa ttgtttctct cttttagatt ccaacctttg 540

gaactg 546

<210> 136

<400> 136

000

<210> 137

<400> 137

000

<210> 138

<400> 138

000

<210> 139

<400> 139

000

<210> 140

<400> 140

000

<210> 141

<400> 141

000

<210> 142

<400> 142

000

<210> 143

<400> 143

000

<210> 144

<400> 144

000

<210> 145

<400> 145

000

<210> 146

<400> 146

000

<210> 147

<400> 147

000

<210> 148

<400> 148

000

<210> 149

<400> 149

000

<210> 150

<400> 150

000

<210> 151

<211> 1530

<212> DNA

<213> Homo sapiens

<400> 151

atgtccatgg gggcaccgcg gtccctcctc ctggccctgg ctgctggcct ggccgttgcc 60

cgtccgccca acatcgtgct gatctttgcc gacgacctcg gctatgggga cctgggctgc 120

tatgggcacc ccagctctac cactcccaac ctggaccagc tggcggcggg agggctgcgg 180

ttcacagact tctacgtgcc tgtgtctctg tgcacaccct ctagggccgc cctcctgacc 240

ggccggctcc cggttcggat gggcatgtac cctggcgtcc tggtgcccag ctcccggggg 300

ggcctgcccc tggaggaggt gaccgtggcc gaagtcctgg ctgcccgagg ctacctcaca 360

ggaatggccg gcaagtggca ccttggggtg gggcctgagg gggccttcct gcccccccat 420

cagggcttcc atcgatttct aggcatcccg tactcccacg accagggccc ctgccagaac 480

ctgacctgct tcccgccggc cactccttgc gacggtggct gtgaccaggg cctggtcccc 540

atcccactgt tggccaacct gtccgtggag gcgcagcccc cctggctgcc cggactagag 600

gcccgctaca tggctttcgc ccatgacctc atggccgacg cccagcgcca ggatcgcccc 660

ttcttcctgt actatgcctc tcaccacacc cactaccctc agttcagtgg gcagagcttt 720

gcagagcgtt caggccgcgg gccatttggg gactccctga tggagctgga tgcagctgtg 780

gggaccctga tgacagccat aggggacctg gggctgcttg aagagacgct ggtcatcttc 840

actgcagaca atggacctga gaccatgcgt atgtcccgag gcggctgctc cggtctcttg 900

cggtgtggaa agggaacgac ctacgagggc ggtgtccgag agcctgcctt ggccttctgg 960

ccaggtcata tcgctcccgg cgtgacccac gagctggcca gctccctgga cctgctgcct 1020

accctggcag ccctggctgg ggccccactg cccaatgtca ccttggatgg ctttgacctc 1080

agccccctgc tgctgggcac aggcaagagc cctcggcagt ctctcttctt ctacccgtcc 1140

tacccagacg aggtccgtgg ggtttttgct gtgcggactg gaaagtacaa ggctcacttc 1200

ttcacccagg gctctgccca cagtgatacc actgcagacc ctgcctgcca cgcctccagc 1260

tctctgactg ctcatgagcc cccgctgctc tatgacctgt ccaaggaccc tggtgagaac 1320

tacaacctgc tggggggtgt ggccggggcc accccagagg tgctgcaagc cctgaaacag 1380

cttcagctgc tcaaggccca gttagacgca gctgtgacct tcggccccag ccaggtggcc 1440

cggggcgagg accccgccct gcagatctgc tgtcatcctg gctgcacccc ccgcccagct 1500

tgctgccatt gccgatcc ccatgcctga 1530

<210> 152

<211> 1653

<212> DNA

<213> Homo sapiens

<400> 152

atgccgccac cccggaccgg ccgaggcctt ctctggctgg gtctggttct gagctccgtc 60

tgcgtcgccc tcggatccga aacgcaggcc aactcgacca cagatgctct gaacgttctt 120

ctcatcatcg tggatgacct gcgcccctcc ctgggctgtt atggggataa gctggtgagg 180

tccccaaata ttgaccaact ggcatcccac agcctcctct tccagaatgc ctttgcgcag 240

caagcagtgt gcgccccgag ccgcgtttct ttcctcactg gcaggagacc tgacaccacc 300

cgcctgtacg acttcaactc ctactggagg gtgcacgctg gaaacttctc caccatcccc 360

cagtacttca aggagaatgg ctatgtgacc atgtcggtgg gaaaagtctt tcaccctggg 420

atatcttcta accataccga tgattctccg tatagctggt cttttccacc ttatcatcct 480

tcctctgaga agtatgaaaa cactaagaca tgtcgagggc cagatggaga actccatgcc 540

aacctgcttt gccctgtgga tgtgctggat gttcccgagg gcaccttgcc tgacaaacag 600

agcactgagc aagccataca gttgttggaa aagatgaaaa cgtcagccag tcctttcttc 660

ctggccgttg ggtatcataa gccacacatc cccttcagat accccaagga atttcagaag 720

ttgtatccct tggagaacat caccctggcc cccgatcccg aggtccctga tggcctaccc 780

cctgtggcct acaacccctg gatggacatc aggcaacggg aagacgtcca agccttaaac 840

atcagtgtgc cgtatggtcc aattcctgtg gactttcagc ggaaaatccg ccagagctac 900

tttgcctctg tgtcatattt ggatacacag gtcggccgcc tcttgagtgc tttggacgat 960

cttcagctgg ccaacagcac catcattgca tttacctcgg atcatgggtg ggctctaggt 1020

gaacatggag aatgggccaa atacagcaat tttgatgttg ctacccatgt tcccctgata 1080

ttctatgttc ctggaaggac ggcttcactt ccggaggcag gcgagaagct tttcccttac 1140

ctcgaccctt ttgattccgc ctcacagttg atggagccag gcaggcaatc catggacctt 1200

gtggaacttg tgtctctttt tcccacgctg gctggacttg caggactgca ggttccacct 1260

cgctgccccg ttccttcatt tcacgttgag ctgtgcagag aaggcaagaa ccttctgaag 1320

cattttcgat tccgtgactt ggaagaggat ccgtacctcc ctggtaatcc ccgtgaactg 1380

attgcctata gccagtatcc ccggccttca gacatccctc agtggaattc tgacaagccg 1440

agtttaaaag atataaagat catgggctat tccatacgca ccatagacta taggtatact 1500

gtgtgggttg gcttcaatcc tgatgaattt ctagctaact tttctgacat ccatgcaggg 1560

gaactgtatt ttgtggattc tgacccattg caggatcaca atatgtataa tgattcccaa 1620

ggtggagatc ttttccagtt gttgatgcct tga 1653

<210> 153

<400> 153

000

<210> 154

<400> 154

000

<210> 155

<400> 155

000

<210> 156

<400> 156

000

<210> 157

<400> 157

000

<210> 158

<400> 158

000

<210> 159

<400> 159

000

<210> 160

<400> 160

000

<210> 161

<400> 161

000

<210> 162

<400> 162

000

<210> 163

<400> 163

000

<210> 164

<400> 164

000

<210> 165

<400> 165

000

<210> 166

<400> 166

000

<210> 167

<400> 167

000

<210> 168

<400> 168

000

<210> 169

<400> 169

000

<210> 170

<400> 170

000

<210> 171

<400> 171

000

<210> 172

<400> 172

000

<210> 173

<400> 173

000

<210> 174

<400> 174

000

<210> 175

<400> 175

000

<210> 176

<400> 176

000

<210> 177

<400> 177

000

<210> 178

<400> 178

000

<210> 179

<400> 179

000

<210> 180

<400> 180

000

<210> 181

<400> 181

000

<210> 182

<400> 182

000

<210> 183

<400> 183

000

<210> 184

<400> 184

000

<210> 185

<400> 185

000

<210> 186

<400> 186

000

<210> 187

<400> 187

000

<210> 188

<400> 188

000

<210> 189

<400> 189

000

<210> 190

<400> 190

000

<210> 191

<400> 191

000

<210> 192

<400> 192

000

<210> 193

<400> 193

000

<210> 194

<400> 194

000

<210> 195

<400> 195

000

<210> 196

<400> 196

000

<210> 197

<400> 197

000

<210> 198

<400> 198

000

<210> 199

<400> 199

000

<210> 200

<400> 200

000

<210> 201

<400> 201

000

<210> 202

<400> 202

000

<210> 203

<400> 203

000

<210> 204

<400> 204

000

<210> 205

<400> 205

000

<210> 206

<400> 206

000

<210> 207

<400> 207

000

<210> 208

<400> 208

000

<210> 209

<400> 209

000

<210> 210

<400> 210

000

<210> 211

<400> 211

000

<210> 212

<400> 212

000

<210> 213

<400> 213

000

<210> 214

<400> 214

000

<210> 215

<400> 215

000

<210> 216

<400> 216

000

<210> 217

<400> 217

000

<210> 218

<400> 218

000

<210> 219

<400> 219

000

<210> 220

<400> 220

000

<210> 221

<400> 221

000

<210> 222

<400> 222

000

<210> 223

<400> 223

000

<210> 224

<400> 224

000

<210> 225

<400> 225

000

<210> 226

<400> 226

000

<210> 227

<400> 227

000

<210> 228

<400> 228

000

<210> 229

<400> 229

000

<210> 230

<400> 230

000

<210> 231

<400> 231

000

<210> 232

<400> 232

000

<210> 233

<400> 233

000

<210> 234

<400> 234

000

<210> 235

<400> 235

000

<210> 236

<400> 236

000

<210> 237

<400> 237

000

<210> 238

<400> 238

000

<210> 239

<400> 239

000

<210> 240

<400> 240

000

<210> 241

<400> 241

000

<210> 242

<400> 242

000

<210> 243

<400> 243

000

<210> 244

<400> 244

000

<210> 245

<400> 245

000

<210> 246

<400> 246

000

<210> 247

<400> 247

000

<210> 248

<400> 248

000

<210> 249

<400> 249

000

<210> 250

<400> 250

000

<210> 251

<400> 251

000

<210> 252

<400> 252

000

<210> 253

<400> 253

000

<210> 254

<400> 254

000

<210> 255

<400> 255

000

<210> 256

<400> 256

000

<210> 257

<400> 257

000

<210> 258

<400> 258

000

<210> 259

<400> 259

000

<210> 260

<400> 260

000

<210> 261

<400> 261

000

<210> 262

<400> 262

000

<210> 263

<400> 263

000

<210> 264

<400> 264

000

<210> 265

<400> 265

000

<210> 266

<400> 266

000

<210> 267

<400> 267

000

<210> 268

<400> 268

000

<210> 269

<400> 269

000

<210> 270

<400> 270

000

<210> 271

<400> 271

000

<210> 272

<400> 272

000

<210> 273

<400> 273

000

<210> 274

<400> 274

000

<210> 275

<400> 275

000

<210> 276

<400> 276

000

<210> 277

<400> 277

000

<210> 278

<400> 278

000

<210> 279

<400> 279

000

<210> 280

<400> 280

000

<210> 281

<400> 281

000

<210> 282

<400> 282

000

<210> 283

<400> 283

000

<210> 284

<400> 284

000

<210> 285

<400> 285

000

<210> 286

<400> 286

000

<210> 287

<400> 287

000

<210> 288

<400> 288

000

<210> 289

<400> 289

000

<210> 290

<400> 290

000

<210> 291

<400> 291

000

<210> 292

<400> 292

000

<210> 293

<400> 293

000

<210> 294

<400> 294

000

<210> 295

<400> 295

000

<210> 296

<400> 296

000

<210> 297

<400> 297

000

<210> 298

<400> 298

000

<210> 299

<400> 299

000

<210> 300

<400> 300

000

<210> 301

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 301

gcgcgctcgc tcgctc 16

<210> 302

<211> 8

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 302

actgaggc 8

<210> 303

<211> 22

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 303

cgggcgacca aaggtcgccc ga 22

<210> 304

<211> 10

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 304

cgcccgggcg 10

<210> 305

<211> 8

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 305

gcctcagt 8

<210> 306

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 306

gagcgagcga gcgcgc 16

<210> 307

<400> 307

000

<210> 308

<211> 317

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 308

ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 60

agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 120

tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 180

ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 240

aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag 300

gcctaggctt ttgcaaa 317

<210> 309

<211> 576

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 309

tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa 60

cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata 120

atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag 180

tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc 240

cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta 300

tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg 360

cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt 420

ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca 480

aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag 540

gtctatataa gcagagctgg tttagtgaac cgtcag 576

<210> 310

<211> 1313

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 310

ggagccgaga gtaattcata caaaaggagg gatcgccttc gcaaggggag agcccaggga 60

ccgtccctaa attctcacag acccaaatcc ctgtagccgc cccacgacag cgcgaggagc 120

atgcgcccag ggctgagcgc gggtagatca gagcacacaa gctcacagtc cccggcggtg 180

gggggagggg cgcgctgagc gggggccagg gagctggcgc ggggcaaact gggaaagtgg 240

tgtcgtgtgc tggctccgcc ctcttcccga gggtggggga gaacggtata taagtgcggt 300

agtcgccttg gacgttcttt ttcgcaacgg gtttgccgtc agaacgcagg tgagtggcgg 360

gtgtggcttc cgcgggcccc ggagctggag ccctgctctg agcgggccgg gctgatatgc 420

gagtgtcgtc cgcagggttt agctgtgagc attcccactt cgagtggcgg gcggtgcggg 480

ggtgagagtg cgaggcctag cggcaacccc gtagcctcgc ctcgtgtccg gcttgaggcc 540

tagcgtggtg tccgccgccg cgtgccactc cggccgcact atgcgttttt tgtccttgct 600

gccctcgatt gccttccagc agcatgggct aacaaaggga gggtgtgggg ctcactctta 660

aggagcccat gaagcttacg ttggatagga atggaagggc aggaggggcg actggggccc 720

gcccgccttc ggagcacatg tccgacgcca cctggatggg gcgaggcctg tggctttccg 780

aagcaatcgg gcgtgagttt agcctacctg ggccatgtgg ccctagcact gggcacggtc 840

tggcctggcg gtgccgcgtt cccttgcctc ccaacaaggg tgaggccgtc ccgcccggca 900

ccagttgctt gcgcggaaag atggccgctc ccggggccct gttgcaagga gctcaaaatg 960

gaggacgcgg cagcccggtg gagcgggcgg gtgagtcacc cacacaaagg aagagggcct 1020

tgcccctcgc cggccgctgc ttcctgtgac cccgtggtct atcggccgca tagtcacctc 1080

gggcttctct tgagcaccgc tcgtcgcggc ggggggaggg gatctaatgg cgttggagtt 1140

tgttcacatt tggtgggtgg agactagtca ggccagcctg gcgctggaag tcattcttgg 1200

aatttgcccc tttgagtttg gagcgaggct aattctcaag cctcttagcg gttcaaaggt 1260

attttctaaa cccgtttcca ggtgttgtga aagccaccgc taattcaaag caa 1313

<210> 311

<400> 311

000

<210> 312

<400> 312

000

<210> 313

<211> 19

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic peptide

<400> 313

Met Asp Trp Thr Trp Arg Ile Leu Phe Leu Val Ala Ala Ala Thr Gly

1 5 10 15

Ala His Ser

<210> 314

<211> 19

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic peptide

<400> 314

Met Leu Pro Ser Gln Leu Ile Gly Phe Leu Leu Leu Trp Val Pro Ala

1 5 10 15

Ser Arg Gly

<210> 315

<211> 7

<212> PROTEIN

<213> Simian Virus 40

<400> 315

Pro Lys Lys Lys Arg Lys Val

1 5

<210> 316

<211> 16

<212> PROTEIN

<213> Unknown

<220>

<223> Description of unknown:

Double NLS sequence of nucleoplasmin

<400> 316

Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys Lys

1 5 10 15

<210> 317

<211> 9

<212> PROTEIN

<213> Unknown

<220>

<223> Description of unknown:

C-myc NLS sequence

<400> 317

Pro Ala Ala Lys Arg Val Lys Leu Asp

1 5

<210> 318

<211> 11

<212> PROTEIN

<213> Unknown

<220>

<223> Description of unknown:

C-myc NLS sequence

<400> 318

Arg Gln Arg Arg Asn Glu Leu Lys Arg Ser Pro

1 5 10

<210> 319

<211> 38

<212> PROTEIN

<213> Homo sapiens

<400> 319

Asn Gln Ser Ser Asn Phe Gly Pro Met Lys Gly Gly Asn Phe Gly Gly

1 5 10 15

Arg Ser Ser Gly Pro Tyr Gly Gly Gly Gly Gln Tyr Phe Ala Lys Pro

20 25 30

Arg Asn Gln Gly Gly Tyr

35

<210> 320

<211> 42

<212> PROTEIN

<213> Unknown

<220>

<223> Description of unknown:

IBB domain of importin alpha sequence

<400> 320

Arg Met Arg Ile Glx Phe Lys Asn Lys Gly Lys Asp Thr Ala Glu Leu

1 5 10 15

Arg Arg Arg Arg Val Glu Val Ser Val Glu Leu Arg Lys Ala Lys Lys

20 25 30

Asp Glu Gln Ile Leu Lys Arg Arg Asn Val

35 40

<210> 321

<211> 8

<212> PROTEIN

<213> Unknown

<220>

<223> Description of unknown:

Myoma T protein sequence

<400> 321

Val Ser Arg Lys Arg Pro Arg Pro

1 5

<210> 322

<211> 8

<212> PROTEIN

<213> Unknown

<220>

<223> Description of unknown:

Myoma T protein sequence

<400> 322

Pro Pro Lys Lys Ala Arg Glu Asp

1 5

<210> 323

<211> 8

<212> PROTEIN

<213> Homo sapiens

<400> 323

Pro Gln Pro Lys Lys Lys Pro Leu

1 5

<210> 324

<211> 12

<212> PROTEIN

<213> Mus musculus

<400> 324

Ser Ala Leu Ile Lys Lys Lys Lys Lys Met Ala Pro

1 5 10

<210> 325

<211> 5

<212> PROTEIN

<213> Influenza virus

<400> 325

Asp Arg Leu Arg Arg

1 5

<210> 326

<211> 7

<212> PROTEIN

<213> Influenza virus

<400> 326

Pro Lys Gln Lys Lys Arg Lys

1 5

<210> 327

<211> 10

<212> PROTEIN

<213> Hepatitis delta virus

<400> 327

Arg Lys Leu Lys Lys Lys Ile Lys Lys Leu

1 5 10

<210> 328

<211> 10

<212> PROTEIN

<213> Mus musculus

<400> 328

Arg Glu Lys Lys Lys Phe Leu Lys Arg Arg

1 5 10

<210> 329

<211> 20

<212> PROTEIN

<213> Homo sapiens

<400> 329

Lys Arg Lys Gly Asp Glu Val Asp Gly Val Asp Glu Val Ala Lys Lys

1 5 10 15

Lys Ser Lys Lys

20

<210> 330

<211> 17

<212> PROTEIN

<213> Homo sapiens

<400> 330

Arg Lys Cys Leu Gln Ala Gly Met Asn Leu Glu Ala Arg Lys Thr Lys

1 5 10 15

Lys

<210> 331

<400> 331

000

<210> 332

<400> 332

000

<210> 333

<400> 333

000

<210> 334

<400> 334

000

<210> 335

<400> 335

000

<210> 336

<400> 336

000

<210> 337

<400> 337

000

<210> 338

<400> 338

000

<210> 339

<400> 339

000

<210> 340

<400> 340

000

<210> 341

<400> 341

000

<210> 342

<400> 342

000

<210> 343

<400> 343

000

<210> 344

<400> 344

000

<210> 345

<400> 345

000

<210> 346

<400> 346

000

<210> 347

<400> 347

000

<210> 348

<400> 348

000

<210> 349

<400> 349

000

<210> 350

<400> 350

000

<210> 351

<400> 351

000

<210> 352

<400> 352

000

<210> 353

<400> 353

000

<210> 354

<400> 354

000

<210> 355

<400> 355

000

<210> 356

<400> 356

000

<210> 357

<400> 357

000

<210> 358

<400> 358

000

<210> 359

<400> 359

000

<210> 360

<400> 360

000

<210> 361

<400> 361

000

<210> 362

<400> 362

000

<210> 363

<400> 363

000

<210> 364

<400> 364

000

<210> 365

<400> 365

000

<210> 366

<400> 366

000

<210> 367

<400> 367

000

<210> 368

<400> 368

000

<210> 369

<400> 369

000

<210> 370

<400> 370

000

<210> 371

<400> 371

000

<210> 372

<400> 372

000

<210> 373

<400> 373

000

<210> 374

<400> 374

000

<210> 375

<400> 375

000

<210> 376

<400> 376

000

<210> 377

<400> 377

000

<210> 378

<400> 378

000

<210> 379

<400> 379

000

<210> 380

<400> 380

000

<210> 381

<400> 381

000

<210> 382

<400> 382

000

<210> 383

<400> 383

000

<210> 384

<400> 384

000

<210> 385

<400> 385

000

<210> 386

<400> 386

000

<210> 387

<400> 387

000

<210> 388

<400> 388

000

<210> 389

<400> 389

000

<210> 390

<400> 390

000

<210> 391

<400> 391

000

<210> 392

<400> 392

000

<210> 393

<400> 393

000

<210> 394

<400> 394

000

<210> 395

<400> 395

000

<210> 396

<400> 396

000

<210> 397

<400> 397

000

<210> 398

<400> 398

000

<210> 399

<400> 399

000

<210> 400

<400> 400

000

<210> 401

<400> 401

000

<210> 402

<400> 402

000

<210> 403

<400> 403

000

<210> 404

<400> 404

000

<210> 405

<400> 405

000

<210> 406

<400> 406

000

<210> 407

<400> 407

000

<210> 408

<400> 408

000

<210> 409

<400> 409

000

<210> 410

<400> 410

000

<210> 411

<400> 411

000

<210> 412

<400> 412

000

<210> 413

<400> 413

000

<210> 414

<400> 414

000

<210> 415

<400> 415

000

<210> 416

<400> 416

000

<210> 417

<400> 417

000

<210> 418

<400> 418

000

<210> 419

<400> 419

000

<210> 420

<400> 420

000

<210> 421

<400> 421

000

<210> 422

<400> 422

000

<210> 423

<400> 423

000

<210> 424

<400> 424

000

<210> 425

<400> 425

000

<210> 426

<400> 426

000

<210> 427

<400> 427

000

<210> 428

<400> 428

000

<210> 429

<400> 429

000

<210> 430

<400> 430

000

<210> 431

<400> 431

000

<210> 432

<400> 432

000

<210> 433

<400> 433

000

<210> 434

<400> 434

000

<210> 435

<400> 435

000

<210> 436

<400> 436

000

<210> 437

<400> 437

000

<210> 438

<400> 438

000

<210> 439

<400> 439

000

<210> 440

<400> 440

000

<210> 441

<400> 441

000

<210> 442

<400> 442

000

<210> 443

<400> 443

000

<210> 444

<400> 444

000

<210> 445

<400> 445

000

<210> 446

<400> 446

000

<210> 447

<400> 447

000

<210> 448

<400> 448

000

<210> 449

<400> 449

000

<210> 450

<400> 450

000

<210> 451

<400> 451

000

<210> 452

<400> 452

000

<210> 453

<400> 453

000

<210> 454

<400> 454

000

<210> 455

<400> 455

000

<210> 456

<400> 456

000

<210> 457

<400> 457

000

<210> 458

<400> 458

000

<210> 459

<400> 459

000

<210> 460

<400> 460

000

<210> 461

<400> 461

000

<210> 462

<400> 462

000

<210> 463

<400> 463

000

<210> 464

<400> 464

000

<210> 465

<400> 465

000

<210> 466

<400> 466

000

<210> 467

<400> 467

000

<210> 468

<400> 468

000

<210> 469

<400> 469

000

<210> 470

<400> 470

000

<210> 471

<400> 471

000

<210> 472

<400> 472

000

<210> 473

<400> 473

000

<210> 474

<400> 474

000

<210> 475

<400> 475

000

<210> 476

<400> 476

000

<210> 477

<400> 477

000

<210> 478

<400> 478

000

<210> 479

<400> 479

000

<210> 480

<400> 480

000

<210> 481

<400> 481

000

<210> 482

<400> 482

000

<210> 483

<400> 483

000

<210> 484

<400> 484

000

<210> 485

<400> 485

000

<210> 486

<400> 486

000

<210> 487

<400> 487

000

<210> 488

<400> 488

000

<210> 489

<400> 489

000

<210> 490

<400> 490

000

<210> 491

<400> 491

000

<210> 492

<400> 492

000

<210> 493

<400> 493

000

<210> 494

<400> 494

000

<210> 495

<400> 495

000

<210> 496

<400> 496

000

<210> 497

<400> 497

000

<210> 498

<400> 498

000

<210> 499

<400> 499

000

<210> 500

<211> 43

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 500

gcccgctggt ttccagcggg ctgcgggccc gaaacggggcc cgc 43

<210> 501

<211> 28

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 501

cgggcccgtg cgggcccaaa gggcccgc 28

<210> 502

<211> 28

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 502

gcccgggcac gcccgggttt cccgggcg 28

<210> 503

<211> 22

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 503

cgtgcgggcc caaagggccc gc 22

<210> 504

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 504

cgggcgacca aaggtcgccc g 21

<210> 505

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 505

cgcccgggct ttgcccggggc 20

<210> 506

<211> 42

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 506

cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gc 42

<210> 507

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 507

cgggcgacca aaggtcgccc g 21

<210> 508

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 508

cgcccgggct ttgcccggggc 20

<210> 509

<211> 34

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 509

cgggcgacca aaggtcgccc gacgcccggg cggc 34

<210> 510

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 510

cgggcgacca aaggtcgccc g 21

<210> 511

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 511

cgcccgggct ttgcccggggc 20

<210> 512

<211> 30

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 512

cggggcccga cgcccgggct ttgcccggggc 30

<210> 513

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 513

cgggcgacca aaggtcgccc g 21

<210> 514

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 514

cgcccgggct ttgcccggggc 20

<210> 515

<211> 29

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 515

cgggcccgac gcccggggctt tgcccggggc 29

<210> 516

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 516

cgggcgacca aaggtcgccc g 21

<210> 517

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 517

cgcccgggct ttgcccggggc 20

<210> 518

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 518

gcccgggcaa agcccgggcg 20

<210> 519

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 519

cgggcgacct ttggtcgccc g 21

<210> 520

<211> 42

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 520

gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cg 42

<210> 521

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 521

gcccgggcaa agcccgggcg 20

<210> 522

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 522

cgggcgacct ttggtcgccc g 21

<210> 523

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 523

gcccgggcaa agcccgggcg 20

<210> 524

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 524

cgggcgacct ttggtcgccc g 21

<210> 525

<211> 34

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 525

gccgcccggg cgacgggcga cctttggtcg cccg 34

<210> 526

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 526

gcccgggcaa agcccgggcg 20

<210> 527

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 527

cgggcgacct ttggtcgccc g 21

<210> 528

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 528

gcccgggcaa agcccgggcg 20

<210> 529

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 529

cgggcgacct ttggtcgccc g 21

<210> 530

<211> 4470

<212> DNA

<213> Homo sapiens

<400> 530

gtagatgagg aaactgaagt tgaggaatag tgaagagttt gtccaatgtc atagccccgt 60

aatcaacggg acaaaaattt tcttgctgat gggtcaagat ggcatcgtga agtggttgtt 120

caccgtaaac tgtaatacaa tcctgtttat ggatttgttt gcatattttt ccctccatag 180

ggaaaccttt cttccatggc tcaggacaca ctcctggatc gagccaacag gagaactttc 240

tggtaagcat ttggctaact tttttttttt tgagatggag tcttgctgtg tcgcctaggc 300

tggagtgcag tggcgtgatc ttggctcact gcagcctcca cttcccgggt tcaatcaatt 360

ctcctacctc aacttcctga gtagctggga ttacaggcgc ccgccaccac acccggctca 420

tttttgtact tttagtagag acacagtttt gccatgttgg ccaggctggt cttgaattcc 480

tcagctcagg tgatctgcct gccttggcct ctcaaagtgc tgggattaca ggcgtgagcc 540

actgtgcccg gccttggcta acttttcaaa attaaagatt ttgacttgtt acagtcatgt 600

gacatttttt tctttctgtt tgctgagttt ttgataattt atatctctca aagtggagac 660

tttaaaaaag actcatccgt gtgccgtgtt cactgcctgg tatcttagtg tggaccgaag 720

cctaaggacc ctgaaaacag ctgcagatga agatggcaag cacccgctgc aagctggcca 780

ggtacctgga ggacctggag gatgtggact tgaagaaatt taagatgcac ttagaggact 840

atcctcccca gaagggctgc atccccctcc cgaggggtca gacagagaag gcagaccatg 900

tggatctagc cacgctaatg atcgacttca atggggagga gaaggcgtgg gccatggccg 960

tgtggatctt cgctgcgatc aacaggag acctttatga gaaagcaaaa agagatgagc 1020

cgaagtgggg ttcagataat gcacgtgttt cgaatcccac tgtgatatgc caggaagaca 1080

gcattgaaga ggagtggatg ggtttactgg agtacctttc gagaatctct atttgtaaaa 1140

tgaagaaaga ttaccgtaag aagtacagaa agtacgtgag aagcagattc cagtgcattg 1200

aagacaggaa tgcccgtctg ggtgagagtg tgagcctcaa caaacgctac acacgactgc 1260

gtctcatcaa ggagcaccgg agccagcagg agagggagca ggagcttctg gccatcggca 1320

agaccaagac gtgtgagagc cccgtgagtc ccattaagat ggagttgctg tttgaccccg 1380

atgatgagca ttctgagcct gtgcacaccg tggtgttcca gggggcggca gggattggga 1440

aaacaatcct ggccaggaag atgatgttgg actgggcgtc ggggacactc taccaagaca 1500

ggtttgacta tctgttctat atccactgtc gggaggtgag ccttgtgaca cagaggagcc 1560

tggggggacct gatcatgagc tgctgccccg acccaaaccc acccatccac aagatcgtga 1620

gaaaaccctc cagaatcctc ttcctcatgg acggcttcga tgagctgcaa ggtgcctttg 1680

acgagcacat aggaccgctc tgcactgact ggcagaaggc cgagcgggga gacattctcc 1740

tgagcagcct catcagaaag aagctgcttc ccgaggcctc tctgctcatc accacgagac 1800

ctgtggccct ggagaaactg cagcacttgc tggaccatcc tcggcatgtg gagatcctgg 1860

gtttctccga ggccaaaagg aaagagtact tcttcaagta cttctctgat gaggcccaag 1920

ccagggcagc cttcagtctg attcaggaga acgaggtcct cttcaccatg tgcttcatcc 1980

ccctggtctg ctggatcgtg tgcactggac tgaaacagca gatggagagt ggcaagagcc 2040

ttgccgac atccaagacc accaccgcgg tgtacgtctt cttcctttcc agtttgctgc 2100

agccccgggg agggagccag gagcacggcc tctgcgccca cctctggggg ctctgctctt 2160

tggctgcaga tggaatctgg aaccagaaaa tcctgtttga ggagtccgac ctcaggaatc 2220

atggactgca gaaggcggat gtgtctgctt tcctgaggat gaacctgttc caaaaggaag 2280

tggactgcga gaagttctac agcttcatcc acatgacttt ccaggagttc tttgccgcca 2340

tgtactacct gctggaagag gaaaaggaag gaaggacgaa cgttccaggg agtcgtttga 2400

agcttcccag ccgagacgtg acagtccttc tggaaaacta tggcaaattc gaaaaggggt 2460

atttgatttt tgttgtacgt ttcctctttg gcctggtaaa ccaggagagg acctcctact 2520

tggagaagaa attaagttgc aagatctctc agcaaatcag gctggagctg ctgaaatgga 2580

ttgaagtgaa agccaaagct aaaaagctgc agatccagcc cagccagctg gaattgttct 2640

actgtttgta cgagatgcag gaggaggact tcgtgcaaag ggccatggac tatttcccca 2700

agattgagat caatctctcc accagaatgg accacatggt ttcttccttt tgcattgaga 2760

actgtcatcg ggtggagtca ctgtccctgg ggtttctcca taacatgccc aagggaggaag 2820

aggaggagga aaaggaaggc cgacaccttg atatggtgca gtgtgtcctc ccaagctcct 2880

ctcatgctgc ctgttctcat ggattggtga acagccacct cacttccagt ttttgccggg 2940

gcctcttttc agttctgagc accagccaga gtctaactga attggacctc agtgacaatt 3000

ctctggggga cccagggatg agagtgttgt gtgaaacgct ccagcatcct ggctgtaaca 3060

ttcggagatt gtggttgggg cgctgtggcc tctcgcatga gtgctgcttc gacatctcct 3120

tggtcctcag cagcaaccag aagctggtgg agctggacct gagtgacaac gccctcggtg 3180

acttcggaat cagacttctg tgtgtgggac tgaagcacct gttgtgcaat ctgaagaagc 3240

tctggttggt cagctgctgc ctcacatcag catgttgtca ggatcttgca tcagtattga 3300

gcaccagcca ttccctgacc agactctatg tgggggagaa tgccttggga gactcaggag 3360

tcgcaatttt atgtgaaaaa gccaagaatc cacagtgtaa cctgcagaaa ctggggttgg 3420

tgaattctgg ccttacgtca gtctgttgtt cagctttgtc ctcggtactc agcactaatc 3480

agaatctcac gcacctttac ctgcgaggca acactctcgg agacaagggg atcaaactac 3540

tctgtgaggg actcttgcac cccgactgca agcttcaggt gttggaatta gacaactgca 3600

acctcacgtc acactgctgc tgggatcttt ccacacttct gacctccagc cagagcctgc 3660

gaaagctgag cctgggcaac aatgacctgg gcgacctggg ggtcatgatg ttctgtgaag 3720

tgctgaaaca gcagagctgc ctcctgcaga acctggggtt gtctgaaatg tatttcaatt 3780

atgagacaaa aagtgcgtta gaaacacttc aagaagaaaa gcctgagctg accgtcgtct 3840

ttgagccttc ttggtaggag tggaaacggg gctgccagac gccagtgttc tccggtccct 3900

ccagctgggg gccctcaggt ggagagagct gcgatccatc caggccaaga ccacagctct 3960

gtgatccttc cggtggagtg tcggagaaga gagcttgccg acgatgcctt cctgtgcaga 4020

gcttgggcat ctcctttacg cccaggtgag gaagacacca ggacaatgac agcatcgggt 4080

gttgttgtca tcacagcgcc tcagttagag gatgttcctc ttggtgacct catgtaatta 4140

gctcattcaa taaagcactt tctttatttt tctcttctct gtctaacttt ctttttccta 4200

tcttttttct tctttgttct gtttactttt gctcatatca tcattcccgc tatctttcta 4260

ttaactgacc ataacacaga actagttgac tatatattat gttgaaattt tatggcagct 4320

atttatttat ttaaattttt tgtaacagtt ttgttttcta ataagaaaaa tccatgcttt 4380

ttgtagctgg ttgaaaattc aggaatatgt aaaacttttt ggtatttaat taaattgatt 4440

ccttttctta attttaaaaa aaaaaaaaaa 4470

<210> 531

<211> 3845

<212> DNA

<213> Homo sapiens

<400> 531

gttcctgagg ctggcatctg gatgaggaaa ctgaagttga ggaatagtga agagtttgtc 60

caatgtcata gccccgtaat caacgggaca aaaattttct tgctgatggg tcaagatggc 120

atcgtgaagt ggttgttcac cgtaaactgt aatacaatcc tgtttatgga tttgtttgca 180

tatttttccc tccataggga aacctttctt ccatggctca ggacacactc ctggatcgag 240

ccaacaggag aactttctgg taagcatttg gctaactttt ttttttttga gatggagtct 300

tgctgtgtcg cctaggctgg agtgcagtgg cgtgatcttg gctcactgca gcctccactt 360

cccgggttca atcaattctc ctacctcaac ttcctgagta gctgggatta caggcgcccg 420

ccaccacacc cggctcattt ttgtactttt agtagagaca cagttttgcc atgttggcca 480

ggctggtctt gaattcctca gctcaggtga tctgcctgcc ttggcctctc aaagtgctgg 540

gattacaggc gtgagccact gtgcccggcc ttggctaact tttcaaaatt aaagattttg 600

acttgttaca gtcatgtgac atttttttct ttctgtttgc tgagtttttg ataatttata 660

tctctcaaag tggagacttt aaaaaagact catccgtgtg ccgtgttcac tgcctggtat 720

cttagtgtgg accgaagcct aaggaccctg aaaacagctg cagatgaaga tggcaagcac 780

ccgctgcaag ctggccaggt acctggagga cctggaggat gtggacttga agaaatttaa 840

gatgcactta gaggactatc ctccccagaa gggctgcatc cccctcccga ggggtcagac 900

agagaaggca gaccatgtgg atctagccac gctaatgatc gacttcaatg gggaggagaa 960

ggcgtgggcc atggccgtgt ggatcttcgc tgcgatcaac aggagagacc tttatgagaa 1020

agcaaaaaga gatgagccga agtggggttc agataatgca cgtgtttcga atcccactgt 1080

gatatgccag gaagacagca ttgaagagga gtggatgggt ttactggagt acctttcgag 1140

aatctctatt tgtaaaatga agaaagatta ccgtaagaag tacagaaagt acgtgagaag 1200

cagattccag tgcattgaag acaggaatgc ccgtctgggt gagagtgtga gcctcaacaa 1260

acgctacaca cgactgcgtc tcatcaagga gcaccggagc cagcaggaga gggagcagga 1320

gcttctggcc atcggcaaga ccaagacgtg tgagagcccc gtgagtccca ttaagatgga 1380

gttgctgttt gaccccgatg atgagcattc tgagcctgtg cacaccgtgg tgttccaggg 1440

ggcggcaggg attgggaaaa caatcctggc caggaagatg atgttggact gggcgtcggg 1500

gacactctac caagacaggt ttgactatct gttctatatc cactgtcgag aggtgagcct 1560

tgtgacacag aggagcctgg gggacctgat catgagctgc tgccccgacc caaacccacc 1620

catccacaag atcgtgagaa aaccctccag aatcctcttc ctcatggacg gcttcgatga 1680

gctgcaaggt gcctttgacg agcacatagg accgctctgc actgactggc agaaggccga 1740

gcggggagac attctcctga gcagcctcat cagaaagaag ctgcttcccg aggcctctct 1800

gctcatcacc acgagacctg tggccctgga gaaactgcag cacttgctgg accatcctcg 1860

gcatgtggag atcctgggtt tctccgaggc caaaaggaaa gagtacttct tcaagtactt 1920

ctctgatgag gcccaagcca gggcagcctt cagtctgatt caggagaacg aggtcctctt 1980

caccatgtgc ttcatccccc tggtctgctg gatcgtgtgc actggactga aacagcagat 2040

ggagagtggc aagagccttg cccagacatc caagaccacc accgcggtgt acgtcttctt 2100

cctttccagt ttgctgcagc cccggggagg gagccaggag cacggcctct gcgcccacct 2160

ctggggggctc tgctctttgg ctgcagatgg aatctggaac cagaaaatcc tgtttgagga 2220

gtccgacctc aggaatcatg gactgcagaa ggcggatgtg tctgctttcc tgaggatgaa 2280

cctgttccaa aaggaagtgg actgcgagaa gttctacagc ttcatccaca tgactttcca 2340

ggagttcttt gccgccatgt actacctgct ggaagaggaa aaggaaggaa ggacgaacgt 2400

tccagggagt cgtttgaagc ttcccagccg agacgtgaca gtccttctgg aaaactatgg 2460

caaattcgaa aaggggtatt tgatttttgt tgtacgtttc ctctttggcc tggtaaacca 2520

ggagaggacc tcctacttgg agaagaaatt aagttgcaag atctctcagc aaatcaggct 2580

ggagctgctg aaatggattg aagtgaaagc caaagctaaa aagctgcaga tccagcccag 2640

ccagctggaa ttgttctact gtttgtacga gatgcaggag gaggacttcg tgcaaagggc 2700

catggactat ttccccaaga ttgagatcaa tctctccacc agaatggacc acatggtttc 2760

ttccttttgc attgagaact gtcatcgggt ggagtcactg tccctggggt ttctccataa 2820

catgcccaag gaggaagagg aggaggaaaa ggaaggccga caccttgata tggtgcagtg 2880

tgtcctccca agctcctctc atgctgcctg ttctcatggg ttggggcgct gtggcctctc 2940

gcatgagtgc tgcttcgaca tctccttggt cctcagcagc aaccagaagc tggtggagct 3000

ggacctgagt gacaacgccc tcggtgactt cggaatcaga cttctgtgtg tgggactgaa 3060

gcacctgttg tgcaatctga agaagctctg gttggtgaat tctggcctta cgtcagtctg 3120

ttgttcagct ttgtcctcgg tactcagcac taatcagaat ctcacgcacc tttacctgcg 3180

aggcaacact ctcggagaca aggggatcaa actactctgt gagggactct tgcaccccga 3240

ctgcaagctt caggtgttgg aattagacaa ctgcaacctc acgtcacact gctgctggga 3300

tctttccaca cttctgacct ccagccagag cctgcgaaag ctgagcctgg gcaacaatga 3360

cctgggcgac ctggggggtca tgatgttctg tgaagtgctg aaacagcaga gctgcctcct 3420

gcagaacctg gggttgtctg aaatgtattt caattatgag acaaaaagtg cgttagaaac 3480

acttcaagaa gaaaagcctg agctgaccgt cgtctttgag ccttcttggt aggagtggaa 3540

acggggctgc cagacgccag tgttctccgg tccctccagc tgggggccct caggtggaga 3600

gagctgcgat ccatccaggc caagaccaca gctctgtgat ccttccggtg gagtgtcgga 3660

gaagagagct tgccgacgat gccttcctgt gcagagcttg ggcatctcct ttacgccagg 3720

gtgaggaaga caccaggaca atgacagcat cgggtgttgt tgtcatcaca gcgcctcagt 3780

tagaggatgt tcctcttggt gacctcatgt aattagctca ttcaataaag cactttcttt 3840

atttt 3845

<210> 532

<211> 3545

<212> DNA

<213> Homo sapiens

<400> 532

gttcctgagg ctggcatctg gggaaacctt tcttccatgg ctcaggacac actcctggat 60

cgagccaaca ggagaacttt ctgtgtggac cgaagcctaa ggaccctgaa aacagctgca 120

gatgaagatg gcaagcaccc gctgcaagct ggccaggtac ctggaggacc tggaggatgt 180

ggacttgaag aaatttaaga tgcacttaga ggactatcct ccccagaagg gctgcatccc 240

cctcccgagg ggtcagacag agaaggcaga ccatgtggat ctagccacgc taatgatcga 300

cttcaatggg gaggagaagg cgtgggccat ggccgtgtgg atcttcgctg cgatcaacag 360

gagagacctt tatgagaaag caaaaagaga tgagccgaag tggggttcag ataatgcacg 420

tgtttcgaat cccactgtga tatgccagga agacagcatt gaagaggagt ggatgggttt 480

actggagtac ctttcgagaa tctctatttg taaaatgaag aaagattacc gtaagaagta 540

cagaaagtac gtgagaagca gattccagtg cattgaagac aggaatgccc gtctgggtga 600

gagtgtgagc ctcaacaaac gctacacacg actgcgtctc atcaaggagc accggagcca 660

gcaggagagg gagcaggagc ttctggccat cggcaagacc aagacgtgtg agagccccgt 720

gagtcccatt aagatggagt tgctgtttga ccccgatgat gagcattctg agcctgtgca 780

caccgtggtg ttccagggg cggcagggat tgggaaaaca atcctggcca ggaagatgat 840

gttggactgg gcgtcgggga cactctacca agacaggttt gactatctgt tctatatcca 900

ctgtcgagag gtgagccttg tgacacagag gagcctgggg gacctgatca tgagctgctg 960

ccccgaccca aacccaccca tccacaagat cgtgagaaaa ccctccagaa tcctcttcct 1020

catggacggc ttcgatgagc tgcaaggtgc ctttgacgag cacataggac cgctctgcac 1080

tgactggcag aaggccgagc ggggagacat tctcctgagc agcctcatca gaaagaagct 1140

gcttcccgag gcctctctgc tcatcaccac gagacctgtg gccctggaga aactgcagca 1200

cttgctggac catcctcggc atgtggagat cctgggtttc tccgaggcca aaaggaaaga 1260

gtacttcttc aagtacttct ctgatgaggc ccaagccagg gcagccttca gtctgattca 1320

ggagaacgag gtcctcttca ccatgtgctt catccccctg gtctgctgga tcgtgtgcac 1380

tggactgaaa cagcagatgg agagtggcaa gagccttgcc cagacatcca agaccaccac 1440

cgcggtgtac gtcttcttcc tttccagttt gctgcagccc cggggaggga gccaggagca 1500

cggcctctgc gcccacctct gggggctctg ctctttggct gcagatggaa tctggaacca 1560

gaaaatcctg tttgaggagt ccgacctcag gaatcatgga ctgcagaagg cggatgtgtc 1620

tgctttcctg aggatgaacc tgttccaaaa ggaagtggac tgcgagaagt tctacagctt 1680

catccacatg actttccagg agttctttgc cgccatgtac tacctgctgg aagaggaaaa 1740

ggaaggaagg acgaacgttc cagggagtcg tttgaagctt cccagccgag acgtgacagt 1800

ccttctggaa aactatggca aattcgaaaa ggggtatttg atttttgttg tacgtttcct 1860

ctttggcctg gtaaaccagg agaggacctc ctacttggag aagaaattaa gttgcaagat 1920

ctctcagcaa atcaggctgg agctgctgaa atggattgaa gtgaaagcca aagctaaaaa 1980

gctgcagatc cagcccagcc agctggaatt gttctactgt ttgtacgaga tgcaggagga 2040

ggacttcgtg caaagggcca tggactattt ccccaagatt gagatcaatc tctccaccag 2100

aatggaccac atggtttctt ccttttgcat tgagaactgt catcgggtgg agtcactgtc 2160

cctggggttt ctccataaca tgcccaagga ggaagaggag gaggaaaagg aaggccgaca 2220

ccttgatatg gtgcagtgtg tcctcccaag ctcctctcat gctgcctgtt ctcatggatt 2280

ggtgaacagc cacctcactt ccagtttttg ccggggcctc ttttcagttc tgagcaccag 2340

ccagagtcta actgaattgg acctcagtga caattctctg ggggacccag ggatgagagt 2400

gttgtgtgaa acgctccagc atcctggctg taacattcgg agattgtggt tggggcgctg 2460

tggcctctcg catgagtgct gcttcgacat ctccttggtc ctcagcagca accagaagct 2520

ggtggagctg gacctgagtg acaacgccct cggtgacttc ggaatcagac ttctgtgtgt 2580

gggactgaag cacctgttgt gcaatctgaa gaagctctgg ttggtcagct gctgcctcac 2640

atcagcatgt tgtcaggatc ttgcatcagt attgagcacc agccattccc tgaccagact 2700

ctatgtgggg gagaatgcct tgggagactc aggagtcgca attttatgtg aaaaagccaa 2760

gaatccacag tgtaacctgc agaaactggg gttggtgaat tctggcctta cgtcagtctg 2820

ttgttcagct ttgtcctcgg tactcagcac taatcagaat ctcacgcacc tttacctgcg 2880

aggcaacact ctcggagaca aggggatcaa actactctgt gagggactct tgcaccccga 2940

ctgcaagctt caggtgttgg aattagacaa ctgcaacctc acgtcacact gctgctggga 3000

tctttccaca cttctgacct ccagccagag cctgcgaaag ctgagcctgg gcaacaatga 3060

cctgggcgac ctggggggtca tgatgttctg tgaagtgctg aaacagcaga gctgcctcct 3120

gcagaacctg gggttgtctg aaatgtattt caattatgag acaaaaagtg cgttagaaac 3180

acttcaagaa gaaaagcctg agctgaccgt cgtctttgag ccttcttggt aggagtggaa 3240

acggggctgc cagacgccag tgttctccgg tccctccagc tgggggccct caggtggaga 3300

gagctgcgat ccatccaggc caagaccaca gctctgtgat ccttccggtg gagtgtcgga 3360

gaagagagct tgccgacgat gccttcctgt gcagagcttg ggcatctcct ttacgccagg 3420

gtgaggaaga caccaggaca atgacagcat cgggtgttgt tgtcatcaca gcgcctcagt 3480

tagaggatgt tcctcttggt gacctcatgt aattagctca ttcaataaag cactttcttt 3540

atttt 3545

<210> 533

<211> 4015

<212> DNA

<213> Homo sapiens

<400> 533

gttcctgagg ctggcatctg gatgaggaaa ctgaagttga ggaatagtga agagtttgtc 60

caatgtcata gccccgtaat caacgggaca aaattttctt gctgatgggt caagatggca 120

tcgtgaagtg gttgttcacc gtaaactgta atacaatcct gtttatggat ttgtttgcat 180

atttttccct ccatagggaa acctttcttc catggctcag gacacactcc tggatcgagc 240

caacaggaga actttctggt aagcatttgg ctaacttttt tttttttgag atggagtctt 300

gctgtgtcgc ctaggctgga gtgcagtggc gtgatcttgg ctcactgcag cctccacttc 360

ccgggttcaa tcaattctcc tacctcaact tcctgagtag ctgggattac aggcgcccgc 420

caccacaccc ggctcatttt tgtactttta gtagagacac agttttgcca tgttggccag 480

gctggtcttg aattcctcag ctcaggtgat ctgcctgcct tggcctctca aagtgctggg 540

attacaggcg tgagccactg tgcccggcct tggctaactt ttcaaaatta aagattttga 600

cttgttacag tcatgtgaca tttttttctt tctgtttgct gagtttttga taatttatat 660

ctctcaaagt ggagacttta aaaaagactc atccgtgtgc cgtgttcact gcctggtatc 720

ttagtgtgga ccgaagccta aggaccctga aaacagctgc agatgaagat ggcaagcacc 780

cgctgcaagc tggccaggta cctggaggac ctggaggatg tggacttgaa gaaatttaag 840

atgcacttag aggactatcc tccccagaag ggctgcatcc ccctcccgag gggtcagaca 900

gagaaggcag accatgtgga tctagccacg ctaatgatcg acttcaatgg ggaggagaag 960

gcgtgggcca tggccgtgtg gatcttcgct gcgatcaaca ggagagacct ttatgagaaa 1020

gcaaaaagag atgagccgaa gtggggttca gataatgcac gtgtttcgaa tcccactgtg 1080

atatgccagg aagacagcat tgaagaggag tggatgggtt tactggagta cctttcgaga 1140

atctctattt gtaaaatgaa gaaagattac cgtaagaagt acagaaagta cgtgagaagc 1200

agattccagt gcattgaaga caggaatgcc cgtctgggtg agagtgtgag cctcaacaaa 1260

cgctacacac gactgcgtct catcaaggag caccggagcc agcaggagag ggagcaggag 1320

cttctggcca tcggcaagac caagacgtgt gagagccccg tgagtcccat taagatggag 1380

ttgctgtttg accccgatga tgagcattct gagcctgtgc acaccgtggt gttccagggg 1440

gcggcaggga ttgggaaaac aatcctggcc aggaagatga tgttggactg ggcgtcgggg 1500

acactctacc aagacaggtt tgactatctg ttctatatcc actgtcgaga ggtgagcctt 1560

gtgacacaga ggagcctggg ggacctgatc atgagctgct gccccgaccc aaacccaccc 1620

atccacaaga tcgtgagaaa accctccaga atcctcttcc tcatggacgg cttcgatgag 1680

ctgcaaggtg cctttgacga gcacatagga ccgctctgca ctgactggca gaaggccgag 1740

cggggagaca ttctcctgag cagcctcatc agaaagaagc tgcttcccga ggcctctctg 1800

ctcatcacca cgagacctgt ggccctggag aaactgcagc acttgctgga ccatcctcgg 1860

catgtggaga tcctgggttt ctccgaggcc aaaaggaaag agtacttctt caagtacttc 1920

tctgatgagg cccaagccag ggcagccttc agtctgattc aggagaacga ggtcctcttc 1980

accatgtgct tcatccccct ggtctgctgg atcgtgtgca ctggactgaa acagcagatg 2040

gagagtggca agagccttgc ccagacatcc aagaccacca ccgcggtgta cgtcttcttc 2100

ctttccagtt tgctgcagcc ccggggaggg agccaggagc acggcctctg cgcccacctc 2160

tggggggctct gctctttggc tgcagatgga atctggaacc agaaaatcct gtttgaggag 2220

tccgacctca ggaatcatgg actgcagaag gcggatgtgt ctgctttcct gaggatgaac 2280

ctgttccaaa aggaagtgga ctgcgagaag ttctacagct tcatccacat gactttccag 2340

gagttctttg ccgccatgta ctacctgctg gaagaggaaa aggaaggaag gacgaacgtt 2400

ccagggagtc gtttgaagct tcccagccga gacgtgacag tccttctgga aaactatggc 2460

aaattcgaaa aggggtattt gatttttgtt gtacgtttcc tctttggcct ggtaaaccag 2520

gagaggacct cctacttgga gaagaaatta agttgcaaga tctctcagca aatcaggctg 2580

gagctgctga aatggattga agtgaaagcc aaagctaaaa agctgcagat ccagcccagc 2640

cagctggaat tgttctactg tttgtacgag atgcaggagg aggacttcgt gcaaagggcc 2700

atggactatt tccccaagat tgagatcaat ctctccacca gaatggacca catggtttct 2760

tccttttgca ttgagaactg tcatcgggtg gagtcactgt ccctggggtt tctccataac 2820

atgcccaagg aggaagagga gggaggaaaag gaaggccgac accttgatat ggtgcagtgt 2880

gtcctcccaa gctcctctca tgctgcctgt tctcatggat tggtgaacag ccacctcact 2940

tccagttttt gccggggcct cttttcagtt ctgagcacca gccagagtct aactgaattg 3000

gacctcagtg acaattctct gggggaccca gggatgagag tgttgtgtga aacgctccag 3060

catcctggct gtaacattcg gagattgtgg ttggggcgct gtggcctctc gcatgagtgc 3120

tgcttcgaca tctccttggt cctcagcagc aaccagaagc tggtggagct ggacctgagt 3180

gacaacgccc tcggtgactt cggaatcaga cttctgtgtg tgggactgaa gcacctgttg 3240

tgcaatctga agaagctctg gttggtgaat tctggcctta cgtcagtctg ttgttcagct 3300

ttgtcctcgg tactcagcac taatcagaat ctcacgcacc tttacctgcg aggcaacact 3360

ctcggagaca aggggatcaa actactctgt gagggactct tgcaccccga ctgcaagctt 3420

caggtgttgg aattagacaa ctgcaacctc acgtcacact gctgctggga tctttccaca 3480

cttctgacct ccagccagag cctgcgaaag ctgagcctgg gcaacaatga cctgggcgac 3540

ctggggggtca tgatgttctg tgaagtgctg aaacagcaga gctgcctcct gcagaacctg 3600

gggttgtctg aaatgtattt caattatgag acaaaaagtg cgttagaaac acttcaagaa 3660

gaaaagcctg agctgaccgt cgtctttgag ccttcttggt aggagtggaa acggggctgc 3720

cagacgccag tgttctccgg tccctccagc tggggggccct caggtggaga gagctgcgat 3780

ccatccaggc caagaccaca gctctgtgat ccttccggtg gagtgtcgga gaagagagct 3840

tgccgacgat gccttcctgt gcagagcttg ggcatctcct ttacgccagg gtgaggaaga 3900

caccaggaca atgacagcat cgggtgttgt tgtcatcaca gcgcctcagt tagaggatgt 3960

tcctcttggt gacctcatgt aattagctca ttcaataaag cactttcttt atttt 4015

<210> 534

<211> 4016

<212> DNA

<213> Homo sapiens

<400> 534

gttcctgagg ctggcatctg gatgaggaaa ctgaagttga ggaatagtga agagtttgtc 60

caatgtcata gccccgtaat caacgggaca aaaattttct tgctgatggg tcaagatggc 120

atcgtgaagt ggttgttcac cgtaaactgt aatacaatcc tgtttatgga tttgtttgca 180

tatttttccc tccataggga aacctttctt ccatggctca ggacacactc ctggatcgag 240

ccaacaggag aactttctgg taagcatttg gctaactttt ttttttttga gatggagtct 300

tgctgtgtcg cctaggctgg agtgcagtgg cgtgatcttg gctcactgca gcctccactt 360

cccgggttca atcaattctc ctacctcaac ttcctgagta gctgggatta caggcgcccg 420

ccaccacacc cggctcattt ttgtactttt agtagagaca cagttttgcc atgttggcca 480

ggctggtctt gaattcctca gctcaggtga tctgcctgcc ttggcctctc aaagtgctgg 540

gattacaggc gtgagccact gtgcccggcc ttggctaact tttcaaaatt aaagattttg 600

acttgttaca gtcatgtgac atttttttct ttctgtttgc tgagtttttg ataatttata 660

tctctcaaag tggagacttt aaaaaagact catccgtgtg ccgtgttcac tgcctggtat 720

cttagtgtgg accgaagcct aaggaccctg aaaacagctg cagatgaaga tggcaagcac 780

ccgctgcaag ctggccaggt acctggagga cctggaggat gtggacttga agaaatttaa 840

gatgcactta gaggactatc ctccccagaa gggctgcatc cccctcccga ggggtcagac 900

agagaaggca gaccatgtgg atctagccac gctaatgatc gacttcaatg gggaggagaa 960

ggcgtgggcc atggccgtgt ggatcttcgc tgcgatcaac aggagagacc tttatgagaa 1020

agcaaaaaga gatgagccga agtggggttc agataatgca cgtgtttcga atcccactgt 1080

gatatgccag gaagacagca ttgaagagga gtggatgggt ttactggagt acctttcgag 1140

aatctctatt tgtaaaatga agaaagatta ccgtaagaag tacagaaagt acgtgagaag 1200

cagattccag tgcattgaag acaggaatgc ccgtctgggt gagagtgtga gcctcaacaa 1260

acgctacaca cgactgcgtc tcatcaagga gcaccggagc cagcaggaga gggagcagga 1320

gcttctggcc atcggcaaga ccaagacgtg tgagagcccc gtgagtccca ttaagatgga 1380

gttgctgttt gaccccgatg atgagcattc tgagcctgtg cacaccgtgg tgttccaggg 1440

ggcggcaggg attgggaaaa caatcctggc caggaagatg atgttggact gggcgtcggg 1500

gacactctac caagacaggt ttgactatct gttctatatc cactgtcgag aggtgagcct 1560

tgtgacacag aggagcctgg gggacctgat catgagctgc tgccccgacc caaacccacc 1620

catccacaag atcgtgagaa aaccctccag aatcctcttc ctcatggacg gcttcgatga 1680

gctgcaaggt gcctttgacg agcacatagg accgctctgc actgactggc agaaggccga 1740

gcggggagac attctcctga gcagcctcat cagaaagaag ctgcttcccg aggcctctct 1800

gctcatcacc acgagacctg tggccctgga gaaactgcag cacttgctgg accatcctcg 1860

gcatgtggag atcctgggtt tctccgaggc caaaaggaaa gagtacttct tcaagtactt 1920

ctctgatgag gcccaagcca gggcagcctt cagtctgatt caggagaacg aggtcctctt 1980

caccatgtgc ttcatccccc tggtctgctg gatcgtgtgc actggactga aacagcagat 2040

ggagagtggc aagagccttg cccagacatc caagaccacc accgcggtgt acgtcttctt 2100

cctttccagt ttgctgcagc cccggggagg gagccaggag cacggcctct gcgcccacct 2160

ctggggggctc tgctctttgg ctgcagatgg aatctggaac cagaaaatcc tgtttgagga 2220

gtccgacctc aggaatcatg gactgcagaa ggcggatgtg tctgctttcc tgaggatgaa 2280

cctgttccaa aaggaagtgg actgcgagaa gttctacagc ttcatccaca tgactttcca 2340

ggagttcttt gccgccatgt actacctgct ggaagaggaa aaggaaggaa ggacgaacgt 2400

tccagggagt cgtttgaagc ttcccagccg agacgtgaca gtccttctgg aaaactatgg 2460

caaattcgaa aaggggtatt tgatttttgt tgtacgtttc ctctttggcc tggtaaacca 2520

ggagaggacc tcctacttgg agaagaaatt aagttgcaag atctctcagc aaatcaggct 2580

ggagctgctg aaatggattg aagtgaaagc caaagctaaa aagctgcaga tccagcccag 2640

ccagctggaa ttgttctact gtttgtacga gatgcaggag gaggacttcg tgcaaagggc 2700

catggactat ttccccaaga ttgagatcaa tctctccacc agaatggacc acatggtttc 2760

ttccttttgc attgagaact gtcatcgggt ggagtcactg tccctggggt ttctccataa 2820

catgcccaag gaggaagagg aggaggaaaa ggaaggccga caccttgata tggtgcagtg 2880

tgtcctccca agctcctctc atgctgcctg ttctcatggg ttggggcgct gtggcctctc 2940

gcatgagtgc tgcttcgaca tctccttggt cctcagcagc aaccagaagc tggtggagct 3000

ggacctgagt gacaacgccc tcggtgactt cggaatcaga cttctgtgtg tgggactgaa 3060

gcacctgttg tgcaatctga agaagctctg gttggtcagc tgctgcctca catcagcatg 3120

ttgtcaggat cttgcatcag tattgagcac cagccattcc ctgaccagac tctatgtggg 3180

ggagaatgcc ttgggagact caggagtcgc aattttatgt gaaaaagcca agaatccaca 3240

gtgtaacctg cagaaactgg ggttggtgaa ttctggcctt acgtcagtct gttgttcagc 3300

tttgtcctcg gtactcagca ctaatcagaa tctcacgcac ctttacctgc gaggcaacac 3360

tctcggagac aaggggatca aactactctg tgagggactc ttgcaccccg actgcaagct 3420

tcaggtgttg gaattagaca actgcaacct cacgtcacac tgctgctggg atctttccac 3480

acttctgacc tccagccaga gcctgcgaaa gctgagcctg ggcaacaatg acctgggcga 3540

cctgggggtc atgatgttct gtgaagtgct gaaacagcag agctgcctcc tgcagaacct 3600

ggggttgtct gaaatgtatt tcaattatga gacaaaaagt gcgttagaaa cacttcaaga 3660

agaaaagcct gagctgaccg tcgtctttga gccttcttgg taggagtgga aacggggctg 3720

cgacgcca gtgttctccg gtccctccag ctggggggccc tcaggtggag agagctgcga 3780

tccatccagg ccaagaccac agctctgtga tccttccggt ggagtgtcgg agaagagagc 3840

ttgccgacga tgccttcctg tgcagagctt gggcatctcc tttacgccag ggtgaggaag 3900

acaccaggac aatgacagca tcgggtgttg ttgtcatcac agcgcctcag ttagaggatg 3960

ttcctcttgg tgacctcatg taattagctc attcaataaa gcactttctt tatttt 4016

<210> 535

<400> 535

000

<210> 536

<400> 536

000

<210> 537

<400> 537

000

<210> 538

<211> 14

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic peptide

<400> 538

Asn Glu Ala Tyr Val His Asp Ala Pro Val Arg Ser Leu Asn

1 5 10

<210> 539

<211> 1036

<212> PROTEIN

<213> Homo sapiens

<400> 539

Met Lys Met Ala Ser Thr Arg Cys Lys Leu Ala Arg Tyr Leu Glu Asp

1 5 10 15

Leu Glu Asp Val Asp Leu Lys Lys Phe Lys Met His Leu Glu Asp Tyr

20 25 30

Pro Pro Gln Lys Gly Cys Ile Pro Leu Pro Arg Gly Gln Thr Glu Lys

35 40 45

Ala Asp His Val Asp Leu Ala Thr Leu Met Ile Asp Phe Asn Gly Glu

50 55 60

Glu Lys Ala Trp Ala Met Ala Val Trp Ile Phe Ala Ala Ile Asn Arg

65 70 75 80

Arg Asp Leu Tyr Glu Lys Ala Lys Arg Asp Glu Pro Lys Trp Gly Ser

85 90 95

Asp Asn Ala Arg Val Ser Asn Pro Thr Val Ile Cys Gln Glu Asp Ser

100 105 110

Ile Glu Glu Glu Trp Met Gly Leu Leu Glu Tyr Leu Ser Arg Ile Ser

115 120 125

Ile Cys Lys Met Lys Lys Asp Tyr Arg Lys Lys Tyr Arg Lys Tyr Val

130 135 140

Arg Ser Arg Phe Gln Cys Ile Glu Asp Arg Asn Ala Arg Leu Gly Glu

145 150 155 160

Ser Val Ser Leu Asn Lys Arg Tyr Thr Arg Leu Arg Leu Ile Lys Glu

165 170 175

His Arg Ser Gln Gln Glu Arg Glu Gln Glu Leu Leu Ala Ile Gly Lys

180 185 190

Thr Lys Thr Cys Glu Ser Pro Val Ser Pro Ile Lys Met Glu Leu Leu

195 200 205

Phe Asp Pro Asp Asp Glu His Ser Glu Pro Val His Thr Val Val Phe

210 215 220

Gln Gly Ala Ala Gly Ile Gly Lys Thr Ile Leu Ala Arg Lys Met Met

225 230 235 240

Leu Asp Trp Ala Ser Gly Thr Leu Tyr Gln Asp Arg Phe Asp Tyr Leu

245 250 255

Phe Tyr Ile His Cys Arg Glu Val Ser Leu Val Thr Gln Arg Ser Leu

260 265 270

Gly Asp Leu Ile Met Ser Cys Cys Pro Asp Pro Asn Pro Pro Ile His

275 280 285

Lys Ile Val Arg Lys Pro Ser Arg Ile Leu Phe Leu Met Asp Gly Phe

290 295 300

Asp Glu Leu Gln Gly Ala Phe Asp Glu His Ile Gly Pro Leu Cys Thr

305 310 315 320

Asp Trp Gln Lys Ala Glu Arg Gly Asp Ile Leu Leu Ser Ser Leu Ile

325 330 335

Arg Lys Lys Leu Leu Pro Glu Ala Ser Leu Leu Ile Thr Thr Arg Pro

340 345 350

Val Ala Leu Glu Lys Leu Gln His Leu Leu Asp His Pro Arg His Val

355 360 365

Glu Ile Leu Gly Phe Ser Glu Ala Lys Arg Lys Glu Tyr Phe Phe Lys

370 375 380

Tyr Phe Ser Asp Glu Ala Gln Ala Arg Ala Ala Phe Ser Leu Ile Gln

385 390 395 400

Glu Asn Glu Val Leu Phe Thr Met Cys Phe Ile Pro Leu Val Cys Trp

405 410 415

Ile Val Cys Thr Gly Leu Lys Gln Gln Met Glu Ser Gly Lys Ser Leu

420 425 430

Ala Gln Thr Ser Lys Thr Thr Thr Ala Val Tyr Val Phe Phe Leu Ser

435 440 445

Ser Leu Leu Gln Pro Arg Gly Gly Ser Gln Glu His Gly Leu Cys Ala

450 455 460

His Leu Trp Gly Leu Cys Ser Leu Ala Ala Asp Gly Ile Trp Asn Gln

465 470 475 480

Lys Ile Leu Phe Glu Glu Ser Asp Leu Arg Asn His Gly Leu Gln Lys

485 490 495

Ala Asp Val Ser Ala Phe Leu Arg Met Asn Leu Phe Gln Lys Glu Val

500 505 510

Asp Cys Glu Lys Phe Tyr Ser Phe Ile His Met Thr Phe Gln Glu Phe

515 520 525

Phe Ala Ala Met Tyr Tyr Leu Leu Glu Glu Glu Lys Glu Gly Arg Thr

530 535 540

Asn Val Pro Gly Ser Arg Leu Lys Leu Pro Ser Arg Asp Val Thr Val

545 550 555 560

Leu Leu Glu Asn Tyr Gly Lys Phe Glu Lys Gly Tyr Leu Ile Phe Val

565 570 575

Val Arg Phe Leu Phe Gly Leu Val Asn Gln Glu Arg Thr Ser Tyr Leu

580 585 590

Glu Lys Lys Leu Ser Cys Lys Ile Ser Gln Gln Ile Arg Leu Glu Leu

595 600 605

Leu Lys Trp Ile Glu Val Lys Ala Lys Ala Lys Lys Leu Gln Ile Gln

610 615 620

Pro Ser Gln Leu Glu Leu Phe Tyr Cys Leu Tyr Glu Met Gln Glu Glu

625 630 635 640

Asp Phe Val Gln Arg Ala Met Asp Tyr Phe Pro Lys Ile Glu Ile Asn

645 650 655

Leu Ser Thr Arg Met Asp His Met Val Ser Ser Phe Cys Ile Glu Asn

660 665 670

Cys His Arg Val Glu Ser Leu Ser Leu Gly Phe Leu His Asn Met Pro

675 680 685

Lys Glu Glu Glu Glu Glu Glu Lys Glu Gly Arg His Leu Asp Met Val

690 695 700

Gln Cys Val Leu Pro Ser Ser Ser His Ala Ala Cys Ser His Gly Leu

705 710 715 720

Val Asn Ser His Leu Thr Ser Ser Phe Cys Arg Gly Leu Phe Ser Val

725 730 735

Leu Ser Thr Ser Gln Ser Leu Thr Glu Leu Asp Leu Ser Asp Asn Ser

740 745 750

Leu Gly Asp Pro Gly Met Arg Val Leu Cys Glu Thr Leu Gln His Pro

755 760 765

Gly Cys Asn Ile Arg Arg Leu Trp Leu Gly Arg Cys Gly Leu Ser His

770 775 780

Glu Cys Cys Phe Asp Ile Ser Leu Val Leu Ser Ser Asn Gln Lys Leu

785 790 795 800

Val Glu Leu Asp Leu Ser Asp Asn Ala Leu Gly Asp Phe Gly Ile Arg

805 810 815

Leu Leu Cys Val Gly Leu Lys His Leu Leu Cys Asn Leu Lys Lys Leu

820 825 830

Trp Leu Val Ser Cys Cys Leu Thr Ser Ala Cys Cys Gln Asp Leu Ala

835 840 845

Ser Val Leu Ser Thr Ser His Ser Leu Thr Arg Leu Tyr Val Gly Glu

850 855 860

Asn Ala Leu Gly Asp Ser Gly Val Ala Ile Leu Cys Glu Lys Ala Lys

865 870 875 880

Asn Pro Gln Cys Asn Leu Gln Lys Leu Gly Leu Val Asn Ser Gly Leu

885 890 895

Thr Ser Val Cys Cys Ser Ala Leu Ser Ser Val Leu Ser Thr Asn Gln

900 905 910

Asn Leu Thr His Leu Tyr Leu Arg Gly Asn Thr Leu Gly Asp Lys Gly

915 920 925

Ile Lys Leu Leu Cys Glu Gly Leu Leu His Pro Asp Cys Lys Leu Gln

930 935 940

Val Leu Glu Leu Asp Asn Cys Asn Leu Thr Ser His Cys Cys Trp Asp

945 950 955 960

Leu Ser Thr Leu Leu Thr Ser Ser Gln Ser Leu Arg Lys Leu Ser Leu

965 970 975

Gly Asn Asn Asp Leu Gly Asp Leu Gly Val Met Met Phe Cys Glu Val

980 985 990

Leu Lys Gln Gln Ser Cys Leu Leu Gln Asn Leu Gly Leu Ser Glu Met

995 1000 1005

Tyr Phe Asn Tyr Glu Thr Lys Ser Ala Leu Glu Thr Leu Gln Glu

1010 1015 1020

Glu Lys Pro Glu Leu Thr Val Val Phe Glu Pro Ser Trp

1025 1030 1035

<210> 540

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<220>

<223> Description of the combined DNA/RNA molecule: synthetic

oligonucleotide

<400> 540

gugcauugaa gacaggaaut t 21

<210> 541

<211> 21

<212> DNA

<213> Homo sapiens

<400> 541

ggctgtaaca ttcggagatt g 21

<210> 542

<211> 21

<212> DNA

<213> Homo sapiens

<400> 542

tcatcattcc cgctatcttt from 21

<210> 543

<211> 21

<212> DNA

<213> Homo sapiens

<400> 543

ccgtaagaag tacagaaagt a 21

<210> 544

<211> 21

<212> DNA

<213> Homo sapiens

<400> 544

gagactcagg agtcgcaatt t 21

<210> 545

<211> 21

<212> DNA

<213> Homo sapiens

<400> 545

cctcatgtaa ttagctcatt c 21

<210> 546

<211> 21

<212> DNA

<213> Homo sapiens

<400> 546

gtggatctag ccacgctaat g 21

<210> 547

<211> 21

<212> DNA

<213> Homo sapiens

<400> 547

ccacagtgta acctgcagaa a 21

<210> 548

<211> 21

<212> DNA

<213> Homo sapiens

<400> 548

ccagccagag tctaactgaa t 21

<210> 549

<211> 21

<212> DNA

<213> Homo sapiens

<400> 549

gcgttagaaa cacttcaaga a 21

<210> 550

<211> 21

<212> DNA

<213> Homo sapiens

<400> 550

gctggaattg ttctactgtt t 21

<210> 551

<211> 21

<212> DNA

<213> Homo sapiens

<400> 551

ccacatgact ttccaggagt t 21

<210> 552

<211> 21

<212> DNA

<213> Homo sapiens

<400> 552

ggctgtaaca ttcggagatt g 21

<210> 553

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 553

ccggggctgt aacattcgga gattgctcga gcaatctccg aatgttacag cctttttg 58

<210> 554

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 554

aattcaaaaa ggctgtaaca ttcggagatt gctcgagcaa tctccgaatg ttacagcc 58

<210> 555

<211> 21

<212> DNA

<213> Homo sapiens

<400> 555

tcatcattcc cgctatcttt from 21

<210> 556

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 556

ccggtcatca ttcccgctat ctttcctcga ggaaagatag cgggaatgat gatttttg 58

<210> 557

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 557

aattcaaaaa tcatcattcc cgctatcttt cctcgaggaa agatagcggg aatgatga 58

<210> 558

<211> 21

<212> DNA

<213> Homo sapiens

<400> 558

ccgtaagaag tacagaaagt a 21

<210> 559

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 559

ccggccgtaa gaagtacaga aagtactcga gtactttctg tacttcttac ggtttttg 58

<210> 560

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 560

aattcaaaaa ccgtaagaag tacagaaagt actcgagtac tttctgtact tcttacgg 58

<210> 561

<211> 21

<212> DNA

<213> Homo sapiens

<400> 561

gagactcagg agtcgcaatt t 21

<210> 562

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 562

ccgggagact caggagtcgc aatttctcga gaaattgcga ctcctgagtc tctttttg 58

<210> 563

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 563

aattcaaaaa gagactcagg agtcgcaatt tctcgagaaa ttgcgactcc tgagtctc 58

<210> 564

<211> 21

<212> DNA

<213> Homo sapiens

<400> 564

cctcatgtaa ttagctcatt c 21

<210> 565

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 565

ccggcctcat gtaattagct cattcctcga ggaatgagct aattacatga ggtttttg 58

<210> 566

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 566

aattcaaaaa cctcatgtaa ttagctcatt cctcgaggaa tgagctaatt acatgagg 58

<210> 567

<211> 21

<212> DNA

<213> Homo sapiens

<400> 567

gtggatctag ccacgctaat g 21

<210> 568

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 568

ccgggtggat ctagccacgc taatgctcga gcattagcgt ggctagatcc actttttg 58

<210> 569

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 569

aattcaaaaa gtggatctag ccacgctaat gctcgagcat tagcgtggct agatccac 58

<210> 570

<211> 21

<212> DNA

<213> Homo sapiens

<400> 570

ccacagtgta acctgcagaa a 21

<210> 571

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 571

aattcaaaaa ccacagtgta acctgcagaa actcgagttt ctgcaggtta cactgtgg 58

<210> 572

<211> 21

<212> DNA

<213> Homo sapiens

<400> 572

ccagccagag tctaactgaa t 21

<210> 573

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 573

ccggccagcc agagtctaac tgaatctcga gattcagtta gactctggct ggtttttg 58

<210> 574

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 574

aattcaaaaa ccagccagag tctaactgaa tctcgagatt cagttagact ctggctgg 58

<210> 575

<211> 21

<212> DNA

<213> Homo sapiens

<400> 575

gcgttagaaa cacttcaaga a 21

<210> 576

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 576

ccgggcgtta gaaacacttc aagaactcga gttcttgaag tgtttctaac gctttttg 58

<210> 577

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 577

aattcaaaaa gcgttagaaa cacttcaaga actcgagttc ttgaagtgtt tctaacgc 58

<210> 578

<211> 21

<212> DNA

<213> Homo sapiens

<400> 578

gctggaattg ttctactgtt t 21

<210> 579

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 579

ccgggctgga attgttctac tgtttctcga gaaacagtag aacaattcca gctttttg 58

<210> 580

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 580

aattcaaaaa gctggaattg ttctactgtt tctcgagaaa cagtagaaca attccagc 58

<210> 581

<211> 21

<212> DNA

<213> Homo sapiens

<400> 581

ccacatgact ttccaggagt t 21

<210> 582

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 582

ccggccacat gactttccag gagttctcga gaactcctgg aaagtcatgt ggtttttg 58

<210> 583

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 583

aattcaaaaa ccacatgact ttccaggagt tctcgagaac tcctggaaag tcatgtgg 58

<210> 584

<211> 21

<212> DNA

<213> Homo sapiens

<400> 584

gaaagccaaa gctaagaagt a 21

<210> 585

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 585

ccgggaaagc caaagctaag aagtactcga gtacttctta gctttggctt tctttttg 58

<210> 586

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 586

aattcaaaaa gaaagccaaa gctaagaagt actcgagtac ttcttagctt tggctttc 58

<210> 587

<211> 23

<212> RNA

<213> Homo sapiens

<400> 587

ucuuugguua ucuagcugua uga 23

<210> 588

<211> 23

<212> RNA

<213> Homo sapiens

<400> 588

ucuuugguua ucuagcugua uga 23

<210> 589

<211> 22

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 589

tggggtattt gacaaactga ca 22

<210> 590

<211> 63

<212> RNA

<213> Caenorhabditis brenneri

<400> 590

ucgcccaucc cguuguucca auauuccaac aacaagugau uauugagcaa ugcgcaugug 60

cgg 63

<210> 591

<211> 97

<212> RNA

<213> Caenorhabditis briggsae

<400> 591

aagcauuuuu cugucccgcg caucccuuug uuccaauauu caaaccagua gaaagauuau 60

ugagcaaugc gcaugugcgg gacagauuga auagcug 97

<210> 592

<211> 98

<212> RNA

<213> Caenorhabditis elegans

<400> 592

auauagcauc uuucugucuc gcccaucccg uugcuccaau auucuaacaa caagugauua 60

uugagcaaug cgcaugugcg ggauagacug auggcugc 98

<210> 593

<211> 101

<212> RNA

<213> Caenorhabditis remanei

<400> 593

ugaagcgucu cucugucccg cucauccugu uguuccaaua uuccaacagc ccagugauua 60

uugagcaaug cgcaugugcg ggacagauug uaugcugcca u 101

<210> 594

<211> 22

<212> RNA

<213> Homo sapiens

<400> 594

aguucuucag uggcaagcuu ua 22

<210> 595

<211> 85

<212> RNA

<213> Homo sapiens

<400> 595

ggcugagccg caguaguucu ucaguggcaa gcuuuauguc cugacccagc uaaagcugcc 60

aguugaagaa cuguugcccu cugcc 85

<210> 596

<211> 21

<212> RNA

<213> Mus musculus

<400> 596

gugcauugua guugcauugc a 21

<210> 597

<211> 69

<212> RNA

<213> Mus musculus

<400> 597

cuguggugca uuguaguugc auugcauguu cuggcaauac cugugcaaug uuuccacagu 60

gcaucacgg 69

<210> 598

<211> 343

<212> PROTEIN

<213> Homo sapiens

<400> 598

Met Glu Ser Lys Tyr Lys Glu Ile Leu Leu Leu Thr Gly Leu Asp Asn

1 5 10 15

Ile Thr Asp Glu Glu Leu Asp Arg Phe Lys Phe Phe Leu Ser Asp Glu

20 25 30

Phe Asn Ile Ala Thr Gly Lys Leu His Thr Ala Asn Arg Ile Gln Val

35 40 45

Ala Thr Leu Met Ile Gln Asn Ala Gly Ala Val Ser Ala Val Met Lys

50 55 60

Thr Ile Arg Ile Phe Gln Lys Leu Asn Tyr Met Leu Leu Ala Lys Arg

65 70 75 80

Leu Gln Glu Glu Lys Glu Lys Val Asp Lys Gln Tyr Lys Ser Val Thr

85 90 95

Lys Pro Lys Pro Leu Ser Gln Ala Glu Met Ser Pro Ala Ala Ser Ala

100 105 110

Ala Ile Arg Asn Asp Val Ala Lys Gln Arg Ala Ala Pro Lys Val Ser

115 120 125

Pro His Val Lys Pro Glu Gln Lys Gln Met Val Ala Gln Gln Glu Ser

130 135 140

Ile Arg Glu Gly Phe Gln Lys Arg Cys Leu Pro Val Met Val Leu Lys

145 150 155 160

Ala Lys Lys Pro Phe Thr Phe Glu Thr Gln Glu Gly Lys Gln Glu Met

165 170 175

Phe His Ala Thr Val Ala Thr Glu Lys Glu Phe Phe Phe Val Lys Val

180 185 190

Phe Asn Thr Leu Leu Lys Asp Lys Phe Ile Pro Lys Arg Ile Ile Ile

195 200 205

Ile Ala Arg Tyr Tyr Arg His Ser Gly Phe Leu Glu Val Asn Ser Ala

210 215 220

Ser Arg Val Leu Asp Ala Glu Ser Asp Gln Lys Val Asn Val Pro Leu

225 230 235 240

Asn Ile Ile Arg Lys Ala Gly Glu Thr Pro Lys Ile Asn Thr Leu Gln

245 250 255

Thr Gln Pro Leu Gly Thr Ile Val Asn Gly Leu Phe Val Val Gln Lys

260 265 270

Val Thr Glu Lys Lys Lys Asn Ile Leu Phe Asp Leu Ser Asp Asn Thr

275 280 285

Gly Lys Met Glu Val Leu Gly Val Arg Asn Glu Asp Thr Met Lys Cys

290 295 300

Lys Glu Gly Asp Lys Val Arg Leu Thr Phe Phe Thr Leu Ser Lys Asn

305 310 315 320

Gly Glu Lys Leu Gln Leu Thr Ser Gly Val His Ser Thr Ile Lys Val

325 330 335

Ile Lys Ala Lys Lys Lys Th

340

<210> 599

<400> 599

000

<210> 600

<211> 1558

<212> DNA

<213> Homo sapiens

<400> 600

atagacattt tcttctgtgg ctgctagtga gaacccaaac cagctcagcc aattagagct 60

ccagttgtca ctcctaccca cactgggcct gggggtgaag ggaagtgttt attaggggta 120

catgtgaagc cgtccagaag tgtcagagtc tttgtagctt tgaaagtcac ctaggttatt 180

tgggcatgct ctcctgagtc ctctgctagt taagctctct gaaaagaagg tggcagaccc 240

ggtttgctga tcgccccagg gatcaggagg ctgatcccaa agttgtcaga tggagagtaa 300

atacaaggag atactcttgc taacaggcct ggataacatc actgatgagg aactggatag 360

gtttaagttc tttctttcag acgagtttaa tattgccaca ggcaaactac atactgcaaa 420

cagaatacaa gtagctacct tgatgattca aaatgctggg gcggtgtctg cagtgatgaa 480

gaccattcgt atttttcaga agttgaatta tatgcttttg gcaaaacgtc ttcaggagga 540

gaaggagaaa gttgataagc aatacaaatc ggtaacaaaa ccaaagccac taagtcaagc 600

tgaaatgagt cctgctgcat ctgcagccat cagaaatgat gtcgcaaagc aacgtgctgc 660

accaaaagtc tctcctcatg ttaagcctga acagaaacag atggtggccc agcaggaatc 720

tatcagagaa gggtttcaga agcgctgttt gccagttatg gtactgaaag caaagaagcc 780

cttcacgttt gagacccaag aaggcaagca ggagatgttt catgctacag tggctacaga 840

aaaggaattc ttctttgtaa aagtttttaa tacactgctg aaagataaat tcattccaaa 900

gagaataatt ataatagcaa gatattatcg gcacagtggt ttcttagagg taaatagcgc 960

ctcacgtgtg ttagatgctg aatctgacca aaaggttaat gtcccgctga acattatcag 1020

aaaagctggt gaaaccccga agatcaacac gcttcaaact cagccccttg gaacaattgt 1080

gaatggtttg tttgtagtcc agaaggtaac agaaaagaag aaaaacatat tatttgacct 1140

aagtgacaac actgggaaaa tggaagtact gggggttaga aacgaggaca caatgaaatg 1200

taaggaagga gataaggttc gacttacatt cttcacactg tcaaaaaatg gagaaaaact 1260

acagctgaca tctggagttc atagcaccat aaaggttatt aaggccaaaa aaaaaacata 1320

gagaagtaaa aaggaccaat tcaagccaac tggtctaagc agcatttaat tgaagaatat 1380

gtgatacagc ctcttcaatc agattgtaag ttacctgaaa gctgcagttc acaggctcct 1440

ctctccacca aattaggata gaataattgc tggataaaca aattcagaat atcaacagat 1500

gatcacaata aacatctgtt tctcattcaa aaaaaaaaaa aaaaaaaaaa aaaaaaaa 1558

<210> 601

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 601

cccgaagatc aacacgcttc a 21

<210> 602

<211> 24

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 602

ttagggttag ggttagggtt aggg 24

<210> 603

<211> 24

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 603

ttcaaattca aattcaaatt caaa 24

<210> 604

<211> 6

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 604

ttaggg 6

<210> 605

<211> 21

<212> DNA

<213> Homo sapiens

<400> 605

agccactaag tcaagctgaa a 21

<210> 606

<211> 21

<212> DNA

<213> Homo sapiens

<400> 606

ccaactggtc taagcagcat t 21

<210> 607

<211> 21

<212> DNA

<213> Homo sapiens

<400> 607

gaaacgagga cacaatgaaa t 21

<210> 608

<211> 21

<212> DNA

<213> Homo sapiens

<400> 608

gccactaagt caagctgaaa t 21

<210> 609

<211> 21

<212> DNA

<213> Homo sapiens

<400> 609

ctggagttca tagcaccata a 21

<210> 610

<211> 21

<212> DNA

<213> Homo sapiens

<400> 610

cccgctgaac attatcagaa a 21

<210> 611

<211> 1393

<212> DNA

<213> Homo sapiens

<400> 611

atactttcag tttcagtcac acaagaaggg aggagagaaa agccatggcc gacaaggtcc 60

tgaaggagaa gagaaagctg tttatccgtt ccatgggtga aggtacaata aatggcttac 120

tggatgaatt attacagaca agggtgctga acaaggaaga gatggagaaa gtaaaacgtg 180

aaaatgctac agttatggat aagacccgag ctttgattga ctccgttatt ccgaaagggg 240

cacaggcatg ccaaatttgc atcacataca tttgtgaaga agacagttac ctggcaggga 300

cgctgggact ctcagcagat caaacatctg gaaattacct taatatgcaa gactctcaag 360

gagtactttc ttcctttcca gctcctcagg cagtgcagga caacccagct atgcccacat 420

cctcaggctc agaagggaat gtcaagcttt gctccctaga agaagctcaa aggatatgga 480

aacaaaagtc ggcagagatt tatccaataa tggacaagtc aagccgcaca cgtcttgctc 540

tcattatctg caatgaagaa tttgacagta ttcctagaag aactggagct gaggttgaca 600

tcacaggcat gacaatgctg ctacaaaatc tggggtacag cgtagatgtg aaaaaaaatc 660

tcactgcttc ggacatgact acagagctgg aggcatttgc acaccgccca gagcacaaga 720

cctctgacag cacgttcctg gtgttcatgt ctcatggtat tcgggaaggc atttgtggga 780

agaaacactc tgagcaagtc ccagatatac tacaactcaa tgcaatcttt aacatgttga 840

ataccaagaa ctgcccaagt ttgaaggaca aaccgaaggt gatcatcatc caggcctgcc 900

gtggtgacag ccctggtgtg gtgtggttta aagattcagt aggagtttct ggaaacctat 960

ctttaccaac tacagaagag tttgaggatg atgctattaa gaaagcccac atagagaagg 1020

attttatcgc tttctgctct tccacaccag ataatgtttc ttggagacat cccacaatgg 1080

gctctgtttt tattggaaga ctcattgaac atatgcaaga atatgcctgt tcctgtgatg 1140

tggaggaaat tttccgcaag gttcgatttt catttgagca gccagatggt agagcgcaga 1200

tgcccaccac tgaaagagtg actttgacaa gatgtttcta cctcttccca ggacattaaa 1260

ataaggaaac tgtatgaatg tctgtgggca ggaagtgaag agatccttct gtaaaggttt 1320

ttggaattat gtctgctgaa taataaactt ttttgaaata ataaatctgg tagaaaaatg 1380

aaaaaaaaaa aaa 1393

<210> 612

<211> 404

<212> PROTEIN

<213> Homo sapiens

<400> 612

Met Ala Asp Lys Val Leu Lys Glu Lys Arg Lys Leu Phe Ile Arg Ser

1 5 10 15

Met Gly Glu Gly Thr Ile Asn Gly Leu Leu Asp Glu Leu Leu Gln Thr

20 25 30

Arg Val Leu Asn Lys Glu Glu Met Glu Lys Val Lys Arg Glu Asn Ala

35 40 45

Thr Val Met Asp Lys Thr Arg Ala Leu Ile Asp Ser Val Ile Pro Lys

50 55 60

Gly Ala Gln Ala Cys Gln Ile Cys Ile Thr Tyr Ile Cys Glu Glu Asp

65 70 75 80

Ser Tyr Leu Ala Gly Thr Leu Gly Leu Ser Ala Asp Gln Thr Ser Gly

85 90 95

Asn Tyr Leu Asn Met Gln Asp Ser Gln Gly Val Leu Ser Ser Phe Pro

100 105 110

Ala Pro Gln Ala Val Gln Asp Asn Pro Ala Met Pro Thr Ser Ser Gly

115 120 125

Ser Glu Gly Asn Val Lys Leu Cys Ser Leu Glu Glu Ala Gln Arg Ile

130 135 140

Trp Lys Gln Lys Ser Ala Glu Ile Tyr Pro Ile Met Asp Lys Ser Ser

145 150 155 160

Arg Thr Arg Leu Ala Leu Ile Ile Cys Asn Glu Glu Phe Asp Ser Ile

165 170 175

Pro Arg Arg Thr Gly Ala Glu Val Asp Ile Thr Gly Met Thr Met Leu

180 185 190

Leu Gln Asn Leu Gly Tyr Ser Val Asp Val Lys Lys Asn Leu Thr Ala

195 200 205

Ser Asp Met Thr Thr Glu Leu Glu Ala Phe Ala His Arg Pro Glu His

210 215 220

Lys Thr Ser Asp Ser Thr Phe Leu Val Phe Met Ser His Gly Ile Arg

225 230 235 240

Glu Gly Ile Cys Gly Lys Lys His Ser Glu Gln Val Pro Asp Ile Leu

245 250 255

Gln Leu Asn Ala Ile Phe Asn Met Leu Asn Thr Lys Asn Cys Pro Ser

260 265 270

Leu Lys Asp Lys Pro Lys Val Ile Ile Ile Gln Ala Cys Arg Gly Asp

275 280 285

Ser Pro Gly Val Val Trp Phe Lys Asp Ser Val Gly Val Ser Gly Asn

290 295 300

Leu Ser Leu Pro Thr Thr Glu Glu Phe Glu Asp Asp Ala Ile Lys Lys

305 310 315 320

Ala His Ile Glu Lys Asp Phe Ile Ala Phe Cys Ser Ser Thr Pro Asp

325 330 335

Asn Val Ser Trp Arg His Pro Thr Met Gly Ser Val Phe Ile Gly Arg

340 345 350

Leu Ile Glu His Met Gln Glu Tyr Ala Cys Ser Cys Asp Val Glu Glu

355 360 365

Ile Phe Arg Lys Val Arg Phe Ser Phe Glu Gln Pro Asp Gly Arg Ala

370 375 380

Gln Met Pro Thr Thr Glu Arg Val Thr Leu Thr Arg Cys Phe Tyr Leu

385 390 395 400

Phe Pro Gly His

<210> 613

<211> 21

<212> DNA

<213> Homo sapiens

<400> 613

cacacgtctt gctctcatta t 21

<210> 614

<211> 21

<212> DNA

<213> Homo sapiens

<400> 614

ctacaactca atgcaatcttt 21

<210> 615

<211> 21

<212> DNA

<213> Homo sapiens

<400> 615

ccagatatac tacaactcaa t 21

<210> 616

<211> 21

<212> DNA

<213> Homo sapiens

<400> 616

gaagagtttg aggatgatgc t 21

<210> 617

<211> 21

<212> DNA

<213> Homo sapiens

<400> 617

ccatgggtga aggtacaata a 21

<210> 618

<211> 21

<212> DNA

<213> Homo sapiens

<400> 618

gctttgattg actccgttat t 21

<210> 619

<211> 21

<212> DNA

<213> Homo sapiens

<400> 619

gaaggtacaa taaatggctt a 21

<210> 620

<211> 21

<212> DNA

<213> Homo sapiens

<400> 620

cacacgtctt gctctcatta t 21

<210> 621

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 621

ccggcacacg tcttgctctc attatctcga gataatgaga gcaagacgtg tgtttttg 58

<210> 622

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 622

aattcaaaaa cacacgtctt gctctcatta tctcgagata atgagagcaa gacgtgtg 58

<210> 623

<211> 21

<212> DNA

<213> Homo sapiens

<400> 623

ctacaactca atgcaatcttt 21

<210> 624

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 624

ccggctacaa ctcaatgcaa tctttctcga gaaagattgc attgagttgt agtttttg 58

<210> 625

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 625

aattcaaaaa ctacaactca atgcaatctt tctcgagaaa gattgcattg agttgtag 58

<210> 626

<211> 21

<212> DNA

<213> Homo sapiens

<400> 626

ccagatatac tacaactcaa t 21

<210> 627

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 627

ccggccagat atactacaac tcaatctcga gattgagttg tagtatatct ggtttttg 58

<210> 628

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 628

aattcaaaaa ccagatatac tacaactcaa tctcgagatt gagttgtagt atatctgg 58

<210> 629

<211> 21

<212> DNA

<213> Homo sapiens

<400> 629

tgtatgaatg tctgctgggc a 21

<210> 630

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 630

ccggtgtatg aatgtctgct gggcactcga gtgcccagca gacattcata catttttg 58

<210> 631

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 631

aattcaaaaa tgtatgaatg tctgctgggc actcgagtgc ccagcagaca ttcataca 58

<210> 632

<211> 21

<212> DNA

<213> Homo sapiens

<400> 632

caaggacctg aaggagaaga a 21

<210> 633

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 633

ccggcaagga cctgaaggag aagaactcga gttcttctcc ttcaggtcct tgtttttg 58

<210> 634

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 634

aattcaaaaa caaggacctg aaggagaaga actcgagttc ttctccttca ggtccttg 58

<210> 635

<211> 21

<212> DNA

<213> Homo sapiens

<400> 635

caatgtctgt ggggaggaaga a 21

<210> 636

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 636

ccggcaatgt ctgtgggagg aagaactcga gttcttcctc cccagacat tgtttttg 58

<210> 637

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 637

aattcaaaaa caatgtctgt gggaggaaga actcgagttc ttcctcccac agacattg 58

<210> 638

<211> 21

<212> DNA

<213> Homo sapiens

<400> 638

caaggtcctg tagggagaag a 21

<210> 639

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 639

ccggcaaggt cctgtaggga gaagactcga gtcttctccc tacaggacct tgtttttg 58

<210> 640

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 640

aattcaaaaa caaggtcctg tagggagaag actcgagtct tctccctaca ggaccttg 58

<210> 641

<211> 21

<212> DNA

<213> Homo sapiens

<400> 641

caaggtcctg tagggagaag a 21

<210> 642

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 642

ccggcaaggt cctgtaggga gaagactcga gtcttctccc tacaggacct tgtttttg 58

<210> 643

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 643

aattcaaaaa caaggtcctg tagggagaag actcgagtct tctccctaca ggaccttg 58

<210> 644

<211> 21

<212> DNA

<213> Homo sapiens

<400> 644

acaagcccaa ggtgatcatt a 21

<210> 645

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 645

ccggacaagc ccaaggtgat cattactcga gtaatgatca ccttgggctt gttttttg 58

<210> 646

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 646

aattcaaaaa acaagcccaa ggtgatcatt actcgagtaa tgatcacctt gggcttgt 58

<210> 647

<211> 21

<212> DNA

<213> Homo sapiens

<400> 647

caaggacttg aaggagaaga a 21

<210> 648

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 648

ccggcaagga cttgaaggag aagaactcga gttcttctcc ttcaagtcct tgtttttg 58

<210> 649

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 649

aattcaaaaa caaggacttg aaggagaaga actcgagttc ttctccttca agtccttg 58

<210> 650

<211> 21

<212> DNA

<213> Homo sapiens

<400> 650

cccaagtttg aagtacaagt a 21

<210> 651

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 651

ccggcccaag tttgaagtac aagtactcga gtacttgtac ttcaaacttg ggtttttg 58

<210> 652

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 652

aattcaaaaa cccaagtttg aagtacaagt actcgagtac ttgtacttca aacttggg 58

<210> 653

<211> 21

<212> DNA

<213> Homo sapiens

<400> 653

cccaggacat gataataaga t 21

<210> 654

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 654

ccggcccagg acatgataat aagatctcga gatcttatta tcatgtcctg ggtttttg 58

<210> 655

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 655

aattcaaaaa cccaggacat gataataaga tctcgagatc ttattatcat gtcctggg 58

<210> 656

<211> 21

<212> DNA

<213> Homo sapiens

<400> 656

gaatttgaca gtttcctgcc a 21

<210> 657

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 657

ccgggaattt gacagtttcc tgccactcga gtggcaggaa actgtcaaat tctttttg 58

<210> 658

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 658

aattcaaaaa gaatttgaca gtttcctgcc actcgagtgg caggaaactg tcaaattc 58

<210> 659

<211> 21

<212> DNA

<213> Homo sapiens

<400> 659

cccaagtttg aggtcaaagt t 21

<210> 660

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 660

ccggcccaag tttgaggtca aagttctcga gaactttgac ctcaaacttg ggtttttg 58

<210> 661

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 661

aattcaaaaa cccaagtttg aggtcaaagt tctcgagaac tttgacctca aacttggg 58

<210> 662

<211> 21

<212> DNA

<213> Homo sapiens

<400> 662

cgacaagatg ttctccctca a 21

<210> 663

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 663

ccggcgacaa gatgttctcc ctcaactcga gttgagggag aacatcttgt cgtttttg 58

<210> 664

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 664

aattcaaaaa cgacaagatg ttctccctca actcgagttg agggagaaca tcttgtcg 58

<210> 665

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 665

aaaggttaat gtcccgctga a 21

<210> 666

<211> 21

<212> DNA

<213> Homo sapiens

<400> 666

agccactaag tcaagctgaa a 21

<210> 667

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 667

ccggagccac taagtcaagc tgaaactcga gtttcagctt gacttagtgg cttttttg 58

<210> 668

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 668

aattcaaaaa agccactaag tcaagctgaa actcgagttt cagcttgact tagtggct 58

<210> 669

<211> 21

<212> DNA

<213> Homo sapiens

<400> 669

ccaactggtc taagcagcat t 21

<210> 670

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 670

ccggccaact ggtctaagca gcattctcga gaatgctgct tagaccagtt ggtttttg 58

<210> 671

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 671

aattcaaaaa ccaactggtc taagcagcat tctcgagaat gctgcttaga ccagttgg 58

<210> 672

<211> 21

<212> DNA

<213> Homo sapiens

<400> 672

gaaacgagga cacaatgaaa t 21

<210> 673

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 673

ccgggaaacg aggacacaat gaaatctcga gatttcattg tgtcctcgtt tctttttg 58

<210> 674

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 674

aattcaaaaa gaaacgagga cacaatgaaa tctcgagatt tcattgtgtc ctcgtttc 58

<210> 675

<211> 21

<212> DNA

<213> Homo sapiens

<400> 675

gccactaagt caagctgaaa t 21

<210> 676

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 676

ccgggccact aagtcaagct gaaatctcga gatttcagct tgacttagtg gctttttg 58

<210> 677

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 677

aattcaaaaa gccactaagt caagctgaaa tctcgagatt tcagcttgac ttagtggc 58

<210> 678

<211> 21

<212> DNA

<213> Homo sapiens

<400> 678

ctggagttca tagcaccata a 21

<210> 679

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 679

ccggctggag ttcatagcac cataactcga gttatggtgc tatgaactcc agtttttg 58

<210> 680

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 680

aattcaaaaa ctggagttca tagcaccata actcgagtta tggtgctatg aactccag 58

<210> 681

<211> 21

<212> DNA

<213> Homo sapiens

<400> 681

cccgctgaac attatcagaa a 21

<210> 682

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 682

ccggcccgct gaacattatc agaaactcga gtttctgata atgttcagcg ggtttttg 58

<210> 683

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 683

aattcaaaaa cccgctgaac attatcagaa actcgagttt ctgataatgt tcagcggg 58

<210> 684

<211> 21

<212> DNA

<213> Homo sapiens

<400> 684

agccactaag tcaagctgaa a 21

<210> 685

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 685

ccggagccac taagtcaagc tgaaactcga gtttcagctt gacttagtgg cttttttg 58

<210> 686

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 686

aattcaaaaa agccactaag tcaagctgaa actcgagttt cagcttgact tagtggct 58

<210> 687

<211> 21

<212> DNA

<213> Homo sapiens

<400> 687

ccaactggtc taagcagcat t 21

<210> 688

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 688

ccggccaact ggtctaagca gcattctcga gaatgctgct tagaccagtt ggtttttg 58

<210> 689

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 689

aattcaaaaa ccaactggtc taagcagcat tctcgagaat gctgcttaga ccagttgg 58

<210> 690

<211> 21

<212> DNA

<213> Homo sapiens

<400> 690

gaaacgagga cacaatgaaa t 21

<210> 691

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 691

ccgggaaacg aggacacaat gaaatctcga gatttcattg tgtcctcgtt tctttttg 58

<210> 692

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 692

aattcaaaaa gaaacgagga cacaatgaaa tctcgagatt tcattgtgtc ctcgtttc 58

<210> 693

<211> 21

<212> DNA

<213> Homo sapiens

<400> 693

gccactaagt caagctgaaa t 21

<210> 694

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 694

ccgggccact aagtcaagct gaaatctcga gatttcagct tgacttagtg gctttttg 58

<210> 695

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 695

aattcaaaaa gccactaagt caagctgaaa tctcgagatt tcagcttgac ttagtggc 58

<210> 696

<211> 21

<212> DNA

<213> Homo sapiens

<400> 696

ctggagttca tagcaccata a 21

<210> 697

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 697

ccggctggag ttcatagcac cataactcga gttatggtgc tatgaactcc agtttttg 58

<210> 698

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 698

aattcaaaaa ctggagttca tagcaccata actcgagtta tggtgctatg aactccag 58

<210> 699

<211> 21

<212> DNA

<213> Homo sapiens

<400> 699

cccgctgaac attatcagaa a 21

<210> 700

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 700

ccggcccgct gaacattatc agaaactcga gtttctgata atgttcagcg ggtttttg 58

<210> 701

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 701

aattcaaaaa cccgctgaac attatcagaa actcgagttt ctgataatgt tcagcggg 58

<210> 702

<211> 21

<212> DNA

<213> Homo sapiens

<400> 702

agccactaag tcaagctgaa a 21

<210> 703

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 703

ccggagccac taagtcaagc tgaaactcga gtttcagctt gacttagtgg cttttttg 58

<210> 704

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 704

aattcaaaaa agccactaag tcaagctgaa actcgagttt cagcttgact tagtggct 58

<210> 705

<211> 21

<212> DNA

<213> Homo sapiens

<400> 705

ccaactggtc taagcagcat t 21

<210> 706

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 706

ccggccaact ggtctaagca gcattctcga gaatgctgct tagaccagtt ggtttttg 58

<210> 707

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 707

aattcaaaaa ccaactggtc taagcagcat tctcgagaat gctgcttaga ccagttgg 58

<210> 708

<211> 21

<212> DNA

<213> Homo sapiens

<400> 708

gaaacgagga cacaatgaaa t 21

<210> 709

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 709

ccgggaaacg aggacacaat gaaatctcga gatttcattg tgtcctcgtt tctttttg 58

<210> 710

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 710

aattcaaaaa gaaacgagga cacaatgaaa tctcgagatt tcattgtgtc ctcgtttc 58

<210> 711

<211> 21

<212> DNA

<213> Homo sapiens

<400> 711

gccactaagt caagctgaaa t 21

<210> 712

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 712

ccgggccact aagtcaagct gaaatctcga gatttcagct tgacttagtg gctttttg 58

<210> 713

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 713

aattcaaaaa gccactaagt caagctgaaa tctcgagatt tcagcttgac ttagtggc 58

<210> 714

<211> 21

<212> DNA

<213> Homo sapiens

<400> 714

ctggagttca tagcaccata a 21

<210> 715

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 715

ccggctggag ttcatagcac cataactcga gttatggtgc tatgaactcc agtttttg 58

<210> 716

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 716

aattcaaaaa ctggagttca tagcaccata actcgagtta tggtgctatg aactccag 58

<210> 717

<211> 21

<212> DNA

<213> Homo sapiens

<400> 717

cccgctgaac attatcagaa a 21

<210> 718

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 718

ccggcccgct gaacattatc agaaactcga gtttctgata atgttcagcg ggtttttg 58

<210> 719

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 719

aattcaaaaa cccgctgaac attatcagaa actcgagttt ctgataatgt tcagcggg 58

<210> 720

<211> 21

<212> DNA

<213> Homo sapiens

<400> 720

agccactaag tcaagctgaa a 21

<210> 721

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 721

ccggagccac taagtcaagc tgaaactcga gtttcagctt gacttagtgg cttttttg 58

<210> 722

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 722

aattcaaaaa agccactaag tcaagctgaa actcgagttt cagcttgact tagtggct 58

<210> 723

<211> 21

<212> DNA

<213> Homo sapiens

<400> 723

ccaactggtc taagcagcat t 21

<210> 724

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 724

ccggccaact ggtctaagca gcattctcga gaatgctgct tagaccagtt ggtttttg 58

<210> 725

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 725

aattcaaaaa ccaactggtc taagcagcat tctcgagaat gctgcttaga ccagttgg 58

<210> 726

<211> 21

<212> DNA

<213> Homo sapiens

<400> 726

gaaacgagga cacaatgaaa t 21

<210> 727

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 727

ccgggaaacg aggacacaat gaaatctcga gatttcattg tgtcctcgtt tctttttg 58

<210> 728

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 728

aattcaaaaa gaaacgagga cacaatgaaa tctcgagatt tcattgtgtc ctcgtttc 58

<210> 729

<211> 21

<212> DNA

<213> Homo sapiens

<400> 729

gccactaagt caagctgaaa t 21

<210> 730

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 730

ccgggccact aagtcaagct gaaatctcga gatttcagct tgacttagtg gctttttg 58

<210> 731

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 731

aattcaaaaa gccactaagt caagctgaaa tctcgagatt tcagcttgac ttagtggc 58

<210> 732

<211> 21

<212> DNA

<213> Homo sapiens

<400> 732

ctggagttca tagcaccata a 21

<210> 733

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 733

ccggctggag ttcatagcac cataactcga gttatggtgc tatgaactcc agtttttg 58

<210> 734

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 734

aattcaaaaa ctggagttca tagcaccata actcgagtta tggtgctatg aactccag 58

<210> 735

<211> 21

<212> DNA

<213> Homo sapiens

<400> 735

cccgctgaac attatcagaa a 21

<210> 736

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 736

ccggcccgct gaacattatc agaaactcga gtttctgata atgttcagcg ggtttttg 58

<210> 737

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 737

aattcaaaaa cccgctgaac attatcagaa actcgagttt ctgataatgt tcagcggg 58

<210> 738

<211> 21

<212> DNA

<213> Homo sapiens

<400> 738

agccactaag tcaagctgaa a 21

<210> 739

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 739

ccggagccac taagtcaagc tgaaactcga gtttcagctt gacttagtgg cttttttg 58

<210> 740

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 740

aattcaaaaa agccactaag tcaagctgaa actcgagttt cagcttgact tagtggct 58

<210> 741

<211> 21

<212> DNA

<213> Homo sapiens

<400> 741

ccaactggtc taagcagcat t 21

<210> 742

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 742

ccggccaact ggtctaagca gcattctcga gaatgctgct tagaccagtt ggtttttg 58

<210> 743

<211> 21

<212> DNA

<213> Homo sapiens

<400> 743

gaaacgagga cacaatgaaa t 21

<210> 744

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 744

ccgggaaacg aggacacaat gaaatctcga gatttcattg tgtcctcgtt tctttttg 58

<210> 745

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 745

aattcaaaaa gaaacgagga cacaatgaaa tctcgagatt tcattgtgtc ctcgtttc 58

<210> 746

<211> 21

<212> DNA

<213> Homo sapiens

<400> 746

gccactaagt caagctgaaa t 21

<210> 747

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 747

ccgggccact aagtcaagct gaaatctcga gatttcagct tgacttagtg gctttttg 58

<210> 748

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 748

aattcaaaaa gccactaagt caagctgaaa tctcgagatt tcagcttgac ttagtggc 58

<210> 749

<211> 21

<212> DNA

<213> Homo sapiens

<400> 749

ctggagttca tagcaccata a 21

<210> 750

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 750

ccggctggag ttcatagcac cataactcga gttatggtgc tatgaactcc agtttttg 58

<210> 751

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 751

aattcaaaaa ctggagttca tagcaccata actcgagtta tggtgctatg aactccag 58

<210> 752

<211> 21

<212> DNA

<213> Homo sapiens

<400> 752

cccgctgaac attatcagaa a 21

<210> 753

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 753

ccggcccgct gaacattatc agaaactcga gtttctgata atgttcagcg ggtttttg 58

<210> 754

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 754

aattcaaaaa cccgctgaac attatcagaa actcgagttt ctgataatgt tcagcggg 58

<210> 755

<211> 21

<212> DNA

<213> Homo sapiens

<400> 755

gattgtttca acacaagagt a 21

<210> 756

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 756

ccgggattgt ttcaacacaa gagtactcga gtactcttgt gttgaaacaa tctttttg 58

<210> 757

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 757

aattcaaaaa gattgtttca acacaagagt actcgagtac tcttgtgttg aaacaatc 58

<210> 758

<211> 21

<212> DNA

<213> Homo sapiens

<400> 758

agccactaag tcaagctgaa a 21

<210> 759

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 759

ccggagccac taagtcaagc tgaaactcga gtttcagctt gacttagtgg cttttttg 58

<210> 760

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 760

aattcaaaaa agccactaag tcaagctgaa actcgagttt cagcttgact tagtggct 58

<210> 761

<211> 21

<212> DNA

<213> Homo sapiens

<400> 761

ccaactggtc taagcagcat t 21

<210> 762

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 762

ccggccaact ggtctaagca gcattctcga gaatgctgct tagaccagtt ggtttttg 58

<210> 763

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 763

aattcaaaaa ccaactggtc taagcagcat tctcgagaat gctgcttaga ccagttgg 58

<210> 764

<211> 21

<212> DNA

<213> Homo sapiens

<400> 764

gaaacgagga cacaatgaaa t 21

<210> 765

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 765

ccgggaaacg aggacacaat gaaatctcga gatttcattg tgtcctcgtt tctttttg 58

<210> 766

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 766

aattcaaaaa gaaacgagga cacaatgaaa tctcgagatt tcattgtgtc ctcgtttc 58

<210> 767

<211> 21

<212> DNA

<213> Homo sapiens

<400> 767

gccactaagt caagctgaaa t 21

<210> 768

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 768

ccgggccact aagtcaagct gaaatctcga gatttcagct tgacttagtg gctttttg 58

<210> 769

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 769

aattcaaaaa gccactaagt caagctgaaa tctcgagatt tcagcttgac ttagtggc 58

<210> 770

<400> 770

000

<210> 771

<400> 771

000

<210> 772

<400> 772

000

<210> 773

<400> 773

000

<210> 774

<400> 774

000

<210> 775

<400> 775

000

<210> 776

<400> 776

000

<210> 777

<400> 777

000

<210> 778

<400> 778

000

<210> 779

<400> 779

000

<210> 780

<211> 21

<212> DNA

<213> Homo sapiens

<400> 780

ctggagttca tagcaccata a 21

<210> 781

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 781

ccggctggag ttcatagcac cataactcga gttatggtgc tatgaactcc agtttttg 58

<210> 782

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 782

aattcaaaaa ctggagttca tagcaccata actcgagtta tggtgctatg aactccag 58

<210> 783

<211> 21

<212> DNA

<213> Homo sapiens

<400> 783

cccgctgaac attatcagaa a 21

<210> 784

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 784

ccggcccgct gaacattatc agaaactcga gtttctgata atgttcagcg ggtttttg 58

<210> 785

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 785

aattcaaaaa cccgctgaac attatcagaa actcgagttt ctgataatgt tcagcggg 58

<210> 786

<211> 21

<212> DNA

<213> Homo sapiens

<400> 786

agccactaag tcaagctgaa a 21

<210> 787

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 787

ccggagccac taagtcaagc tgaaactcga gtttcagctt gacttagtgg cttttttg 58

<210> 788

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 788

aattcaaaaa agccactaag tcaagctgaa actcgagttt cagcttgact tagtggct 58

<210> 789

<211> 21

<212> DNA

<213> Homo sapiens

<400> 789

ccaactggtc taagcagcat t 21

<210> 790

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 790

ccggccaact ggtctaagca gcattctcga gaatgctgct tagaccagtt ggtttttg 58

<210> 791

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 791

aattcaaaaa ccaactggtc taagcagcat tctcgagaat gctgcttaga ccagttgg 58

<210> 792

<211> 21

<212> DNA

<213> Homo sapiens

<400> 792

gaaacgagga cacaatgaaa t 21

<210> 793

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 793

ccgggaaacg aggacacaat gaaatctcga gatttcattg tgtcctcgtt tctttttg 58

<210> 794

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 794

aattcaaaaa gaaacgagga cacaatgaaa tctcgagatt tcattgtgtc ctcgtttc 58

<210> 795

<211> 21

<212> DNA

<213> Homo sapiens

<400> 795

gccactaagt caagctgaaa t 21

<210> 796

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 796

ccgggccact aagtcaagct gaaatctcga gatttcagct tgacttagtg gctttttg 58

<210> 797

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 797

aattcaaaaa gccactaagt caagctgaaa tctcgagatt tcagcttgac ttagtggc 58

<210> 798

<211> 21

<212> DNA

<213> Homo sapiens

<400> 798

ctggagttca tagcaccata a 21

<210> 799

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 799

ccggctggag ttcatagcac cataactcga gttatggtgc tatgaactcc agtttttg 58

<210> 800

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 800

aattcaaaaa ctggagttca tagcaccata actcgagtta tggtgctatg aactccag 58

<210> 801

<211> 21

<212> DNA

<213> Homo sapiens

<400> 801

cccgctgaac attatcagaa a 21

<210> 802

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 802

ccggcccgct gaacattatc agaaactcga gtttctgata atgttcagcg ggtttttg 58

<210> 803

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 803

aattcaaaaa cccgctgaac attatcagaa actcgagttt ctgataatgt tcagcggg 58

<210> 804

<211> 6

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 804

ggttga 6

<210> 805

<211> 4

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 805

agtt 4

<210> 806

<211> 6

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 806

ggttgg 6

<210> 807

<211> 6

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 807

agttgg 6

<210> 808

<211> 6

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 808

agttga 6

<210> 809

<211> 6

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 809

rrttrr 6

<210> 810

<400> 810

000

<210> 811

<400> 811

000

<210> 812

<400> 812

000

<210> 813

<400> 813

000

<210> 814

<400> 814

000

<210> 815

<400> 815

000

<210> 816

<400> 816

000

<210> 817

<400> 817

000

<210> 818

<400> 818

000

<210> 819

<400> 819

000

<210> 820

<400> 820

000

<210> 821

<400> 821

000

<210> 822

<400> 822

000

<210> 823

<400> 823

000

<210> 824

<400> 824

000

<210> 825

<400> 825

000

<210> 826

<400> 826

000

<210> 827

<400> 827

000

<210> 828

<400> 828

000

<210> 829

<400> 829

000

<210> 830

<400> 830

000

<210> 831

<400> 831

000

<210> 832

<400> 832

000

<210> 833

<400> 833

000

<210> 834

<400> 834

000

<210> 835

<400> 835

000

<210> 836

<400> 836

000

<210> 837

<400> 837

000

<210> 838

<400> 838

000

<210> 839

<400> 839

000

<210> 840

<400> 840

000

<210> 841

<400> 841

000

<210> 842

<400> 842

000

<210> 843

<400> 843

000

<210> 844

<400> 844

000

<210> 845

<400> 845

000

<210> 846

<400> 846

000

<210> 847

<400> 847

000

<210> 848

<400> 848

000

<210> 849

<400> 849

000

<210> 850

<400> 850

000

<210> 851

<400> 851

000

<210> 852

<400> 852

000

<210> 853

<400> 853

000

<210> 854

<400> 854

000

<210> 855

<400> 855

000

<210> 856

<400> 856

000

<210> 857

<400> 857

000

<210> 858

<400> 858

000

<210> 859

<400> 859

000

<210> 860

<400> 860

000

<210> 861

<400> 861

000

<210> 862

<400> 862

000

<210> 863

<400> 863

000

<210> 864

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 864

tcctggaggg gttgt 15

<210> 865

<400> 865

000

<210> 866

<400> 866

000

<210> 867

<400> 867

000

<210> 868

<400> 868

000

<210> 869

<400> 869

000

<210> 870

<400> 870

000

<210> 871

<400> 871

000

<210> 872

<400> 872

000

<210> 873

<400> 873

000

<210> 874

<400> 874

000

<210> 875

<400> 875

000

<210> 876

<400> 876

000

<210> 877

<400> 877

000

<210> 878

<400> 878

000

<210> 879

<400> 879

000

<210> 880

<400> 880

000

<210> 881

<400> 881

000

<210> 882

<211> 131

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic polypeptide

<400> 882

Met Ala Ala Pro Arg Gly Arg Pro Lys Lys Asp Leu Thr Met Glu Asp

1 5 10 15

Leu Thr Ala Lys Ile Ser Gln Leu Thr Val Glu Asn Arg Glu Leu Arg

20 25 30

Lys Ala Leu Gly Ser Thr Ala Asp Pro Arg Asp Arg Pro Leu Thr Ala

35 40 45

Thr Glu Lys Glu Ala Gln Leu Thr Ala Thr Val Gly Ala Leu Ser Ala

50 55 60

Ala Ala Ala Lys Lys Ile Glu Ala Arg Val Arg Thr Ile Phe Ser Lys

65 70 75 80

Val Val Thr Gln Lys Gln Val Asp Asp Ala Leu Lys Gly Leu Ser Leu

85 90 95

Arg Ile Asp Val Cys Met Ser Asp Gly Gly Thr Ala Lys Pro Pro Pro

100 105 110

Gly Ala Asn Asn Arg Arg Arg Arg Gly Ala Ser Thr Thr Arg Ala Gly

115 120 125

Val Asp Asp

130

<210> 883

<211> 1129

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic polypeptide

<400> 883

Met Ala Pro Pro Gly Met Arg Leu Arg Ser Gly Arg Ser Thr Gly Ala

1 5 10 15

Pro Leu Thr Arg Gly Ser Cys Arg Lys Arg Asn Arg Ser Pro Glu Arg

20 25 30

Cys Asp Leu Gly Asp Asp Leu His Leu Gln Pro Arg Arg Lys His Val

35 40 45

Ala Asp Ser Val Asp Gly Arg Glu Cys Gly Pro His Thr Leu Pro Ile

50 55 60

Pro Gly Ser Pro Thr Val Phe Thr Ser Gly Leu Pro Ala Phe Val Ser

65 70 75 80

Ser Pro Thr Leu Pro Val Ala Pro Ile Pro Ser Pro Ala Pro Ala Thr

85 90 95

Pro Leu Pro Pro Pro Ala Leu Leu Pro Pro Val Thr Thr Ser Ser Ser

100 105 110

Pro Ile Pro Pro Ser His Pro Val Ser Pro Gly Thr Thr Asp Thr His

115 120 125

Ser Pro Ser Pro Ala Leu Pro Pro Thr Gln Ser Pro Glu Ser Ser Gln

130 135 140

Arg Pro Pro Leu Ser Ser Pro Thr Gly Arg Pro Asp Ser Ser Thr Pro

145 150 155 160

Met Arg Pro Pro Pro Ser Gln Gln Thr Thr Pro Pro His Ser Pro Thr

165 170 175

Thr Pro Pro Pro Glu Pro Pro Ser Lys Ser Ser Pro Asp Ser Leu Ala

180 185 190

Pro Ser Thr Leu Arg Ser Leu Arg Lys Arg Arg Leu Ser Ser Pro Gln

195 200 205

Gly Pro Ser Thr Leu Asn Pro Ile Cys Gln Ser Pro Pro Val Ser Pro

210 215 220

Pro Arg Cys Asp Phe Ala Asn Arg Ser Val Tyr Pro Pro Trp Ala Thr

225 230 235 240

Glu Ser Pro Ile Tyr Val Gly Ser Ser Ser Asp Gly Asp Thr Pro Pro

245 250 255

Arg Gln Pro Pro Thr Ser Pro Ile Ser Ile Gly Ser Ser Ser Pro Ser

260 265 270

Glu Gly Ser Trp Gly Asp Asp Thr Ala Met Leu Val Leu Leu Ala Glu

275 280 285

Ile Ala Glu Glu Ala Ser Lys Asn Glu Lys Glu Cys Ser Glu Asn Asn

290 295 300

Gln Ala Gly Glu Asp Asn Gly Asp Asn Glu Ile Ser Lys Glu Ser Gln

305 310 315 320

Val Asp Lys Asp Asp Asn Asp Asn Lys Asp Asp Glu Glu Glu Gln Glu

325 330 335

Thr Asp Glu Glu Asp Glu Glu Asp Asp Glu Glu Asp Asp Glu Glu Asp

340 345 350

Asp Glu Glu Asp Asp Glu Glu Asp Asp Glu Glu Asp Asp Glu Glu Asp

355 360 365

Asp Glu Glu Glu Asp Glu Glu Glu Asp Glu Glu Glu Asp Glu Glu Glu

370 375 380

Asp Glu Glu Glu Glu Glu Asp Glu Glu Asp Asp Asp Asp Glu Asp Asn

385 390 395 400

Glu Asp Glu Glu Asp Asp Glu Glu Glu Asp Lys Lys Glu Asp Glu Glu

405 410 415

Asp Gly Gly Asp Gly Asn Lys Thr Leu Ser Ile Gln Ser Ser Gln Gln

420 425 430

Gln Gln Glu Pro Gln Gln Gln Glu Pro Gln Gln Gln Glu Pro Gln Gln

435 440 445

Gln Glu Pro Gln Gln Gln Glu Pro Gln Gln Gln Glu Pro Gln Gln Gln

450 455 460

Glu Pro Gln Gln Gln Glu Pro Gln Gln Arg Glu Pro Gln Gln Arg Glu

465 470 475 480

Pro Gln Gln Arg Glu Pro Gln Gln Arg Glu Pro Gln Gln Arg Glu Pro

485 490 495

Gln Gln Arg Glu Pro Gln Gln Arg Glu Pro Gln Gln Arg Glu Pro Gln

500 505 510

Gln Arg Glu Pro Gln Gln Arg Glu Pro Gln Gln Arg Glu Pro Gln Gln

515 520 525

Arg Glu Pro Gln Gln Gln Glu Pro Gln Gln Gln Glu Pro Gln Gln Gln

530 535 540

Glu Pro Gln Gln Gln Glu Pro Gln Gln Gln Glu Pro Gln Gln Gln Glu

545 550 555 560

Pro Gln Gln Gln Glu Pro Gln Gln Gln Glu Pro Gln Gln Gln Glu Pro

565 570 575

Gln Gln Gln Glu Pro Gln Gln Gln Glu Pro Gln Gln Gln Glu Pro Gln

580 585 590

Gln Gln Asp Glu Gln Gln Gln Asp Glu Gln Gln Gln Asp Glu Gln Gln

595 600 605

Gln Asp Glu Gln Gln Gln Asp Glu Gln Gln Gln Asp Glu Gln Gln Gln

610 615 620

Asp Glu Gln Gln Gln Asp Glu Gln Glu Gln Gln Asp Glu Gln Gln Gln

625 630 635 640

Asp Glu Gln Gln Gln Gln Asp Glu Gln Glu Gln Gln Glu Glu Gln Glu

645 650 655

Gln Gln Glu Glu Gln Gln Gln Asp Glu Gln Gln Gln Asp Glu Gln Gln

660 665 670

Gln Asp Glu Gln Gln Gln Asp Glu Gln Glu Gln Gln Asp Glu Gln Gln

675 680 685

Gln Asp Glu Gln Gln Gln Gln Asp Glu Gln Glu Gln Gln Glu Glu Gln

690 695 700

Glu Gln Gln Glu Glu Gln Glu Gln Gln Glu Glu Gln Glu Gln Gln Glu

705 710 715 720

Glu Gln Glu Gln Glu Leu Glu Glu Gln Glu Gln Glu Leu Glu Glu Gln

725 730 735

Glu Gln Glu Leu Glu Glu Gln Glu Gln Glu Leu Glu Glu Gln Glu Gln

740 745 750

Glu Leu Glu Glu Gln Glu Gln Glu Leu Glu Glu Gln Glu Gln Glu Leu

755 760 765

Glu Glu Gln Glu Gln Glu Leu Glu Glu Gln Glu Gln Glu Leu Glu Glu

770 775 780

Gln Glu Gln Glu Leu Glu Glu Gln Glu Gln Glu Leu Glu Glu Gln Glu

785 790 795 800

Gln Glu Leu Glu Glu Gln Glu Gln Glu Leu Glu Glu Gln Glu Gln Glu

805 810 815

Leu Glu Glu Gln Glu Gln Glu Gln Glu Leu Glu Glu Val Glu Glu Gln

820 825 830

Glu Gln Glu Gln Glu Glu Gln Glu Leu Glu Glu Val Glu Glu Gln Glu

835 840 845

Gln Glu Gln Glu Glu Gln Glu Glu Gln Glu Leu Glu Glu Val Glu Glu

850 855 860

Gln Glu Glu Gln Glu Leu Glu Glu Val Glu Glu Gln Glu Glu Gln Glu

865 870 875 880

Leu Glu Glu Val Glu Glu Gln Glu Gln Gln Gly Val Glu Gln Gln Glu

885 890 895

Gln Glu Thr Val Glu Glu Pro Ile Ile Leu His Gly Ser Ser Ser Glu

900 905 910

Asp Glu Met Glu Val Asp Tyr Pro Val Val Ser Thr His Glu Gln Ile

915 920 925

Ala Ser Ser Pro Pro Gly Asp Asn Thr Pro Asp Asp Asp Pro Gln Pro

930 935 940

Gly Pro Ser Arg Glu Tyr Arg Tyr Val Leu Arg Thr Ser Pro Pro His

945 950 955 960

Arg Pro Gly Val Arg Met Arg Arg Val Pro Val Thr His Pro Lys Lys

965 970 975

Pro His Pro Arg Tyr Gln Gln Pro Pro Val Pro Tyr Arg Gln Ile Asp

980 985 990

Asp Cys Pro Ala Lys Ala Arg Pro Gln His Ile Phe Tyr Arg Arg Phe

995 1000 1005

Leu Gly Lys Asp Gly Arg Arg Asp Pro Lys Cys Gln Trp Lys Phe

1010 1015 1020

Ala Val Ile Phe Trp Gly Asn Asp Pro Tyr Gly Leu Lys Lys Leu

1025 1030 1035

Ser Gln Ala Phe Gln Phe Gly Gly Val Lys Ala Gly Pro Val Ser

1040 1045 1050

Cys Leu Pro His Pro Gly Pro Asp Gln Ser Pro Ile Thr Tyr Cys

1055 1060 1065

Val Tyr Val Tyr Cys Gln Asn Lys Asp Thr Ser Lys Lys Val Gln

1070 1075 1080

Met Ala Arg Leu Ala Trp Glu Ala Ser His Pro Leu Ala Gly Asn

1085 1090 1095

Leu Gln Ser Ser Ile Val Lys Phe Lys Lys Pro Leu Pro Leu Thr

1100 1105 1110

Gln Pro Gly Glu Asn Gln Gly Pro Gly Asp Ser Pro Gln Glu Met

1115 1120 1125

Th

<210> 884

<211> 160

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic polypeptide

<400> 884

Met Ala Pro Pro Gly Met Arg Leu Arg Ser Gly Arg Ser Thr Gly Ala

1 5 10 15

Pro Leu Thr Arg Gly Ser Cys Arg Lys Arg Asn Arg Ser Pro Glu Arg

20 25 30

Cys Asp Leu Gly Asp Asp Leu His Leu Gln Pro Arg Arg Lys His Val

35 40 45

Ala Asp Ser Val Asp Gly Arg Glu Cys Gly Pro His Thr Leu Pro Ile

50 55 60

Pro Gly Ser Pro Thr Val Phe Thr Ser Gly Leu Pro Ala Phe Val Ser

65 70 75 80

Ser Pro Thr Leu Pro Val Ala Pro Ile Pro Ser Pro Ala Pro Ala Thr

85 90 95

Pro Leu Pro Pro Pro Ala Leu Leu Pro Pro Val Thr Thr Ser Ser Ser

100 105 110

Pro Ile Pro Pro Ser His Pro Val Ser Pro Gly Thr Thr Asp Thr His

115 120 125

Ser Pro Ser Pro Ala Leu Pro Pro Thr Gln Ser Pro Glu Ser Ser Gln

130 135 140

Arg Pro Pro Leu Ser Ser Pro Thr Gly Arg Pro Asp Ser Ser Thr Pro

145 150 155 160

<210> 885

<211> 84

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 885

ggccagtgtt gagaggcgga gacttgggca attgctggac gctgccctgg gcattgcact 60

tgtctcggtc tgacagtgcc ggcc 84

<210> 886

<211> 80

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 886

ctggggggctc caaagtgctg ttcgtgcagg tagtgtgatt acccaaccta ctgctgagct 60

agcacttccc gagcccccgg 80

<210> 887

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<220>

<221> modified_base

<222> (4)..(8)

<223> a, c, t, g, unknown or other

<220>

<221> other_feature

<222> (4)..(8)

<223> This region may include 3-5 nucleotides

<220>

<221> other_feature

<222> (9)..(13)

<223> This region may include 3-5 nucleotides

<400> 887

cctnnnnngg gggrr 15

<210> 888

<211> 6

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 888

ttaggg 6

<210> 889

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 889

tcctggcggg gaagt 15

<210> 890

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 890

tcctggaggg gaagt 15

<210> 891

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 891

gggggggggg gggggggggg 20

<210> 892

<211> 24

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 892

ttagggttag ggttagggtt aggg 24

<210> 893

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 893

ctcctattgg gggtttccta t 21

<210> 894

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 894

tcctggaggg gttgt 15

<210> 895

<211> 24

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 895

cctggatggg aattcccatc cagg 24

<210> 896

<211> 24

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 896

ttcccatcca ggcctggatg ggaa 24

<210> 897

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 897

tgcttgcaag cttgcaagca 20

<210> 898

<211> 15

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 898

tcctggatgggaagt 15

<210> 899

<211> 12

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 899

cctggatggg aa 12

<210> 900

<211> 24

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 900

cttaccgctg cacctggatg ggaa 24

<210> 901

<211> 24

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 901

cctggatggg aacttaccgc tgca 24

<210> 902

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 902

tgctcctgga ggggttgt 18

<210> 903

<211> 26

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 903

ggtggtggtg gttgtggtgg tggtgg 26

<210> 904

<211> 19

<212> RNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 904

gaaggcccau auagagaaa 19

<210> 905

<211> 143

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 905

ttgcccactc cctctctgcg cgctcgctcg ctcggtgggg cctgcggacc aaaggtccgc 60

agacggcaga ggtctcctct gccggcccca ccgagcgagc gacgcgcgca gagagggagt 120

gggcaactcc atcactaggg taa 143

<210> 906

<211> 143

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 906

ttaccctagt gatggagttg cccactccct ctctgcgcgc gtcgctcgct cggtggggcc 60

ggcagaggag acctctgccg tctgcggacc tttggtccgc aggccccacc gagcgagcga 120

gcgcgcagag agggagtggg caa 143

<210> 907

<211> 141

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 907

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc t 141

<210> 908

<211> 141

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 908

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc 120

gagcgcgcag ctgcctgcag g 141

<210> 909

<211> 144

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 909

ttggccactc cctctatgcg cactcgctcg ctcggtgggg cctggcgacc aaaggtcgcc 60

agacggacgt gggtttccac gtccggcccc accgagcgag cgagtgcgca tagagggagt 120

ggccaactcc atcactagag gtat 144

<210> 910

<211> 144

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 910

atacctctag tgatggagtt ggccactccc tctatgcgca ctcgctcgct cggtggggcc 60

ggacgtggaa acccacgtcc gtctggcgac ctttggtcgc caggccccac cgagcgagcg 120

agtgcgcata gagggagtgg ccaa 144

<210> 911

<211> 127

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 911

ttggccactc cctctatgcg cgctcgctca ctcactcggc cctggagacc aaaggtctcc 60

agactgccgg cctctggccg gcagggccga gtgagtgagc gagcgcgcat agagggagtg 120

gccaact 127

<210> 912

<211> 127

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 912

agttggccac attagctatg cgcgctcgct cactcactcg gccctggaga ccaaaggtct 60

ccagactgcc ggcctctggc cggcagggcc gagtgagtga gcgagcgcgc atagagggag 120

tggccaa 127

<210> 913

<211> 166

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 913

tcccccctgt cgcgttcgct cgctcgctgg ctcgtttggg ggggcgacgg ccagaggggcc 60

gtcgtctggc agctctttga gctgccaccc ccccaaacga gccagcgagc gagcgaacgc 120

gacagggggg agagtgccac actctcaagc aagggggttt tgtaag 166

<210> 914

<211> 166

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 914

cttacaaaac ccccttgctt gagagtgtgg cactctcccc cctgtcgcgt tcgctcgctc 60

gctggctcgt ttggggggggt ggcagctcaa agagctgcca gacgacggcc ctctggccgt 120

cgccccccca aacgagccag cgagcgagcg aacgcgacag ggggga 166

<210> 915

<211> 144

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 915

ttgcccactc cctctaatgc gcgctcgctc gctcggtggg gcctgcggac caaaggtccg 60

cagacggcag aggtctcctc tgccggcccc accgagcgag cgagcgcgca tagagggagt 120

gggcaactcc atcactaggg gtat 144

<210> 916

<211> 144

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 916

atacccctag tgatggagtt gcccactccc tctatgcgcg ctcgctcgct cggtggggcc 60

ggcagaggag acctctgccg tctgcggacc tttggtccgc aggccccacc gagcgagcga 120

gcgcgcatta gagggagtgg gcaa 144

<210> 917

<211> 120

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 917

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

cgcacgcccg ggtttcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg 120

<210> 918

<211> 122

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 918

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca 120

gg 122

<210> 919

<211> 129

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 919

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgcccgg gcgcctcagt gagcgagcga gcgcgcagct 120

gcctgcagg 129

<210> 920

<211> 101

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 920

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ctttgcctca gtgagcgagc gagcgcgcag ctgcctgcag g 101

<210> 921

<211> 139

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 921

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgaca aagtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga 120

gcgcgcagct gcctgcagg 139

<210> 922

<211> 137

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 922

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgaaa atcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc 120

gcgcagctgc ctgcagg 137

<210> 923

<211> 135

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 923

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgaaa cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc 120

gcagctgcct gcagg 135

<210> 924

<211> 133

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 924

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcaaag cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc 120

agctgcctgc agg 133

<210> 925

<211> 139

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 925

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgcccgg gtttcccggg cggcctcagt gagcgagcga 120

gcgcgcagct gcctgcagg 139

<210> 926

<211> 137

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 926

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgcccgg tttccgggcg gcctcagtga gcgagcgagc 120

gcgcagctgc ctgcagg 137

<210> 927

<211> 135

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 927

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgcccgt ttcgggcggc ctcagtgagc gagcgagcgc 120

gcagctgcct gcagg 135

<210> 928

<211> 133

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 928

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgccctt tgggcggcct cagtgagcga gcgagcgcgc 120

agctgcctgc agg 133

<210> 929

<211> 131

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 929

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgccttt ggcggcctca gtgagcgagc gagcgcgcag 120

ctgcctgcag g 131

<210> 930

<211> 129

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 930

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgctttg cggcctcagt gagcgagcga gcgcgcagct 120

gcctgcagg 129

<210> 931

<211> 127

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 931

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgtttcg gcctcagtga gcgagcgagc gcgcagctgc 120

ctgcagg 127

<210> 932

<211> 122

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 932

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacggcctc agtgagcgag cgagcgcgca gctgcctgca 120

gg 122

<210> 933

<211> 130

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 933

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc 120

tgcctgcagg 130

<210> 934

<211> 120

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 934

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggaaacc cgggcgtgcg 60

cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct 120

<210> 935

<211> 122

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 935

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgt cgggcgacct ttggtcgccc 60

ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc 120

ct 122

<210> 936

<211> 122

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 936

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc 120

ct 122

<210> 937

<211> 129

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 937

cctgcaggca gctgcgcgct cgctcgctca ctgaggcgcc cgggcgtcgg gcgacctttg 60

gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta 120

ggggttcct 129

<210> 938

<211> 101

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 938

cctgcaggca gctgcgcgct cgctcgctca ctgaggcaaa gcctcagtga gcgagcgagc 60

gcgcagagag ggagtggcca actccatcac taggggttcc t 101

<210> 939

<211> 139

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 939

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacttt gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac 120

tccatcacta ggggttcct 139

<210> 940

<211> 137

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 940

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgatttt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc 120

catcactagg ggttcct 137

<210> 941

<211> 135

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 941

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgtttcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca 120

tcactagggg ttcct 135

<210> 942

<211> 133

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 942

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggctttgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc 120

actaggggtt cct 133

<210> 943

<211> 139

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 943

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggaaacc cgggcgtcgg 60

gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac 120

tccatcacta ggggttcct 139

<210> 944

<211> 137

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 944

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccggaaaccg ggcgtcgggc 60

gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc 120

catcactagg ggttcct 137

<210> 945

<211> 135

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 945

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgaaacggg cgtcgggcga 60

cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca 120

tcactagggg ttcct 135

<210> 946

<211> 133

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 946

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccaaagggcg tcgggcgacc 60

tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc 120

actaggggtt cct 133

<210> 947

<211> 131

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 947

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc caaaggcgtc gggcgacctt 60

tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac 120

taggggttcct 131

<210> 948

<211> 129

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 948

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc aaagcgtcgg gcgacctttg 60

gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta 120

ggggttcct 129

<210> 949

<211> 127

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 949

cctgcaggca gctgcgcgct cgctcgctca ctgaggccga aacgtcgggc gacctttggt 60

cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg 120

ggttcct 127

<210> 950

<211> 19

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic peptide

<400> 950

Met Asp Trp Thr Trp Arg Ile Leu Phe Leu Val Ala Ala Ala Thr Gly

1 5 10 15

Ala His Ser

<210> 951

<211> 19

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic peptide

<400> 951

Met Leu Pro Ser Gln Leu Ile Gly Phe Leu Leu Leu Trp Val Pro Ala

1 5 10 15

Ser Arg Gly

<210> 952

<211> 141

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 952

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc 120

gagcgcgcag ctgcctgcag g 141

<210> 953

<211> 141

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 953

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc t 141

<210> 954

<211> 130

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 954

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc 120

tgcctgcagg 130

<210> 955

<211> 130

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 955

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 60

ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 120

aggggttcct 130

<210> 956

<211> 143

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 956

ttgcccactc cctctctgcg cgctcgctcg ctcggtgggg cctgcggacc aaaggtccgc 60

agacggcaga ggtctcctct gccggcccca ccgagcgagc gacgcgcgca gagagggagt 120

gggcaactcc atcactaggg taa 143

<210> 957

<211> 144

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 957

ttggccactc cctctatgcg cactcgctcg ctcggtgggg cctggcgacc aaaggtcgcc 60

agacggacgt gggtttccac gtccggcccc accgagcgag cgagtgcgca tagagggagt 120

ggccaactcc atcactagag gtat 144

<210> 958

<211> 127

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 958

ttggccactc cctctatgcg cgctcgctca ctcactcggc cctggagacc aaaggtctcc 60

agactgccgg cctctggccg gcagggccga gtgagtgagc gagcgcgcat agagggagtg 120

gccaact 127

<210> 959

<211> 166

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 959

tcccccctgt cgcgttcgct cgctcgctgg ctcgtttggg ggggcgacgg ccagaggggcc 60

gtcgtctggc agctctttga gctgccaccc ccccaaacga gccagcgagc gagcgaacgc 120

gacagggggg agagtgccac actctcaagc aagggggttt tgtaag 166

<210> 960

<211> 144

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 960

ttgcccactc cctctaatgc gcgctcgctc gctcggtggg gcctgcggac caaaggtccg 60

cagacggcag aggtctcctc tgccggcccc accgagcgag cgagcgcgca tagagggagt 120

gggcaactcc atcactaggg gtat 144

<210> 961

<211> 143

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 961

ttaccctagt gatggagttg cccactccct ctctgcgcgc gtcgctcgct cggtggggcc 60

ggcagaggag acctctgccg tctgcggacc tttggtccgc aggccccacc gagcgagcga 120

gcgcgcagag agggagtggg caa 143

<210> 962

<211> 144

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 962

atacctctag tgatggagtt ggccactccc tctatgcgca ctcgctcgct cggtggggcc 60

ggacgtggaa acccacgtcc gtctggcgac ctttggtcgc caggccccac cgagcgagcg 120

agtgcgcata gagggagtgg ccaa 144

<210> 963

<211> 127

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 963

agttggccac attagctatg cgcgctcgct cactcactcg gccctggaga ccaaaggtct 60

ccagactgcc ggcctctggc cggcagggcc gagtgagtga gcgagcgcgc atagagggag 120

tggccaa 127

<210> 964

<211> 166

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 964

cttacaaaac ccccttgctt gagagtgtgg cactctcccc cctgtcgcgt tcgctcgctc 60

gctggctcgt ttggggggggt ggcagctcaa agagctgcca gacgacggcc ctctggccgt 120

cgccccccca aacgagccag cgagcgagcg aacgcgacag ggggga 166

<210> 965

<211> 144

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 965

atacccctag tgatggagtt gcccactccc tctatgcgcg ctcgctcgct cggtggggcc 60

ggcagaggag acctctgccg tctgcggacc tttggtccgc aggccccacc gagcgagcga 120

gcgcgcatta gagggagtgg gcaa 144

<210> 966

<211> 120

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 966

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

cgcacgcccg ggtttcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg 120

<210> 967

<211> 122

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 967

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca 120

gg 122

<210> 968

<211> 129

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 968

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgcccgg gcgcctcagt gagcgagcga gcgcgcagct 120

gcctgcagg 129

<210> 969

<211> 101

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 969

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ctttgcctca gtgagcgagc gagcgcgcag ctgcctgcag g 101

<210> 970

<211> 139

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 970

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgaca aagtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga 120

gcgcgcagct gcctgcagg 139

<210> 971

<211> 137

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 971

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgaaa atcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc 120

gcgcagctgc ctgcagg 137

<210> 972

<211> 135

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 972

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgaaa cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc 120

gcagctgcct gcagg 135

<210> 973

<211> 133

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 973

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcaaag cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc 120

agctgcctgc agg 133

<210> 974

<211> 139

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 974

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgcccgg gtttcccggg cggcctcagt gagcgagcga 120

gcgcgcagct gcctgcagg 139

<210> 975

<211> 137

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 975

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgcccgg tttccgggcg gcctcagtga gcgagcgagc 120

gcgcagctgc ctgcagg 137

<210> 976

<211> 135

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 976

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgcccgt ttcgggcggc ctcagtgagc gagcgagcgc 120

gcagctgcct gcagg 135

<210> 977

<211> 133

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 977

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgccctt tgggcggcct cagtgagcga gcgagcgcgc 120

agctgcctgc agg 133

<210> 978

<211> 131

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 978

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgccttt ggcggcctca gtgagcgagc gagcgcgcag 120

ctgcctgcag g 131

<210> 979

<211> 129

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 979

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgctttg cggcctcagt gagcgagcga gcgcgcagct 120

gcctgcagg 129

<210> 980

<211> 127

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 980

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgtttcg gcctcagtga gcgagcgagc gcgcagctgc 120

ctgcagg 127

<210> 981

<211> 122

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 981

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacggcctc agtgagcgag cgagcgcgca gctgcctgca 120

gg 122

<210> 982

<211> 130

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 982

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc 120

tgcctgcagg 130

<210> 983

<211> 120

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 983

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggaaacc cgggcgtgcg 60

cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct 120

<210> 984

<211> 122

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 984

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgt cgggcgacct ttggtcgccc 60

ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc 120

ct 122

<210> 985

<211> 122

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 985

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc 120

ct 122

<210> 986

<211> 129

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 986

cctgcaggca gctgcgcgct cgctcgctca ctgaggcgcc cgggcgtcgg gcgacctttg 60

gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta 120

ggggttcct 129

<210> 987

<211> 101

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 987

cctgcaggca gctgcgcgct cgctcgctca ctgaggcaaa gcctcagtga gcgagcgagc 60

gcgcagagag ggagtggcca actccatcac taggggttcc t 101

<210> 988

<211> 139

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 988

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgacttt gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac 120

tccatcacta ggggttcct 139

<210> 989

<211> 137

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 989

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgatttt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc 120

catcactagg ggttcct 137

<210> 990

<211> 135

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 990

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggcgtttcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca 120

tcactagggg ttcct 135

<210> 991

<211> 133

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 991

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60

gggctttgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc 120

actaggggtt cct 133

<210> 992

<211> 139

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 992

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggaaacc cgggcgtcgg 60

gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac 120

tccatcacta ggggttcct 139

<210> 993

<211> 137

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 993

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccggaaaccg ggcgtcgggc 60

gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc 120

catcactagg ggttcct 137

<210> 994

<211> 135

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 994

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgaaacggg cgtcgggcga 60

cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca 120

tcactagggg ttcct 135

<210> 995

<211> 133

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 995

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccaaagggcg tcgggcgacc 60

tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc 120

actaggggtt cct 133

<210> 996

<211> 131

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 996

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc caaaggcgtc gggcgacctt 60

tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac 120

taggggttcc t 131

<210> 997

<211> 129

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 997

cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc aaagcgtcgg gcgacctttg 60

gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta 120

ggggttcct 129

<210> 998

<211> 127

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 998

cctgcaggca gctgcgcgct cgctcgctca ctgaggccga aacgtcgggc gacctttggt 60

cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg 120

ggttcct 127

<210> 999

<211> 122

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 999

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgggcgacc aaaggtcgcc cgacggcctc agtgagcgag cgagcgcgca gctgcctgca 120

gg 122

<210> 1000

<211> 12

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic oligonucleotide

<400> 1000

cgatcgttcg at 12

<210> 1001

<211> 12

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic oligonucleotide

<400> 1001

atcgaaccat cg 12

<210> 1002

<211> 12

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1002

atcgaacgat cg 12

<210> 1003

<211> 165

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1003

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60

ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc 120

gagcgcgcag agagggagtg gccaactcca tcactagggg ttcct 165

<210> 1004

<211> 140

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1004

cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccgcc 60

cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg 120

cgcagagaga tcactagggg 140

<210> 1005

<211> 91

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1005

gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg 60

tcgcccggcc tcagtgagcg agcgagcgcg c 91

<210> 1006

<211> 91

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1006

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc 60

ccgggcggcc tcagtgagcg agcgagcgcg c 91

<210> 1007

<211> 1662

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1007

gccgccacca tggaagacgc caaaaacata aagaaaggcc cggcgccatt ctatccgctg 60

gaagatggaa ccgctggaga gcaactgcat aaggctatga agagatacgc cctggttcct 120

ggaacaattg cttttacaga tgcacatatc gaggtggaca tcacttacgc tgagtacttc 180

gaaatgtccg ttcggttggc agaagctatg aaacgatatg ggctgaatac aaatcacaga 240

atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg cgcgttattt 300

atcggagttg cagttgcgcc cgcgaacgac atttataatg aacgtgaatt gctcaacagt 360

atgggcattt cgcagcctac cgtggtgttc gtttccaaaa aggggttgca aaaaattttg 420

aacgtgcaaa aaaagctccc aatcatccaa aaaattatta tcatggattc taaaacggat 480

taccagggat ttcagtcgat gtacacgttc gtcacatctc atctacctcc cggttttaat 540

gaatacgatt ttgtgccaga gtccttcgat agggacaaga caattgcact gatcatgaac 600

tcctctggat ctactggtct gcctaaaggt gtcgctctgc ctcatagaac tgcctgcgtg 660

agattctcgc atgccagaga tcctattttt ggcaatcaaa tcattccgga tactgcgatt 720

ttaagtgttg ttccattcca tcacggtttt ggaatgttta ctacactcgg atatttgata 780

tgtggatttc gagtcgtctt aatgtataga tttgaagaag agctgtttct gaggagcctt 840

caggattaca agattcaaag tgcgctgctg gtgccaaccc tattctcctt cttcgccaaa 900

agcactctga ttgacaaata cgatttatct aatttacacg aaattgcttc tggtggcgct 960

cccctctcta aggaagtcgg ggaagcggtt gccaagaggt tccatctgcc aggtatcagg 1020

caaggatatg ggctcactga gactacatca gctattctga ttacacccga gggggatgat 1080

aaaccgggcg cggtcggtaa agttgttcca ttttttgaag cgaaggttgt ggatctggat 1140

accgggaaaa cgctgggcgt taatcaaaga ggcgaactgt gtgtgagagg tcctatgatt 1200

atgtccggtt atgtaaacaa tccggaagcg accaacgcct tgattgacaa ggatggatgg 1260

ctacattctg gagacatagc ttactgggac gaagacgaac acttcttcat cgttgaccgc 1320

ctgaagtctc tgattaagta caaaggctat caggtggctc ccgctgaatt ggaatccatc 1380

ttgctccaac accccaacat cttcgacgca ggtgtcgcag gtcttcccga cgatgacgcc 1440

ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa agacgatgac ggaaaaagag 1500

atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg aggagttgtg 1560

tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa aatcagagag 1620

atcctcataa aggccaagaa gggcggaaag atcgccgtgt aa 1662

<210> 1008

<211> 453

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic polypeptide

<220>

<221>MOD_RES

<222> (1)..(1)

<223> Any amino acid

<400> 1008

Xaa Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Phe Ser Ser Tyr

20 25 30

Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Val Ile Trp Phe Asp Gly Thr Lys Lys Tyr Tyr Thr Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Thr Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Arg Gly Ile Gly Ala Arg Arg Gly Pro Tyr Tyr Met Asp

100 105 110

Val Trp Gly Lys Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys

115 120 125

Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly

130 135 140

Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro

145 150 155 160

Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr

165 170 175

Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val

180 185 190

Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn

195 200 205

Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro

210 215 220

Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu

225 230 235 240

Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp

245 250 255

Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp

260 265 270

Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly

275 280 285

Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn

290 295 300

Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp

305 310 315 320

Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro

325 330 335

Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu

340 345 350

Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn

355 360 365

Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile

370 375 380

Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr

385 390 395 400

Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys

405 410 415

Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys

420 425 430

Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu

435 440 445

Ser Leu Ser Pro Gly

450

<210> 1009

<211> 214

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of artificial sequence: synthetic polypeptide

<400> 1009

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<210> 1010

<211> 1310

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1010

ggagccgaga gtaattcata caaaaggagg gatcgccttc gcaaggggag agcccaggga 60

ccgtccctaa attctcacag acccaaatcc ctgtagccgc cccacgacag cgcgaggagc 120

atgcgctcag ggctgagcgc ggggagagca gagcacacaa gctcatagac cctggtcgtg 180

ggggggagga ccggggagct ggcgcggggc aaactgggaa agcggtgtcg tgtgctggct 240

ccgccctctt cccgagggtg ggggagaacg gtatataagt gcggcagtcg ccttggacgt 300

tctttttcgc aacgggtttg ccgtcagaac gcaggtgagg ggcgggtgtg gcttccgcgg 360

gccgccgagc tggaggtcct gctccgagcg ggccggggccc cgctgtcgtc ggcggggatt 420

agctgcgagc attcccgctt cgagttgcgg gcggcgcggg aggcagagtg cgaggcctag 480

cggcaacccc gtagcctcgc ctcgtgtccg gcttgaggcc tagcgtggtg tccgcgccgc 540

cgccgcgtgc tactccggcc gcactctggt cttttttttt tttgttgttg ttgccctgct 600

gccttcgatt gccgttcagc aataggggct aacaaaggga gggtgcgggg cttgctcgcc 660

cggagcccgg agaggtcatg gttggggagg aatggaggga caggagtggc ggctggggcc 720

cgcccgcctt cggagcacat gtccgacgcc acctggatgg ggcgaggcct ggggtttttc 780

ccgaagcaac caggctgggg ttagcgtgcc gaggccatgt ggccccagca cccggcacga 840

tctggcttgg cggcgccgcg ttgccctgcc tccctaacta gggtgaggcc atcccgtccg 900

gcaccagttg cgtgcgtgga aagatggccg ctcccggggcc ctgttgcaag gagctcaaaa 960

tggagaggacgc ggcagcccgg tggagcgggc gggtgagtca cccacacaaa ggaagagggc 1020

ctggtccctc accggctgct gcttcctgtg accccgtggt cctatcggcc gcaatagtca 1080

cctcgggctt ttgagcacgg ctagtcgcgg cggggggagg ggatgtaatg gcgttggagt 1140

ttgttcacat ttggtgggtg gagactagtc aggccagcct ggcgctggaa gtcatttttg 1200

gaatttgtcc ccttgagttt tgagcggagc taattctcgg gcttcttagc ggttcaaagg 1260

tatcttttaa accctttttt aggtgttgtg aaaaccaccg ctaattcaaa 1310

<210> 1011

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1011

gcgcgctcgc tcgctc 16

<210> 1012

<211> 6

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1012

ggttga 6

<210> 1013

<211> 4

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1013

agtt 4

<210> 1014

<211> 6

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1014

ggttgg 6

<210> 1015

<211> 6

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1015

agttgg 6

<210> 1016

<211> 6

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1016

agttga 6

<210> 1017

<211> 6

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1017

rrttrr 6

<210> 1018

<211> 581

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1018

gagcatctta ccgccattta ttcccatatt tgttctgttt ttcttgattt gggtatacat 60

ttaaatgtta ataaaacaaa atggtggggc aatcatttac atttttaggg atatgtaatt 120

actagttcag gtgtattgcc acaagacaaa catgttaaga aactttcccg ttatttacgc 180

tctgttcctg ttaatcaacc tctggattac aaaatttgtg aaagattgac tgatattctt 240

aactatgttg ctccttttac gctgtgtgga tatgctgctt tatagcctct gtatctagct 300

attgcttccc gtacggcttt cgttttctcc tccttgtata aatcctggtt gctgtctctt 360

ttagaggagt tgtggcccgt tgtccgtcaa cgtggcgtgg tgtgctctgt gtttgctgac 420

gcaaccccca ctggctgggg cattgccacc acctgtcaac tcctttctgg gactttcgct 480

ttccccctcc cgatcgccac ggcagaactc atcgccgcct gccttgcccg ctgctggaca 540

ggggctaggt tgctgggcac tgataattcc gtggtgttgt c 581

<210> 1019

<211> 225

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1019

tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct 60

ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct 120

gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg 180

ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggc 225

<210> 1020

<211> 8

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1020

actgaggc 8

<210> 1021

<211> 8

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1021

gcctcagt 8

<210> 1022

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1022

gagcgagcga gcgcgc 16

<210> 1023

<211> 1923

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1023

tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta 60

ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc 120

aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg 180

gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 240

gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 300

agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 360

ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga 420

cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg 480

gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg 540

cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta 600

attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgcgccag gcggggcggg 660

gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg 720

cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg 780

aagcgcgcgg cgggcgggag tcgctgcgac gctgccttcg ccccgtgccc cgctccgccg 840

ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc 900

gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt 960

ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc gggggggagc 1020

ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg cccgcgctgc 1080

ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg 1140

aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag 1200

gctgcgtgcg gggtgtgtgc gtggggggggt gagcaggggg tgtgggcgcg gcggtcgggc 1260

tgtaaccccc ccctgcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg 1320

gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg 1380

ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga ggggcgcggc 1440

ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg 1500

taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct 1560

gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg cgccggcagg 1620

aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc 1680

cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg 1740

ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tttagccttc 1800

ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca tcatttgtcg 1860

acagaattcc tcgaagatcc gaaggggttc aagcttggca ttccggtact gttggtaaag 1920

cca 1923

<210> 1024

<211> 1272

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1024

aggctcagag gcacacagga gtttctgggc tcaccctgcc cccttccaac ccctcagttc 60

ccatcctcca gcagctgttt gtgtgctgcc tctgaagtcc acactgaaca aacttcagcc 120

tactcatgtc cctaaaatgg gcaaacattg caagcagcaa acagcaaaca cacagccctc 180

cctgcctgct gaccttggag ctggggcaga ggtcagagac ctctctgggc ccatgccacc 240

tccaacatcc actcgacccc ttggaatttc ggtggagagg agcagaggtt gtcctggcgt 300

ggtttaggta gtgtgagagg gtccgggttc aaaaccactt gctgggtggg gagtcgtcag 360

taagtggcta tgccccgacc ccgaagcctg tttccccatc tgtacaatgg aaatgataaa 420

gacgcccatc tgatagggtt tttgtggcaa ataaacattt ggtttttttg ttttgttttg 480

ttttgttttt tgagatggag gtttgctctg tcgcccaggc tggagtgcag tgacacaatc 540

tcatctcacc acaaccttcc cctgcctcag cctcccaagt agctgggatt acaagcatgt 600

gccaccacac ctggctaatt ttctattttt agtagagacg ggtttctcca tgttggtcag 660

cctcagcctc ccaagtaact gggattacag gcctgtgcca ccacacccgg ctaatttttt 720

ctatttttga cagggacggg gtttcaccat gttggtcagg ctggtctaga ggtaccggat 780

cttgctacca gtggaacagc cactaaggat tctgcagtga gagcagaggg ccagctaagt 840

ggtactctcc cagagactgt ctgactcacg ccaccccctc caccttggac acaggacgct 900

gtggtttctg agccaggtac aatgactcct ttcggtaagt gcagtggaag ctgtacactg 960

cccaggcaaa gcgtccgggc agcgtaggcg ggcgactcag atcccagcca gtggacttag 1020

cccctgtttg ctcctccgat aactggggtg accttggtta atattcacca gcagcctccc 1080

ccgttgcccc tctggatcca ctgcttaaat acggacgagg acagggccct gtctcctcag 1140

cttcaggcac caccactgac ctgggacagt gaatccggac tctaaggtaa atataaaatt 1200

tttaagtgta taatgtgtta aactactgat tctaattgtt tctctctttt agattccaac 1260

ctttggaact ga 1272

<210> 1025

<211> 1177

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1025

ggctcagagg ctcagaggca cacaggagtt tctgggctca ccctgccccc ttccaacccc 60

tcagttccca tcctccagca gctgtttgtg tgctgcctct gaagtccaca ctgaacaaac 120

ttcagcctac tcatgtccct aaaatgggca aacattgcaa gcagcaaaca gcaaacacac 180

agccctccct gcctgctgac cttggagctg gggcagaggt cagagacctc tctgggccca 240

tgccacctcc aacatccact cgaccccttg gaatttcggt ggagaggagc agaggttgtc 300

ctggcgtggt ttaggtagtg tgagagggtc cgggttcaaa accacttgct gggtggggag 360

tcgtcagtaa gtggctatgc cccgaccccg aagcctgttt ccccatctgt acaatggaaa 420

tgataaagac gcccatctga tagggttttt gtggcaaata aacatttggt ttttttgttt 480

tgttttgttt tgttttttga gatggaggtt tgctctgtcg cccaggctgg agtgcagtga 540

cacaatctca tctcaccaca accttcccct gcctcagcct cccaagtagc tgggattaca 600

agcatgtgcc accacacctg gctaattttc tatttttagt agagacgggt ttctccatgt 660

tggtcagcct cagcctccca agtaactggg attacaggcc tgtgccacca cacccggcta 720

attttttcta tttttgacag ggacggggtt tcaccatgtt ggtcaggctg gtctagaggt 780

accggatctt gctaccagtg gaacagccac taaggattct gcagtgagag cagaggcca 840

gctaagtggt actctcccag agactgtctg actcacgcca ccccctccac cttggacaca 900

ggacgctgtg gtttctgagc caggtacaat gactcctttc ggtaagtgca gtggaagctg 960

tacactgccc aggcaaagcg tccgggcagc gtaggcgggc gactcagatc ccagccagtg 1020

gacttagccc ctgtttgctc ctccgataac tggggtgacc ttggttaata ttcaccagca 1080

gcctcccccg ttgcccctct ggatccactg cttaaatacg gacgaggaca gggccctgtc 1140

tcctcagctt caggcaccac cactgacctg ggacagt 1177

<210> 1026

<211> 547

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1026

ccctaaaatg ggcaaacatt gcaagcagca aacagcaaac acacagccct ccctgcctgc 60

tgaccttgga gctggggcag aggtcagaga cctctctggg cccatgccac ctccaacatc 120

cactcgaccc cttggaattt ttcggtggag aggagcagag gttgtcctgg cgtggtttag 180

gtagtgtgag aggggaatga ctcctttcgg taagtgcagt ggaagctgta cactgcccag 240

gcaaagcgtc cgggcagcgt aggcgggcga ctcagatccc agccagtgga cttagcccct 300

gtttgctcct ccgataactg gggtgacctt ggttaatatt caccagcagc ctcccccgtt 360

gcccctctgg atccactgct taaatacgga cgaggacagg gccctgtctc ctcagcttca 420

ggcaccacca ctgacctggg acagtgaatc cggactctaa ggtaaatata aaatttttaa 480

gtgtataatg tgttaaacta ctgattctaa ttgtttctct cttttagatt ccaacctttg 540

gaactga 547

<210> 1027

<211> 556

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1027

ccctaaaatg ggcaaacatt gcaagcagca aacagcaaac acacagccct ccctgcctgc 60

tgaccttgga gctggggcag aggtcagaga cctctctggg cccatgccac ctccaacatc 120

cactcgaccc cttggaattt cggtggagag gagcagaggt tgtcctggcg tggtttaggt 180

agtgtgagag gggaatgact cctttcggta agtgcagtgg aagctgtaca ctgcccaggc 240

aaagcgtccg ggcagcgtag gcgggcgact cagatcccag ccagtggact tagcccctgt 300

ttgctcctcc gataactggg gtgaccttgg ttaatattca ccagcagcct cccccgttgc 360

ccctctggat ccactgctta aatacggacg aggacactcg agggccctgt ctcctcagct 420

tcaggcacca ccactgacct gggacagtga atccggacat cgattctaag gtaaatataa 480

aatttttaag tgtataattt gttaaactac tgattctaat tgtttctctc ttttagattc 540

caacctttgg aactga 556

<210> 1028

<211> 1179

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1028

ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 60

ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 120

gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 180

gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc 240

gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt 300

acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 360

gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 420

cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 480

ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 540

caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 600

gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 660

gcgcggccac cgagaatcgg acggggggtag tctcaagctg gccggcctgc tctggtgcct 720

ggtctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 780

gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 840

acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 900

tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 960

tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 1020

gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 1080

ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 1140

gtggttcaaa gtttttttct tccatttcag gtgtcgtga 1179

<210> 1029

<211> 141

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1029

aataaacgat aacgccgttg gtggcgtgag gcatgtaaaa ggttacatca ttatcttgtt 60

cgccatccgg ttggtataaa tagacgttca tgttggtttt tgtttcagtt gcaagttggc 120

tgcggcgcgc gcagcacctt t 141

<210> 1030

<211> 317

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1030

ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 60

agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 120

tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 180

ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 240

aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag 300

gcctaggctt ttgcaaa 317

<210> 1031

<211> 241

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1031

gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag 60

ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga 120

aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat 180

atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga 240

c 241

<210> 1032

<211> 215

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1032

gaacgctgac gtcatcaacc cgctccaagg aatcgcgggc ccagtgtcac taggcgggaa 60

cacccagcgc gcgtgcgccc tggcaggaag atggctgtga gggacagggg agtggcgccc 120

tgcaatattt gcatgtcgct atgtgttctg ggaaatcacc ataaacgtga aatgtctttg 180

gatttgggaa tcgtataaga actgtatgag accac 215

<210> 1033

<211> 546

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1033

ccctaaaatg ggcaaacatt gcaagcagca aacagcaaac acacagccct ccctgcctgc 60

tgaccttgga gctggggcag aggtcagaga cctctctggg cccatgccac ctccaacatc 120

cactcgaccc cttggaattt ttcggtggag aggagcagag gttgtcctgg cgtggtttag 180

gtagtgtgag aggggaatga ctcctttcgg taagtgcagt ggaagctgta cactgcccag 240

gcaaagcgtc cgggcagcgt aggcgggcga ctcagatccc agccagtgga cttagcccct 300

gtttgctcct ccgataactg gggtgacctt ggttaatatt caccagcagc ctcccccgtt 360

gcccctctgg atccactgct taaatacgga cgaggacagg gccctgtctc ctcagcttca 420

ggcaccacca ctgacctggg acagtgaatc cggactctaa ggtaaatata aaatttttaa 480

gtgtataatg tgttaaacta ctgattctaa ttgtttctct cttttagatt ccaacctttg 540

gaactg 546

<210> 1034

<211> 576

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1034

tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa 60

cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata 120

atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag 180

tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc 240

cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta 300

tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg 360

cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt 420

ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca 480

aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag 540

gtctatataa gcagagctgg tttagtgaac cgtcag 576

<210> 1035

<211> 150

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1035

ataaacgata acgccgttgg tggcgtgagg catgtaaaag gttacatcat tatcttgttc 60

gccatccggt tggtataaat agacgttcat gttggttttt gtttcagttg caagttggct 120

gcggcgcgcg cagcaccttt gcggccatct 150

<210> 1036

<211> 1313

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1036

ggagccgaga gtaattcata caaaaggagg gatcgccttc gcaaggggag agcccaggga 60

ccgtccctaa attctcacag acccaaatcc ctgtagccgc cccacgacag cgcgaggagc 120

atgcgcccag ggctgagcgc gggtagatca gagcacacaa gctcacagtc cccggcggtg 180

gggggagggg cgcgctgagc gggggccagg gagctggcgc ggggcaaact gggaaagtgg 240

tgtcgtgtgc tggctccgcc ctcttcccga gggtggggga gaacggtata taagtgcggt 300

agtcgccttg gacgttcttt ttcgcaacgg gtttgccgtc agaacgcagg tgagtggcgg 360

gtgtggcttc cgcgggcccc ggagctggag ccctgctctg agcgggccgg gctgatatgc 420

gagtgtcgtc cgcagggttt agctgtgagc attcccactt cgagtggcgg gcggtgcggg 480

ggtgagagtg cgaggcctag cggcaacccc gtagcctcgc ctcgtgtccg gcttgaggcc 540

tagcgtggtg tccgccgccg cgtgccactc cggccgcact atgcgttttt tgtccttgct 600

gccctcgatt gccttccagc agcatgggct aacaaaggga gggtgtgggg ctcactctta 660

aggagcccat gaagcttacg ttggatagga atggaagggc aggaggggcg actggggccc 720

gcccgccttc ggagcacatg tccgacgcca cctggatggg gcgaggcctg tggctttccg 780

aagcaatcgg gcgtgagttt agcctacctg ggccatgtgg ccctagcact gggcacggtc 840

tggcctggcg gtgccgcgtt cccttgcctc ccaacaaggg tgaggccgtc ccgcccggca 900

ccagttgctt gcgcggaaag atggccgctc ccggggccct gttgcaagga gctcaaaatg 960

gaggacgcgg cagcccggtg gagcgggcgg gtgagtcacc cacacaaagg aagagggcct 1020

tgcccctcgc cggccgctgc ttcctgtgac cccgtggtct atcggccgca tagtcacctc 1080

gggcttctct tgagcaccgc tcgtcgcggc ggggggaggg gatctaatgg cgttggagtt 1140

tgttcacatt tggtgggtgg agactagtca ggccagcctg gcgctggaag tcattcttgg 1200

aatttgcccc tttgagtttg gagcgaggct aattctcaag cctcttagcg gttcaaaggt 1260

attttctaaa cccgtttcca ggtgttgtga aagccaccgc taattcaaag caa 1313

<210> 1037

<211> 213

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1037

taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta 60

tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag 120

ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt 180

tttaaagcaa gtaaaacctc tacaaatgtg gta 213

<210> 1038

<211> 7

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic peptide

<400> 1038

Pro Lys Lys Lys Arg Lys Val

1 5

<210> 1039

<211> 19

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic peptide

<400> 1039

Met Asp Trp Thr Trp Arg Ile Leu Phe Leu Val Ala Ala Ala Thr Gly

1 5 10 15

Ala His Ser

<210> 1040

<211> 19

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic peptide

<400> 1040

Met Leu Pro Ser Gln Leu Ile Gly Phe Leu Leu Leu Trp Val Pro Ala

1 5 10 15

Ser Arg Gly

<210> 1041

<211> 7

<212> PROTEIN

<213> Simian Virus 40

<400> 1041

Pro Lys Lys Lys Arg Lys Val

1 5

<210> 1042

<211> 21

<212> DNA

<213> Simian Virus 40

<400> 1042

cccaagaaga agaggaaggt g 21

<210> 1043

<211> 16

<212> PROTEIN

<213> Unknown

<220>

<223> Description of unknown:

Double NLS sequence of nucleoplasmin

<400> 1043

Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys Lys

1 5 10 15

<210> 1044

<211> 9

<212> PROTEIN

<213> Unknown

<220>

<223> Description of unknown:

C-myc NLS sequence

<400> 1044

Pro Ala Ala Lys Arg Val Lys Leu Asp

1 5

<210> 1045

<211> 11

<212> PROTEIN

<213> Unknown

<220>

<223> Description of unknown:

C-myc NLS sequence

<400> 1045

Arg Gln Arg Arg Asn Glu Leu Lys Arg Ser Pro

1 5 10

<210> 1046

<211> 38

<212> PROTEIN

<213> Homo sapiens

<400> 1046

Asn Gln Ser Ser Asn Phe Gly Pro Met Lys Gly Gly Asn Phe Gly Gly

1 5 10 15

Arg Ser Ser Gly Pro Tyr Gly Gly Gly Gly Gln Tyr Phe Ala Lys Pro

20 25 30

Arg Asn Gln Gly Gly Tyr

35

<210> 1047

<211> 42

<212> PROTEIN

<213> Unknown

<220>

<223> Description of unknown:

IBB domain of importin alpha sequence

<400> 1047

Arg Met Arg Ile Glx Phe Lys Asn Lys Gly Lys Asp Thr Ala Glu Leu

1 5 10 15

Arg Arg Arg Arg Val Glu Val Ser Val Glu Leu Arg Lys Ala Lys Lys

20 25 30

Asp Glu Gln Ile Leu Lys Arg Arg Asn Val

35 40

<210> 1048

<211> 8

<212> PROTEIN

<213> Unknown

<220>

<223> Description of unknown:

Myoma T protein sequence

<400> 1048

Val Ser Arg Lys Arg Pro Arg Pro

1 5

<210> 1049

<211> 8

<212> PROTEIN

<213> Unknown

<220>

<223> Description of unknown:

Myoma T protein sequence

<400> 1049

Pro Pro Lys Lys Ala Arg Glu Asp

1 5

<210> 1050

<211> 8

<212> PROTEIN

<213> Homo sapiens

<400> 1050

Pro Gln Pro Lys Lys Lys Pro Leu

1 5

<210> 1051

<211> 12

<212> PROTEIN

<213> Mus musculus

<400> 1051

Ser Ala Leu Ile Lys Lys Lys Lys Lys Met Ala Pro

1 5 10

<210> 1052

<211> 70

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1052

gcgcgctcgc tcgctcactg aggccgcccg ggaaacccgg gcgtgcgcct cagtgagcga 60

gcgagcgcgc 70

<210> 1053

<211> 70

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1053

gcgcgctcgc tcgctcactg aggcgcacgc ccgggtttcc cgggcggcct cagtgagcga 60

gcgagcgcgc 70

<210> 1054

<211> 72

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1054

gcgcgctcgc tcgctcactg aggccgtcgg gcgacctttg gtcgcccggc ctcagtgagc 60

gagcgagcgc gc 72

<210> 1055

<211> 72

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1055

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacggc ctcagtgagc 60

gagcgagcgc gc 72

<210> 1056

<211> 72

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1056

gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggc ctcagtgagc 60

gagcgagcgc gc 72

<210> 1057

<211> 72

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1057

gcgcgctcgc tcgctcactg aggccgacgc ccgggctttg cccgggcggc ctcagtgagc 60

gagcgagcgc gc 72

<210> 1058

<211> 83

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1058

gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg ctttgcccgg 60

cctcagtgag cgagcgagcg cgc 83

<210> 1059

<211> 83

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1059

gcgcgctcgc tcgctcactg aggccgggca aagcccgacg cccgggcttt gcccgggcgg 60

cctcagtgag cgagcgagcg cgc 83

<210> 1060

<211> 77

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1060

gcgcgctcgc tcgctcactg aggccgaaac gtcgggcgac ctttggtcgc ccggcctcag 60

tgagcgagcg agcgcgc 77

<210> 1061

<211> 77

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1061

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgtt tcggcctcag 60

tgagcgagcg agcgcgc 77

<210> 1062

<211> 51

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1062

gcgcgctcgc tcgctcactg aggcaaagcc tcagtgagcg agcgagcgcg c 51

<210> 1063

<211> 51

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1063

gcgcgctcgc tcgctcactg aggctttgcc tcagtgagcg agcgagcgcg c 51

<210> 1064

<211> 80

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1064

gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct 60

cagtgagcga gcgagcgcgc 80

<210> 1065

<211> 80

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1065

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggcggcct 60

cagtgagcga gcgagcgcgc 80

<210> 1066

<211> 79

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1066

gcgcgctcgc tcgctcactg aggcgcccgg gcgtcgggcg acctttggtc gcccggcctc 60

agtgagcgag cgagcgcgc 79

<210> 1067

<211> 79

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1067

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggcgcctc 60

agtgagcgag cgagcgcgc 79

<210> 1068

<211> 5

<212> PROTEIN

<213> Influenza virus

<400> 1068

Asp Arg Leu Arg Arg

1 5

<210> 1069

<211> 7

<212> PROTEIN

<213> Influenza virus

<400> 1069

Pro Lys Gln Lys Lys Arg Lys

1 5

<210> 1070

<211> 10

<212> PROTEIN

<213> Hepatitis delta virus

<400> 1070

Arg Lys Leu Lys Lys Lys Ile Lys Lys Leu

1 5 10

<210> 1071

<211> 10

<212> PROTEIN

<213> Mus musculus

<400> 1071

Arg Glu Lys Lys Lys Phe Leu Lys Arg Arg

1 5 10

<210> 1072

<211> 20

<212> PROTEIN

<213> Homo sapiens

<400> 1072

Lys Arg Lys Gly Asp Glu Val Asp Gly Val Asp Glu Val Ala Lys Lys

1 5 10 15

Lys Ser Lys Lys

20

<210> 1073

<211> 17

<212> PROTEIN

<213> Homo sapiens

<400> 1073

Arg Lys Cys Leu Gln Ala Gly Met Asn Leu Glu Ala Arg Lys Thr Lys

1 5 10 15

Lys

<210> 1074

<211> 8

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1074

gtttaaac 8

<210> 1075

<211> 8

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1075

ttaattaa 8

<210> 1076

<211> 141

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1076

aataaacgat aacgccgttg gtggcgtgag gcatgtaaaa ggttacatca ttatcttgtt 60

cgccatccgg ttggtataaa tagacgttca tgttggtttt tgtttcagtt gcaagttggc 120

tgcggcgcgc gcagcacctt t 141

<210> 1077

<211> 317

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1077

ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 60

agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 120

tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 180

ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 240

aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag 300

gcctaggctt ttgcaaa 317

<210> 1078

<211> 72

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1078

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacggc ctcagtgagc 60

gagcgagcgc gc 72

<210> 1079

<211> 60

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1079

gagacagaca cactcctgct atgggtactg ctgctctggg ttccaggttc cactggtgac 60

<210> 1080

<211> 1260

<212> DNA

<213> Adeno-associated virus - 2

<400> 1080

atggagctgg tcgggtggct cgtggacaag gggattacct cggagaagca gtggatccag 60

gaggaccagg cctcatacat ctccttcaat gcggcctcca actcgcggtc ccaaatcaag 120

gctgccttgg acaatgcggg aaagattatg agcctgacta aaaccgcccc cgactacctg 180

gtgggccagc agcccgtgga ggacatttcc agcaatcgga tttataaaat tttggaacta 240

aacgggtacg atccccaata tgcggcttcc gtctttctgg gatgggccac gaaaaagttc 300

ggcaagagga acaccatctg gctgtttggg cctgcaacta ccgggaagac caacatcgcg 360

gaggccatag cccacactgt gcccttctac gggtgcgtaa actggaccaa tgagaacttt 420

cccttcaacg actgtgtcga caagatggtg atctggtggg aggaggggaa gatgaccgcc 480

aaggtcgtgg agtcggccaa agccattctc ggaggaagca aggtgcgcgt ggaccagaaa 540

tgcaagtcct cggcccagat agacccgact cccgtgatcg tcacctccaa caccaacatg 600

tgcgccgtga ttgacgggaa ctcaacgacc ttcgaacacc agcagccgtt gcaagaccgg 660

atgttcaaat ttgaactcac ccgccgtctg gatcatgact ttgggaaggt caccaagcag 720

gaagtcaaag actttttccg gtgggcaaag gatcacgtgg ttgaggtgga gcatgaattc 780

tacgtcaaaa agggtggagc caagaaaaga cccgccccca gtgacgcaga tataagtgag 840

cccaaacggg tgcgcgagtc agttgcgcag ccatcgacgt cagacgcgga agcttcgatc 900

aactacgcag acaggtacca aaacaaatgt tctcgtcacg tgggcatgaa tctgatgctg 960

tttccctgca gacaatgcga gagaatgaat cagaattcaa atatctgctt cactcacgga 1020

cagaaagact gtttagagtg ctttcccgtg tcagaatctc aacccgtttc tgtcgtcaaa 1080

aaggcgtatc agaaactgtg ctacattcat catatcatgg gaaaggtgcc agacgcttgc 1140

actgcctgcg atctggtcaa tgtggatttg gatgactgca tctttgaaca ataaatgatt 1200

taaatcaggt atggctgccg atggttatct tccagattgg ctcgaggaca ctctctctga 1260

<210> 1081

<211> 1932

<212> DNA

<213> Adeno-associated virus - 2

<400> 1081

atgccggggt tttacgagat tgtgattaag gtccccagcg accttgacga gcatctgccc 60

ggcatttctg acagctttgt gaactgggtg gccgagaagg aatgggagtt gccgccagat 120

tctgacatgg atctgaatct gattgagcag gcacccctga ccgtggccga gaagctgcag 180

cgcgactttc tgacggaatg gcgccgtgtg agtaaggccc cggaggccct tttctttgtg 240

caatttgaga agggagagag ctacttccac atgcacgtgc tcgtggaaac caccggggtg 300

aaatccatgg ttttgggacg tttcctgagt cagattcgcg aaaaactgat tcagagaatt 360

taccgcggga tcgagccgac tttgccaaac tggttcgcgg tcacaaagac cagaaatggc 420

gccggaggcg ggaacaaggt ggtggatgag tgctacatcc ccaattactt gctccccaaa 480

acccagcctg agctccagtg ggcgtggact aatatggaac agtatttaag cgcctgtttg 540

aatctcacgg agcgtaaacg gttggtggcg cagcatctga cgcacgtgtc gcagacgcag 600

gagcagaaca aagagaatca gaatcccaat tctgatgcgc cggtgatcag atcaaaaact 660

tcagccaggt acatggagct ggtcgggtgg ctcgtggaca aggggattac ctcggagaag 720

cagtggatcc aggaggacca ggcctcatac atctccttca atgcggcctc caactcgcgg 780

tcccaaatca aggctgcctt ggacaatgcg ggaaagatta tgagcctgac taaaaccgcc 840

cccgactacc tggtggggcca gcagcccgtg gaggacatt ccagcaatcg gatttataaa 900

attttggaac taaacgggta cgatccccaa tatgcggctt ccgtctttct gggatgggcc 960

acgaaaaagt tcggcaagag gaacaccatc tggctgtttg ggcctgcaac taccgggaag 1020

accaacatcg cggaggccat agcccacact gtgcccttct acgggtgcgt aaactggacc 1080

aatgagaact ttcccttcaa cgactgtgtc gacaagatgg tgatctggtg ggaggagggg 1140

aagatgaccg ccaaggtcgt ggagtcggcc aaagccattc tcggaggaag caaggtgcgc 1200

gtggaccaga aatgcaagtc ctcggcccag atagacccga ctcccgtgat cgtcacctcc 1260

aacaccaaca tgtgcgccgt gattgacggg aactcaacga ccttcgaaca ccagcagccg 1320

ttgcaagacc ggatgttcaa atttgaactc acccgccgtc tggatcatga ctttgggaag 1380

gtcaccaagc aggaagtcaa agactttttc cggtgggcaa aggatcacgt ggttgaggtg 1440

gagcatgaat tctacgtcaa aaagggtgga gccaagaaaa gacccgcccc cagtgacgca 1500

gatataagtg agcccaaacg ggtgcgcgag tcagttgcgc agccatcgac gtcagacgcg 1560

gaagcttcga tcaactacgc agacaggtac caaaacaaat gttctcgtca cgtgggcatg 1620

aatctgatgc tgtttccctg cagacaatgc gagagaatga atcagaattc aaatatctgc 1680

ttcactcacg gacagaaaga ctgtttagag tgctttcccg tgtcagaatc tcaacccgtt 1740

tctgtcgtca aaaaggcgta tcagaaactg tgctacattc atcatatcat gggaaaggtg 1800

cgacgctt gcactgcctg cgatctggtc aatgtggatt tggatgactg catctttgaa 1860

caataaatga tttaaatcag gtatggctgc cgatggttat cttccagatt ggctcgagga 1920

cactctctct ga 1932

<210> 1082

<211> 1876

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1082

cgcagccacc atggcggggt tttacgagat tgtgattaag gtccccagcg accttgacgg 60

gcatctgccc ggcatttctg acagctttgt gaactgggtg gccgagaagg aatgggagtt 120

gccgccagat tctgacatgg atctgaatct gattgagcag gcacccctga ccgtggccga 180

gaagctgcag cgcgactttc tgacggaatg gcgccgtgtg agtaaggccc cggaggccct 240

tttctttgtg caatttgaga agggagagag ctacttccac atgcacgtgc tcgtggaaac 300

caccggggtg aaatccatgg ttttgggacg tttcctgagt cagattcgcg aaaaactgat 360

tcagagaatt taccgcggga tcgagccgac tttgccaaac tggttcgcgg tcacaaagac 420

cagaaatggc gccggaggcg ggaacaaggt ggtggatgag tgctacatcc ccaattactt 480

gctccccaaa acccagcctg agctccagtg ggcgtggact aatatggaac agtatttaag 540

cgcctgtttg aatctcacgg agcgtaaacg gttggtggcg cagcatctga cgcacgtgtc 600

gcagacgcag gagcagaaca aagagaatca gaatcccaat tctgatgcgc cggtgatcag 660

atcaaaaact tcagccaggt acatggagct ggtcgggtgg ctcgtggaca aggggattac 720

ctcggagaag cagtggatcc aggaggacca ggcctcatac atctccttca atgcggcctc 780

caactcgcgg tcccaaatca aggctgcctt ggacaatgcg ggaaagatta tgagcctgac 840

taaaaccgcc cccgactacc tggtgggcca gcagcccgtg gaggacatt ccagcaatcg 900

gatttataaa attttggaac taaacgggta cgatccccaa tatgcggctt ccgtctttct 960

gggatgggcc acgaaaaagt tcggcaagag gaacaccatc tggctgtttg ggcctgcaac 1020

taccgggaag accaacatcg cggaggccat agcccacact gtgcccttct acgggtgcgt 1080

aaactggacc aatgagaact ttcccttcaa cgactgtgtc gacaagatgg tgatctggtg 1140

ggaggagggg aagatgaccg ccaaggtcgt ggagtcggcc aaagccattc tcggaggaag 1200

caaggtgcgc gtggaccaga aatgcaagtc ctcggcccag atagacccga ctcccgtgat 1260

cgtcacctcc aacaccaaca tgtgcgccgt gattgacggg aactcaacga ccttcgaaca 1320

ccagcagccg ttgcaagacc ggatgttcaa atttgaactc acccgccgtc tggatcatga 1380

ctttgggaag gtcaccaagc aggaagtcaa agactttttc cggtgggcaa aggatcacgt 1440

ggttgaggtg gagcatgaat tctacgtcaa aaagggtgga gccaagaaaa gacccgcccc 1500

cagtgacgca gatataagtg agcccaaacg ggtgcgcgag tcagttgcgc agccatcgac 1560

gtcagacgcg gaagcttcga tcaactacgc agacaggtac caaaacaaat gttctcgtca 1620

cgtgggcatg aatctgatgc tgtttccctg cagacaatgc gagagaatga atcagaattc 1680

aaatatctgc ttcactcacg gacagaaaga ctgtttagag tgctttcccg tgtcagaatc 1740

tcaacccgtt tctgtcgtca aaaaggcgta tcagaaactg tgctacattc atcatatcat 1800

gggaaaggtg cgacgctt gcactgcctg cgatctggtc aatgtggatt tggatgactg 1860

catctttgaa caataa 1876

<210> 1083

<211> 1194

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1083

atggagctgg tcgggtggct cgtggacaag gggattacct cggagaagca gtggatccag 60

gaggaccagg cctcatacat ctccttcaat gcggcctcca actcgcggtc ccaaatcaag 120

gctgccttgg acaatgcggg aaagattatg agcctgacta aaaccgcccc cgactacctg 180

gtgggccagc agcccgtgga ggacatttcc agcaatcgga tttataaaat tttggaacta 240

aacgggtacg atccccaata tgcggcttcc gtctttctgg gatgggccac gaaaaagttc 300

ggcaagagga acaccatctg gctgtttggg cctgcaacta ccgggaagac caacatcgcg 360

gaggccatag cccacactgt gcccttctac gggtgcgtaa actggaccaa tgagaacttt 420

cccttcaacg actgtgtcga caagatggtg atctggtggg aggaggggaa gatgaccgcc 480

aaggtcgtgg agtcggccaa agccattctc ggaggaagca aggtgcgcgt ggaccagaaa 540

tgcaagtcct cggcccagat agacccgact cccgtgatcg tcacctccaa caccaacatg 600

tgcgccgtga ttgacgggaa ctcaacgacc ttcgaacacc agcagccgtt gcaagaccgg 660

atgttcaaat ttgaactcac ccgccgtctg gatcatgact ttgggaaggt caccaagcag 720

gaagtcaaag actttttccg gtgggcaaag gatcacgtgg ttgaggtgga gcatgaattc 780

tacgtcaaaa agggtggagc caagaaaaga cccgccccca gtgacgcaga tataagtgag 840

cccaaacggg tgcgcgagtc agttgcgcag ccatcgacgt cagacgcgga agcttcgatc 900

aactacgcag accgctacca aaacaaatgt tctcgtcacg tgggcatgaa tctgatgctg 960

tttccctgca gacaatgcga gagaatgaat cagaattcaa atatctgctt cactcacgga 1020

cagaaagact gtttagagtg ctttcccgtg tcagaatctc aacccgtttc tgtcgtcaaa 1080

aaggcgtatc agaaactgtg ctacattcat catatcatgg gaaaggtgcc agacgcttgc 1140

actgcctgcg atctggtcaa tgtggatttg gatgactgca tctttgaaca ataa 1194

<210> 1084

<211> 1876

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1084

cgcagccacc atggcggggt tttacgagat tgtgattaag gtccccagcg accttgacgg 60

gcatctgccc ggcatttctg acagctttgt gaactgggtg gccgagaagg aatgggagtt 120

gccgccagat tctgacatgg atctgaatct gattgagcag gcacccctga ccgtggccga 180

gaagctgcag cgcgactttc tgacggaatg gcgccgtgtg agtaaggccc cggaggccct 240

tttctttgtg caatttgaga agggagagag ctacttccac atgcacgtgc tcgtggaaac 300

caccggggtg aaatccatgg ttttgggacg tttcctgagt cagattcgcg aaaaactgat 360

tcagagaatt taccgcggga tcgagccgac tttgccaaac tggttcgcgg tcacaaagac 420

cagaaatggc gccggaggcg ggaacaaggt ggtggatgag tgctacatcc ccaattactt 480

gctccccaaa acccagcctg agctccagtg ggcgtggact aatatggaac agtatttaag 540

cgcctgtttg aatctcacgg agcgtaaacg gttggtggcg cagcatctga cgcacgtgtc 600

gcagacgcag gagcagaaca aagagaatca gaatcccaat tctgatgcgc cggtgatcag 660

atcaaaaact tcagccaggt acatggagct ggtcgggtgg ctcgtggaca aggggattac 720

ctcggagaag cagtggatcc aggaggacca ggcctcatac atctccttca atgcggcctc 780

caactcgcgg tcccaaatca aggctgcctt ggacaatgcg ggaaagatta tgagcctgac 840

taaaaccgcc cccgactacc tggtgggcca gcagcccgtg gaggacatt ccagcaatcg 900

gatttataaa attttggaac taaacgggta cgatccccaa tatgcggctt ccgtctttct 960

gggatgggcc acgaaaaagt tcggcaagag gaacaccatc tggctgtttg ggcctgcaac 1020

taccgggaag accaacatcg cggaggccat agcccacact gtgcccttct acgggtgcgt 1080

aaactggacc aatgagaact ttcccttcaa cgactgtgtc gacaagatgg tgatctggtg 1140

ggaggagggg aagatgaccg ccaaggtcgt ggagtcggcc aaagccattc tcggaggaag 1200

caaggtgcgc gtggaccaga aatgcaagtc ctcggcccag atagacccga ctcccgtgat 1260

cgtcacctcc aacaccaaca tgtgcgccgt gattgacggg aactcaacga ccttcgaaca 1320

ccagcagccg ttgcaagacc ggatgttcaa atttgaactc acccgccgtc tggatcatga 1380

ctttgggaag gtcaccaagc aggaagtcaa agactttttc cggtgggcaa aggatcacgt 1440

ggttgaggtg gagcatgaat tctacgtcaa aaagggtgga gccaagaaaa gacccgcccc 1500

cagtgacgca gatataagtg agcccaaacg ggtgcgcgag tcagttgcgc agccatcgac 1560

gtcagacgcg gaagcttcga tcaactacgc agacaggtac caaaacaaat gttctcgtca 1620

cgtgggcatg aatctgatgc tgtttccctg cagacaatgc gagagaatga atcagaattc 1680

aaatatctgc ttcactcacg gacagaaaga ctgtttagag tgctttcccg tgtcagaatc 1740

tcaacccgtt tctgtcgtca aaaaggcgta tcagaaactg tgctacattc atcatatcat 1800

gggaaaggtg cgacgctt gcactgcctg cgatctggtc aatgtggatt tggatgactg 1860

catctttgaa caataa 1876

<210> 1085

<211> 51

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1085

ctaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg c 51

<210> 1086

<211> 65

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1086

ctaggactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct 60

cagtc 65

<210> 1087

<211> 67

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1087

ggactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag 60

tcctgca 67

<210> 1088

<211> 41

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1088

gtgcgggcga ccaaaggtcg cccgacgccc gggcgcactc a 41

<210> 1089

<211> 56

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1089

ggactgaggc cgggcgacca aaggtcgccc gacgcccggg cggcctcagt cctgca 56

<210> 1090

<211> 54

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1090

ctaggactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc agtc 54

<210> 1091

<211> 48

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1091

ggactgaggc cgggcgacca aaggtcgccc gacggcctca gtcctgca 48

<210> 1092

<211> 46

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1092

ctaggactga ggccgtcggg cgacctttgg tcgcccggcc tcagtc 46

<210> 1093

<211> 67

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1093

ggactgaggc ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcgcctcag 60

tcctgca 67

<210> 1094

<211> 47

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1094

atacctaggc acgcgtgtta ctagttatta atagtaatca attacgg 47

<210> 1095

<211> 29

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1095

atacctaggg gccgcacgcg tgttactag 29

<210> 1096

<211> 42

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1096

atacactcag tgcctgcagg cacgtggtcc ggagatccag ac 42

<210> 1097

<211> 3754

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1097

cctaggtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct 60

tgtagttaat gattaacccg ccatgctact tatcgcggcc gctcaatatt ggccattagc 120

catattattc attggttata tagcataaat caatattggc tattggccat tgcatacgtt 180

gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac cgccatgttg 240

gcattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc 300

atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa 360

cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac 420

tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca 480

agtgtatcat atgccaagtc cgccccctat tgacgtcaat gacggtaaat ggcccgcctg 540

gcattatgcc cagtacatga ccttacggga ctttcctact tggcagtaca tctacgtatt 600

agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc 660

ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat 720

gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg 780

cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc 840

ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg 900

agtcgctgcg acgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc 960

cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc 1020

cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa 1080

gccttgaggg gctccgggag ggccctttgt gcggggggga gcggctcggg gggtgcgtgc 1140

gtgtgtgtgt gcgtggggag cgccgcgtgc ggcccgcgct gcccggcggc tgtgagcgct 1200

gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg 1260

gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt 1320

gcgtgggggg gtgagcaggg ggtgtgggcg cggcggtcgg gctgtaaccc ccccctgcac 1380

ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt 1440

ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg 1500

gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc 1560

ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg 1620

cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc 1680

ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag 1740

ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg 1800

cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg 1860

accggcggct ctagagcctc tgctaaccat gttttagcct tcttcttttt cctacagctc 1920

ctgggcaacg tgctggttat tgtgctgtct catcatttgt cgacagaatt cctcgaagat 1980

ccgaaggggt tcaagcttgg cattccggta ctgttggtaa agccagttta aacgccgcca 2040

ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg 2100

acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct 2160

acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca 2220

ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga 2280

agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct 2340

tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc 2400

tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc 2460

acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga 2520

acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg 2580

ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc 2640

actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg 2700

tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt 2760

aattaattaa gagcatctta ccgccattta ttcccatatt tgttctgttt ttcttgattt 2820

gggtatacat ttaaatgtta ataaaacaaa atggtggggc aatcatttac atttttaggg 2880

atatgtaatt actagttcag gtgtattgcc acaagacaaa catgttaaga aactttcccg 2940

ttatttacgc tctgttcctg ttaatcaacc tctggattac aaaatttgtg aaagattgac 3000

tgatattctt aactatgttg ctccttttac gctgtgtgga tatgctgctt tatagcctct 3060

gtatctagct attgcttccc gtacggcttt cgttttctcc tccttgtata aatcctggtt 3120

gctgtctctt ttagaggagt tgtggcccgt tgtccgtcaa cgtggcgtgg tgtgctctgt 3180

gtttgctgac gcaaccccca ctggctgggg cattgccacc acctgtcaac tcctttctgg 3240

gactttcgct ttccccctcc cgatcgccac ggcagaactc atcgccgcct gccttgcccg 3300

ctgctggaca ggggctaggt tgctgggcac tgataattcc gtggtgttgt ctgtgccttc 3360

tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc 3420

cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg 3480

tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa 3540

tagcaggcat gctggggatg cggtgggctc tatggctcta gagcatggct acgtagataa 3600

gtagcatggc gggttaatca ttaactacac ctgcagcagg aacccctagt gatggagttg 3660

gccactccct ctctgcgcgc tcgctcgctc cctgcaggac tgaggccggg cgaccaaagg 3720

tcgcccgacg cccgggcggc ctcagtcctg cagg 3754

<210> 1098

<211> 8418

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1098

ggcagctgcg cgctcgctcg ctcacctagg ccgcccgggc aaagcccggg cgtcgggcga 60

cctttggtcg cccggcctag gtgagcgagc gagcgcgcag agagggagtg gccaactcca 120

tcactagggg ttccttgtag ttaatgatta acccgccatg ctacttatcg cggccgctca 180

atattggcca ttagccatat tattcattgg ttatatagca taaatcaata ttggctattg 240

gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat 300

atgaccgcca tgttggcatt gattattgac tagttattaa tagtaatcaa ttacggggtc 360

attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc 420

tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt 480

aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca 540

cttggcagta catcaagtgt atcatatgcc aagtccgccc cctattgacg tcaatgacgg 600

taaatggccc gcctggcatt atgcccagta catgacctta cgggactttc ctacttggca 660

gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt 720

cactctcccc atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt 780

attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccaggcg gggcggggcg 840

gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc 900

gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag 960

cgcgcggcgg gcgggagtcg ctgcgacgct gccttcgccc cgtgccccgc tccgccgccg 1020

cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg 1080

acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc 1140

tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg ggggagcggc 1200

tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggccc gcgctgcccg 1260

gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg 1320

ggagcgcggc cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct 1380

gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcggcg gtcgggctgt 1440

aacccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg 1500

ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg 1560

gtgccgggcg gggcggggcc gcctcggggcc ggggagggct cgggggaggg gcgcggcggc 1620

ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa 1680

tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg 1740

aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag 1800

gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag 1860

cctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc 1920

ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgtttt agccttcttc 1980

tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca tttgtcgaca 2040

gaattcctcg aagatccgaa ggggttcaag cttggcattc cggtactgtt ggtaaagcca 2100

gtttaaacgc cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca 2160

tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg 2220

agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc 2280

ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct 2340

accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc 2400

aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt 2460

tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg 2520

gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg 2580

ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg 2640

gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc 2700

tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga 2760

agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg 2820

acgagctgta caagtaatta attaagagca tcttaccgcc atttattccc atatttgttc 2880

tgtttttctt gatttgggta tacatttaaa tgttaataaa acaaaatggt ggggcaatca 2940

tttacatttt tagggatatg taattactag ttcaggtgta ttgccacaag acaaacatgt 3000

taagaaactt tcccgttatt tacgctctgt tcctgttaat caacctctgg attacaaaat 3060

ttgtgaaaga ttgactgata ttcttaacta tgttgctcct tttacgctgt gtggatatgc 3120

tgctttatag cctctgtatc tagctattgc ttcccgtacg gctttcgttt tctcctcctt 3180

gtataaatcc tggttgctgt ctcttttaga ggagttgtgg cccgttgtcc gtcaacgtgg 3240

cgtggtgtgc tctgtgtttg ctgacgcaac ccccactggc tggggcattg ccaccacctg 3300

tcaactcctt tctgggactt tcgctttccc cctcccgatc gccacggcag aactcatcgc 3360

cgcctgcctt gcccgctgct ggacaggggc taggttgctg ggcactgata attccgtggt 3420

gttgtctgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 3480

gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 3540

ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 3600

ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg ctctagagca 3660

tggctacgta gataagtagc atggcgggtt aatcattaac tacacctgca gcaggaaccc 3720

ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctccctgc aggactgagg 3780

ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag tcctgcaggg agcgagcgag 3840

cgcgcagctg cctgcacggg cgcgccggta ccgggagatg ggggaggcta actgaaacac 3900

ggaagggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa 3960

cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct 4020

gtcgataccc caccgagacc ccattgggac caatacgccc gcgtttcttc cttttcccca 4080

ccccaacccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaagc 4140

cctgccatag ccactacggg tacgtaggcc aaccactaga actatagcta gagtcctggg 4200

cgaacaaacg atgctcgcct tccagaaaac cgaggatgcg aaccacttca tccggggtca 4260

gcaccaccgg caagcgccgc gacggccgag gtctaccgat ctcctgaagc cagggcagat 4320

ccgtgcacag caccttgccg tagaagaaca gcaaggccgc caatgcctga cgatgcgtgg 4380

agaccgaaac cttgcgctcg ttcgccagcc aggacagaaa tgcctcgact tcgctgctgc 4440

ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagttgacat 4500

aagcctgttc ggttcgtaaa ctgtaatgca agtagcgtat gcgctcacgc aactggtcca 4560

gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg 4620

actgtttttt tgtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg 4680

gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagcaacga tgttacgcag 4740

cagggcagtc gccctaaaac aaagttaggt ggctcaagta tgggcatcat tcgcacatgt 4800

aggctcggcc ctgaccaagt caaatccatg cgggctgctc ttgatctttt cggtcgtgag 4860

ttcggagacg tagccaccta ctcccaacat cagccggact ccgattacct cgggaacttg 4920

ctccgtagta agacattcat cgcgcttgct gccttcgacc aagaagcggt tgttggcgct 4980

ctcgcggctt acgttctgcc caggtttgag cagccgcgta gtgagatcta tatctatgat 5040

ctcgcagtct ccggcgagca ccggaggcag ggcattgcca ccgcgctcat caatctcctc 5100

aagcatgagg ccaacgcgct tggtgcttat gtgatctacg tgcaagcaga ttacggtgac 5160

gatcccgcag tggctctcta tacaaagttg ggcatacggg aagaagtgat gcactttgat 5220

atcgacccaa gtaccgccac ctaacaattc gttcaagccg agatcggctt cccggccgcg 5280

gagttgttcg gtaaattgtc acaacgccgc gaatatagtc tttaccatgc ccttggccac 5340

gcccctcttt aatacgacgg gcaatttgca cttcagaaaa tgaagagttt gctttagcca 5400

taacaaaagt ccagtatgct ttttcacagc ataactggac tgatttcagt ttacaactat 5460

tctgtctagt ttaagacttt attgtcatag tttagatcta ttttgttcag tttaagactt 5520

tattgtccgc ccacacccgc ttacgcaggg catccattta ttactcaacc gtaaccgatt 5580

ttgccaggtt acgcggctgg tctgcggtgt gaaataccgc acagatgcgt aaggagaaaa 5640

taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 5700

ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 5760

gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 5820

gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 5880

cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 5940

ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 6000

tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg 6060

gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 6120

tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 6180

ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 6240

ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct 6300

ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 6360

accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 6420

tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 6480

cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 6540

taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 6600

caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 6660

gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 6720

gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 6780

ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 6840

attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 6900

gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 6960

tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 7020

agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 7080

gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 7140

actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 7200

tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 7260

attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 7320

tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 7380

tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 7440

aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 7500

tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 7560

cgcacatttc cccgaaaagt gccacctgaa attgtaaacg ttaatatttt gttaaaattc 7620

gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc 7680

ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag 7740

agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc 7800

gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa 7860

gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg 7920

aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt 7980

gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc 8040

gcgtcccatt cgccattcag gctgcaaata agcgttgata ttcagtcaat tacaaacatt 8100

aataacgaag agatgacaga aaaattttca ttctgtgaca gagaaaaagt agccgaagat 8160

gacggtttgt cacatggagt tggcaggatg tttgattaaa aacataacag gaagaaaaat 8220

gccccgctgt gggcggacaa aatagttggg aactgggagg ggtggaaatg gagtttttaa 8280

ggattattta gggaagagtg acaaaataga tgggaactgg gtgtagcgtc gtaagctaat 8340

acgaaaatta aaaatgacaa aatagtttgg aactagatt cacttatctg gttcggatct 8400

cctagtgagc tccctgca 8418

<210> 1099

<211> 225

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1099

tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct 60

ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct 120

gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg 180

ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggc 225

<210> 1100

<211> 1177

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1100

ggctcagagg ctcagaggca cacaggagtt tctgggctca ccctgccccc ttccaacccc 60

tcagttccca tcctccagca gctgtttgtg tgctgcctct gaagtccaca ctgaacaaac 120

ttcagcctac tcatgtccct aaaatgggca aacattgcaa gcagcaaaca gcaaacacac 180

agccctccct gcctgctgac cttggagctg gggcagaggt cagagacctc tctgggccca 240

tgccacctcc aacatccact cgaccccttg gaatttcggt ggagaggagc agaggttgtc 300

ctggcgtggt ttaggtagtg tgagagggtc cgggttcaaa accacttgct gggtggggag 360

tcgtcagtaa gtggctatgc cccgaccccg aagcctgttt ccccatctgt acaatggaaa 420

tgataaagac gcccatctga tagggttttt gtggcaaata aacatttggt ttttttgttt 480

tgttttgttt tgttttttga gatggaggtt tgctctgtcg cccaggctgg agtgcagtga 540

cacaatctca tctcaccaca accttcccct gcctcagcct cccaagtagc tgggattaca 600

agcatgtgcc accacacctg gctaattttc tatttttagt agagacgggt ttctccatgt 660

tggtcagcct cagcctccca agtaactggg attacaggcc tgtgccacca cacccggcta 720

attttttcta tttttgacag ggacggggtt tcaccatgtt ggtcaggctg gtctagaggt 780

accggatctt gctaccagtg gaacagccac taaggattct gcagtgagag cagaggcca 840

gctaagtggt actctcccag agactgtctg actcacgcca ccccctccac cttggacaca 900

ggacgctgtg gtttctgagc caggtacaat gactcctttc ggtaagtgca gtggaagctg 960

tacactgccc aggcaaagcg tccgggcagc gtaggcgggc gactcagatc ccagccagtg 1020

gacttagccc ctgtttgctc ctccgataac tggggtgacc ttggttaata ttcaccagca 1080

gcctcccccg ttgcccctct ggatccactg cttaaatacg gacgaggaca gggccctgtc 1140

tcctcagctt caggcaccac cactgacctg ggacagt 1177

<210> 1101

<211> 1326

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1101

ctgcagggcc cactagtgga gccgagagta attcatacaa aaggagggat cgccttcgca 60

aggggagagc cccaggaccg tccctaaatt ctcacagacc caaatccctg tagccgcccc 120

acgacagcgc gaggagcatg cgcccagggc tgagcgcggg tagatcagag cacacaagct 180

cacagtcccc ggcggtgggg ggaggggcgc gctgagcggg ggccagggag ctggcgcggg 240

gcaaactggg aaagtggtgt cgtgtgctgg ctccgccctc ttcccgaggg tggggggagaa 300

cggtatataa gtgcggtagt cgccttggac gttctttttc gcaacgggtt tgccgtcaga 360

acgcaggtga gtggcgggtg tggcttccgc gggccccgga gctggagccc tgctctgagc 420

gggccgggct gatatgcgag tgtcgtccgc agggtttagc tgtgagcatt cccacttcga 480

gtggcgggcg gtgcgggggt gagagtgcga ggcctagcgg caaccccgta gcctcgcctc 540

gtgtccggct tgaggcctag cgtggtgtcc gccgccgcgt gccactccgg ccgcactatg 600

cgttttttgt ccttgctgcc ctcgattgcc ttccagcagc atgggctaac aaagggaggg 660

tgtggggctc actcttaagg agcccatgaa gcttacgttg gataggaatg gaagggcagg 720

aggggcgact ggggcccgcc cgccttcgga gcacatgtcc gacgccacct ggatggggcg 780

aggcctgtgg ctttccgaag caatcgggcg tgagtttagc ctacctgggc catgtggccc 840

tagcactggg cacggtctgg cctggcggtg ccgcgttccc ttgcctccca acaagggtga 900

ggccgtcccg cccggcacca gttgcttgcg cggaaagatg gccgctcccg gggccctgtt 960

gcaaggagct caaaatggag gacgcggcag cccggtggag cgggcgggtg agtcaccac 1020

acaaaggaag agggccttgc ccctcgccgg ccgctgcttc ctgtgacccc gtggtctatc 1080

ggccgcatag tcacctcggg cttctcttga gcaccgctcg tcgcggcggg gggaggggat 1140

ctaatggcgt tggagtttgt tcacatttgg tgggtggaga ctagtcaggc cagcctggcg 1200

ctggaagtca ttcttggaat ttgccccttt gagtttggag cgaggctaat tctcaagcct 1260

cttagcggtt caaaggtatt ttctaaaccc gtttccaggt gttgtgaaag ccaccgctaa 1320

ttcaaa 1326

<210> 1102

<211> 573

<212> DNA

<213> Mus musculus

<400> 1102

gtaagagttt tatgtttttt catctctgct tgtatttttc tagtaatgga agcctggtat 60

tttaaaatag ttaaattttc ctttagtgct gatttctaga ttattattac tgttgttgtt 120

gttattattg tcattatttg catctgagaa cccttaggtg gttatattat tgatatattt 180

ttggtatctt tgatgacaat aatgggggat tttgaaagct tagctttaaa tttcttttaa 240

ttaaaaaaaa atgctaggca gaatgactca aattacgttg gatacagttg aatttattac 300

ggtctcatag ggcctgcctg ctcgaccatg ctatactaaa aattaaaagt gtgtgttact 360

aattttataa atggagtttc catttatatt tacctttatt tcttatttac cattgtctta 420

gtagatattt acaaacatga cagaaacact aaatcttgag tttgaatgca cagatataaa 480

cacttaacgg gttttaaaaa taataatgtt ggtgaaaaaa tataactttg agtgtagcag 540

agaggaacca ttgccacctt cagattttcc tgt 573

<210> 1103

<211> 1993

<212> DNA

<213> Mus musculus

<400> 1103

acgatcggga actggcatct tcagggagta gcttaggtca gtgaagagaa gaacaaaaag 60

cagcatatta cagttagttg tcttcatcaa tctttaaata tgttgtgtgg tttttctctc 120

cctgtttcca cagacaagag tgagatcgcc catcggtata atgatttggg agaacaacat 180

ttcaaaggcc tgtaagttat aatgctgaaa gcccacttaa tatttctggt agtattagtt 240

aaagttttaa aacacctttt tccaccttga gtgtgagaat tgtagagcag tgctgtccag 300

tagaaatgtg tgcattgaca gaaagactgt ggatctgtgc tgagcaatgt ggcagccaga 360

gatcacaagg ctatcaagca ctttgcacat ggcaagtgta actgagaagc acacattcaa 420

ataatagtta attttaattg aatgtatcta gccatgtgtg gctagtagct cctttcctgg 480

agagagaatc tggagcccac atctaacttg ttaagtctgg aatcttattt tttatttctg 540

gaaaggtcta tgaactatag ttttgggggc agctcactta ctaactttta atgcaataag 600

atctcatggt atcttgagaa cattattttg tctctttgta gtactgaaac cttatacatg 660

tgaagtaagg ggtctatact taagtcacat ctccaacctt agtaatgttt taatgtagta 720

aaaaaatgag taattaattt atttttagaa ggtcaatagt atcatgtatt ccaaataaca 780

gaggtatatg gttagaaaag aaacaattca aaggacttat ataatatcta gccttgacaa 840

tgaataaatt tagagagtag tttgcctgtt tgcctcatgt tcataaatct attgacacat 900

atgtgcatct gcacttcagc atggtagaag tccatattcc tttgcttgga aaggcaggtg 960

ttcccattac gcctcagaga atagctgacg ggaagaggct ttctagatag ttgtatgaaa 1020

gatatacaaa atctcgcagg tatacacagg catgatttgc tggttgggag agccacttgc 1080

ctcatactga ggtttttgtg tctgcttttc agagtcctga ttgccttttc ccagtatctc 1140

cagaaatgct catacgatga gcatgccaaa ttagtgcagg aagtaacaga ctttgcaaag 1200

acgtgtgttg ccgatgagtc tgccgccaac tgtgacaaat cccttgtgag taccttctga 1260

ttttgtggat ctactttcct gctttctgga actctgtttc aaagccaatc atgactccat 1320

cacttaaggc cccgggaaca ctgtggcaga gggcagcaga gagattgata aagccaggt 1380

gatgggaatt ttctgtggga ctccatttca tagtaattgc agaagctaca atacactcaa 1440

aaagtctcac cacatgactg cccaaatggg agcttgacag tgacagtgac agtagatatg 1500

ccaaagtgga tgagggaaag accacaagag ctaaaccctg taaaaagaac tgtaggcaac 1560

taaggaatgc agagagaaga agttgccttg gaagagcata ccaactgcct ctccaatacc 1620

aatggtcatc cctaaaacat acgtatgaat aacatgcaga ctaagcaggc tacatttagg 1680

aatatacatg tatttacata aatgtatatg catgtaacaa caatgaatga aaactgaggt 1740

catggatctg aaagagagca agggggctta catgagagg tttggaggga ggggttggag 1800

ggagggaggt attattcttt agttttacag ggaacgtagt aaaaacatag gcttctccca 1860

aaggagcaga gcccatgagg agctgtgcaa ggttccccag cttgatttta cctgctcctc 1920

aaattccctt gatttgtttt tattataatg actttactcc tagcttttag tgtcagatag 1980

aaaacatgga agg 1993

<210> 1104

<211> 1350

<212> DNA

<213> Homo sapiens

<400> 1104

taggaggctg aggcaggagg atcgcttgag cccaggagtt cgagaccagc ctgggcaaca 60

tagtgtgatc ttgtatctat aaaaataaac aaaattagct tggtgtggtg gcgcctgtag 120

tccccagcca cttggagggg tgaggtgaga ggattgcttg agcccgggat ggtccaggct 180

gcagtgagcc atgatcgtgc cactgcactc cagcctgggc gacagagtga gaccctgtct 240

cacaacaaca acaacaacaa caaaaaggct gagctgcacc atgcttgacc cagtttctta 300

aaattgttgt caaagcttca ttcactccat ggtgctatag agcacaagat tttatttggt 360

gagatggtgc tttcatgaat tcccccaaca gagccaagct ctccatctag tggacaggga 420

agctagcagc aaaccttccc ttcactacaa aacttcattg cttggccaaa aagagagtta 480

attcaatgta gacatctatg taggcaatta aaaacctatt gatgtataaa acagtttgca 540

ttcatggagg gcaactaaat acattctagg actttataaa agatcacttt ttatttatgc 600

acagggtgga acaagatgga ttatcaagtg tcaagtccaa tctatgacat caattattat 660

acatcggagc cctgccaaaa aatcaatgtg aagcaaatcg cagcccgcct cctgcctccg 720

ctctactcac tggtgttcat ctttggtttt gtgggcaaca tgctggtcat cctcatcctg 780

ataaactgca aaaggctgaa gagcatgact gacatctacc tgctcaacct ggccatctct 840

gacctgtttt tccttcttac tgtccccttc tgggctcact atgctgccgc ccagtgggac 900

tttggaaata caatgtgtca actcttgaca gggctctatt ttataggctt cttctctgga 960

atcttcttca tcatcctcct gacaatcgat aggtacctgg ctgtcgtcca tgctgtgttt 1020

gctttaaaag ccaggacggt cacctttggg gtggtgacaa gtgtgatcac ttgggtggtg 1080

gctgtgtttg cgtctctccc aggaatcatc tttaccagat ctcaaaaaga aggtcttcat 1140

tacacctgca gctctcattt tccatacagt cagtatcaat tctggaagaa tttccagaca 1200

ttaaagatag tcatcttggg gctggtcctg ccgctgcttg tcatggtcat ctgctactcg 1260

ggaatcctaa aaactctgct tcggtgtcga aatgagaaga agaggcacag ggctgtgagg 1320

cttatcttca ccatcatgat tgtttatttt 1350

<210> 1105

<211> 1223

<212> DNA

<213> Homo sapiens

<400> 1105

tgacagagac tcttgggatg acgcactgct gcatcaaccc catcatctat gcctttgtcg 60

gggagaagtt cagaaactac ctcttagtct tcttccaaaa gcacattgcc aaacgcttct 120

gcaaatgctg ttctattttc cagcaagagg ctcccgagcg agcaagctca gtttacaccc 180

gatccactgg ggagcaggaa atatctgtgg gcttgtgaca cggactcaag tgggctggtg 240

acccagtcag agttgtgcac atggcttagt tttcatacac agcctgggct gggggtgggg 300

tggggagaggt cttttttaaa aggaagttac tgttatagag ggtctaagat tcatccattt 360

atttggcatc tgtttaaagt agattagatc ttttaagccc atcaattata gaaagccaaa 420

tcaaaatatg ttgatgaaaa atagcaacct ttttatctcc ccttcacatg catcaagtta 480

ttgacaaact ctcccttcac tccgaaagtt ccttatgtat atttaaaaga aagcctcaga 540

gaattgctga ttcttgagtt tagtgatctg aacagaaata ccaaaattat ttcagaaatg 600

tacaactttt tacctagtac aaggcaacat ataggttgta aatgtgttta aaacaggtct 660

ttgtcttgct atggggagaa aagacatgaa tatgattagt aaagaaatga cacttttcat 720

gtgtgatttc ccctccaagg tatggttaat aagtttcact gacttagaac caggcgagag 780

acttgtggcc tgggagagct ggggaagctt cttaaatgag aaggaatttg agttggatca 840

tctattgctg gcaaagacag aagcctcact gcaagcactg catgggcaag cttggctgta 900

gaaggagaca gagctggttg ggaagacatg gggaggaagg acaaggctag atcatgaaga 960

accttgacgg cattgctccg tctaagtcat gagctgagca gggagatcct ggttggtgtt 1020

gcagaaggtt tactctgtgg ccaaaggagg gtcaggaagg atgagcattt agggcaagga 1080

gaccaccaac agccctcagg tcagggtgag gatggcctct gctaagctca aggcgtgagg 1140

atgggaagga gggaggtatt cgtaaggatg ggaaggaggg aggtattcgt gcagcatatg 1200

aggatgcaga gtcagcagaa ctg 1223

<210> 1106

<211> 215

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1106

gaacgctgac gtcatcaacc cgctccaagg aatcgcgggc ccagtgtcac taggcgggaa 60

cacccagcgc gcgtgcgccc tggcaggaag atggctgtga gggacagggg agtggcgccc 120

tgcaatattt gcatgtcgct atgtgttctg ggaaatcacc ataaacgtga aatgtctttg 180

gatttgggaa tcttataagt tctgtatgag accac 215

<210> 1107

<211> 141

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1107

cctgcaggca gctgcgcgct cgctcgctca cctaggccgc ccgggcaaag cccgggcgtc 60

gggcgacctt tggtcgcccg gcctaggtga gcgagcgagc gcgcagagag ggagtggcca 120

actccatcac taggggttcc t 141

<210> 1108

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1108

gcgcgctcgc tcgctcacc 19

<210> 1109

<211> 22

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1109

ctaggtgagc gagcgagcgc gc 22

<210> 1110

<211> 75

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1110

cctgcaggac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg 60

cctcagtcct gcagg 75

<210> 1111

<211> 130

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1111

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagt 60

gcgggcgacc aaaggtcgcc cgacgcccgg gcgcactcag tgagcgagcg agcgcgcagc 120

tgcctgcagg 130

<210> 1112

<211> 142

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1112

cctgcaggca gctgcgcgct cgctcgctcc ctaggactga ggccgcccgg gcgtcgggcg 60

acctttggtc gcccggcctc agtcctaggg agcgagcgag cgcgcagaga gggagtggcc 120

aactccatca ctaggggttc ct 142

<210> 1113

<211> 80

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1113

gcgcgctcgc tcgctcactg agtgcgggcg accaaaggtc gcccgacgcc cgggcgcact 60

cagtgagcga gcgagcgcgc 80

<210> 1114

<211> 21

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1114

gcgcgctcgc tcgctcactg a 21

<210> 1115

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1115

gtgagcgagc gagcgcgc 18

<210> 1116

<211> 89

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1116

gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgactttgtc 60

gcccggcctc agtgagcgag cgagcgcgc 89

<210> 1117

<211> 89

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1117

gcgcgctcgc tcgctcactg aggccgggcg acaaagtcgc ccgacgcccg ggctttgccc 60

gggcggcctc agtgagcgag cgagcgcgc 89

<210> 1118

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1118

gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgattttcgc 60

ccggcctcag tgagcgagcg agcgcgc 87

<210> 1119

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1119

gcgcgctcgc tcgctcactg aggccgggcg aaaatcgccc gacgcccggg ctttgcccgg 60

gcggcctcag tgagcgagcg agcgcgc 87

<210> 1120

<211> 85

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1120

gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgtttcgccc 60

ggcctcagtg agcgagcgag cgcgc 85

<210> 1121

<211> 85

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1121

gcgcgctcgc tcgctcactg aggccgggcg aaacgcccga cgcccgggct ttgcccgggc 60

ggcctcagtg agcgagcgag cgcgc 85

<210> 1122

<211> 89

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1122

gcgcgctcgc tcgctcactg aggccgcccg ggaaacccgg gcgtcgggcg acctttggtc 60

gcccggcctc agtgagcgag cgagcgcgc 89

<210> 1123

<211> 89

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1123

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggtttccc 60

gggcggcctc agtgagcgag cgagcgcgc 89

<210> 1124

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1124

gcgcgctcgc tcgctcactg aggccgcccg gaaaccgggc gtcgggcgac ctttggtcgc 60

ccggcctcag tgagcgagcg agcgcgc 87

<210> 1125

<211> 87

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1125

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cggtttccgg 60

gcggcctcag tgagcgagcg agcgcgc 87

<210> 1126

<211> 85

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1126

gcgcgctcgc tcgctcactg aggccgcccg aaacgggcgt cgggcgacct ttggtcgccc 60

ggcctcagtg agcgagcgag cgcgc 85

<210> 1127

<211> 85

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1127

gcgcgctcgc tcgctcactg aggccggggcg accaaaggtc gcccgacgcc cgtttcgggc 60

ggcctcagtg agcgagcgag cgcgc 85

<210> 1128

<211> 83

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1128

gcgcgctcgc tcgctcactg aggccgccca aagggcgtcg ggcgaccttt ggtcgcccgg 60

cctcagtgag cgagcgagcg cgc 83

<210> 1129

<211> 83

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1129

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc ctttgggcgg 60

cctcagtgag cgagcgagcg cgc 83

<210> 1130

<211> 81

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1130

gcgcgctcgc tcgctcactg aggccgccaa aggcgtcggg cgacctttgg tcgcccggcc 60

tcagtgagcg agcgagcgcg c 81

<210> 1131

<211> 81

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1131

gcgcgctcgc tcgctcactg aggccggggcg accaaaggtc gcccgacgcc tttggcggcc 60

tcagtgagcg agcgagcgcg c 81

<210> 1132

<211> 79

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1132

gcgcgctcgc tcgctcactg aggccgcaaa gcgtcgggcg acctttggtc gcccggcctc 60

agtgagcgag cgagcgcgc 79

<210> 1133

<211> 79

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1133

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgct ttgcggcctc 60

agtgagcgag cgagcgcgc 79

<210> 1134

<211> 81

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1134

ctgcgcgctc gctcgctcac tgaggccgaa acgtcgggcg acctttggtc gcccggcctc 60

agtgagcgag cgagcgcgca g 81

<210> 1135

<211> 81

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1135

ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg tttcggcctc 60

agtgagcgag cgagcgcgca g 81

<210> 1136

<211> 72

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1136

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacggc ctcagtgagc 60

gagcgagcgc gc 72

<210> 1137

<211> 80

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1137

gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggcggcct 60

cagtgagcga gcgagcgcgc 80

<210> 1138

<211> 79

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1138

gcgcgctcgc tcgctcactg aggcgcccgg gcgtcgggcg acctttggtc gcccggcctc 60

agtgagcgag cgagcgcgc 79

<210> 1139

<211> 48

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1139

ggagtcaaag ttctgtttgc cctgatctgc atcgctgtgg ccgaggcc 48

<210> 1140

<211> 99

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1140

attcatacca acttgaagaa aaagttcagc ctcttcatcc tggtctttct cctgttcgca 60

gtcatctgtg tttggaagaa agggagcgac tatgaggcc 99

<210> 1141

<211> 588

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1141

gcccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt 60

gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc 120

ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag 180

gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac 240

aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc 300

tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc 360

acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca 420

aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt 480

gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg 540

gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaacc 588

<210> 1142

<211> 31

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1142

gcccgggcgt cgggcgacct ttggtcgccc g 31

<210> 1143

<211> 31

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1143

gcccgggcgt cgggcgacct ttggtcgccc g 31

<210> 1144

<211> 2218

<212> DNA

<213> Homo sapiens

<400> 1144

cagggcagcc ttcagtctga ttcaggagaa cgaggtcctc ttcaccatgt gcttcatccc 60

cctggtctgc tggatcgtgt gcactggact gaaacagcag atggagagtg gcaagagcct 120

tgcccagaca tccaagacct ccaccgcggt gtacgtcttc ttcctttcca gtttgctgca 180

gccccgggga gggagccagg agcacggcct ctgcgcccac ctctgggggc tctgctcttt 240

ggctgcagat ggaatctgga accagaaaat cctgtttgaa gagtccgacc tcaggaatca 300

tggactgcag aaggcggatg tgtctgcttt cctgaggatg aacctgttcc aaaaggaagt 360

ggactgcgag aagttctaca gcttcatcca catgactttc caggagttct ttgccgccat 420

gtactacctg ctggaagagg aaaaggaagg aaggacgaac gttccaggga gtcgtttgaa 480

gcttcccagc cgagacgtga cagtccttct ggaaaactat ggcaaattcg aaaaggggta 540

tttgattttt gttgtacgtt tcctctttgg cctggtaaac caggagagga cctcctactt 600

ggagaagaaa ttaagttgca tgatctctca gcaaatcagg ctggagctgc tgaaatggat 660

tgaagtgaaa gccaaagcta aaaagctgca tgatcagccc agccagctgg aattgttcta 720

ctgtttgtac gagatgcagg aggaggactt cgtgcaaagg gccatggact atttccccaa 780

gattgagatc aatctctcca ccagaatgga ccacatggtt tcctcctttt gcattgagaa 840

ctgtcatcgg gtggagtcac tgtccctggg gtttctccat aacatgccca aggaggaaga 900

ggagggaggaa aaggaaggcc gacaccttga tatggtgcag tgtgtcctcc caagctcctc 960

tcatgctgcc tgttctcatg ggttggggcg ctgtggcctc tcccatgagt gctgcttcga 1020

catctccttg gtcctcagca gcaaccagaa gctggtggag ctggacctga gtgacaacgc 1080

cctcggtgac ttcggaatca gacttctgtg tgtgggactg aagcacctgt tgtgcaatct 1140

gaagaagctc tggttggtga attctgcctt acgtcagtct gttgttcagc tttgtcctcg 1200

gtactcagca ctaatcagaa tctcacgcac ctttactgcg aggcaacact ctcggagaca 1260

agggatcaaa ctactctgtg agggactctt gcaccccgac tgcaagcttc aggtgttgga 1320

attagacaac tgcaacctca cgtcacactg ctgctgggat ctttccacac ttctgacctc 1380

cagccagagc ctgcgaaagc tgagcctggg caacaatgac ctgggcgacc tggggggtcat 1440

gatgttctgt gaagtgctga aacagcagag ctgcctcctg cagaacctgg ggttgtctga 1500

aatgtatttc aattatgaga caaaaagtgc gttagaaaca cttcaagaag aaaagcctga 1560

gctgaccgtc gtctttgagc cttcttggta ggagtggaaa cggggctgcc agacgccagt 1620

gttctccggt ccctccagct gggggccctc aggtggagag agctgcgatc catccaggcc 1680

aagaccacag ctctgtgatc cttccggtgg agtgtcggag aagagagctt gccgacgatg 1740

ccttcctgtg cagagcttgg gcatctcctt tacgccaggg tgaggaagac accaccacaa 1800

tgacagcatc gggtgttgtt gtcatcacag cgcctcagtt agaggatgtt cctctggtga 1860

cctcatgtaa ttagctcatt caataaagca ctttctttat ttttctcttc tctgtctaac 1920

cttctttttc ctatcttttt ttcttctttg ttctgtttac ttttgctcat atcatcattc 1980

ccgctatctt tctattaact gaccataaca cagaactagt tgactatata ttatgttgaa 2040

attttatggc agctatttat ttatttaaat tttttgtaat agttttgttt tctaataaga 2100

aaaatccatg ctttttgtag ctggttgaaa attcaggaat atgtaaaact ttttggtatt 2160

taattaaatt gattcctttt cttaatttta aaaaaaaaaa aaaaaaaaaa aaaaaaaa 2218

<210> 1145

<211> 58

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1145

ccggccacag tgtaacctgc agaaactcga gtttctgcag gttacactgt ggtttttg 58

<210> 1146

<211> 16

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1146

gcgcgctcgc tcgctc 16

<210> 1147

<211> 518

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1147

tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta 60

ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc 120

aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg 180

gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 240

gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 300

agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 360

ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga 420

cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg 480

gcagtacatc tacgtattag tcatcgctat taccatgg 518

<210> 1148

<211> 777

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1148

aggctcagag gcacacagga gtttctgggc tcaccctgcc cccttccaac ccctcagttc 60

ccatcctcca gcagctgttt gtgtgctgcc tctgaagtcc acactgaaca aacttcagcc 120

tactcatgtc cctaaaatgg gcaaacattg caagcagcaa acagcaaaca cacagccctc 180

cctgcctgct gaccttggag ctggggcaga ggtcagagac ctctctgggc ccatgccacc 240

tccaacatcc actcgacccc ttggaatttc ggtggagagg agcagaggtt gtcctggcgt 300

ggtttaggta gtgtgagagg gtccgggttc aaaaccactt gctgggtggg gagtcgtcag 360

taagtggcta tgccccgacc ccgaagcctg tttccccatc tgtacaatgg aaatgataaa 420

gacgcccatc tgatagggtt tttgtggcaa ataaacattt ggtttttttg ttttgttttg 480

ttttgttttt tgagatggag gtttgctctg tcgcccaggc tggagtgcag tgacacaatc 540

tcatctcacc acaaccttcc cctgcctcag cctcccaagt agctgggatt acaagcatgt 600

gccaccacac ctggctaatt ttctattttt agtagagacg ggtttctcca tgttggtcag 660

cctcagcctc ccaagtaact gggattacag gcctgtgcca ccacacccgg ctaatttttt 720

ctatttttga cagggacggg gtttcaccat gttggtcagg ctggtctaga ggtaccg 777

<210> 1149

<211> 427

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1149

gagtcaatgg gaaaaaccca ttggagccaa gtacactgac tcaataggga ctttccatg 60

ggttttgccc agtacataag gtcaataggg ggtgagtcaa caggaaagtc ccattggagc 120

caagtacatt gagtcaatag ggactttcca atgggttttg cccagtacat aaggtcaatg 180

ggaggtaagc caatgggttt ttcccattac tgacatgtat actgagtcat tagggacttt 240

ccaatgggtt ttgcccagta cataaggtca ataggggtga atcaacagga aagtcccatt 300

ggagccaagt acactgagtc aatagggact ttccattggg ttttgcccag tacaaaaggt 360

caatagggg tgagtcaatg ggtttttccc attattggca catacataag gtcaataggg 420

gtgacta 427

<210> 1150

<211> 83

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1150

cgggggaggc tgctggtgaa tattaaccaa ggtcacccca gttatcggag gagcaaacag 60

gggctaagtc cacacgcgtg gta 83

<210> 1151

<211> 777

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1151

aggctcagag gcacacagga gtttctgggc tcaccctgcc cccttccaac ccctcagttc 60

ccatcctcca gcagctgttt gtgtgctgcc tctgaagtcc acactgaaca aacttcagcc 120

tactcatgtc cctaaaatgg gcaaacattg caagcagcaa acagcaaaca cacagccctc 180

cctgcctgct gaccttggag ctggggcaga ggtcagagac ctctctgggc ccatgccacc 240

tccaacatcc actcgacccc ttggaatttc ggtggagagg agcagaggtt gtcctggcgt 300

ggtttaggta gtgtgagagg gtccgggttc aaaaccactt gctgggtggg gagtcgtcag 360

taagtggcta tgccccgacc ccgaagcctg tttccccatc tgtacaatgg aaatgataaa 420

gacgcccatc tgatagggtt tttgtggcaa ataaacattt ggtttttttg ttttgttttg 480

ttttgttttt tgagatggag gtttgctctg tcgcccaggc tggagtgcag tgacacaatc 540

tcatctcacc acaaccttcc cctgcctcag cctcccaagt agctgggatt acaagcatgt 600

gccaccacac ctggctaatt ttctattttt agtagagacg ggtttctcca tgttggtcag 660

cctcagcctc ccaagtaact gggattacag gcctgtgcca ccacacccgg ctaatttttt 720

ctatttttga cagggacggg gtttcaccat gttggtcagg ctggtctaga ggtactg 777

<210> 1152

<211> 66

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1152

gtttgctgct tgcaatgttt gcccatttta gggtggacac aggacgctgt ggtttctgag 60

ccaggg 66

<210> 1153

<211> 212

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1153

ggaggggtgg agtcgtgacc cctaaaatgg gcaaacattg caagcagcaa acagcaaaca 60

cacagccctc cctgcctgct gaccttggag ctggggcaga ggtcagagac ctctctgggc 120

ccatgccacc tccaacatcc actcgacccc ttggaatttc ggtggagagg agcagaggtt 180

gtcctggcgt ggtttaggta gtgtgagagg gg 212

<210> 1154

<211> 330

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1154

aggctcagag gcacacagga gtttctgggc tcaccctgcc cccttccaac ccctcagttc 60

ccatcctcca gcagctgttt gtgtgctgcc tctgaagtcc acactgaaca aacttcagcc 120

tactcatgtc cctaaaatgg gcaaacattg caagcagcaa acagcaaaca cacagccctc 180

cctgcctgct gaccttggag ctggggcaga ggtcagagac ctctctgggc ccatgccacc 240

tccaacatcc actcgacccc ttggaatttc ggtggagagg agcagaggtt gtcctggcgt 300

ggtttaggta gtgtgagagg ggtacccggg 330

<210> 1155

<211> 194

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1155

ccctaaaatg ggcaaacatt gcaagcagca aacagcaaac acacagccct ccctgcctgc 60

tgaccttgga gctggggcag aggtcagaga cctctctggg cccatgccac ctccaacatc 120

cactcgaccc cttggaattt ttcggtggag aggagcagag gttgtcctgg cgtggtttag 180

gtagtgtgag agg 194

<210> 1156

<211> 240

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1156

gggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg ctgaaagaac 60

cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga 120

agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc 180

ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccacta 240

<210> 1157

<211> 73

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1157

cgggggaggc tgctggtgaa tattaaccaa ggtcacccca gttatcggag gagcaaacag 60

gggctaagtc cac 73

<210> 1158

<211> 100

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1158

aggttaattt ttaaaaagca gtcaaaagtc caagtggccc ttggcagcat ttactctctc 60

tgtttgctct ggttaataat ctcaggagca caaacattcc 100

<210> 1159

<211> 296

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1159

gttacataac ttatggtaaa tggcctgcct ggctgactgc ccaatgaccc ctgcccaatg 60

atgtcaataa tgatgtatgt tcccatgtaa tgccaatagg gactttccat tgatgtcaat 120

gggtggagta tttatggtaa ctgcccactt ggcagtacat caagtgtatc atatgccaag 180

tatgccccct attgatgtca atgatggtaa atggcctgcc tggcattatg cccagtacat 240

gaccttatgg gactttccta cttggcagta catctatgta ttagtcattg ctatta 296

<210> 1160

<211> 235

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1160

ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc 60

agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa 120

gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc 180

cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtccc 235

<210> 1161

<211> 1127

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1161

ggagtcgctg cgacgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc 60

gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc 120

tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga 180

aagccttgag gggctccggg agggcccttt gtgcgggggg gagcggctcg gggggtgcgt 240

gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggcccgcg ctgcccggcg gctgtgagcg 300

ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg 360

gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt 420

gtgcgtgggg gggtgagcag ggggtgtggg cgcggcggtc gggctgtaac ccccccctgc 480

acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc 540

gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg 600

cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg 660

gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagagg 720

cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca 780

ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg 840

agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc 900

cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg 960

tgaccggcgg ctctagagcc tctgctaacc atgttttagc cttcttcttt ttcctacagc 1020

tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt gtcgacagaa ttcctcgaag 1080

atccgaaggg gttcaagctt ggcattccgg tactgttggt aaagcca 1127

<210> 1162

<211> 93

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1162

ctctaaggta aatataaaat ttttaagtgt ataatgtgtt aaactactga ttctaattgt 60

ttctctcttt tagattccaa cctttggaac tga 93

<210> 1163

<211> 54

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1163

gccctgtctc ctcagcttca ggcaccacca ctgacctggg acagtgaatc cgga 54

<210> 1164

<211> 173

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1164

ctgccttctc cctcctgtga gtttggtaag tcactgactg tctatgcctg ggaaagggtg 60

ggcaggagat ggggcagtgc aggaaaagtg gcactatgaa ccctgcagcc ctagacaatt 120

gtactaacct tcttctcttt cctctcctga caggttggtg tacagtagct tcc 173

<210> 1165

<211> 91

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1165

aagaggtaag ggtttaaggg atggttggtt ggtggggtat taatgtttaa ttacctggag 60

cacctgcctg aaatcacttt ttttcaggtt g 91

<210> 1166

<211> 54

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1166

gccctgtctc ctcagcttca ggcaccacca ctgacctggg acagtgaata atta 54

<210> 1167

<211> 147

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1167

gccctgtctc ctcagcttca ggcaccacca ctgacctggg acagtgaatc cggactctaa 60

ggtaaatata aaatttttaa gtgtataatg tgttaaacta ctgattctaa ttgtttctct 120

cttttagatt ccaacctttg gaactga 147

<210> 1168

<211> 147

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1168

gccctgtctc ctcagcttca ggcaccacca ctgacctggg acagtgaata attactctaa 60

ggtaaatata aaatttttaa gtgtataatg tgttaaacta ctgattctaa ttgtttctct 120

cttttagatt ccaacctttg gaactga 147

<210> 1169

<211> 48

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1169

tcctcagctt caggcaccac cactgacctg ggacagtgaa tcgccacc 48

<210> 1170

<211> 128

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1170

gctagcaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc 60

cttgcgtgcc ttgaattact gacactgaca tccacttttt ctttttctcc acaggtttaa 120

acgccacc 128

<210> 1171

<211> 98

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1171

aagaggtaag ggtttaagtt atcgttagtt cgtgcaccat taatgtttaa ttacctggag 60

cacctgcctg aaatcatttt tttttcaggt tggctagt 98

<210> 1172

<211> 172

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1172

gcttagtgct gagcacatcc agtgggtaaa gttccttaaa atgctctgca aagaaattgg 60

gacttttcat taaatcagaa attttacttt tttcccctcc tgggagctaa agatatttta 120

gagaagaatt aaccttttgc ttctccagtt gaacatttgt agcaataagt ca 172

<210> 1173

<211> 160

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1173

gccctgtctc ctcagcttca ggcaccacca ctgacctggg acagtgaatc cggactctaa 60

ggtaaatata aaatttttaa gtgtataatg tgttaaacta ctgattctaa ttgtttctct 120

cttttagatt ccaacctttg gaactgaatt ctagaccacc 160

<210> 1174

<211> 29

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1174

accactttca caatctgcta gcaaaggtt 29

<210> 1175

<211> 133

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1175

gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga 60

cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc 120

tttctctcca cag 133

<210> 1176

<211> 341

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1176

tgggcaggaa ctgggcactg tgcccagggc atgcactgcc tccacgcagc aaccctcaga 60

gtcctgagct gaaccaagaa ggaggagggg gtcgggcctc cgaggaaggc ctagccgctg 120

ctgctgccag gaattccagg ttggaggggc ggcaacctcc tgccagcctt caggccactc 180

tcctgtgcct gccagaagag acagagcttg aggagagctt gaggagca ggaaagcctc 240

ccccgttgcc cctctggatc cactgcttaa atacggacga ggacagggcc ctgtctcctc 300

agcttcaggc accaccactg acctgggaca gtgaatcgac a 341

<210> 1177

<211> 316

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1177

tctagagaag ctttattgcg gtagtttatc acagttaaat tgctaacgca gtcagtgctt 60

ctgacacaac agtctcgaac ttaagctgca gtgactctct taaggtagcc ttgcagaagt 120

tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta aggagaccaa 180

tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt 240

cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccagtt caattacagc 300

tcttaaggcc ctgcag 316

<210> 1178

<211> 76

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1178

caaagtccag gcccctctgc tgcagcgccc gcgcgtccag aggccctgcc agacacgcgc 60

gaggttcgag gctgag 76

<210> 1179

<211> 127

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1179

agaatgatga aaaccgaggt tggaaaaggt tgtgaaacct tttaactctc cacagtggag 60

tccattattt cctctggctt cctcaaattc atattcacag ggtcgttggc tgtgggttgc 120

aattacc 127

<210> 1180

<211> 80

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1180

atagcagagc aatcaccacc aagcctggaa taactgcaag ggctctgctg acatcttcct 60

gaggtgccaa ggaaatgagg 80

<210> 1181

<211> 208

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1181

gggtcaccac cacctccaca gcacagacag acactcagga gccagccagc caggtaagtt 60

tagtcttttt gtcttttatt tcaggtcccg gatccggtgg tggtgcaaat caaagaactg 120

ctcctcagtg gatgttgcct ttacttctag gcctgtacgg aagtgttact tctgctctaa 180

aagctgcgga attgtacccg cggccgcg 208

<210> 1182

<211> 159

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1182

aagcttctgc cttctccctc ctgtgagttt ggtaagtcac tgactgtcta tgcctgggaa 60

agggtgggca ggagatgggg cagtgcagga aaagtggcac tatgaaccct gcagccctag 120

acaattgtac taaccttctt ctctttcctc tcctgacag 159

<210> 1183

<211> 36

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1183

cgcgcctagc agtgtcccag ccgggttcgt gtcgcc 36

<210> 1184

<211> 141

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1184

acgccgcctg ggtcccagtc cccgtcccat cccccggcgg cctaggcagc gtttccagcc 60

ccgagaactt tgttcttttt gtcccgcccc ctgcgcccaa ccgcctgcgc cgccttccgg 120

cccgagttct ggagactcaa c 141

<210> 1185

<211> 110

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1185

gttggatgaa accttcctcc tactgcacag cccgcccccc tacagccccg gtccccacgc 60

ctagaagaca gcggaactaa gaaaagaaga ggcctgtgga cagaacaatc 110

<210> 1186

<211> 164

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1186

ggtggggcgg ggttgagtcg gaaccacaat agccaggcga agaaactaca actcccagg 60

cgtcccggag caggccaacg ggactacggg aagcagcggg cagcggcccg cgggaggcac 120

ctcggagatc tgggtgcaaa agcccagggt taggaaccgt aggc 164

<210> 1187

<211> 127

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1187

ggccaccgga attaaccctt cagggctggg ggccgcgcta tgccccgccc cctccccagc 60

cccagacacg gaccccgcag gagatgggtg cccccatccg cacactgtcc tttggccacc 120

ggacatc 127

<210> 1188

<211> 341

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1188

tgggcaggaa ctgggcactg tgcccagggc atgcactgcc tccacgcagc aaccctcaga 60

gtcctgagct gaaccaagaa ggaggagggg gtcgggcctc cgaggaaggc ctagccgctg 120

ctgctgccag gaattccagg ttggaggggc ggcaacctcc tgccagcctt caggccactc 180

tcctgtgcct gccagaagag acagagcttg aggagagctt gaggagca ggaaagcctc 240

ccccgttgcc cctctggatt cactgcttaa atacggacga ggacagggcc ctgtctcctc 300

agcttcaggc accaccactg acctgggaca gtgaatcgac a 341

<210> 1189

<211> 77

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1189

tccataaagt aggaaacact acacgattcc ataaagtagg aaacactaca tcactccata 60

aagtaggaaa cactaca 77

<210> 1190

<211> 88

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1190

tgaaagatgg atttccaagg ttaattcatt ggaattgaaa attaacagag atctagagct 60

gaattcctgc agccagggg atcagcct 88

<210> 1191

<211> 395

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1191

taaaatacag catagcaaaa ctttaacctc caaatcaagc ctctacttga atccttttct 60

gagggatgaa taaggcatag gcatcagggg ctgttgccaa tgtgcattag ctgtttgcag 120

cctcaccttc tttcatggag tttaagatat agtgtatttt cccaaggttt gaactagctc 180

ttcatttctt tatgttttaa atgcactgac ctcccacatt ccctttttag taaaatattc 240

agaaataatt taaatacatc attgcaatga aaataaatgt tttttattag gcagaatcca 300

gatgctcaag gcccttcata atatccccca gtttagtagt tggacttagg gaacaaagga 360

acctttaata gaaattggac agcaagaaag cgagc 395

<210> 1192

<211> 800

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1192

agtcaatatg ttcaccccaa aaaagctgtt tgttaacttg ccaacctcat tctaaaatgt 60

atatagaagc ccaaaagaca ataacaaaaa tattcttgta gaacaaaatg ggaaagaatg 120

ttccactaaa tatcaagatt tagagcaaag catgagatgt gtggggatag acagtgaggc 180

tgataaaata gagtagagct cagaaacaga cccattgata tatgtaagtg acctatgaaa 240

aaaatatggc attttacaat gggaaaatga tggtcttttt cttttttaga aaaacaggga 300

aatatattta tatgtaaaaa ataaaaggga acccatatgt cataccatac acacaaaaaa 360

attccagtga attataagtc taaatggaga aggcaaaact ttaaatcttt tagaaaataa 420

tatagaagca tgccatcaag acttcagtgt agagaaaaat ttcttatgac tcaaagtcct 480

aaccacaaag aaaagattgt taattagatt gcatgaatat taagacttat ttttaaaatt 540

aaaaaaccat taagaaaagt caggccatag aatgacagaa aatatttgca acaccccagt 600

aaagagaatt gtaatatgca gattataaaa agaagtctta caaatcagta aaaaataaaa 660

ctagacaaaa atttgaacag atgaaagaga aactctaaat aatcattaca catgagaaac 720

tcaatctcag aaatcagaga actatcattg catatacact aaattagaga aatattaaaa 780

ggctaagtaa catctgtggc 800

<210> 1193

<211> 407

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1193

aattatctct aaggcatgtg aactggctgt cttggttttc atctgtactt catctgctac 60

ctctgtgacc tgaaacatat ttataattcc attaagctgt gcatatgata gatttatcat 120

atgtattttc cttaaaggat ttttgtaaga actaattgaa ttgatacctg taaagtcttt 180

atcacactac ccaataaata ataaatctct ttgttcagct ctctgtttct ataaatatgt 240

accagtttta ttgtttttag tggtagtgat tttattctct ttctatatat atacacacac 300

atgtgtgcat tcataaatat atacaatttt tatgaataaa aaattattag caatcaatat 360

tgaaaaccac tgatttttgt ttatgtgagc aaacagcaga ttaaaag 407

<210> 1194

<211> 186

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1194

catcacattt aaaagcatct cagcctacca tgagaataag agaaagaaaa tgaagatcaa 60

aagcttattc atctgttttt ctttttcgtt ggtgtaaagc caacaccctg tctaaaaaac 120

ataaatttct ttaatcattt tgcctctttt ctctgtgctt caattaataa aaaatggaaa 180

gaatct 186

<210> 1195

<211> 395

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1195

taaaatacag catagcaaaa ctttaacctc caaatcaagc ctctacttga atccttttct 60

gagggatgaa taaggcatag gcatcagggg ctgttgccaa tgtgcattag ctgtttgcag 120

cctcaccttc tttcatggag tttaagatat agtgtatttt cccaaggttt gaactagctc 180

ttcatttctt tatgttttaa atgcactgac ctcccacatt ccctttttag taaaatattc 240

agaaataatt taaatacatc attgcaatga aaataaatgt tttttattag gcagaatcca 300

gatgctcaag gcccttcata atatccccca gtttagtagt tggacttagg gaacaaagga 360

acctttaata gaaattggac agcaagaaag ccagc 395

<210> 1196

<211> 580

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1196

gagcatctta ccgccattta ttcccatatt tgttctgttt ttcttgattt gggtatacat 60

ttaaatgtta ataaaacaaa atggtggggc aatcatttac atttttaggg atatgtaatt 120

actagttcag gtgtattgcc acaagacaaa catgttaaga aactttcccg ttatttacgc 180

tctgttcctg ttaatcaacc tctggattac aaaatttgtg aaagattgac tgatattctt 240

aactatgttg ctccttttac gctgtgtgga tatgctgctt tatagcctct gtatctagct 300

attgcttccc gtacggcttt cgttttctcc tccttgtata aatcctggtt gctgtctctt 360

ttagaggagt tgtggcccgt tgtccgtcaa cgtggcgtgg tgtgctctgt gtttgctgac 420

gcaaccccca ctggctgggg cattgccacc acctgtcaac tcctttctgg gactttcgct 480

ttccccctcc cgatcgccac ggcagaactc atcgccgcct gccttgcccg ctgctggaca 540

ggggctaggt tgctgggcac tgataattcc gtggtgttgt 580

<210> 1197

<211> 64

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1197

cctcgccccg gacctgccct cccgccaggt gcacccacct gcaataaatg cagcgaagcc 60

ggga 64

<210> 1198

<211> 247

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1198

gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 60

gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 120

cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa 180

ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 240

tccgtgg 247

<210> 1199

<211> 144

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1199

aaatacatca ttgcaatgaa aataaatgtt ttttattagg cagaatccag atgctcaagg 60

cccttcataa tatcccccag tttagtagtt ggacttaggg aacaaaggaa cctttaatag 120

aaattggaca gcaagaaagc gagc 144

<210> 1200

<211> 62

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1200

gagcatctta ccgccattta ttcccatatt tgttctgttt ttcttgattt gggtatacat 60

tt 62

<210> 1201

<211> 49

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1201

aataaaagat ctttattttc attagatctg tgtgttggtt ttttgtgtg 49

<210> 1202

<211> 54

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1202

gcggccgcaa taaaagatca gagctctaga gatctgtgtg ttggtttttt gtgt 54

<210> 1203

<211> 74

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1203

ggatccaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgttttc 60

ctgtaacgat cggg 74

<210> 1204

<211> 143

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1204

ctcgatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct 60

gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg 120

tgtgggaggt tttttaaact agt 143

<210> 1205

<211> 228

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1205

ctactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccttg ccttccttga 60

ccctggaagg tgccactccc actngtccttt cctaataaaa tgaggaaatt gcatcacatt 120

gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg 180

attgggaaga caatagcagg catgctgggg atgcagtggg ctctatgg 228

<210> 1206

<211> 222

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1206

cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa 60

aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca 120

ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt 180

gggaggtttt ttaaagcaag taaaacctct acaaatgtgg ta 222

<210> 1207

<211> 226

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1207

ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa 60

aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc 120

aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg 180

tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatgg 226

<210> 1208

<211> 129

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1208

gttaacaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 60

aaaaaaaaaa tgcatccccc cccccccccc cccccccccc ccccccaaag gctcttttca 120

gagccacca 129

<210> 1209

<211> 232

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1209

gcggccgcgg ggatccagac atgataagat acattgatga gtttggacaa accacaacta 60

gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa 120

ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg 180

ttcaggggga ggtgtgggag gttttttagt cgaccatgct ggggagagat ct 232

<210> 1210

<211> 135

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1210

gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga 60

aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc 120

tgcaataaac aagtt 135

<210> 1211

<211> 49

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1211

cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttc 49

<210> 1212

<211> 226

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1212

ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa 60

cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg 120

ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 180

tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgg 226

<210> 1213

<211> 416

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1213

catcacattt aaaagcatct cagcctacca tgagaataag agaaagaaaa tgaagatcaa 60

aagcttattc atctgttttt ctttttcgtt ggtgtaaagc caacaccctg tctaaaaaac 120

ataaatttct ttaatcattt tgcctctttt ctctgtgctt caattaataa aaaatggaaa 180

gaatctaata gagtggtaca gcactgttat ttttcaaaga tgtgttgcta tcctgaaaat 240

tctgtaggtt ctgtggaagt tccagtgttc tctcttattc cacttcggta gaggatttct 300

agtttcttgt gggctaatta aataaatcat taatactctt ctaagttatg gattataaac 360

attcaaaata atattttgac attatgataa ttctgaataa aagaacaaaa accatg 416

<210> 1214

<211> 415

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1214

atcacattta aaagcatctc agcctaccat gagaataaga gaaagaaaat gaagatcaaa 60

agcttattca tctgtttttc tttttcgttg gtgtaaagcc aacaccctgt ctaaaaaaca 120

taaatttctt taatcatttt gcctcttttc tctgtgcttc aattaataaa aaatggaaag 180

aatctaatag agtggtacag cactgttatt tttcaaagat gtgttgctat cctgaaaatt 240

ctgtaggttc tgtggaagtt ccagtgttct ctcttattcc acttcggtag aggatttcta 300

gtttcttgtg ggctaattaa ataaatcatt aatactcttc taagttatgg attataaaca 360

ttcaaaataa tattttgaca ttatgataat tctgaataaa agaacaaaaa ccatg 415

<210> 1215

<211> 122

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1215

taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta 60

tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag 120

tt 122

<210> 1216

<211> 133

<212> DNA

<213> Artificial sequence

<220>

<223> Description of an artificial sequence: synthetic polynucleotide

<400> 1216

tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat 60

aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg 120

gaggtttttt aaa 133

<210> 1217

<211> 64

<212> DNA

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic

oligonucleotide

<400> 1217

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 60

aaaa 64

<210> 1218

<211> 4

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic peptide

<400> 1218

Tyr Val Ala Asp

1

<210> 1219

<211> 4

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic peptide

<400> 1219

Tyr Val His Asp

1

<210> 1220

<211> 4

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Description of the artificial sequence: synthetic peptide

<400> 1220

Trp Glu His Asp

1

<---

Claims (139)

1. A method for inhibiting an immune response in which at least one transgene is expressed in a cell, characterized in that said method comprises: introducing into said cell at least one immune response inhibitor, wherein the immune response inhibitor is a Toll-like receptor (TLR) inhibitor, an NLRP3 inflammasome inhibitor, an AIM2 inflammasome inhibitor, a caspase-1 inhibitor, or a cyclo-GMP-AMP synthase (cGAS) inhibitor; and a non-viral capsidless DNA vector with covalently closed ends (ccDNA vector), wherein said ccDNA vector comprises said at least one transgene located between two flanking inverted terminal repeats (ITR). 2. The method according to claim 1, characterized in that said immune response is an innate immune response. 3. The method according to claim 1 or 2, characterized in that said immune response inhibitor is an innate immune response inhibitor. 4. The method according to any one of claims 1-3, characterized in that said ccDNA vector additionally encodes said at least one immune response inhibitor. 5. The method according to any one of paragraphs 1-4, characterized in that said immune response inhibitor is administered separately, in different compositions, from said ccDNA vector. 6. The method according to claim 5, characterized in that said immune response inhibitor is administered at the same time as said ccDNA vector. 7. The method according to claim 5, characterized in that said immune response inhibitor is administered before the administration of said ccDNA vector. 8. The method according to claim 5, characterized in that said immune response inhibitor is administered after the administration of said ccDNA vector. 9. The method according to any one of paragraphs 1-4, characterized in that said immune response inhibitor and said ccDNA vector are administered at the same time in the same composition. 10. The method according to claim 1, characterized in that said TLR inhibitor is a TLR9 inhibitor. 11. The method according to claim 1, characterized in that said TLR inhibitor is rapamycin or a rapamycin analogue. 12. The method according to claim 1, characterized in that said AIM2 inflammasome inhibitor is A151 of SEQ ID NO: 602 or a variant thereof that inhibits the AIM2 inflammasome or caspase-1. 13. The method according to claim 1, characterized in that said NLRP3 inflammasome inhibitor, AIM2 inflammasome inhibitor, or caspase-1 inhibitor is an siRNA specific for the NLRP3 inflammasome, AIM2 inflammasome, or caspase-1, respectively. 14. The method according to claim 1, characterized in that said NLRP3 inflammasome inhibitor, AIM2 inflammasome inhibitor, or caspase-1 inhibitor is a siRNA specific for the NLRP3 inflammasome, AIM2 inflammasome, or caspase-1, respectively. 15. The method according to claim 1, characterized in that said NLRP3 inflammasome inhibitor, AIM2 inflammasome inhibitor, or caspase-1 inhibitor is an antibody or antigen-binding fragment that binds the NLRP3 inflammasome, the AIM2 inflammasome, or caspase-1, respectively. 16. The method according to claim 1, characterized in that said NLRP3 inflammasome inhibitor is selected from the group consisting of: MCC950, 16673-34-0, Bay11-7082, glibenclamide, and isoliquiritigenin. 17. The method according to claim 1, characterized in that said AIM2 inflammasome inhibitor is selected from the group consisting of: A151 (SEQ ID NO: 602), miR-223, and a monoclonal antibody to human ASC. 18. The method according to claim 1, characterized in that said caspase-1 inhibitor is selected from the group consisting of: belnakazan, shikonin, acetylshikonin, pralnakazan, Z-VAD-FMK, Z-WEHD-FMK; Ac-YVAD-cmk, Ac-YVAD-CHO and parthenolide. 19. The method according to claim 1, characterized in that said cGAS inhibitor is the Kaposi's sarcoma-associated herpesvirus ORF52 protein of SEQ ID NO: 882 or a variant thereof that inhibits cGAS, or an ortholog of gammaherpesvirus ORF52. 20. The method according to claim 1, characterized in that said cGAS inhibitor is a cytoplasmic isoform of the latency-associated nuclear antigen (LANA) of the herpesvirus associated with Kaposi's sarcoma, from SEQ ID NO: 883. 21. The method according to claim 20, characterized in that said cytoplasmic isoform of LANA of the herpesvirus associated with Kaposi's sarcoma is LANAΔ161 SEQ ID NO: 884. 22. The method according to claim 1, characterized in that said cGAS inhibitor is a siRNA specific for cGAS. 23. The method according to claim 22, characterized in that said miRNA specific for cGAS is selected from the group consisting of miR-25 SEQ ID NO: 885 and miR-93 SEQ ID NO: 886. 24. The method according to claim 1, characterized in that said cGAS inhibitor is a siRNA specific for cGAS. 25. The method according to claim 1, characterized in that said cGAS inhibitor is an antibody or antigen-binding fragment that binds cGAS. 26. The method according to claim 1, characterized in that said cGAS inhibitor is an antimalarial drug. 27. The method according to claim 26, characterized in that said antimalarial drug is an antimalarial drug based on aminoquinoline or aminoacridine. 28. The method according to claim 27, characterized in that said antimalarial drug is selected from the group consisting of quinacrine (QC), 9-amino-6-chloro-2-methoxyacridine (AMCA), hydroxychloroquine (HCQ) and chloroquine (CQ). 29. The method according to claim 1, characterized in that said cGAS inhibitor is a low molecular weight compound that binds to the catalytic pocket of cGAS. 30. The method according to claim 29, characterized in that said low molecular weight compound that binds to the catalytic pocket of cGAS is selected from the group consisting of RU166365, RU281332, RU320521, RU320519, RU320461, RU320462, RU320520, RU320467, and RU320582. 31. The method according to claim 30, characterized in that said low molecular weight compound that binds to the catalytic pocket of cGAS is RU320521. 32. The method according to claim 29, characterized in that said low molecular weight compound that binds to the catalytic pocket of cGAS is PF-06928215. 33. The method according to claim 10, characterized in that said TLR9 inhibitor is an oligonucleotide that inhibits TLR9. 34. The method according to claim 33, characterized in that said oligonucleotide sequence that inhibits TLR9 has one or more of the following features: (i) three consecutive G nucleotides at the 3' end; (ii) a CC(T) triplet at the 5' end; and (iii) the distance between the three consecutive G nucleotides at the 3' end and the CC(T) triplet at the 5' end is 3-5 nucleotides in length. 35. The method according to claim 33, characterized in that said oligonucleotide sequence that inhibits TLR9 contains the sequence TTAGGGn SEQ ID NO: 888. 36. The method of claim 33, wherein said TLR9 inhibitory oligonucleotide sequence is selected from the group consisting of ODN-2088 TCCTGGCGGGGAAGT, SEQ ID NO: 889, ODN-2114 TCCTGGAGGGGAAGT, SEQ ID NO: 890, poly-G GGGGGGGGGGGGGGGGGGGGG, SEQ ID NO: 891, ODN-A151 TTAGGGTTAGGGTTAGGGTTAGGG, SEQ ID NO: 892, G-ODN CTCC-TATTGGGGGTTTCCTAT, SEQ ID NO: 893, IRS-869 TCCTGGAGGGGTTGT, SEQ ID NO: 894, INH-1 CCTGGATGGGAATTCCCATCCAGG, SEQ ID NO: 895, INH-4 TTCCCATCCAGGCCTGGATGGGAA, SEQ ID NO: 896, IRS-661 TGCTTGCAAGCTT-GCAAGCA, SEQ ID NO: 897, 4024 TCCTGGATGGGAAGT, SEQ ID NO: 898, 4084F CCTGGATGGGAA, SEQ ID NO: 899, INH-13 CTTACCGCTGCACCTGGATGGGAA, SEQ ID NO: 900, INH-18 CCTGGATGGGAACTTACCGCTGCA, SEQ ID NO: 901, and IRS-954 TGCTCCTGGAGGGGTTGT, SEQ ID NO: 902, and AS1411 GGTGGTGGTGGTTGTGGTGGTGGTGG, SEQ ID NO: 903. 37. The method according to claim 10, characterized in that said TLR9 inhibitor is a miRNA specific for TLR9. 38. The method according to claim 10, characterized in that said TLR9 inhibitor is a siRNA specific for TLR9. 39. The method according to claim 10, characterized in that said TLR9 inhibitor is an antibody or antigen-binding fragment that binds TLR9. 40. The method according to any one of claims 1-39, characterized in that said at least one transgene is functionally located between two flanking wild-type inverted terminal repeats (WT-ITR). 41. The method according to any one of claims 1-39, characterized in that said at least one transgene is located between two flanking mutant inverted terminal repeats (mutant ITRs). 42. The method according to any one of claims 1-39, characterized in that said at least one transgene is located between two flanking inverted terminal repeats, wherein one flanking ITR is a WT-ITR, and one flanking ITR is a mutant ITR. 43. The method according to any one of claims 1-42, characterized in that one or both of said flanking ITRs are derived from a virus selected from the group consisting of parvovirus, dependovirus and adeno-associated virus (AAV). 44. The method according to any one of paragraphs 1-43, characterized in that said flanking ITRs are symmetric or asymmetric ITRs. 45. The method according to claim 44, characterized in that said flanking ITRs are symmetrical or substantially symmetrical ITRs. 46. The method according to claim 44, characterized in that said flanking ITRs are asymmetric ITRs. 47. The method according to any one of claims 1-39, characterized in that one of said flanking ITRs is a wild-type ITR, or both of said flanking ITRs are wild-type ITRs. 48. The method according to any one of claims 1-39, characterized in that said flanking ITRs are from different viral serotypes. 49. The method according to any one of claims 1-39, characterized in that at least one of said flanking ITRs is altered relative to the sequence of the wild-type AAB ITR by deletion, addition and/or substitution, which affects the overall three-dimensional conformation of said ITR. 50. The method of claim 49, wherein one or both of said flanking ITRs are from an AAB serotype selected from the group consisting of AAB1, AAB2, AAB3, AAB4, AAB5, AAB6, AAB7, AAB8, AAB9, AAB10, AAB11, and AAB12. 51. The method according to any one of paragraphs 1-50, characterized in that one or both of said ITRs are synthetic. 52. The method according to claim 50 or 51, characterized in that one of said flanking ITRs is not a wild-type ITR, or both of said flanking ITRs are not a wild-type ITR. 53. The method according to any one of claims 1-50, characterized in that one or both of said flanking ITRs are modified by deletion, insertion and/or substitution in at least one of the ITR regions selected from the group consisting of A, A', B, B', C, C', D and D' regions. 54. The method according to claim 53, characterized in that said deletion, insertion and/or substitution results in the removal of all or part of the stem-loop structure formed by said regions A, A', B, B', C and/or C'. 55. The method of claim 53, wherein one or both of said flanking ITRs is modified by deletion, insertion and/or substitution, resulting in the removal of all or part of the stem-loop structure formed by said regions B and B'. 56. The method of claim 53, wherein one or both of said flanking ITRs is modified by deletion, insertion and/or substitution, resulting in the removal of all or part of the stem-loop structure formed by said C and C' regions. 57. The method of claim 53, wherein one or both of said flanking ITRs is modified by deletion, insertion and/or substitution, resulting in the removal of all or part of the stem-loop structure formed by said regions B and B', and/or all or part of the stem-loop structure formed by said regions C and C'. 58. The method according to any one of claims 1-50, characterized in that one or both of said flanking ITRs comprises one stem-loop structure in a region that, in a wild-type ITR, comprises a first stem-loop structure formed by said regions B and B', and a second stem-loop structure formed by said regions C and C'. 59. The method according to any one of claims 1-50, characterized in that one or both of said flanking ITRs comprises one stem and two loops in a region that, in a wild-type ITR, comprises a first stem-loop structure formed by said regions B and B', and a second stem-loop structure formed by said regions C and C'. 60. The method according to any one of claims 1-50, characterized in that one or both of said flanking ITRs comprises one stem and one loop in a region that, in a wild-type ITR, comprises a first stem-loop structure formed by said regions B and B', and a second stem-loop structure formed by said regions C and C'. 61. The method according to any one of claims 1-60, characterized in that both flanking ITRs are modified so that this results in overall three-dimensional symmetry when said ITRs are inverted relative to each other. 62. The method according to any one of paragraphs 1-61, characterized in that said ccDNA vector contains at least one regulatory switch. 63. The method according to claim 62, characterized in that said at least one regulatory switch is selected from the group consisting of a binary regulatory switch, a small molecule regulatory switch, a regulatory switch "with an access code", a regulatory switch based on a nucleic acid, a post-transcriptional regulatory switch, a radiation-controlled or ultrasound-controlled regulatory switch, a hypoxia-mediated regulatory switch, an inflammatory response regulatory switch, a shift-activated regulatory switch, and a kill switch. 64. The method according to any one of paragraphs 1-63, characterized in that said ccDNA vector and/or immune response inhibitor are encapsulated. 65. The method according to any one of paragraphs 1-63, characterized in that the said ccDNA vector and/or immune response inhibitor are located in a nanocarrier. 66. The method according to claim 65, characterized in that said nanocarrier contains a lipid nanoparticle (LNP). 67. The method according to claim 1, characterized in that said ecDNA, when cleaved with a restriction enzyme having one recognition site on said ecDNA vector and analyzed on a non-denaturing gel, demonstrates the presence of characteristic bands of linear and continuous DNA compared to control linear and discontinuous DNA. 68. The method according to any of the preceding claims, characterized in that said ccDNA vector comprises at least one promoter linked to said at least one transgene. 69. The method according to claim 68, characterized in that said promoter is selected from the group consisting of the CAG promoter, the HAAT promoter, the human EF1-α promoter or a fragment thereof, and the rat EF1-α promoter. 70. The method according to any of the preceding claims, characterized in that said ccDNA vector further comprises an enhancer comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1147-1160. 71. The method according to any of the preceding claims, characterized in that said ccDNA vector comprises a 5'-untranslated region (UTR) and/or intron sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1161-1188. 72. The method according to any one of the preceding claims, characterized in that said ccDNA vector comprises a 3'-UTR comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 8 and 1189-1200. 73. The method according to any one of the preceding claims, characterized in that said ccDNA vector comprises at least one poly(A) sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 9 and 1201-1216. 74. The method according to any of the preceding claims, characterized in that increasing the amount of said ccDNA vector that is introduced into the cell increases the expression of said at least one transgene in said cell. 75. The method according to any one of the preceding claims, characterized in that said at least one transgene encodes a therapeutic transgene, and wherein the target level of expression of said transgene is a therapeutically effective amount. 76. The method according to any of the preceding claims, characterized in that said at least one transgene, when transcribed or translated, modifies an abnormal amount of an endogenous protein in the subject. 77. The method of any one of the preceding claims, wherein said at least one transgene, when transcribed or translated, modifies an abnormal function or activity of an endogenous protein or pathway in the subject. 78. The method according to claim 76 or 77, characterized in that said subject is a human patient in need of treatment. 79. The method according to any of the preceding claims, characterized in that said at least one transgene encodes or contains a nucleotide molecule selected from the group consisting of RNAi, siRNA, miRNA, lncRNA and an antisense oligo- or polynucleotide. 80. The method according to any one of paragraphs 1-79, characterized in that said at least one transgene encodes a protein. 81. The method according to claim 80, characterized in that said protein is a marker protein. 82. The method according to any one of claims 1-79, characterized in that said at least one transgene encodes an agonist or antagonist of an endogenous protein or pathway associated with said disease or disorder. 83. The method according to any one of paragraphs 1-79, characterized in that said at least one transgene encodes an antibody. 84. The method according to any one of the preceding claims, further comprising administering an additional immune response inhibitor. 85. The method according to any one of claims 1 to 84, characterized in that said ccDNA vector is obtained by a method comprising the steps of: (a) incubating a population of insect cells bearing an ccDNA expression construct in the presence of at least one Rep protein, wherein said ccDNA expression construct encodes said ccDNA vector under effective conditions and for a time sufficient to induce production of said ccDNA vector in said insect cells; and (b) isolating said ccDNA vector from said insect cells. 86. The method according to claim 85, characterized in that said expression construct of ecDNA is selected from the group consisting of ecDNA plasmid, ecDNA bacmid, and ecDNA baculovirus. 87. The method of claim 85, characterized in that said insect cells express at least one Rep protein. 88. The method according to claim 87, characterized in that said at least one Rep protein is derived from a virus selected from the group consisting of parvovirus, dependovirus, and adeno-associated virus (AAV). 89. The method according to claim 88, characterized in that said at least one Rep protein is from a serotype AAB selected from the group consisting of AAB1, AAB2, AAB3, AAB4, AAB5, AAB6, AAB7, AAB8, AAB9, AAB10, AAB11 and AAB12. 90. A composition for inhibiting an immune response, comprising at least one immune response inhibitor, wherein the immune response inhibitor is a Toll-like receptor (TLR) inhibitor, an NLRP3 inflammasome inhibitor, an AIM2 inflammasome inhibitor, a caspase-1 inhibitor, or a cyclo-GMP-AMP synthase (cGAS) inhibitor; and a non-viral capsidless DNA vector with covalently closed ends (ccDNA vector), wherein said ccDNA vector comprises at least one transgene located between two flanking inverted terminal repeats (ITRs). 91. The composition according to claim 90, characterized in that said ccDNA vector additionally codes said at least one immune response inhibitor. 92. The composition according to claim 90 or 91, characterized in that said immune response is an innate immune response. 93. The composition according to claim 90 or 92, characterized in that said at least one transgene is located between two flanking wild-type inverted terminal repeats (WT-ITR). 94. The composition according to claim 90 or 92, characterized in that said at least one transgene is located between two flanking mutant inverted terminal repeats (mutant ITRs). 95. The composition according to claim 90 or 92, characterized in that said at least one transgene is located between two flanking inverted terminal repeats (ITRs), wherein one of the flanking ITRs is a WT-ITR and the other flanking ITR is a mutant ITR. 96. The composition according to claim 90 or 92, characterized in that one or both of said flanking ITRs are derived from a virus selected from the group consisting of parvovirus, dependovirus, and adeno-associated virus (AAV). 97. The composition according to claim 96, characterized in that said flanking ITRs are symmetrical or asymmetrical ITRs. 98. The composition according to claim 90 or 92, characterized in that both of said ITRs are wild-type ITRs. 99. The composition according to claim 90 or 92, characterized in that said flanking ITRs originate from different viral serotypes. 100. The composition according to claim 90 or 92, characterized in that at least one of said flanking ITRs is altered relative to the sequence of the wild-type AAB ITR by deletion, addition and/or substitution, which affects the overall three-dimensional conformation of said ITR. 101. The composition according to claim 90 or 92, characterized in that one or both of said ITRs are from a serotype AAB selected from the group consisting of AAB1, AAB2, AAB3, AAB4, AAB5, AAB6, AAB7, AAB8, AAB9, AAB10, AAB11 and AAB12. 102. The composition according to any one of claims 90 or 92-101, characterized in that one or more of said flanking ITRs are synthetic. 103. The composition according to claim 90 or 92, characterized in that both of said flanking ITRs are not wild-type ITRs. 104. The composition according to claim 90 or 92, characterized in that one or both of said flanking ITRs is modified by deletion, insertion and/or substitution in at least one of the ITR regions selected from the group consisting of regions A, A', B, B', C, C', D and D'. 105. The composition according to claim 104, characterized in that said deletion, insertion and/or substitution results in the removal of all or part of the stem-loop structure formed by said regions A, A', B, B', C and/or C'. 106. The composition according to claim 90 or 92, characterized in that one or both of said flanking ITRs is modified by deletion, insertion and/or substitution, which results in the removal of all or part of the stem-loop structure formed by said regions B and B'. 107. The composition of claim 90 or 92, wherein one or both of said flanking ITRs is modified by deletion, insertion and/or substitution, resulting in the removal of all or part of the stem-loop structure formed by said C and C' regions. 108. The composition according to claim 90 or 92, characterized in that one or both of said flanking ITRs is modified by deletion, insertion and/or substitution, which results in the removal of all or part of the stem-loop structure formed by said regions B and B', and/or all or part of the stem-loop structure formed by said regions C and C'. 109. The composition of claim 90 or 92, wherein one or both of said flanking ITRs comprises one stem-loop structure in a region that, in a wild-type ITR, comprises a first stem-loop structure formed by said regions B and B' and a second stem-loop structure formed by said regions C and C'. 110. The composition of claim 90 or 92, wherein one or both of said flanking ITRs comprises one stem and two loops in a region that, in a wild-type ITR, comprises a first stem-loop structure formed by said regions B and B' and a second stem-loop structure formed by said regions C and C'. 111. The composition of claim 90 or 92, wherein one or both of said flanking ITRs comprises one stem and one loop in a region that, in a wild-type ITR, comprises a first stem-loop structure formed by said regions B and B' and a second stem-loop structure formed by said regions C and C'. 112. The composition according to claim 90 or 92, characterized in that both flanking ITRs are modified so that this results in overall three-dimensional symmetry when said ITRs are inverted relative to each other. 113. The composition according to any one of claims 90 and 92-112, characterized in that said flanking ITRs are based on sequences from a virus selected from the group consisting of parvovirus, dependovirus and adeno-associated virus (AAV). 114. A composition according to any one of paragraphs 90 and 92-113, characterized in that said ccDNA vector includes at least one regulatory switch. 115. The composition of claim 114, wherein at least one regulatory switch is selected from the group consisting of a binary regulatory switch, a low-molecular regulatory switch, an “access code” regulatory switch, a nucleic acid-based regulatory switch, a post-transcriptional regulatory switch, a radiation-controlled or ultrasound-controlled regulatory switch, a hypoxia-mediated regulatory switch, an inflammatory response regulatory switch, a shift-activated regulatory switch, and a safety switch. 116. A composition according to any one of paragraphs 90 and 92-115, characterized in that said ccDNA vector is encapsulated. 117. A composition according to any one of paragraphs 90 and 92-116, characterized in that said ccDNA vector is located in a nanocarrier. 118. The composition according to claim 117, characterized in that said nanocarrier contains a lipid nanoparticle (LNP). 119. A composition according to any one of claims 90 and 92-118, characterized in that said ecDNA, when cleaved with a restriction enzyme having one recognition site on said ecDNA vector, and when analyzed on a non-denaturing gel, demonstrates the presence of characteristic bands of linear and continuous DNA compared to a control linear and discontinuous DNA. 120. The composition according to any one of claims 90 and 92-119, characterized in that said ccDNA vector contains an enhancer containing a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1147-1160. 121. The composition according to any one of claims 90 and 92-120, characterized in that said ccDNA vector contains a 5'-UTR and/or intron sequence containing a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1161-1188. 122. The composition according to any one of claims 90 and 92-121, characterized in that said ccDNA vector contains a 3'-UTR containing a nucleic acid sequence selected from the group consisting of SEQ ID NO: 8 and 1189-1200. 123. The composition according to any one of claims 90 and 92-122, characterized in that said ccDNA vector contains at least one poly(A) sequence containing a nucleic acid sequence selected from the group consisting of SEQ ID NO: 9 and 1201-1216. 124. A composition according to any one of claims 90 and 92-123, characterized in that said ccDNA vector contains at least one promoter associated with said at least one transgene. 125. The composition according to any one of claims 90 and 92-124, characterized in that at least one of the flanking ITRs comprises a functional end resolution site and a Rep binding site, and wherein one of said flanking ITRs comprises a deletion, insertion and/or substitution relative to the other flanking ITR. 126. A composition according to any one of claims 90 and 92-125, characterized in that said at least one transgene is codon optimized for expression in a eukaryotic cell. 127. The composition according to any one of claims 90 and 92-126, further comprising a pharmaceutically acceptable carrier. 128. The composition according to claim 90, characterized in that said TLR inhibitor is a TLR9 inhibitor. 129. The composition according to claim 90, characterized in that said TLR inhibitor is rapamycin or a rapamycin analogue. 130. A method for expressing an immune response inhibitor in a cell, characterized in that said method comprises bringing said cell into contact with a composition according to any one of paragraphs 90 and 92-129. 131. The method according to item 130, characterized in that said cell is in vitro or in vivo conditions. 132. A pharmaceutical composition for inhibiting an immune response in which a transgene is expressed in a cell, comprising a composition according to any one of claims 90 and 92-129. 133. The pharmaceutical composition according to claim 132, characterized in that said immune response inhibitor is an innate immune response inhibitor. 134. The pharmaceutical composition according to item 133, characterized in that said inhibitor of the innate immune response is manufactured separately from said ccDNA vector. 135. A cell for expressing an immune response inhibitor, comprising a composition according to any one of claims 90 and 92-129. 136. A kit for inhibiting an immune response, in which at least one transgene is expressed in a cell, comprising a composition according to any one of claims 90 and 92-129, a pharmaceutical composition according to any one of claims 132-134 or a cell according to claim 135, and a pharmaceutically acceptable carrier.