Adenovirus vector and its use

Abstract

The present invention provides chimeric adenoviral vectors. The provided chimeric adenoviral vectors can be used to induce a protective immune response and / or express a transgene in a subject in need thereof.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Application No. 62 / 909,853, filed October 3, 2019, the disclosure of which is incorporated herein by reference in its entirety. Technical Field

[0003] The present invention relates to biotechnology, and in particular to the field and use of adenoviral vectors, such as replication-defective adenoviral vectors, for delivering antigens and inducing an immune response in a host.

[0004] References to electronically submitted sequence listings

[0005] This application contains a sequence listing, which was submitted electronically via EFS-Web as an ASCII formatted sequence listing, with the file name "004852.120WO1 Sequence Listing," a creation date of August 13, 2020, and a size of 262 kb. The sequence listing submitted via EFS-Web is part of this specification and is incorporated herein by reference in its entirety. Background Art

[0006] Recombinant adenoviral vectors are widely used in gene therapy applications and vaccines. AdV-5 vector-based vaccines have been shown to induce effective and protective immune responses in various animal models (see, for example, WO2001 / 02607; WO2002 / 22080; Shiver et al., Nature 415:331 (2002); Letvin et al., Ann. Rev. Immunol. 20:73 (2002); Shiver and Emini, Ann. Rev. Med. 55:355 (2004)). However, the effectiveness of recombinant vaccines based on AdV-5 vectors may be limited by the high seroprevalence of AdV-5-specific neutralizing antibodies (NAbs) in the human population. The presence of anti-AdV-5 immunity has been shown to greatly inhibit the immunogenicity of AdV-5-based vaccines in studies in mice, rhesus monkeys, and humans.

[0007] A promising strategy to circumvent the presence of pre-existing immunity in individuals previously infected or treated with the most common human adenoviruses (e.g., AdV-5) involves developing recombinant vectors from adenovirus serotypes that do not encounter such pre-existing immunity. One such strategy is based on the use of chimeric adenoviruses that include replacing native capsid protein sequences (e.g., hexon and / or fiber protein sequences) with those of an adenovirus with low (or no) seroprevalence.

[0008] Therefore, there is a need in the art for alternative adenoviral vectors that can be produced in large quantities and do not encounter pre-existing immunity in the host, yet are immunogenic and capable of inducing a strong immune response against the antigen encoded by the heterologous nucleic acid inserted into the vector. Summary of the Invention

[0009] Provided herein is an isolated nucleic acid sequence encoding a chimeric adenoviral capsid or a functional derivative thereof. The chimeric adenoviral capsid or a functional derivative thereof may comprise, for example, a fiber polypeptide having an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 11; a hexon polypeptide having an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 12; and a penton polypeptide having an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 13. In certain embodiments, the fiber polypeptide sequence comprises the amino acid sequence of SEQ ID NO: 11. In certain embodiments, the hexon polypeptide sequence comprises the amino acid sequence of SEQ ID NO: 12. In certain embodiments, the penton polypeptide comprises the amino acid sequence of SEQ ID NO: 13.

[0010] Also provided herein are vectors comprising the isolated nucleic acid described herein.In certain embodiments, the vector is an adenoviral vector.

[0011] In certain embodiments, the adenoviral vector further comprises an E1 deletion. In certain embodiments, the adenoviral vector further comprises an E3 deletion. In certain embodiments, the adenoviral vector is a chimeric adenoviral vector comprising one or more adenoviral nucleic acid sequences from at least one of human adenovirus-4, human adenovirus-5, human adenovirus-26, or human adenovirus-35. The adenoviral vector may comprise, for example, human adenovirus-5 (HAdV-5) E4 orf6. In certain embodiments, the adenoviral vector may comprise, for example, a nucleic acid sequence selected from SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10.

[0012] In certain embodiments, the adenoviral vector further comprises at least one transgene. In certain embodiments, the at least one transgene is located at the E1 deletion, the E3 deletion, and / or adjacent to the right inverted terminal repeat (rITR).

[0013] The present invention also provides a recombinant cell comprising the adenoviral vector described herein. The present invention also provides a method for producing the adenoviral vector. The method comprises (a) growing the recombinant cell described herein under conditions for producing the adenoviral vector; and (b) isolating the adenoviral vector from the recombinant cell.

[0014] The present invention also provides a pharmaceutical composition comprising the adenoviral vector described herein and a pharmaceutically acceptable carrier.

[0015] The present invention also provides a method for inducing an immune response in a subject in need thereof, comprising administering to the subject a pharmaceutical composition described herein.

[0016] The present invention also provides a method of producing a pharmaceutical composition, the method comprising combining the adenoviral vector described herein with a pharmaceutically acceptable carrier.

[0017] The present invention also provides methods for expressing a transgene in a subject in need thereof. The methods comprise (a) identifying a subject in whom expression of a transgene is desired; (b) contacting the subject with an adenoviral vector comprising a transgene as described herein; and (c) expressing the transgene in the subject. In certain embodiments, expression of the transgene in a subject in need thereof treats or prevents a disease or condition. In certain embodiments, contacting the subject with the vector may comprise, for example, isolating cells from the subject and contacting the cells with the vector. The subject may be, for example, a human subject. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The foregoing summary, as well as the following detailed description of preferred embodiments of the application, will be better understood when read in conjunction with the accompanying drawings. It should be understood, however, that this application is not limited to the precise embodiments shown in the drawings.

[0019] Figure 1 The complete genome map of the Ad20-42-42 wild-type virus (SEQ ID NO: 1) is shown.

[0020] Figure 2 Schematic diagram showing the construction of adenoviral vector genome.

[0021] Figure 3 Schematic diagram showing the multiple cloning site (MCS) in the linker plasmid.

[0022] Figure 4 A schematic diagram showing the linker plasmid pAdApt20-42-42.Empty.

[0023] Figure 5 Schematic diagram of the intermediate plasmid pBR.Ad20-42-42.SbfI final interm is shown.

[0024] Figure 6 A schematic diagram of plasmid pBrAd20-42-42Srfl-rITR.dE3.5orf6 is shown on the right.

[0025] Figures 7A-7B The cellular immune response induced by Ad20-42-42.FLuc is shown. Figure 7A Schematic diagram showing the immunization schedule. Figure 7B Shown are graphs demonstrating the cellular immune response induced by Ad20-42-42.FLuc.

[0026] Figure 8 It was shown that no significant cross-neutralization was observed between Ad20-42-42 and Ad26 vectors.

[0027] Figure 9 Shown is a graph displaying the seroprevalence results of different adenoviral constructs in human subjects.

[0028] Figure 10 A graph showing the transduction potential of Ad20-42-42, HAd5 and HAd35 vectors is shown. Luciferase expression is expressed as relative light units (RLU) per milligram (mg) of protein.

[0029] Figure 11 Representative LacZ staining of cells transduced with Ad5LacZ and Ad20-42-42LacZ is shown. Transductions were performed with three vector doses: 10,000, 5000, and 1,000 viral particles (VP) per cell, with the addition of FX.

[0030] Figures 12A-12B Shown are graphs showing HAdV5 and Ad20-42-42 transduction of CHO cells expressing or lacking CAR or sialic acid and TCl cells expressing or lacking desmoglein 2 (DSG2).

[0031] Figures 12C-12D Shown are graphs demonstrating HAdV5 and Ad20-42-42 transduction of CHO cells lacking (negative) or expressing different isoforms of CD46 (BC1, BC2, C1, C2).

[0032] Figure 13 Distribution of luciferase-encoding HAdV5 and Ad20-42-42 in mice is shown. Mice were pretreated without (CL-; upper right and lower right panels) or with (CL+; upper left and lower left panels) sodium clodronate and injected intravenously with each vector. Control animals were injected with phosphate-buffered saline (PBS). Luciferase expression is from low to high.

[0033] Figure 14A-D shows the quantification of AdV vectors (HAdV-5 and Ad20-42-42, respectively). Figure 14A and 14C ) or (CL+; Figure 14B and 14D ) DNA copy number after intravenous transduction of mice in the presence of sodium clodronate. Vector genomes were quantified by qPCR. DETAILED DESCRIPTION

[0034] The present disclosure is based, at least in part, on the isolation of chimeric adenoviral vectors comprising a chimeric capsid polypeptide or a functional derivative thereof, wherein the chimeric capsid polypeptide comprises a fiber and hexon polypeptide from a first adenovirus (e.g., human adenovirus-42) and a penton polypeptide from a second adenovirus (e.g., human adenovirus-20). The adenoviral vectors are capable of eliciting an immune response while maintaining a low seroprevalence. The adenoviral vectors can be formulated into vaccines for inducing protective immunity against a specific antigen of interest. The adenoviral vectors can also be constructed to express a transgene of interest in a subject in need thereof.

[0035] Various publications, articles, and patents are cited or described in the Background and throughout the specification; each of these references is incorporated herein by reference in its entirety. The discussion of documents, provisions, materials, devices, articles, and the like included in this specification is solely for the purpose of providing a context for the present invention. Such discussion is not an admission that any or all of these materials form part of the prior art with respect to any invention disclosed or claimed.

[0036] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the invention belongs. Otherwise, certain terms used herein have the meaning as defined in the specification.

[0037] It must be noted that as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0038] Unless otherwise indicated, any numerical value, such as a concentration or concentration range described herein, should be understood to be modified by the term "about" in all cases. Thus, numerical values generally include ±10% of the stated value. For example, a concentration of 1 mg / mL includes 0.9 mg / mL-1.1 mg / mL. Similarly, a concentration range of 1%-10% (w / v) includes 0.9% (w / v)-11% (w / v). As used herein, the use of numerical ranges explicitly includes all possible subranges, all individual numerical values within that range, including integers and fractions of values within such ranges, unless the context clearly indicates otherwise.

[0039] Unless otherwise indicated, the term "at least" preceding a series of elements should be understood to refer to every element in the series. Those skilled in the art will recognize or be able to ascertain, using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. The present invention is intended to encompass such equivalents.

[0040] As used herein, the terms "comprises," "comprising," "includes," "including," "has," "having," "contains," or "containing," or any other variation thereof, will be understood to mean the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers, and are intended to be non-exclusive or open-ended. For example, a composition, mixture, process, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, mixture, process, method, article, or apparatus. Furthermore, unless expressly stated to the contrary, "or" refers to an inclusive or and not an exclusive or. For example, condition A or B is satisfied by any of: A being true (or present) and B being false (or absent), A being false (or absent) and B being true (or present), and both A and B being true (or present).

[0041] As used herein, the term "and / or" in conjunction with multiple listed elements is understood to encompass both individual and combined options. For example, when two elements are connected by "and / or," the first option refers to the applicability of the first element without the second element. The second option refers to the applicability of the second element without the first element. The third option refers to the applicability of the first and second elements together. Any of these options is understood to fall within the meaning and therefore meets the requirements of the term "and / or" as used herein. The concurrent applicability of more than one option is also understood to fall within the meaning and therefore meets the requirements of the term "and / or."

[0042] As used herein, the term "consists of" or variations such as "consist of" or "consisting of" used throughout the specification and claims means that any recited integer or group of integers is included, but additional integers or groups of integers may not be added to the specified method, structure, or composition.

[0043] As used herein, the term "consists essentially of," or variations such as "consist essentially of," as used throughout the specification and claims, means that any recited integer or group of integers is included, and optionally includes, any recited integer or group of integers that does not materially change the basic or novel characteristic of the specified method, structure, or composition. See MPEP §2111.03.

[0044] As used herein, "subject" refers to any animal, preferably a mammal, and most preferably a human. As used herein, the term "mammal" encompasses any mammal. Examples of mammals include, but are not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, and the like, more preferably humans.

[0045] It should also be understood that when referring to dimensions or features of components of the preferred invention, the terms "about," "approximately," "generally," "substantially," and similar terms used herein indicate that the described dimensions / features are not strict boundaries or parameters and do not exclude minor variations that are functionally the same or similar, as would be understood by one of ordinary skill in the art. At a minimum, such references including numerical parameters will include not changing the least significant digit using mathematical and industrial principles accepted in the art (e.g., rounding, measurement or other systematic errors, manufacturing tolerances, etc.).

[0046] The term "identical" or percent "identity" in the context of two or more nucleic acid or polypeptide sequences (e.g., hexon and fiber polypeptides and polynucleotides encoding them) means that two or more sequences or subsequences are the same or have a specified percentage of amino acid residues or nucleotides that are identical when compared and aligned for maximum correspondence, as measured using one of the following sequence comparison algorithms or by visual inspection.

[0047] For sequence comparison, there is typically one sequence that acts as a reference sequence to which test sequences are compared. When using a sequence comparison algorithm, the test and reference sequences are input into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. The sequence comparison algorithm then calculates the percent sequence identity for the test sequences relative to the reference sequence, based on the specified program parameters.

[0048] Optimal alignment of sequences for comparison can be performed, for example, by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), the search for similarity method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444 (1988), by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, WI), or by visual inspection (see generally, Current Protocols in Molecular Biology, FM Ausubel et al., eds., Current Protocols, a joint venture between Greene Publishing Associates, Inc. and John Wiley & Sons, Inc., (1995 Supplement) (Ausubel)).

[0049] Examples of algorithms suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al. (1990) J. Mol. Biol. 215:403-410 and Altschul et al. (1997) Nucleic Acids Res. 25:3389-3402, respectively. Software for performing BLAST analysis is publicly available through the National Center for Biotechnology Information. This algorithm involves first identifying high-scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence that either match or satisfy a positive threshold score T when aligned with a sequence of the same length in a database sequence. T is called the neighborhood word score threshold (Altschul et al, supra). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are then extended in both directions along each sequence as far as the cumulative alignment score can be increased.

[0050] For nucleotide sequences, the cumulative score is calculated using the parameters M (reward score for a pair of matching residues; always > 0) and N (penalty score for mismatching residues; always < 0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction is stopped when: the cumulative alignment score drops by the amount X from its maximum achieved value; the cumulative score becomes zero or lower due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses a word length (W) of 11, an expectation (E) of 10, M=5, N=-4, and a comparison of both chains as defaults. For amino acid sequences, the BLASTP program uses a word length (W) of 3, an expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff & Henikoff, Proc. Natl. Acad. Sci. USA 89:10915 (1989)) as defaults.

[0051] In addition to calculating percent sequence identity, the BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, e.g., Karlin & Altschul, Proc. Nat'l. Acad. Sci. USA 90:5873-5787 (1993)). One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability that a match between two nucleotide or amino acid sequences would occur by chance. For example, a nucleic acid is considered similar to a reference sequence if the smallest sum probability in a comparison of the test nucleic acid to the reference nucleic acid is less than about 0.1, more preferably less than about 0.01, and most preferably less than about 0.001.

[0052] A further indication that two nucleic acid sequences or polypeptides are substantially identical is that the polypeptide encoded by the first nucleic acid is immunologically cross-reactive with the polypeptide encoded by the second nucleic acid, as described below. Thus, a polypeptide is typically substantially identical to a second polypeptide, for example, where the two peptides differ only in conservative substitutions. Another indication that two nucleic acid sequences are substantially identical is that the two molecules hybridize to each other under stringent conditions.

[0053] As used herein, the term "polynucleotide," synonymously referred to as "nucleic acid molecule," "nucleotide," or "nucleic acid," refers to any polyribonucleotide or polydeoxyribonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA. "Polynucleotide" includes, but is not limited to, single-stranded and double-stranded DNA, DNA that is a mixture of single-stranded and double-stranded regions, single-stranded and double-stranded RNA, RNA that is a mixture of single-stranded and double-stranded regions, hybrid molecules comprising DNA and RNA, which may be single-stranded or, more commonly, double-stranded or a mixture of single-stranded and double-stranded regions. In addition, "polynucleotide" refers to triple-stranded regions comprising RNA or DNA or RNA and DNA. The term polynucleotide also includes DNA or RNA containing one or more modified bases, as well as DNA or RNA with a backbone modified for stability or other reasons. "Modified" bases include, for example, tritylated bases and unusual bases such as inosine. Various modifications can be made to DNA and RNA; thus, "polynucleotide" encompasses chemically, enzymatically, or metabolically modified forms of polynucleotides commonly found in nature, as well as chemical forms of DNA and RNA that are unique to viruses and cells. "Polynucleotide" also encompasses relatively short nucleic acid chains often referred to as oligonucleotides.

[0054] As used herein, the term "vector" is a replicon into which another nucleic acid segment is operably inserted so as to effect the replication or expression of that segment.

[0055] As used herein, the term "host cell" refers to a cell that contains a nucleic acid molecule of the present invention. A "host cell" can be any type of cell, for example, a primary cell, a cell in culture, or a cell from a cell line. In one embodiment, a "host cell" is a cell transfected with a nucleic acid molecule of the present invention. In another embodiment, a "host cell" is a progeny or potential progeny of such a transfected cell. The progeny of a cell may or may not be identical to the parent cell, for example, due to mutations or environmental influences that may occur in subsequent generations or due to integration of the nucleic acid molecule into the host cell genome.

[0056] As used herein, the term "expression" refers to the biosynthesis of a gene product. The term encompasses the transcription of a gene into RNA. The term also encompasses the translation of RNA into one or more polypeptides, and further encompasses all naturally occurring post-transcriptional and post-translational modifications. The expressed polypeptide may be within the cytoplasm of the host cell, enter the extracellular environment, such as the growth medium of a cell culture, or be anchored to the cell membrane.

[0057] As used herein, the terms "peptide," "polypeptide," or "protein" may refer to molecules comprising amino acids and may be recognized as proteins by those skilled in the art. Conventional single-letter or three-letter codes for amino acid residues are used herein. The terms "peptide," "polypeptide," and "protein" are used interchangeably herein to refer to amino acid polymers of any length. The polymer may be linear or branched, it may contain modified amino acids, or it may be interrupted by non-amino acids. The term also encompasses amino acid polymers that have been modified naturally or by intervention; for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification, such as conjugation with a labeling component. Also included in this definition are, for example, polypeptides containing one or more amino acid analogs (including, for example, non-natural amino acids, etc.), as well as other modifications known in the art.

[0058] The peptide sequences described herein are written according to the usual convention, i.e. the N-terminal region of the peptide is on the left and the C-terminal region is on the right. Although isomeric forms of amino acids are known, unless otherwise explicitly stated, the L-form of the amino acid is represented.

[0059] As used herein, the term "protective immunity" or "protective immune response" refers to the ability of a vaccinated subject to control infection with the pathogenic agent for which the vaccine is administered. The pathogenic agent can be, for example, an antigenic gene product or an antigenic protein or fragment thereof. Typically, a subject who has developed a "protective immune response" develops only mild to moderate clinical symptoms or no symptoms at all. Typically, a subject who has a "protective immune response" or "protective immunity" against a substance will not die from infection with the substance.

[0060] The term "adjuvant" is defined as one or more substances that cause stimulation of the immune system. In this context, adjuvants are used to enhance the immune response to the adenoviral vectors of the present invention.

[0061] As used herein, the term "antigenic gene product or fragment thereof" or "antigenic protein" may include proteins or fragments thereof of bacteria, viruses, parasites or fungi. Preferably, the antigenic protein or antigenic gene product is capable of eliciting a protective immune response in a host, for example, inducing an immune response against a disease or infection (e.g., a bacterial, viral, parasitic or fungal disease or infection), and / or generating immunity against a disease or infection in a subject (i.e., vaccination), which protects the subject from the disease or infection.

[0062] As used herein, the term "chimeric" means a gene, nucleic acid, protein, peptide or polypeptide that comprises two or more genes, nucleic acids, proteins, peptides or polypeptides that are not normally associated together. A "chimeric" gene, nucleic acid or protein can be a fusion between two or more unrelated sequences (e.g., two or more different nucleic acids encoding two or more different proteins). A "chimeric" gene, nucleic acid or protein can be a fusion between two or more related sequences (e.g., nucleic acids encoding the same protein, but the nucleic acids are derived from different source materials, i.e., one nucleic acid is derived from one human adenovirus and the other nucleic acid is derived from a second, unrelated human adenovirus).

[0063] Adenoviral vector

[0064] Exposure to certain adenoviruses results in an immune response to certain adenovirus serotypes, which can affect the effectiveness of adenovirus vectors. Since human adenovirus infection is common in humans, the prevalence of neutralizing antibodies against human adenoviruses in the population is very high. Such neutralizing antibodies present in individuals may reduce the effectiveness of gene transfer vectors based on human adenovirus backbones. One of the methods to circumvent the reduction in effectiveness is to replace the epitope on the adenovirus capsid protein that serves as the target of neutralizing antibodies. The target sequence on the capsid protein can be replaced with a protein sequence from other adenoviruses (such as chimeric adenoviruses of multiple human adenoviruses) that have a low prevalence and therefore rare neutralizing antibodies against them in the population.

[0065] "Capsid protein" refers to a protein on the capsid of an adenovirus (e.g., AD20 and / or AD42) or a functional fragment or derivative thereof, which is involved in determining the serotype and / or tropism of the adenovirus. Capsid proteins generally include fiber protein, penton protein, and / or hexon protein. In certain embodiments, the capsid protein is the entire or full-length capsid protein of an adenovirus. In other embodiments, the capsid protein is a fragment or derivative of the full-length capsid protein of an adenovirus. In certain embodiments, the penton, hexon, and fiber encoded by the adenovirus vector of the present invention are derived from different adenovirus backgrounds.

[0066] As used herein, a "chimeric adenoviral capsid" refers to an adenoviral-derived capsid comprising fiber, penton, and / or hexon polypeptides, wherein the fiber, penton, and / or hexon are derived from different adenoviral sources (e.g., Ad42 fiber and hexon polypeptides and Ad20 penton polypeptides).

[0067] "Hexon polypeptide" refers to adenovirus hexon coat protein, functional fragments and derivatives thereof.

[0068] "Fiber polypeptide" refers to adenovirus fiber protein, functional fragments and derivatives thereof.

[0069] "Penton polypeptide" refers to adenovirus penton protein, functional fragments and derivatives thereof.

[0070] One target of neutralizing antibodies against adenoviruses is the major capsid protein, the hexon protein. Replacing the serotype-defining hexon protein or the variable sequences within the hexon protein that bind to neutralizing antibodies with hexon proteins or the variable sequences within the hexon protein from adenoviruses that are rare in the human population could allow the construction of adenoviral vectors that are less susceptible to neutralization by antibodies commonly found in humans.

[0071] Hexon hypervariable regions (HVRs) are regions of the hexon polypeptide that represent the highest variability among different adenovirus serotypes. In general, these HVRs are thought to correspond to the solvent-exposed surface of the hexon protein trimer (within the context of intact viral particles), and, accordingly, are expected to be important determinants in antibody-mediated neutralization of adenoviruses (Roberts et al., Nature 441: 239-43 (2006)). Replacing the hexon HVRs of a given adenovirus vector with the hexon HVRs of an adenovirus with low (or no) seroprevalence in humans represents a possible means of circumventing pre-existing anti-vector humoral immunity in the target population. Therefore, there have been several studies exploring the concept of hexon chimerism, mainly involving hexon sequence replacement within HAdV-5 vectors (Roy et al., J Virol. 72:6875-9 (1998); Gall et al., J Virol. 72:10260-4 (1998); Youil et al., Hum. Gene Ther. 13:311-20 (2002); Wu et al. J Virol. 76:12775-82 (2002); Roy et al., Virology. 333:207-14 (2005); Roberts et al., Nature 441:239-43 (2006); Bradley et al., J Virol. 86:1267-72 (2012); Yu et al., Biochem Biophys Res Commun. 421:170-6(2012); Bruder et al., PLoS One. 7(4):e33920(2012)).

[0072] A second target for neutralizing antibodies against adenoviruses is the fiber protein. Replacing the fiber protein with fiber sequences from rare adenoviruses of human origin, and more preferably replacing variable sequences within the fiber protein, can also allow the construction of adenoviral vectors that are less susceptible to neutralization by antibodies commonly found in humans. Combining these fiber replacements with hexon replacements can confer additional resistance to neutralization by antibodies prevalent in the human population.

[0073] A third target for neutralizing antibodies against adenoviruses is the penton protein. Replacing the penton protein with penton sequences from rare adenoviruses of human origin could also allow for the construction of adenoviral vectors that are less susceptible to neutralization by common human antibodies. Combinations of hexon, fiber, and penton replacements could confer additional resistance to neutralization by antibodies prevalent in the human population.

[0074] The present disclosure provides isolated nucleic acid sequences encoding chimeric adenoviral capsids or functional derivatives thereof. The chimeric adenoviral capsids or functional derivatives thereof can comprise, for example, a fiber polypeptide, a hexon polypeptide, and a penton polypeptide. The fiber and hexon polypeptides can be derived from, for example, a first adenovirus (e.g., human adenovirus-42), and the penton polypeptide can be derived from, for example, a second adenovirus (e.g., human adenovirus-20).

[0075] A "functional derivative" of a polypeptide appropriately refers to a modified form of the polypeptide, e.g., wherein one or more amino acids of the polypeptide may be deleted, inserted, modified, and / or substituted. A derivative of an unmodified adenoviral capsid protein is considered functional if, for example, (a) the adenovirus comprising the derivative capsid protein in its capsid retains substantially the same or lower seroprevalence compared to an adenovirus comprising the unmodified capsid protein; and / or (b) the adenovirus comprising the derivative capsid protein in its capsid retains substantially the same or higher host cell infectivity compared to an adenovirus comprising the unmodified capsid protein; and / or (c) the adenovirus comprising the derivative capsid protein in its capsid retains substantially the same or higher immunogenicity compared to an adenovirus comprising the unmodified capsid protein; and / or (d) the adenovirus comprising the derivative capsid protein in its capsid retains substantially the same or higher levels of transgene productivity compared to an adenovirus comprising the unmodified capsid protein.

[0076] "Adenoviral vector" refers to a recombinant vector derived from at least a portion of an adenoviral gene set or containing at least a portion of an adenoviral genome.

[0077] In preferred embodiments, the chimeric adenoviral capsid can comprise, for example, a fiber polypeptide having an amino acid sequence that is 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%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 11; a hexon polypeptide having an amino acid sequence that is 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%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 12; and a penton polypeptide having an amino acid sequence that is 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%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 13. In certain embodiments, the fiber polypeptide sequence comprises the amino acid sequence of SEQ ID NO: 11. In certain embodiments, the hexon polypeptide sequence comprises the amino acid sequence of SEQ ID NO: 12. In certain embodiments, the penton polypeptide comprises the amino acid sequence of SEQ ID NO: 13.

[0078] In a preferred embodiment, the present invention provides a vector, preferably an adenoviral vector, comprising an isolated nucleic acid disclosed herein. The adenoviral vector comprises an isolated nucleic acid encoding a chimeric adenoviral capsid or a functional derivative thereof, wherein the chimeric adenoviral capsid or a functional derivative thereof comprises: a fiber polypeptide having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 11; a hexon polypeptide having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 12; and a penton polypeptide having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 13.

[0079] Typically, the adenoviral vectors of the present invention comprise a complete recombinant adenoviral genome on, for example, a plasmid, cosmid, or baculovirus vector. The nucleic acid molecules of the present invention may be in the form of RNA or DNA obtained by cloning or synthesis. The DNA may be double-stranded or single-stranded.

[0080] Those of ordinary skill will recognize that elements from multiple serotypes can be combined in a single adenoviral vector, such as human or simian adenovirus. Thus, chimeric adenoviral vectors can be generated that combine desirable properties from different serotypes. Thus, in some embodiments, the chimeric adenoviral vectors of the invention can combine the lack of pre-existing immunity of chimeric hexon and / or fiber polypeptide sequences with the high-level antigen and / or transgene delivery and presentation capabilities of existing adenoviral vectors (e.g., rAd4, rAd5, rAd26, or rAd35).

[0081] Advantages of adenoviral vectors for use as vaccines and / or transgene expression vehicles may include, but are not limited to, ease of handling, good large-scale manufacturability, and an excellent safety record, based on years of experience in research, development, manufacturing, and clinical trials of numerous adenoviral vectors. Adenoviral vectors used as vaccines generally provide a good immune response, including a cellular immune response, to the transgene-encoded protein or transgene-encoded antigenic gene product. Adenoviral vectors according to the present invention can be based on any type of adenovirus, and in certain embodiments, are human adenoviruses, which can be of any group or serotype. In preferred embodiments, the recombinant adenovirus is based on a human adenovirus from groups A, B, C, D, E, F, or G. In other preferred embodiments, the recombinant adenovirus is based on human adenovirus serotypes 5, 11, 26, 34, 35, 48, 49, or 50. In other embodiments, it is a simian adenovirus, such as a chimpanzee or gorilla adenovirus, which can be of any serotype. In certain embodiments, the recombinant adenovirus is based on chimpanzee adenovirus type 1, 3, 7, 8, 21, 22, 23, 24, 25, 26, 27.1, 28.1, 29, 30, 31.1, 32, 33, 34, 35.1, 36, 37.2, 39, 40.1, 41.1, 42.1, 43, 44, 45, 46, 48, 49, 50, 67, or SA7P.

[0082] In a more preferred embodiment, the chimpanzee adenovirus vector of the second composition is ChAdV3. Recombinant chimpanzee adenovirus serotype 3 (ChAd3 or cAd3) is a subgroup C adenovirus with properties similar to those of human adenovirus serotype 5 (Ad5). ChAd3 has demonstrated safety and immunogenicity in human studies evaluating a hepatitis C virus (HCV) vaccine candidate (Barnes E, et al. 2012 Science Translational Medicine 4:115ra1). ChAd3-based vaccines have been reported to induce immune responses comparable to human Ad5-based vaccines. See, for example, Peruzzi D, et al. 2009 Vaccine 27:1293-300 and Quinn KM, et al. 2013 J Immunol 190:2720-35; WO 2005 / 071093; WO 2011 / 0130627, etc.

[0083] Adenoviral vectors, methods for their construction, and methods for their propagation are well known in the art and are described, for example, in U.S. Pat. Nos. 5,559,099, 5,837,511, 5,846,782, 5,851,806, 5,994,106, 5,994,128, 5,965,541, 5,981,225, 6,040,174, 6,020,191, and 6,113,913, and in Thomas Shenk, “Adenoviridae and their Replication,” and M.S. Horwitz, “Adenoviruses,” in Virology, B.N. Fields et al., eds., 3d., Raven Press, Ltd., New York (1996), Chapters 67 and 68, respectively, and in other references cited herein. Generally, construction of adenoviral vectors involves the use of standard molecular biology techniques, such as those described in Sambrook et al., Molecular Cloning, a Laboratory Manual, 2d ed., Cold Spring Harbor Press, Cold Spring Harbor, NY (1989), Watson et al., Recombinant DNA, 2d ed., Scientific American Books (1992), and Ausubel et al., Current Protocols in Molecular Biology, Wiley Interscience Publishers, NY (1995), as well as other references cited herein.

[0084] In certain embodiments, the adenoviral vector comprises an E1 deletion and / or an E3 deletion. The E1 or E3 deletion may include, for example, a complete or partial deletion of a gene, which renders the E1 or E3 gene product functionally defective. Thus, in certain embodiments, the adenovirus is replication-defective, for example, because it contains a deletion in the E1 region of the genome. As known to those skilled in the art, when key regions in the adenoviral genome are deleted, the functions encoded by these regions must be provided in trans, preferably by the producer cell, i.e., when part or all of the E1, E2 and / or E4 regions are deleted from the adenovirus, these must be present in the producer cell, for example, integrated into its genome, or in the form of a so-called helper adenovirus or helper plasmid. Adenoviruses may also have a deletion in the E3 region that is not essential for replication and therefore does not require supplementation of such a deletion. One or more of the E1, E2, E3 and E4 regions may also be inactivated by other means, such as by inserting a transgene of interest (usually linked to a promoter) into the region to be inactivated.

[0085] The producer cells that can be used (sometimes also referred to as "packaging cells" or "complementing cells" in the art and herein) can be any producer cells in which the desired adenovirus can be propagated. For example, the propagation of the recombinant adenoviral vector is carried out in producer cells that complement adenovirus defects. Such producer cells preferably have at least one adenovirus E1 sequence in their genome, so that recombinant adenoviruses with deletions in the E1 region can be complemented. Any producer cell that complements E1 can be used, such as human retinal cells immortalized by E1, such as 911 or PER.C6 cells (see U.S. Patent No. 5,994,128), amniotic fluid transformed by E1 (see EP Patent No. 1230354), A549 cells transformed by E1 (see, for example, WO98 / 39411, U.S. Patent No. 5,891,690), GH329:HeLa (Gao et al, 2000, Hum Gene Ther 11:213-19), 293, etc. In certain embodiments, the producer cells are, for example, HEK293 cells, PER.C6 cells, 911 cells, IT293SF cells, etc. Production of adenoviral vectors in producer cells is reviewed in (Kovesdi et al., 2010, Viruses 2: 1681-703).

[0086] In certain embodiments, the adenoviral vector is a chimeric adenoviral vector comprising one or more human adenoviral nucleic acid sequences. The human adenoviral nucleic acid can be selected from, for example, human adenovirus-4 (Ad-4), human adenovirus-5 (Ad-5), human adenovirus-26 (Ad-26), or human adenovirus-35 (Ad-35). In certain embodiments, the E1-deficient adenoviral vector comprises the E4-orf6 coding sequence of a human Ad5 adenovirus. This allows such adenoviruses to be propagated in well-known complementing cell lines that express the E1 gene of Ad5, such as 293 cells or PER.C6 cells (see, for example, Fallaux et al., 1998, Hum Gene Ther 9: 1909-17, Haveng et al., 2006, J Gen Virol 87: 2135-43, WO 03 / 104467, incorporated herein by reference in their entirety).

[0087] In certain embodiments, the adenoviral vector comprises a transgene. "Transgene" refers to a heterologous nucleic acid, which is a non-naturally occurring nucleic acid in the vector. According to the present invention, the transgene can encode an antigenic gene product or antigenic protein that induces an immune response in a subject. The transgene can also encode a therapeutic protein to treat or prevent a disease in a subject in need. The transgene can be introduced into the vector by, for example, standard molecular biology techniques. The transgene can be cloned into, for example, the deleted E1 or E3 region of the adenoviral vector, or in the region between the E4 region and the rITR. The transgene is generally operably linked to an expression control sequence. In a preferred embodiment, the transgene is inserted into the transgene insertion site.

[0088] If necessary, the chimeric adenoviral capsid sequence and / or its transgene comprising the fiber, hexon and penton polypeptide sequences according to embodiments of the present invention can be codon optimized to ensure correct expression in the treated host (e.g., human). Codon optimization is a technique widely used in the art.

[0089] The transgenic gene can be under the control of (i.e., operably linked to) an adenovirus-derived promoter (e.g., a major late promoter), or can be under the control of a heterologous promoter. Examples of suitable heterologous promoters include CMV promoters and RSV promoters. Preferably, the promoter is located upstream of the heterologous gene of interest in the expression cassette.

[0090] In a preferred embodiment, the adenoviral vector comprises a nucleic acid sequence selected from the group consisting of: SEQ ID NO:8, SEQ ID NO:9, and SEQ ID NO:10.

[0091] Pharmaceutical and / or immunogenic compositions

[0092] In another general aspect, the present invention provides a pharmaceutical composition comprising an isolated polynucleotide of the present invention, an isolated polypeptide of the present invention, a vector of the present invention, an adenoviral vector of the present invention, and / or a host cell of the present invention and a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutical composition" refers to a product comprising an isolated polynucleotide of the present invention, an isolated polypeptide of the present invention, an isolated vector (e.g., an adenoviral vector) of the present invention, and / or a host cell of the present invention together with a pharmaceutically acceptable carrier. The polynucleotides, polypeptides, vectors, and / or host cells of the present invention, and compositions comprising the same, can also be used to manufacture the medicaments for therapeutic applications mentioned herein.

[0093] As used herein, the term "carrier" refers to any excipient, diluent, filler, salt, buffer, stabilizer, solubilizer, oil, lipid, lipid-containing vesicle, microsphere, liposomal encapsulation (liposomal encapsulation) or other materials known in the art for pharmaceutical preparations. It will be appreciated that the characteristics of the carrier, excipient or diluent will depend on the route of administration for a particular application. As used herein, the term "pharmaceutically acceptable carrier" refers to a non-toxic material that does not interfere with the effectiveness of the compositions of the present invention or the biological activity of the compositions of the present invention. According to specific embodiments, in view of the present disclosure, any pharmaceutically acceptable carrier suitable for polynucleotides, polypeptides, vectors and / or host cell pharmaceutical compositions can be used in the present invention.

[0094] The formulation of pharmaceutically active ingredients with pharmaceutically acceptable carriers is known in the art, for example, Remington: The Science and Practice of Pharmacy (e.g., 21st edition (2005) and any later editions). Non-limiting examples of additional ingredients include: buffers, diluents, solvents, tonicity adjusters, preservatives, stabilizers, and chelating agents. One or more pharmaceutically acceptable carriers can be used to formulate the pharmaceutical compositions of the present invention.

[0095] The pharmaceutical composition can be formulated, for example, for expressing a transgene in a subject in need thereof (i.e., a pharmaceutical composition designed to express a transgene in a subject in need thereof). The pharmaceutical composition can be formulated, for example, for expressing an antigenic polypeptide or antigenic fragment thereof (e.g., a pharmaceutical composition for inducing an immune response in a subject in need thereof).

[0096] Pharmaceutical compositions designed to induce an immune response in a subject in need thereof may be referred to as, for example, immunogenic compositions. An immunogenic composition is a composition comprising an immunologically effective amount of a purified or partially purified adenoviral vector for use in the present invention. The composition can be formulated into a vaccine (also referred to as an "immunogenic composition") according to methods well known in the art. Such compositions may include an adjuvant to enhance the immune response. In view of the present disclosure, the optimal ratio of each component in the formulation can be determined by techniques well known to those skilled in the art.

[0097] In view of the present disclosure, immunogenic compositions according to embodiments of the present invention can be prepared using methods known to those skilled in the art. Liquid pharmaceutical compositions generally include a liquid carrier such as water, petroleum, animal or vegetable oils, mineral oils, or synthetic oils. These may include physiological saline, glucose or other sugar solutions, or glycols such as ethylene glycol, propylene glycol, or polyethylene glycol.

[0098] The immunogenic compositions useful in the present invention may include an adjuvant. Adjuvants suitable for co-administration according to the present invention are those that are potentially safe, well tolerated, and effective in subjects, including QS-21, Detox-PC, MPL-SE, MoGM-CSF, TiterMax-G, CRL-1005, GERBU, TERamide, PSC97B, Adjumer, PG-026, GSK-1, AS01, AS03, AS04, AS15, GcMAF, B-alethine, MPC-026, Adjuvax, CpG ODN, Betafectin, alum, and MF59.

[0099] Other adjuvants that can be administered include lectins, growth factors, cytokines and lymphokines such as alpha-interferon, gamma-interferon, platelet-derived growth factor (PDGF), granulocyte colony-stimulating factor (gCSF), granulocyte macrophage colony-stimulating factor (gMCSF), tumor necrosis factor (TNF), epidermal growth factor (EGF), IL-1, IL-2, IL-4, IL-6, IL-8, IL-10 and IL-12, or nucleic acids encoding them.

[0100] The compositions of the present invention may include pharmaceutically acceptable excipients, carriers, buffers, stabilizers, or other materials well known to those skilled in the art. Such materials should be nontoxic and should not interfere with the effectiveness of the active ingredient. The exact nature of the carrier or other materials may depend on the route of administration, for example, intramuscular, subcutaneous, oral, intravenous, cutaneous, intramucosal (e.g., intestinal), intranasal, or intraperitoneal routes.

[0101] Methods for inducing protective immunity and / or expressing transgenes

[0102] Another general aspect of the present invention relates to methods for inducing an immune response and / or expressing a transgene in a subject in need thereof. The methods may include, for example, identifying a subject in need thereof; contacting the subject in need thereof with a pharmaceutical composition and / or immunogenic composition described herein; and inducing an immune response and / or expressing a transgene in the subject in need thereof. In certain embodiments, the methods may include, for example, administering to the subject a vaccine comprising an adenoviral vector described herein and a pharmaceutically acceptable carrier.

[0103] Also provided herein are methods for producing vaccines, comprising combining the adenoviral vectors described herein with a pharmaceutically acceptable carrier.

[0104] Any immunogenic composition according to embodiments of the present invention, including but not limited to those described herein, can be used as a vaccine in the methods of the present invention. Any pharmaceutical composition according to embodiments of the present invention, including but not limited to those described herein, can be used in the methods of the present invention to treat or prevent a disease in a subject in need thereof by expressing a transgene of interest.

[0105] Administration of the immunogenic composition / vaccine / pharmaceutical composition comprising the vector is typically intramuscular or subcutaneous. However, other routes of administration are also contemplated, such as intravenous, cutaneous, intradermal or intranasal. Intramuscular administration of the immunogenic composition can be achieved by injecting a suspension of the adenoviral vector using a needle. Alternatively, the composition can be administered using a needle-free injection device (using, for example, ) or freeze-dried powder containing the vaccine.

[0106] For intravenous, cutaneous or subcutaneous injection, or injection in the affected area, the carrier will be in the form of a parenteral acceptable aqueous solution that is pyrogen-free and has suitable pH, isotonicity and stability. Those skilled in the art are well able to prepare suitable solutions, for example, using isotonic vehicles such as sodium chloride injection, Ringer's injection, lactated Ringer's injection. Preservatives, stabilizers, buffers, antioxidants and / or other additives can be incorporated as needed. Sustained release formulations can also be used.

[0107] Typically, administration will have preventive purposes, to generate an immune response to an antigen of interest (e.g., a bacterium, virus, parasite, and / or fungal pathogen) before infection or symptom development. Administration of an adenoviral vector expressing a transgenic agent of interest may also have preventive purposes in a subject in need thereof. For example, a subject in need thereof may have reduced or disappeared endogenous expression of a gene corresponding to the transgenic agent of interest. Diseases and conditions that can be treated or prevented according to the present invention include those in which an immune response can play a protective or therapeutic role, and / or corrective expression of a transgenic agent results in the normal functioning of cells in a subject in need thereof. In other embodiments, adenoviral vectors may be administered for post-exposure prophylaxis.

[0108] An immunogenic composition containing a chimeric human adenoviral vector is administered to a subject to generate an immune response to the antigen of interest in the subject. An amount of the composition sufficient to induce a detectable immune response is defined as an "immunologically effective dose" or "effective amount" of the composition. The immunogenic compositions of the present invention can induce both humoral and cell-mediated immune responses. In a typical embodiment, the immune response is a protective immune response.

[0109] The pharmaceutical composition can be administered to a subject in need thereof in a therapeutically effective amount to treat or prevent a disease. A therapeutically effective amount refers to an amount of an adenoviral vector expressing a transgene of interest that results in treatment of a disease, disorder, or condition; an amount that prevents or slows the progression of a disease, disorder, or condition; or an amount that reduces or completely alleviates the symptoms associated with a disease, disorder, or condition.

[0110] According to certain embodiments, a therapeutically effective amount is an amount sufficient to achieve one, two, three, four or more of the following: (i) reduce or lessen the severity of the disease, disorder or condition being treated, or symptoms associated therewith; (ii) reduce the duration of the disease, disorder or condition being treated, or symptoms associated therewith; (iii) prevent the progression of the disease, disorder or condition being treated, or symptoms associated therewith; (iv) cause regression of the disease, disorder or condition being treated, or symptoms associated therewith; (v) prevent the development or onset of the disease, disorder or condition being treated, or symptoms associated therewith (vi) prevent recurrence of the disease, disorder or condition to be treated, or symptoms associated therewith; (vii) reduce hospitalization of a subject with the disease, disorder or condition to be treated, or symptoms associated therewith; (viii) reduce the length of hospital stay of a subject with the disease, disorder or condition to be treated, or symptoms associated therewith; (ix) increase survival of a subject with the disease, disorder or condition to be treated, or symptoms associated therewith; (xi) inhibit or reduce the disease, disorder or condition to be treated, or symptoms associated therewith, in a subject; and / or (xii) enhance or improve the prophylactic or therapeutic effect of another therapy.

[0111] As used herein, the terms "treat," "treating," and "treatment" are all intended to refer to the alleviation or reversal of at least one measurable physical parameter associated with a disease, disorder, or condition, which is not necessarily discernible in the subject, but is discernible in the subject. The terms "treat," "treating," and "treatment" may also refer to causing regression of, preventing progression of, or at least slowing the progression of a disease, disorder, or condition. In a specific embodiment, "treat," "treating," and "treatment" refer to alleviating, preventing the development or onset of, or reducing the duration of, one or more symptoms associated with a disease, disorder, or condition. In a specific embodiment, "treat," "treating," and "treatment" refer to preventing the recurrence of a disease, disorder, or condition. In a specific embodiment, "treat," "treating," and "treatment" refer to increasing the survival of a subject having a disease, disorder, or condition. In a specific embodiment, "treat," "treating," and "treatment" refer to eliminating the disease, disorder, or condition in a subject.

[0112] The actual amount administered, as well as the rate and time course of administration, will depend on the nature and severity of the disease being treated. Prescription of treatment, such as decisions regarding dosage, etc., is the purview of general practitioners and other medical doctors, or in a veterinary context, of veterinarians, and generally takes into account the disease to be treated, the condition of the individual patient, the site of delivery, the method of administration, and other factors known to practitioners. Examples of the above techniques and protocols can be found in Remington's Pharmaceutical Sciences, 16th edition, Osol, A. ed., 1980.

[0113] After the adenoviral vectors are produced and such particles are optionally formulated into a composition, the vectors can be administered to a subject, particularly a human or another primate. Administration can be to a human, or to another mammal, for example, a mouse, rat, hamster, guinea pig, rabbit, sheep, goat, pig, horse, cow, donkey, monkey, dog, or cat. Delivery to non-human mammals need not be for therapeutic purposes, but can be used in an experimental setting, for example, to study the mechanisms of the immune response to adenoviral vectors.

[0114] In one exemplary embodiment, the adenoviral vector is administered in a volume of about 100 μl to about 10 ml (containing a concentration of about 104 -10 12 Preferably, the adenoviral vector is administered in a volume of 0.1-2.0 ml. For example, the adenoviral vector can be administered in 100 μl, 500 μl, 1 ml, or 2 ml. More preferably, the adenoviral vector is administered in a volume of 0.5 ml. Optionally, the adenoviral vector can be administered in a volume of about 10 7 vp / ml, 10 8 vp / ml, 10 9 vp / ml, 10 10 vp / ml, 5x10 10 vp / ml, 10 11 vp / ml or 10 12 vp / ml. Typically, adenoviral vectors are administered at a concentration of about 10 9 - About 10 12 The amount of virus particles (vp) administered to a human subject is, more typically, about 10 10 - About 10 12 The initial administration may be followed by a booster, for example, as described above.

[0115] After the initial administration, the vaccine / composition can be boosted or kicked with the same adenoviral vector encoding the antigen of interest and / or transgene of interest or with a different adenoviral vector encoding the same antigen of interest and / or transgene of interest.

[0116] If desired, the composition can be presented in a kit, pack or dispenser, which can contain one or more unit dosage forms containing the active ingredient. The kit can comprise, for example, metal or plastic foil, such as a blister pack. The kit, pack or dispenser can be accompanied by instructions for administration.

[0117] The compositions of the present invention may be administered alone or in combination with other treatment methods, either simultaneously or sequentially, depending on the condition to be treated.

[0118] Implementation Plan

[0119] The present invention also provides the following non-limiting embodiments.

[0120] Embodiment 1 is an isolated nucleic acid sequence encoding a chimeric adenoviral capsid or a functional derivative thereof, wherein the chimeric adenoviral capsid or a functional derivative thereof comprises: a fiber polypeptide having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 11; a hexon polypeptide having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 12; and a penton polypeptide having an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 13.

[0121] Embodiment 2 is the isolated nucleic acid sequence of embodiment 1, wherein the fiber polypeptide sequence comprises the amino acid sequence of SEQ ID NO:11.

[0122] Embodiment 3 is the isolated nucleic acid sequence of embodiment 1 or 2, wherein the hexon polypeptide sequence comprises the amino acid sequence of SEQ ID NO:12.

[0123] Embodiment 4 is the isolated nucleic acid of any one of embodiments 1-3, wherein the penton polypeptide comprises the amino acid sequence of SEQ ID NO:13.

[0124] Embodiment 5 is a vector comprising the isolated nucleic acid of any one of embodiments 1-4.

[0125] Embodiment 6 is the vector of embodiment 5, wherein the vector is an adenoviral vector.

[0126] Embodiment 7 is the vector of embodiment 6, wherein the adenoviral vector further comprises a transgene, optionally, wherein the transgene is a therapeutic transgene.

[0127] Embodiment 8 is the vector of embodiment 6 or 7, wherein the adenoviral vector further comprises an E1 deletion.

[0128] Embodiment 9 is the vector of any one of embodiments 6-8, wherein the adenoviral vector further comprises an E3 deletion.

[0129] Embodiment 10 is the vector of any one of embodiments 6-9, wherein the adenoviral vector is a chimeric adenoviral vector comprising one or more adenoviral nucleic acid sequences from at least one of human adenovirus-4, human adenovirus-5, human adenovirus-26, or human adenovirus-35.

[0130] Embodiment 11 is the vector of embodiment 10, wherein the adenoviral vector comprises human adenovirus-5 (HAdV-5) E4 orf6.

[0131] Embodiment 12 is the vector of any one of embodiments 6-10, wherein the adenoviral vector comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO: 10.

[0132] Embodiment 13 is the vector of any one of embodiments 6-12, wherein the transgene is located at the E1 deletion, the E3 deletion and / or adjacent to the right inverted terminal repeat (rITR).

[0133] Embodiment 14 is a recombinant cell comprising the vector of any one of embodiments 5-12.

[0134] Embodiment 15 is a method for producing a carrier, comprising:

[0135] (a) growing the recombinant cell of embodiment 14 under conditions for producing an adenoviral vector; and

[0136] (b) isolating the vector from the recombinant cell.

[0137] Embodiment 16 is an immunogenic composition comprising the adenoviral vector of any one of embodiments 6 to 13 and a pharmaceutically acceptable carrier.

[0138] Embodiment 17 is a method of inducing an immune response in a subject in need thereof, comprising administering the immunogenic composition of embodiment 16 to the subject.

[0139] Embodiment 18 is a method of producing a vaccine, comprising combining the adenoviral vector of any one of embodiments 6-13 with a pharmaceutically acceptable carrier.

[0140] Embodiment 19 is an adenoviral vector comprising:

[0141] (a) at least one transgene; and

[0142] (b) a nucleic acid encoding a chimeric adenoviral capsid or a functional derivative thereof, wherein the chimeric adenoviral capsid comprises: a fiber polypeptide having an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 11; a hexon polypeptide having an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 12; and a penton polypeptide having an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 13.

[0143] Embodiment 20 is the adenoviral vector of embodiment 19, wherein the fiber polypeptide sequence comprises the amino acid sequence of SEQ ID NO:11.

[0144] Embodiment 21 is the adenoviral vector of embodiment 19 or 20, wherein the hexon polypeptide sequence comprises the amino acid sequence of SEQ ID NO:12.

[0145] Embodiment 22 is the adenoviral vector of any one of embodiments 19-21, wherein the penton polypeptide comprises the amino acid sequence of SEQ ID NO:13.

[0146] Embodiment 23 is the adenoviral vector of any one of embodiments 19-22, wherein the adenoviral vector further comprises an E1 deletion.

[0147] Embodiment 24 is the adenoviral vector of any one of embodiments 19-23, wherein the adenoviral vector further comprises an E3 deletion.

[0148] Embodiment 25 is the adenoviral vector of any one of embodiments 19-24, wherein the adenoviral vector is a chimeric adenoviral vector comprising one or more adenoviral nucleic acid sequences from at least one of human adenovirus-4, human adenovirus-5, human adenovirus-26, or human adenovirus-35.

[0149] Embodiment 26 is the adenoviral vector of embodiment 25, wherein the adenoviral vector comprises human adenovirus-5 (HAdV-5) E4 orf6.

[0150] Embodiment 27 is the adenoviral vector of any one of embodiments 19-26, wherein the adenoviral vector comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO:8, SEQ ID NO:9, and SEQ ID NO:10.

[0151] Embodiment 28 is the adenoviral vector of any one of embodiments 19-27, wherein the transgene is located at the E1 deletion, the E3 deletion, and / or adjacent to the right inverted terminal repeat (rITR).

[0152] Embodiment 29 is the adenoviral vector of any one of embodiments 19-28, wherein the transgene is a therapeutic transgene.

[0153] Embodiment 30 is a recombinant cell comprising the adenoviral vector of any one of embodiments 19-29.

[0154] Embodiment 31 is a method for producing an adenoviral vector, comprising:

[0155] (a) growing the recombinant cell of embodiment 30 under conditions for producing an adenoviral vector; and

[0156] (b) isolating the adenoviral vector from the recombinant cell.

[0157] Embodiment 32 is a pharmaceutical composition comprising the adenoviral vector of any one of embodiments 19-29 and a pharmaceutically acceptable carrier.

[0158] Embodiment 33 is a method of inducing an immune response in a subject in need thereof, comprising administering the pharmaceutical composition of embodiment 32 to the subject.

[0159] Embodiment 34 is a method of producing a vaccine, the method comprising combining the adenoviral vector of any one of embodiments 19-29 with a pharmaceutically acceptable carrier.

[0160] Embodiment 35 is a method of expressing a transgene in a subject in need thereof, the method comprising:

[0161] a. Identify the subject in whom expression of the transgene is desired;

[0162] b. contacting the subject with the vector of embodiment 7 or the adenoviral vector of any one of embodiments 19-29; and

[0163] c. expressing the transgene in the subject.

[0164] Embodiment 36 is the method of embodiment 35, wherein expression of the transgene in the subject in need thereof treats or prevents a disease or disorder.

[0165] Embodiment 37 is the method of embodiment 35, wherein contacting the subject with the vector comprises isolating cells from the subject, and contacting the cells with the vector.

[0166] Embodiment 38 is the method of any one of embodiments 35-37, wherein the subject in need thereof is a human subject.

[0167] Embodiment 39 is the use of the vector of embodiment 7 for treating or preventing a disease or disorder in a subject in need thereof, the method comprising contacting the subject in need thereof with the vector, wherein contacting the subject in need thereof results in expression of a therapeutic transgene.

[0168] Embodiment 40 is the use of embodiment 39, wherein contacting the subject with the vector comprises isolating cells from the subject, and contacting the cells with the vector.

[0169] Embodiment 41 is the use of the adenoviral vector of any one of embodiments 19-29 for treating or preventing a disease or disorder in a subject in need thereof, the method comprising contacting the subject in need thereof with the adenoviral vector, wherein contacting the subject in need thereof results in expression of a therapeutic transgene.

[0170] Embodiment 42 is the use of embodiment 41, wherein contacting the subject with the adenoviral vector comprises isolating cells from the subject, and contacting the cells with the adenoviral vector.

[0171] Example

[0172] Example 1: Generation of E1- and E3-deleted vectors based on the novel adenoviral isolate Ad20-42-42.

[0173] A novel human adenovirus isolate, Ad20-42-42 (SEQ ID NO: 1), was identified and sequenced. This human adenovirus isolate was found to belong phylogenetically to human adenovirus species D (HAdV-D) and to be a natural chimera of HAdV-20 and HAdV-42. The penton gene is derived from HAdV-20, while the hexon and fiber genes are derived from HAdV-42. Figure 1 Shown is the genome map of the Ad20-42-42 human adenovirus isolate.

[0174] Description of the three-plasmid system for the generation of Ad20-42-42-based Ad vectors

[0175] A recombinant adenoviral vector based on Ad20-42-42 was generated by using a three-plasmid system. The plasmid system includes a "linker plasmid" covering the 5' end of the adenoviral genome, in which the E1 region is deleted and replaced with an expression cassette carrying the gene of interest (LacZ, Luc+, or eGFP). The second "middle" plasmid covers the middle part of the adenoviral genome without any modification. The 3' end of the adenoviral sequence is carried by the "right end" plasmid. In the right end construct, the E3 region is deleted, and the native ORF6 / 7 in the E4 region is replaced with the ORF6 / 7 of HAdV-5.

[0176] Table 1: Plasmids used to construct recombinant adenoviral vectors based on Ad20-42-42

[0177]

[0178] The viral vector sequences carried by the plasmids overlapped at ~2000 nucleotide positions to allow for the expression of these sequences in HEK293 or PER. Homologous recombination in cells ( Figure 2 ).

[0179] Ad20-42-42-based Ad vector genome design:The plasmid system was constructed by several steps of standard molecular cloning methods. Each Ad20-42-42-based Ad vector genome was designed to contain an E1 deletion, an E3 deletion, a different transgene insertion site, and replacement of the native E4 open reading frame (orf) 6 and orf6 / 7 with the E4 open reading frame (orf) 6 and orf6 / 7 of human adenovirus-5 (HAdV-5) (GenBank sequence AC_000008, base pairs 32966-34077).

[0180] Connector plasmid

[0181] First, the E1-deleted linker plasmid "pAdApt20-42-42.Empty" was constructed. The expression cassette was placed in the previous position of the deleted E1 region. This cassette is driven by the cytomegalovirus major immediate early promoter (i.e., "CMV promoter") and contains an SV40-derived polyadenylation signal ("SV40 poly A"). The expression cassette is equipped with a multiple cloning site to facilitate the insertion of various genes of interest (LacZ, Luc+, and eGFP). The empty linker plasmid pAdApt20-42-42 was constructed as follows:

[0182] Fragment 1 was generated covering nucleotides (nt) 1-461 of the wild-type adenovirus sequence, and PCR primers were used to introduce a PacI restriction enzyme site on the 5' side and an AvrII site on the 3' side. The PCR product of fragment 1 was double digested with PacI and AvrII.

[0183] Segment 2, which contains nucleotides 3361-5908 of the SV40 polyA and wild-type adenovirus sequence, was generated by preparing two PCR amplicons and then performing assembly PCR on the two products. The first PCR product containing "SV40 polyA" was amplified from a previously constructed plasmid (pAdApt26.Empty; Abbink et al., J. Virol. 81(9):4654-63 (2007)). An XbaI restriction site was introduced at the 5' end of the PCR product, and a reverse primer was used to introduce an Ad20-42-42 homology fragment into the PCR product. This overlap contains a natural XbaI recognition site (present in the Ad20-42-42 genome but intentionally destroyed by changing one base in the recognition site). This PCR fragment is 174 bp long. The second PCR covers the Ad20-42-42 genome from the beginning of pIX (including pIX) to approximately the middle of the polymerase gene. The forward primer for this PCR product overlaps with PCR product 1, which contains the SV40 polyA (with the XbaI recognition site disrupted). A reverse primer introduces a PacI recognition site into this PCR fragment. This PCR fragment is 2574 bp long. Subsequently, an assembly PCR is performed using PCR products 1 and 2 as templates to combine these fragments. The resulting assembly PCR product is 2710 bp in size. This assembly PCR product is then double-digested with XbaI and PacI.

[0184] A previously constructed plasmid (Abbink et al., J. Virol. 81(9):4654-63 (2007)) was digested with XbaI, AvrII, and PacI to obtain the plasmid backbone and CMV promoter (fragments 2108 and 880). A four-point ligation was then performed using pAdApt26.Empty (fragments 2108 and 880) and fragments 1 and 2. This generated the pAdApt20-42-42.Empty plasmid.

[0185] The reporter gene was inserted into the MCS by using unique KpnI, HIndIII or BamHI sites and subsequent ligation of the XbaI site.

[0186] intermediate plasmid

[0187] The intermediate plasmid contains the wild-type adenoviral genome from nt position 2088 to 18494 without any modification. First, two PCR fragments were created. One covers the 5' end of the intermediate fragment, incorporates a PacI site in the forward primer, and the reverse primer is designed slightly downstream of the natural SbfI site in the adenoviral genome (product size: 2273 bp). The second PCR fragment was created with a forward primer designed slightly upstream of another natural SbfI site in the adenoviral genome, and incorporates a PacI site in the reverse primer (product size: 2407 bp). Both PCR fragments were digested with PacI and SbfI restriction enzymes.

[0188] The pBR322 subcloning backbone plasmid (Abbink et al., J. Virol. 81(9):4654-63 (2007)) (2086 bp) flanked by PacI sites was digested with PacI. A three-point ligation was performed using the double-digested PCR product and the PacI-digested, dephosphorylated pBR backbone plasmid. This generated the plasmid pBR.Ad20-42-42.PacI-SbfI. The pBR.Ad20-42-42.PacI-SbfI plasmid obtained in the previous step was cut with the SbfI restriction enzyme. Wild-type Ad20-42-42 genomic DNA was digested with the SbfI restriction enzyme, and an 11,858 nt fragment covering the wild-type genome from the polymerase gene to approximately the middle of the pVI gene was ligated into the pBR.Ad20-42-42.PacI-SbfI plasmid. This generated the "pBR.Ad20-42-42.SbfI final interm" plasmid.

[0189] Right end plasmid

[0190] Construction of pBrAd20-42-42.SrfI-rITR The 5' end of the right end of the Ad-20-42-42 genome (fragment 1, 2098 bp) was amplified by PCR, resulting in a natural SrfI-SbfI fragment within the PCR product. A PacI recognition site was also added to the forward primer. A second fragment covering the 3' end of the right end of the Ad-20-42-42 genome was generated using an SbfI recognition site introduced by the forward primer and a PacI recognition site incorporated into the reverse primer. A natural MluI site was present near the 5' end of the PCR product. Both PCR products were then digested with PacI and SbfI enzymes.

[0191] The pBR322 subcloning backbone plasmid (Abbink et al., J. Virol. 81(9):4654-63 (2007)) flanked by PacI sites was digested with PacI (fragment size 2108 bp). The two PacI and SbfI digested PCR fragments from the previous step were cloned into the pBR backbone to generate the pBrSrfI-SbfI / MluI-rITR plasmid. This plasmid was then digested with SbfI and MluI. The Ad20-42-42 wild-type genomic DNA was digested with SbfI and MluI, and the fragment between genomic nucleotide positions 17742 and 33714 was isolated and cloned into the above plasmid. This generated pBrAd20-42-42SrfI-rITR, which carries the wild-type Ad20-42-42 genome from position 15373 to the last nucleotide of the rITR (35187).

[0192] Deletion of the E3 region: To delete the E3 region, two PCR reactions were performed using the pBrAd20-42-42SrfI-rITR plasmid as a template. The first was designed upstream of the region to be deleted; the forward primer overlapped a native AscI site, while the reverse primer incorporated a SpeI site. The second PCR reaction was designed downstream of the E3 region. The reverse primer overlapped a native EcoRI site in the Ad20-42-42 genome, while the forward primer incorporated a SpeI site. The first product was then double-digested with SpeI and AscI, and the second product with SpeI and EcoRI. The pBrAd20-42-42Srf-rITR plasmid was digested with AscI and EcoRI, and the digested plasmid and PCR product were subjected to a three-point ligation. As a result, the fragment between genomic nucleotide positions 26673 and 30753 of wild-type Ad20-42-42 was detached from the genome, and the sequence was ligated via the introduced SpeI site. This generated the pBrAd20-42-42.SrfI-rITR.dE3 plasmid.

[0193] Replacement of native ORF6 / 7 with homologous regions from HAdV-5: To replace ORF6 / 7, three fragments were amplified by PCR. Two fragments were designed to cover the upstream and downstream regions of the ORF6 / 7 to be replaced. The forward primer of the first PCR had an AscI site, and the reverse primer of the second PCR had an EcoRI site. A third PCR was performed to obtain HAdV-5ORF6 / 7 from a previously generated plasmid (pBr.Ad26.SrfI-rITR.dE3.5orf6 (Abbink et al., J. Virol. 81(9):4654-63 (2007)), which partially overlapped with the other two PCR products. These three PCR products were then subjected to fusion PCR and subsequently double digested with AscI and EcoRI. The pBrAd20-42-42.SrfI-rITR.dE3 plasmid was also digested with AscI and EcoRI, and the digested fusion PCR product was inserted by 2-point ligation to obtain the final right-end plasmid: pBrAd20-42-42Srf-rITR.dE3.5orf6.

[0194] Generation and production of Ad20-42-42-based adenoviral vectors

[0195] Adenoviral vectors Ad20-42-42.LacZ.5ORF6, Ad20-42-42.Luc+.5ORF6, and Ad20-42-42.eGFP.5ORF6, comprising the adenoviral vector genome sequences of SEQ ID NO:8, SEQ ID NO:9, and SEQ ID NO:10, respectively, were generated by transfecting the corresponding plasmids: (1) the linker plasmid "pAdApt20-42-42.LacZ" (SEQ ID NO:3) or "pAdApt20-42-42.Luc+" (SEQ ID NO:4) or "pAdApt20-42-42.eGFP" (SEQ ID NO:5); (2) the intermediate plasmid "pBR.Ad20-42-42.Sbf final interm" (SEQ ID NO:6); Figure 5 ); (3) right end plasmid "pBrAd20-42-42SrfI-rITR.dE3.5orf6" (SEQ ID NO: 7; Figure 6 ).

[0196] Transfection of the three-plasmid system was performed in E1-supplemented HEK293 cells. Prior to transfection into HEK293 cells, HEK293 cells were grown as adherent cultures in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS). The Ad vector genome plasmids were digested with PacI to release the respective adenoviral vector genome segments from the plasmids. Transfections were performed using Lipofectamine transfection reagent (Invitrogen; Carlsbad, CA) according to standard protocols. Following harvesting of the virus rescue transfection, the virus was further amplified by performing several consecutive rounds of infection in HEK293 cell cultures. Viruses were purified from crude virus harvests using a two-step cesium chloride (CsCl) density gradient ultracentrifugation method as previously described (Havenga et al., "Novel replication-incompetent adenoviral B-group vectors: high vector stability and yield in PER.C6 cells," J. Gen. Virol. 87(8):2135-43 (2006)). Viral particle (VP) titers were measured by a spectrophotometric method as previously described (Maizel et al., "The polypeptides of adenovirus: I. Evidence for multiple protein components in the virion and a comparison of types 2, 7A, and 12," Virology, 36(1):115-25 (1968)).

[0197] Example 2: Cellular and humoral immune responses induced by novel adenoviral vectors

[0198] This example describes experiments conducted to evaluate the immunogenicity of novel adenoviral vectors based on Ad20-42-42 generated herein. In these experiments, the ability of the novel vectors to induce humoral and cellular immune responses against vector-encoded (model) antigens in mice following intramuscular immunization was evaluated. The vectors tested expressed firefly luciferase (FLuc) as a model antigen. The vectors were compared side by side with a benchmark vector based on human adenovirus type 26 (HAdV-26, also referred to herein as Ad26). The immune response to the respective antigens was measured using well-known immunological assays such as enzyme-linked immunospot assays (ELISPOT) and enzyme-linked immunosorbent assays (ELISA).

[0199] Cellular immune response induced by Ad20-42-42.FLuc

[0200] The immunogenicity of the novel adenoviral vector Ad20-42-42 was evaluated in Balb / C mice immunized intramuscularly with Ad26.FLuc (benchmark control), Ad20-42-42 vector expressing firefly luciferase (Ad20-42-42.FLuc), or an adenoviral vector lacking the transgene (Ad26.E). Two vector doses were tested: 10 9 and 10 10 Virus particles (vp) per mouse, except for the Ad26.E group, which used 10 10 Two weeks after immunization, the animals were sacrificed and serum and spleen cells were sampled ( Figure 7A ).

[0201] Cellular immune responses to vector-encoded antigens were assessed by FLuc-specific IFN-γ ELISPOT assay. To this end, splenocytes sampled from immunized mice at sacrifice were stimulated overnight with a pool of 15mer overlapping FLuc peptides. Antigen-specific immune responses were determined by measuring the relative number of cells secreting IFN-γ. Figure 7B The results showed that at the highest vaccination dose (10 10 ), the cellular immune response induced by Ad20-42-42 was comparable to that observed with Ad26.Fluc. As expected, no FLuc-specific response was detected against the adenovector lacking firefly luciferase (Ad26.E). Overall, the data show that Ad20-42-42 induces a robust cellular response.

[0202] Example 3: Evaluation of serological cross-neutralization between novel and existing adenoviral vectors

[0203] For its potential utility as a novel adenoviral vaccine vector, the novel Ad20-42-42 adenoviral vector created herein is preferably serologically distinct from existing adenoviral vectors currently being developed as vaccine vectors, such as vectors based on the human adenovirus serotype HAdV-26. Therefore, cross-neutralization experiments were performed between the novel Ad20-42-42 adenoviral vector and existing vectors based on HAdV-26 (Ad26). To this end, mouse antisera raised against these vectors were cross-tested against the two vectors in adenoviral neutralization experiments. 10Mouse antisera for this study were sampled from Balb / C mice two weeks after immunization with 10 vector particles per mouse. Adenovirus neutralization assays were performed as previously described (Spangers et al 2003. J. Clin. Microbiol. 41:5046-5052). Briefly, serum was serially diluted two-fold, starting at a 1:16 dilution, and then premixed with an adenoviral vector expressing firefly luciferase (FLuc) and incubated with A549 cells overnight (multiplicity of infection (MOI) of 500 viral particles per cell). The level of luciferase activity in infected cell lysates measured 24 hours after infection represented vector infection efficiency. The neutralization titer for a given vector was defined as the highest serum dilution that reduced vector infection efficiency by 90%. Neutralization titers were arbitrarily categorized as: <16 (no neutralization), 16.1-200 (mild cross-neutralization), 201-2000 (cross-neutralization), and >2001 (strong cross-neutralization). The results showed no significant cross-neutralization between the vectors tested ( Figure 8 Only slight, one-way cross-neutralization was observed between the vectors Ad20-42-42 and Ad26, with Ad26 antiserum exhibiting a neutralization titer of 23 against Ad20-42-42, while Ad20-42-42 antiserum showed no neutralization against Ad26. Thus, the novel adenoviral vector Ad20-42-42 exhibited low to non-existent cross-neutralization with the human adenoviral vector Ad26.

[0204] Example 4: Seroprevalence of novel adenoviral vectors in the human population

[0205] High levels of pre-existing anti-vector humoral immunity in the vaccine target population may hinder the potential use of novel adenoviral vectors as effective vaccine platforms. Therefore, the seroprevalence of the Ad20-42-42 vector in 103 human serum samples was assessed. Human serum samples were tested for vector neutralization using a CPE-based assay (for wild-type (wt) virus) and a reporter assay (for Luc+-expressing virus).

[0206] Standard adenovirus neutralization assays were performed as described in Example 3 and previously (Spangers et al. 2003. J. Clin. Microbiol. 41: 5046-5052). For the wild-type adenovirus neutralization assay, serum was heat-inactivated at 55°C for 15 minutes and diluted (1 / 2, 1 / 4, 1 / 8, 1 / 16, or 1 / 32). Next, 50 μl of adenovirus stock (diluted to a 50% inhibitory dose (CCID) of 200 cells) was added to the serum. 50)) were added to the wells containing serum. On days 5 to 6, the plates were analyzed for inhibition of CPE using the MTT assay (Promega). Neutralization of serum was scored as positive when protection from CPE was >90%. The percentage of replication inhibition was calculated relative to the positive and negative controls.

[0207] As described above, briefly, serum was serially diluted 2-fold starting from a 1:16 dilution and then premixed with an adenoviral vector expressing firefly luciferase (FLuc) and subsequently incubated with A549 cells overnight (multiplicity of infection of 500 viral particles per cell). The level of luciferase activity in infected cell lysates measured 24 hours after infection represents the vector infection efficiency. The neutralization titer for a given vector was defined as the highest serum dilution that reduced the vector infection efficiency by 90%. Neutralization titers were arbitrarily divided into the following categories: <16 (no neutralization), 16-50, 50-200, 200-500, 500-1000, and >1000.

[0208] The results showed that the Ad20-42-42 adenoviral vector had a considerably lower seroprevalence (20-35% seropositive) in the human subjects studied compared to the HAdV-5 vector tested in the same assay ( Figure 9 ). In addition, positive neutralization titers against the novel Ad20-42-42 vector were generally low, mostly no higher than 200. In contrast, positive neutralization titers against HAdV-5 were found to be in the higher range, reaching over 1000. Although the vector HAdV-35 showed a similar % seropositivity as Ad20-42-42, the titers observed for HAdV-35 were in a higher range than those for Ad20-42-42.

[0209] In summary, the above data indicate that pre-existing humoral anti-vector immunity to the Ad20-42-42 vector can be considered low in the vaccine target population evaluated, suggesting that this vector has the potential to serve as an effective vaccine vector in this population.

[0210] Example 5: Transduction ability of Ad20-42-42 vector in human vascular cells

[0211] The ability to transduce cells of interest and express the encoded protein is an essential characteristic of a vector to be used in gene therapy. The transduction capacity of the novel adenoviral vector Ad20-42-42 in vascular cells was tested using a luciferase assay.

[0212] Briefly, HSVEC (human saphenous vein endothelial cell) cells were transduced in a dose-dependent manner with Ad20-42-42 vectors expressing luciferase and LacZ, and in the presence or absence of hemagglutination factor X (FX) to determine sensitivity to FX-mediated tropism or modification without tropism. HAd5 and HAd35 were used as control vectors. Cells were plated in 96-well plates at a density of 10,000 cells / well. Luciferase was expressed at 37°C for 3 hours with or without the addition of FX, using Ad5Luc, Ad35Luc, and Ad20-42-42Luc+ at 1000, 5000, and 10,000 viral particles (vp) per cell. After a 3-hour incubation, the culture medium was removed, and the cells were cultured in complete medium for 48 hours before analysis of luciferase transgene expression, expressed as relative light units (RLU) per milligram (mg) of protein.

[0213] Statistical significance between groups was calculated using a two-sample, two-tailed t-test, where p < 0.05 was considered statistically significant (p < 0.05*, p < 0.01**). All results represent the average data of several experiments with four replicates per condition ( Figure 10 ).

[0214] The data obtained showed that the transduction potential of Ad20-42-42 was significantly higher than that of HAd5 and HAd35 in the presence of FX. The highest luciferase expression was noted when a dose of 10,000 vp / cell was applied, reaching 1.5x10 9 , while without FX it reaches ~1x10 8 Thus, these results indicate that transduction by Ad20-42-42 is significantly enhanced by FX and that this vector has stronger transduction properties than the two control vectors in the presence of FX.

[0215] The superior transduction profile of Ad20-42-42 compared to the HAd5 control vector was confirmed by LacZ staining of HSVEC cells in the presence of FX using three different vector doses (1000, 5000, and 10,000 vp / cell). The staining was observed using simple light microscopy ( Figure 11 ).

[0216] As expected, the results showed a gradual increase in staining with increasing doses. Figure 11 The staining at 1000, 5000, and 10,000 vp / cell is shown from right to left in the middle. Ad20-42-42-transduced cells showed stronger staining compared to HAd5-transduced cells.

[0217] In summary, the data show that in the presence of FX, Ad20-42-42 is able to transduce vascular cells at a higher rate than a control vector. This makes Ad20-42-42 an excellent candidate gene therapy vector for the treatment of diseases in which endothelial cells play a key role, such as cardiovascular disease or cancer.

[0218] Example 6: Candidate receptors for AdV20-42-42

[0219] An important feature of Ad20-42-42 that distinguishes it from many other vectors is its potential to bind to the coxsackievirus-adenovirus receptor (CAR) and the CD46 cellular receptor, as well as its enhanced sensitivity to FX during transduction, thereby expanding the range of human cells and tissues that can be targeted for gene therapy using the Ad20-42-42 vector.

[0220] To test the ability of Ad20-42-42 to bind to multiple receptors and use it as a vehicle for cell entry, several indicator cell types were used. Chinese hamster ovary (CHO) cells expressing or lacking CAR; CHO cells expressing or lacking sialic acid; and TC1 cells expressing or lacking desmoglein 2 (DSG2) were transduced with Ad20-42-42 and HAd5 as a control vector. Figure 12A and 12B ). Cells were transduced with 10,000 vp / cell and luciferase assays were performed as described above.

[0221] The results obtained from these experiments define CAR as the potential major transmembrane receptor bound by Ad20-42-42 in cells that possess all three receptors on their surface, compared with sialic acid and DSG2.

[0222] Similar experiments were performed to test the ability of this vector to bind to CD46, another receptor commonly used by many adenovirus types for cell entry. For this purpose, CHO cells containing or lacking different isoforms of this receptor (BC1, BC2, C1, and C2) were transduced with Ad20-42-42 and HAd5 as a control vector. The results showed that Ad20-42-42 appeared to predominantly use the C2 receptor isoform during cell infection. On the other hand, K1 cells lacking CD46 were poorly transduced ( Figure 12C and 12D ).

[0223] Statistical significance between groups was calculated using a two-sample, two-tailed t-test, where p < 0.05 was considered statistically significant (p < 0.05*, p < 0.01**). All results represent the average data of several experiments with four replicates per condition. Error bars represent the standard error of the mean (SEM).

[0224] These findings indicate that the novel adenoviral vector, Ad20-42-42, binds to both CAR and CD46 receptors, which are present on many cell types. This broadens the range of cell types that can be transduced by the Ad20-42-42 vector and may facilitate its use in gene therapy.

[0225] Example 7: Biodistribution of Ad20-42-42 in mice

[0226] In vivo experiments were performed with the aim of examining the biodistribution of Ad20-42-42, its effects on the cardiovascular system (CVS), and its role as a gene delivery vehicle.

[0227] Immunocompetent male mice aged 8-10 weeks were used. Six groups of animals were formed (5 animals per virus test group and 3 animals per PBS control group). To remove circulating macrophages and more effectively assess viral transport throughout the body, 200 μl of clodophosphate liposomes (CL) were intravenously (iv) administered to the corresponding groups 48 hours before virus administration.

[0228] The treatment group received a single viral dose (10x10 11 The control group was injected with 100 ml PBS at the same time point.

[0229] Luciferase activity was read out using bioluminescence imaging 48 hours after viral delivery after injecting animals with 0.5 ml of luciferin. Animals were maintained under inhalation anesthesia. Luciferase expression levels were detected at Figure 13 Displayed in.

[0230] After imaging, the animals were sacrificed and their organs (liver, heart, spleen, kidney, intestine, pancreas, and lung) were collected. The vector genome was quantified by qPCR.

[0231] In the control group of animals infected with HAd5, the virus was mainly distributed in the liver and spleen, and in the group not treated with CL, the levels were 2x10 5 and ~3x10 5 bioluminescent units, while in the group pretreated with CL, the distribution was higher, approaching 5x10 5 ( Figures 14A-14B ).

[0232] On the other hand, Ad20-42-42 seemed to have a tropism only for the spleen, and was not detected in other organs. As expected, compared with the group without CL pretreatment (1x10 5) compared to the total DNA copy number when CL was added (~2.5x10 6 )( Figures 14C-14D ).

[0233] Together, these data suggest that Ad20-42-42 has a favorable safety profile, with tropism only observed for the spleen in the study, while DNA copy numbers were undetectable in other organs tested. The Ad20-42-42 vector did not exhibit any tropism for the liver, making it a vector with low liver availability and toxicity, and therefore, more suitable for gene therapy applications where gene transfer to the liver would hinder efficacy in the target tissue.

[0234] Those skilled in the art will appreciate that changes can be made to the embodiments described above without departing from the broad inventive concept thereof. It should therefore be understood that this invention is not limited to the particular embodiments disclosed, but is intended to cover modifications within the spirit and scope of the invention as defined herein. Sequence Listing <110> Janssen Vaccines & Prevention Batavia Biosciences University of Edinburgh Board of Trustees Niklas Arnberg Consulting <120> Adenovirus vector and its use <130> 004852.120WO1 <150> US62 / 909,853 <151> 2019-10-03 <160> 13 <170> PatentIn version 3.5 <210> 1 <211> 35187 <212> DNA <213> Artificial sequence <220> <223> Ad20-42-42 <400> 1 catcatcaat aatatacccc acaaagtaaa caaaagttaa tatgcaaatg agcttttgaa 60 tttagggcgg agccgacgct gattggacga gagaagacga tgcaaatgac gtcacgacgc 120 acggctaacg gtcgccgcgg aggcgtggcc tagcccgga gcaagtcgcg gggctgatga 180 cgtataaaaa agcggacttt agacccgga atggccgatt ttcccgcggc cacggccgga 240 300. the ttctgggcgg atgcaagtga aattaggtca ttttggcgcg aaaactgaat gaggaagtga aaagcgaaaa ataccgatcc cgcccagggc ggaatttta ccgagggccg agagactttg accgattacg tggggggtttc gattgcggtg tttttttcgc gaatttccgc 420 gtccgtgtca aagtccggtg tttatgtcac agatcagctg atccacaggg tatttaaacc 480 agtcgagccc gtcaagaggc cactcttgag tgccagcgag tagatttc tctgagctcc 540 gctcccagag tgtgagaaaa 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gctgcgctta gtcatgtgta 1440 aataaagttg tacaataaaa gtatatgtga cgcatgcaag gtgtggttta tgactcatgg 1500 gcggggctta ggcctatata agtggtaaca cctgggcact caggcacaga cattcaggga 1560 <h2 style=";text-align:left;direction:ltr">gctcctgatg gaggtgtgga ctatccttgg agactttaac aagacacgcc ggcttgtaga 1620<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> ggatagttca gacgggtgct ccgggttctg gagacactgg tttggaactc ctctatctcg 1680<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> cctggtgtac acagttaaga aggattataa cgaggaattt gaaaatcttt ttgccgactg 1740<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> ctctggcctg cttgattctc tgaattttgg ccaccagtcc cttttccagg aaagggtcct 1800<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> ccacagcctt gatttttcca gcccagggcg cactacagcc ggggttgctt ttgtggtttt 1860<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> tctggttgac aaatggagcc agaacaccca actgagcagg ggctacattc tggacttcgc 1920<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> agccatgcac ctgtggagag cctgggtcag gcagcgggga cagagaatct tgaactactg 1980<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> gcttctacag ccagcagctc cgggtcttct tcgtctacac agacaaacat ccatgttgga 2040<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> ggaagaaatg aggcaggcca tggacgagaa cccgaggagc ggcctggacc ctccgtcgga 2100<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> agaggagctg gattgaatca ggtatccagc ctgtacccag agcttagcaa ggtgctgaca 2160<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> tccatggcca ggggagtgaa gagggagagg agcgatgggg gcaataccgg gatgatgacc 2220<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> gagctgacgg ccagcctgat gaatcgcaag cgcccagagc gcattacctg gcacgagcta 2280<h2 style=";text-align:left;direction:ltr"> cagatggagt gcagggatga ggtgggcctg atgcaggata aatatggcct ggagcagata 2340 aaaacccact ggttgaaccc agatgaggat tgggaggagg ccattaagaa atatgccaag 2400 atagccctgc gcccagattg caagtatagg gtgaccaaga ccgtgaatat cagacatgct 2460 tgctacatct cggggaacgg ggcagaggtg gtcatcgata ccctggacaa ggccgccttc 2520 aggtgttgca tgatgggaat gagagccgga gtgatgaata tgaattccat gatcttcatg 2580 aacatgaagt tcaatggaga gaagtttaat ggggtgatgt tcatggccaa cagccacatg 2640 accctgcacg gatgcagttt cttcggcttc aacaatatgt gcgcagaggt ctggggcgct 2700 gctaagatca ggggatgtaa gttttatgga tgctggatgg gcgtggtcgg aagacccaag 2760 agcgagatgt ctgtgaagca gtgtgtgttt gagaaatgct acctgggagt ctctaccgag 2820 ggcaatgcta gagtgagaca ttgctcttcc ctggagacgg gctgcttctg cctggtgaag 2880 ggcacagcct ctctgaagca tatatggtg aagggctgca cggatgagcg catgtacaac 2940 atgctgacat gcgactcggg ggtctgccat atcctgaaga acatccatgt gacctcccac 3000 ccccggaaga agtggccagt gtttgagaat aacctgctga tcaagtgcca tatgcacctg 3060 ggagccagaa ggggcacctt ccagccgtac cagtgcaact ttagccagac caagctgctg 3120 ttggagaacg atgccttctc cagggtgaac ctgaacggca tctttgacat ggatgtctcg 3180 gtgtacaaga tcctgagata cgatgagacc aggtccaggg tgcgcgcttg cgagtgcggg 3240 ggcagacaca ccaggatgca accggtggcc ctggatgtga ccgaggagct gaggcccgac 3300 cacctggtga tggcatgtac cgggaccgag ttcagctcca gcggggagga cacagattag 3360 aggtaggttt gagtagtggg cgtggctaag atgactataa aggtgggtgt cttacgaggg 3420 tctttttgct tttctgcaga catcatgaac gggaccggcg gggccttcga aggggggctt 3480 tttagccctt atttgacaac ccgcctgccg ggatgggccg gagttcgtca gaatgtgatg 3540 ggatcgacgg tggacgggcg tccagtgctt ccagcaaatt cctcgaccat gacctacgcg 3600 accgtgggga actcgtcgct cgacagcacc gccgcagccg cggcagccgc agccgccatg 3660 acagcgacga gactggcttc gagctacatg cccagcagcg gtagcagccc ctctgtgccc 3720 agttccatca tcgccgagga gaaactgctg gccctcctgg ccgagctgga agccctgagc 3780 cgccagctgg ccgccctgac ccagcaggtg tccgagctcc gcgaacagca gcagcagcaa 3840 aataaatgat tcaataaaca cagattctga ttcaaacagc aaagcatctt tattatttat 3900 tttttcgcgc gcggtaggcc ctggtccacc tctcccgatc attgagagtg cggtggattt 3960 tttccaggac ccggtagaga tgggattgga tgttgaggta catgggcatg agcccgtccc 4020 gggggtgaag gtagcaccac tgcatggcct catgctctgg ggtcgtgttg tagatgatcc 4080 agtcatagca ggggcgctgg gcgtggtgct ggatgatgtc tttgaggagg agactgatgg 4140 ccacggggag ccccttggtg taggtgttgg cgaagcggtt gagctgggag ggatgcatgc 4200 ggggggagat gatgtgcagt ttggcctgga tcttgaggtt ggcaatgttg ccgcccagat 4260 cccgccgggg gttcatgttg tgcaggacca ccaggacggt gtagcccgtg cacttgggga 4320 acttatcatg caacttggaa gggaatgcgt ggaagaattt ggagacgccc ttgtgcccgc 4380 ccaggttttc catgcactca tccatgatga tggcgatggg cccgtgggct gcggctttgg 4440 caaagacgtt tctggggtca gagacatcat aattatgctc ctgggtgaga tcatcataag acattttaat gaatttgggg cggaggggtgc cagattgggg gacgatcgtt ccctcgggcc ccggggcga gttcccctca cagatctgca tctcccaggc tttcatctcg gagggggga tcatgtccac ctgcggggcg atgaaaaaaa cggtttccgg ggcgggggtg atgagctgcg 4680 aggagagcag gtttctcaac agctgggact tgccgcaccc ggtcgggccg taggaccc 4740 cgatgacggg ttgcaggtgg tagttcaagg agatgcagct gccgtcttcc cggaggaggg 4800 gggccacctc gttgagcatg tctctgactt ggaggttttc ccggacgagc tcgccgagga 4860 ggcggtcccc gcccagcgag agcagctctt gcagggaagc aaagtttttc aggggcttga 4920. gcccgtcggc catgggcatc ttggcgaggg tctgcgagag gagttccaag cggtcccaga 4980. gctcggtgac gtgctctcg gcatctcgat ccaacagact tcctcgtttc gggggttggg 5040 acgactgcga ctgtagggca cgagacgatg ggcgtccagc gctgccagcg tcatgtcctt 5100 ccagggtctc agggtccgcg tgagcgtggt ctccgtcacg gtgaaggggt gggccccggg 5160 ctgggcgctt gcaagggtgc gcttgagact catcctgctg gtgctgaaac gggcacggtc 5220 ttcgccctgc gcgtcggcga gatagcagtt gaccatgagc tcgtagttga gggcctcggc 5280 ggcgtggccc ttggcgcgga gcttgccctt ggaagagcgc ccgcaggcgg gacagaggag 5340 ggattgcagg gcgtagagct tgggcgcgag aaagacggac tcgggggcga aagcgtccgc 5400 tccgcagtgg gcgcagacgg tctcgcactc gacgagccag gtgagctcgg gatgttcggg 5460 gtcaaaaacc agttttcccc cgttcttttt gatgcgcttc ttacctcgcg tctccatgag 5520 tctgtgtccg cgctcggtga caaacaggct gtctgtgtcc ccgtagacgg acttgatggg 5580 cctgtcctgc aggggcgtcc cgcggtcctc ctcgtagaga aactcggacc actctgagac 5640 gaaggcgcgc gtccacgcca agacaaagga ggccacgtgc gaggggtagc ggtcgttgtc 5700 caccaggggg tccacctttt ccaccgtgtg cagacacatg tccccctcct cagcatccaa 5760 gaaggtgatt ggcttgtagg tgtaggccac gtgaccgggg gtccccgacg ggggggtata 5820 aaagggggcg ggtctgtgct cgtcctcact ctcttccgcg tcgctgtcca cgagcgccag 5880 ctgttggggt aggtattccc tctcgagagc gggcatgacc tcggcactca ggttgtcagt 5940 ttctagaaac gaggaggatt tgatgttggc ctgccctgcc gcaatgcttt tgagtaaact 6000 ctcatccatc tggtcagaaa agactatttt tttattgtca agcttggtgg cgaaggagcc 6060 atagagggcg ttggagagaa gcttggcgat ggatctcatg gtctgatttt tgtcacggtc 6120 ggcgcgctcc ttggccgcga tgtttagctg gacatattcg cgcgcgacac acttccattc 6180 ggggaagacg gtggtgcgct cgtcgggcac gatcctgacg cgccagccgc ggttatgcag 6240 ggtgaccagg tccacgctgg tggccacctc gccgcgcagg ggctcgttgg tccagcagag 6300 tcgcccgcct ttgcgcgaac agaaaggggg cagcacatca agcagatgct cgtcgggggg 6360 gtccgcatcg atggtgaaga taccgggaca gagttccttg tcaaaatagt ctatttttga 6420 ggatgcatca tccaaggcca tctgccactc acgggcggcc attgctcgct cgtaggggtt 6480 gaggggcgga ccccagggca tgggatgcgt gagggcggag gcgtacatgc cgcagatgtc 6540 atagacatag atgggctccg agaggatgcc gatgtaggtg ggataacagc gccccccgcg 6600 gatgctggcg cgcacgtagt catacaactc gtgcgagggg gccaagaagg cggggccgag 6660 attggtgcgc tggggctgct cggcgcggaa gacgatctgg cgaaagatgg cgtgcgagtt 6720 ggaggagatg gtgggccgtt ggaagatgtt aaagtgggcg tggggcaagc ggaccgagtc 6780 gcggatgaag tgcgcgtagg agctttgcag cttggcgacg agctcggcgg tgacgaggac 6840 gtccatggcg cagtagtcca gcgtttcgcg gatgatgtca taacccgtct ctccttttct 6900 ctcccacagc tcgcggttga gggcgtactc ctcgtcatcc ttccagtact cccggagcgg 6960 gaatcctcga tcgtccgcac ggtaagagcc cagcatgtag aaatggttca cggccttgta 7020 gggacagcag cccttctcca cggggagggc gtaagcttga gcggccttgc ggagcgaggt 7080 gtgcgtcagg gcgaaggtgt ccctgaccat gactttcaag aactggtact tgaaatccga 7140 gtcgtcgcag ccgccgtgct cccagagctc gaaatcggtg cgcttcttcg agagggggtt 7200 aggcagagcg aaagtgacgt cattgaagag aatcttgcct gcccgcggca tgaaattgcg 7260 ggtgatgcgg aaagggcccg ggacggaggc tcggttgttg atgacctggg cggcgaggac 7320 gatctcgtcg aagccgttga tgttgtgccc gacgatgtag agttccatga atcgcggggcg 7380. gcctttgatg tgcggcagct ttttgagctc ctcgtaggtg aggtcctcgg ggcattgcag 7440 gccgtgctgc tctagcgccc actcctggag atgtgggttg gcttgcatga aggaagccca 7500 gagctcgcgg gccatgaggg tctggagctc gtcacgaaag aggcggaact gctggcccac 7560 ggccatcttt tcgggtgtga cgcagtagaa ggtgaggggg tcccgctccc agcgatccca 7620 gcgtaagcgc acggcgagat cgcgagcgag ggcgaccagc tctgggtccc ccgagaattt 7680. 7740. catgaccagc atgaagggga cgagctgctt gccgaaggac cccatccagg tgtaggtttc tacatcgtag gtgacaaaga gccgctccgt gcgaggatga gagccgattg ggaagaactg gatttcctgc caccagttgg acgagtggct gttgatgtga tgaagtaga aatcccgccg 7860. gcgaaccgag cactcgtgct gatgcttgta aaagcgtccg cagtactcgc agcgctgcac 7920. gggctgtacc tcatccacga gatacacagc gcgtcccttg aggaggaact tcaggagtgg 7980 cggccctggc tggtggtttt catgttcgcc tgcgtgggac tcaccctggg gctcctcgag 8040 gacggagagg ctgacgagcc cgcgtgggag ccaggtccag atctcggcgc ggcgggggcg 8100 gagagcgaag acgagggcgc gcagttggga gctgtccatg gtgtcgcgga gatccaggtc 8160 cgggggcagg gttctgaggt tgacctcgta gaggcgggtg agggcgtgct tgagatgcag 8220 atggtacttg atttctacgg gtgagttggt ggccgtgtcc acgcattgca tgagcccgta 8280 gctgcgcggg gccacgaccg tgccgcggtg cgcttttaga agcggtgtcg cggacgcgct 8340 cccggcggca gcggcggttc cggtcccgcg ggcaggggcg gcagaggcac gtcggcgtgg 8400 cgctcgggca ggtcccggtg ctgcgccctg agagcgctgg cgtgcgcgac gacgcggcgg 8460 ttgacatcct ggatctgccg cctctgcgtg aagaccacgg gccccgtgac tttgaacctg 8520 aaagacagtt caacagaatc aatctcggcg tcattgacgg cggcctgacg caggatctct 8580 tgcacgtcgc ccgagttgtc ctggtaggcg atctcggaca tgaactgctc gatctcctcc 8640 tcctggagat cgccgcggcc cgcgcgctcg acggtggcgg cgaggtcatt cgagatgcga 8700 cccatgagct gcgagaaggc gcccaggccg ctctcgttcc agacgcggct gtagaccacg 8760 tccccgtcgg cgtcgcgcgc gcgcatgacc acctgcgcga ggttgagctc cacgtgccgc 8820 gcgaagacgg cgtagttgcg caggcgctgg aagaggtagt tgagggtggt ggcgatgtgc 8880 tcggtgacga agaagtacat gatccagcgg cgcagaggca tctcgctgat gtcgccgatg 8940 gcctccagcc tttccatggc ctcgtagaaa tccacagcga agttgaaaaa ctgggcgttg 9000 cgggccgaga ccgtgagctc gtcttccagg agcctgatga gctcggcgat ggtggcgcgc 9060 acctcgcgct cgaaatcccc gggggcctcc tcctcttcct cttcttccat gacgacctct 9120 tcttctattt cttcctctgg gggcggtggt ggtggcgggg cccgacgacg acggcgacgc 9180 accgggagac ggtcgacgaa gcgctcgatc atctccccgc ggcggcgacg catggtttcg 9240 gtgacggcgc gaccccgttc gcgaggacgc agcgtgaaga cgccgccggt catctcccgg 9300 taatggggcg ggtccccgtt gggcagcgat agtgcgctga cgatgcatct tatcaattgc 9360 ggtgtagggg acgtgagcgc gtcgagatcg accggatcgg agaatctttc gaggaaagcg 9420 tctagccaat cgcagtcgca aggtaagctc aaacacgtag cagccctgtg gacgctgtta 9480 gaattgcggt tgctgatgat gtaattgaag taggcgtttt tgaggcggcg gatggtggcg aggaggacca ggtccttgggg tcccgcttgc tggatgcgga gccgctcggc catgccccag 9600 gcctggccct gacaccggct caggttcttg tagtagtcat gcatgagcct ctcgatgtca 9660 tcactggcgg aggcggagtc ttccatgcgg gtgaccccaa cgcccctgag cggctgcacg 9720 agcgccaggt cggcgacgac gcgctcggcg aggatggcct gttgcacgcg ggtgagggtg 9780 tcctggaagt cgtccatgtc gacgaagcgg tggtaggccc ctgtgttgat ggtgtaagtg cagttggcca tgagcgacca gttgacggtc tgcaggccgg gctgcacgac ctcggagtac 9900 ctgagccgcg agaaggcgcg cgagtcgaag acgtagtcgt tgcaggtgcg cacgaggtac 9960. tggtatccga ctaggaagtg cggcggcggc tggcggtaga gcggccagcg ctgggtggcc 10020 ggcgcgcccg gggccaggtc ctcgagcatg aggcggtggt agccgtag gtagcgggac 10080 atccaggtga tgccggcggc ggtggtggag gcgcgcggga actcgcggac gcggttccag 10140 atgttgcgca gcggcagga atagtccatg gtcggcacgg tctggccggt gagacgcgcg 10200. cagtcattga cgctctagag gcaaaaacga aagcggttga gcgggctatt cctccgtagc 10260 ctggcggaac gcaaacgggt taggccgcgt gtgtaccccg gttcgagtcc cctcgaatca 10320 ggctggagcc gcgactaacg tggtattggc actcccgtct cgacccgagc ccgatagctg 10380 ccaggatacg gcggagagcc cttttggcgg ccgagggggg tcgctagact tgaaagcggc 10440 cgaaaaccct gccgggtagt ggctcgcgcc cgtagtctgg agaagcatcg ccagggttga 10500 gtcgcggcag aacccggttc gcggacggcc gcggagagcg ggacttggtc accccgccga 10560 tttaaaaacc cacagccagc cgacttctcc agttacggga gcgagccccc ttttttcttt 10620 ttgccagatg catcccgtcc tgcgccaaat gcgtcccacc ccccctccgg cgaccaccgc 10680 aaccgcggcc gtagcaggcg ccggcgctag ccagccacag ccacagacag agatggactt 10740 ggaagagggc gaagggctgg cgagattggg ggcgccgtct ccggagcgac acccccgcgt 10800 gcagctgcag aaggacgtgc gcccggcgta cgtgcctgcg cagaacctgt tcagggaccg 10860 cagcggggag gagcccgagg agatgcgcga ctgccgcttt cgggcgggca gggagctgcg 10920 cgagggtctg gaccgccagc gcgtgctgcg cgatgaggat ttcgagccga acgagcagac 10980 ggggatcagc cccgcgcgcg cgcacgtggc ggcggccaac ctggtgacgg cctacgagca 11040 gacggtgaag caggagcgca acttccaaaa gagtttcaac aaccacgtgc gcacgctgat 11100 cgcgcgcgag gaggtggccc tgggcctgat gcacctgtgg gacctggcgg aggccatcgt 11160 gcagaacccg gacagcaagc ctctgacggc gcagctgttc ctggtggtgc agcacagtag 11220 ggacaacgag gcgttcaggg aggcgctgct gaacatcgcc gagcccgagg gtcgctggct 11280 gctggagctg atcaacatct tgcagagcat cgtagtgcag gagcgcagcc tgagcctggc 11340 cgagaaagtg gcggcgatca actactcggt gctgagcctg ggcaagtttt acgcgcgcaa 11400 gatttacaag acgccatacg tgcccataga caaggaggtg aagatagaca gcttttacat 11460 gcgcatggcg ctcaaggtgc tgacgctgag cgacgacctg ggcgtgtacc gcaacgaccg 11520 catccacaag gccgtgagcg cgagccggcg gcgcgagctg agcgaccgcg agctgatgct 11580 gagcctgcgc cgggcgctgg tagggggcgc cgccggcggc gaggagtcct acttcgacat 11640 ggggggcggac ctgcattggc agccgagccg gcgcgccttg gaagccgcct acggtccaga 11700 ggacttggat gaggatgagg aagaggaga ggatgcaccc gctgcggggt actgacgcct 11760 ccgtgatgtg tttttagatg tcccagcaag ccccggaccc cgccataagg gcggcgctgc 11820 aaagccagcc gtccggtcta gcatcggacg actgggaggc cgcgatgcaa cgcatcatgg 11880 ccctgacgac ccgcaacccc gagtccttta gacaacagcc gcaggccaac agactttcgg 11940 ccattctgga ggcggtggtc ccctctcgaa ccaaccccac gcacgagaag gtctggcga 12000 tcgtgaacgc gctggcggag aacaaggcca tccgtcccga cgaggccggt ctggtgtaca 12060 acgccctgct ggagcgcgtg ggccgctaca acagcaaa cgtgcagtcc aacctggacc 12120 ggctggtgac ggacgtgcgc gaggccgtgg cgcagcgcga gcggttcaag aacgaaggcc 12180 tgggctcgct ggtggcgctg aacgccttcc tggcgacgca gccggcgaac gtgccgcgcg 12240 ggcaggacga ttataccaac tttataagcg cgctgcggct gatggtgacc gaggtgcccc 12300 agagcgaggt gtaccagtcg ggcccggatt actttttcca aactagcaga cagggcctgc agacggtgaa cctgagccag gcctttaaga acctgcgcgg gctgtggggc gtgcaggcgc 12480. ccgtgggcga ccggtcgacg gtgagcagct tgctgacgcc caactcgcgg ctgctgctgc tgctgatcgc gcccttcacc gacagcggca gcgtgaaccg caactcgtac ctgggccacc tgctgacgct gtaccgcgag gccataggcc aggcacaggt ggacgagcag accttccagg agatcactag cgtgagccgc gcgctggggc cgacagtctt agggccaccc tgaacttctt gctgaccaat agatcccggc agatcgcg ctatcggccg aggaggagcg catcctgaga tatgtgcagc agagcgtagg gctgttcctg atgcaggagg 12780 gggccacccc cagcgccgcg ctggacatga ccgcgcgcaa catggaacct agcatgtacg ctgccaaccg gccgttcatc aataagctga tggactacct gcaccgcgcg gcggccatga actcggacta ctttacaaac gccatcctga acccgcactg gctcccgccg ccggggttct acacgggcga gtacgatatg cccgacccca acgacgggtt cctgtgggac gacgtggaca 13020 gcgcggtgtt ctccccgacc ttgcaaaagc gccaggaggc ggtgcgcacg cccgcgagcg 13080 agggtgcggt gggtcggagc ccctttccta gcttagggag tttgcatagc ttgccgggct 13140 cggtgaacag cggcagggtg agccgaccgc gcttgctggg cgaggacgag tacctgaacg 13200 actcgctgct gcagccgccg cgggtcaaga acgccatggc caataacggg atagagagtc 13260 tggtggacaa actgaaccgc tggaagacct acgctcagga ccatagggac gcgcccgcgc 13320 cgcggcgaca gcgccacgac cggcagcggg gcctggtgtg ggacgacgag gactcggccg 13380 acgatagcag cgtgttggac ttgggcggga gcggtggggc caacccgttc gcgcatctgc 13440 agcccaaact ggggcgacgg atgttttgaa atgcaaaata aaactcacca aggccatagc 13500 gtgcgttctc ttccttgtta gagatgaggc gtgcggtggt gtcttcctct cctcctccct 13560 cgtacgagag cgtgatggcg caggcgaccc tggaggttcc gtttgtgcct ccgcggtata 13620 tggctcctac ggagggcaga aacagcattc gttattcgga gctggctccg ctgtacgaca 13680 ccactcgcgt gtacttggtg gacaacaagt cggcggacat cgcttccctg aactaccaaa 13740 acgaccacag caacttcctg accacggtgg tgcagaacaa cgatttcacc cccgccgagg 13800 ccagcacgca gacgataaat tttgacgagc ggtcgcggtg gggcggtgat ctgaagacca 13860 ttctgcacac caacatgccc aatgtgaacg agtacatgtt caccagcaag tttaaggcgc 13920 gggtgatggt ggctagaaag cacccagaga atgtagctaa agacgatttg agtcaggata 13980 agcttgaata tgagtggttt gagtttaccc tgcccgaggg caacttttcc gagaccatga 14040 ccatagacct gatgaacaac gccatcttgg aaaactactt gcaagtgggg cggcaaaatg 14100 gcgtgctgga gagcgatatc ggagtcaagt ttgacagcag aaatttcaag ctgggctggg 14160 acccggtgac caagttggtg atgccagggg tctacaccta cgaggccttc cacccggacg 14220 tggtgctgct gccgggctgc ggggtggact tcaccgagag ccgcctgagc aacctcctgg 14280 gcattcgcaa gaagcaacct ttccaagagg gcttcagaat catgtatgaa gatctagtag 14340 ggggcaacat cccagccctc ctgaatgtca aggagtatct gaaggataag gaagaagctg 14400 gcacagcaga tgcaaatacc attaaggctc agaatgatgc agtcccaaga ggagataact 14460 atgcatcagc ggcagaagcc aaagcagcag gaaagaaat tgagttgaag gccattttga 14520 aagatgattc aaacagaagc tacaatgtga tcgagggaac cacagacacc ctgtaccgca 14580 gttggtacct gtcctatacc tacgggggacc ccgagaaggg ggtgcagtcg tggacgctgc 14640 tcaccacccc ggacgtcaca tgcggcgcgg agcaagtcta ctggtcgctg ccggacctca 14700 tgcaagaccc cgtcaccttc cgctctaccc agcaagtcag caactacccc gtggtcggcg 14760 ccgagctcat gcccttccgc gccaagagct tttacaacga cctcgccgtc tactcccagc 14820 tcatccgcag ctacacctcc ctcacccag tcttcaaccg cttccccgac aaccagatcc 14880 tctgccgccc gcccgcgccc accatcacca ccgtcagtga aaacgtgcct gctctcacag 14940 atcacgggac gctaccgctg cgcagcagta tccgcggagt ccagcgagtg accgtcactg 15000 acgccccgtcg ccgcacctgt ccctacgtct acaaggccct gggcatagtc gcgccgcgcg 15060 tgctatccag tcgcaccttc taaaaaatgt ctattctcat ctcgcccagc aataacaccg 15120 gctggggtat tactaggccc agcaccatgt acggaggagc caagaagcgc tcccagcagc 15180 accccgtccg cgtccgcggc cacttccgcg ctccctgggg cgcttacaag cgcgggcgga 15240 cttctaccgc cgctgccgtg cgtaccaccg tcgacgacgt catcgactcg gtggtcgccg 15300 acgcgcgcaa ctacacccc gccccctca ccgtggacgc ggtcattgac agcgtggtgg 15360 ccgacgcgcg cgactatgcc agacgcaaga gccggcggcg acggatcgcc aggcgccacc 15420 ggagcacgcc cgccatgcgc gccgcccggg ctctgctgcg ccgcgccaga cgcacaggcc 15480 gccgggccat gatgcgagcc gcgcgccgcg ctgccactgc acccaccccc gcaggcagga 15540 ctcgcagacg agcggccgcc gccgctgccg cggccatctc tagcatgacc agacccaggc 15600 gcggaaacgt gtactgggtg cgcgactccg tcacgggcgt gcgcgtgccc gtgcgcaccc 15660 gtcctcctcg tccctgatct aatgcttgtg tcctcccccg caagcgacga tgtcaaagcg 15720 caaaatcaag gaggagatgc tccaggtcgt cgccccggag atttacggac caacccaggc 15780 ggaccagaaa ccccgcaaaa tcaagcgggt taaaaaaaag gatgaggtgg acgagggggc 15840 agtagagttt gtgcgcgagt tcgctccgcg gcggcgcgta aattggaagg ggcgcagggt 15900 gcagcgcgtg ttgcggcccg gcacggcggt ggtgttcacg cctggcgagc ggtcctcggt 15960 caggagcaag cgtagctatg acgaggtata cggcgacgac gacatcctgg accaggcggc 16020 ggagcgggcg ggcgagtttg cctacgggaa gcggtcgcgc gaagaggagc tgatctcgct 16080 gccgctggac gaaagcaacc ccacgccaag cctgaagccc gtgaccctgc agcaggtgct 16140 gccccaggcg gtgctgctgc cgagccgcgg ggtcaagcgc gagggcgaga acatgtaccc 16200 gaccatgcag atcatggtgc ccaagcgccg gcgcgtggag gacgtgctgg acaccgtgaa 16260 aatggatgtg gagcccgagg tcaaggtgcg ccccatcaag caggtggcgc cgggcctggg 16320 cgtgcagacc gtggacattc agatccccac cgacatggat gtcgacaaaa aaccctcgac 16380 cagcatcgag gtgcagaccg acccctggct cccagcctcc actgctaccg cctccacttc 16440 taccgtcgcc acggccaccg agcctcccag gaggcgaaga tggggccctg ccaaccggct 16500 gatgcccaac tacgtgttgc atccttccat catcccgacg ccgggctacc gcggcactcg 16560 gtactacgcc agccgcaggc gcccagccag caaacgccgc cgccgcaccg ccacccgccg 16620 ccgtctggcc cccgcccgcg tgcgccgcgt gaccacgcgc cggggccgct cgctcgttct 16680 gcccaccgtg cgctaccacc ccagcatcct ttaatccgtg tgctgtgata ctgttgcaga 16740 gagatggctc tcacttgccg cctgcgcatc cccgtcccga attaccgagg aagatcccgc 16800 cgcaggagag gcatggcagg cagtggcctg aaccgccgcc ggcggcgggc catgcgcagg 16860 cgcctgagtg gcggctttct acccgcgctc atccccataa tcgccgcggc catcggcacg 16920 atcccgggca tagcttccgt tgcgctgcag gcgtcgcagc gccgttgatg tgcgaataaa 16980 gcctctttag actctgacac acctggtcct gtatattttt agaatggaag acatcaattt 17040 tgcgtccctg gctccgcggc acggcacgcg gccgttcatg ggcacctgga acgagatcgg 17100 caccagccag ctgaacgggg gcgccttcaa ttggagcagt gtctggagcg ggcttaaaaa 17160 tttcggctcg acgctccgga cctatgggaa caaggcctgg aatagtagca cggggcagtt 17220 gttaagggaa aagctcaaag accagaactt ccagcagaag gtggtggacg ggctggcctc 17280 gggcattaac ggggtggtgg acatcgcgaa ccaggccgtg cagcgcgaga taaacagccg 17340 cctggacccg cgcccgccca cggtggtgga gatggaagat gcaactcttc cgccgcccaa 17400 aggcgagaag cggccgcggc ccgacgcgga ggagacgatc ctgcaggtgg acgagccgcc 17460 ctcgtacgag gaggccgtga aggccggcat gcccaccacg cgcatcatcg cgccgctggc 17520 cacgggtgta atgaaacccg ccacccttga cctgcctcca ccacccacgc ccgctccacc 17580 gaaggcagct ccggttgtgc agccccctcc tgtggcgacc gccgtgcgcc gcgtccccgc 17640 ccgccgccag gcccagaact ggcagagcac gctgcacagt atcgtgggcc tgggagtgaa 17700 aagtctgaag cgccgccgat gctattgaga gagaggaaag aggacactaa agggagagct 17760 taacttgtat gtgccttacc gccagagaac gcgcgaagat ggccaccccc tcgatgatgc 17820 cgcagtgggc gtacatgcac atcgccgggc aggacgcctc ggagtacctg agcccgggtc 17880 tggtgcagtt tgcccgcgcc accgacacgt acttcagcct gggcaacaag tttaggaacc 17940 ccacggtggc tcccacccac gatgtgacca cggaccggtc ccagcgtctg acgctgcgct 18000 tcgtgcccgt ggatcgcgag gacaccacgt actcgtacaa ggcgcgctc actctggccg 18060 tgggcgacaa ccgggtgcta gacatggcca gcacttactt tgacatccgc ggcgtcctgg 18120 accgcggccc cagcttcaaa ccctactcgg gcacggccta caacagtctg gcccccaagg 18180 gcgcccccaa ctccagccag tgggctgaca aagaaagggt gaatggaggc ggtaacacca 18240 aagatgttac aaagacattt ggtgtggcag ccatgggtgg agaagacatt acggaaaagg 18300 gtctgaagat tggaactgat acaacagaaa atgaaccaat atttgcagac aaaaatttc 18360 agcctgaacc ccaagtggga gaggaaaact ggcaggaaac atttgttttc tatggaggaa 18420 gagctcttaa aaaagaaacc aaaatgaagc cttgctatgg ttcatttgcc agaccgacta 18480 atgaaaaagg tggccaggct aaatttataa ttggagataa tggtcaaccc actgaaaacc 18540 atgacataac aatggctttc tttgacactc ctggtggtac aatcacagga ggtactggtg 18600 gtccccaaga tgaacttaaa gctgacattg ttatgtacac tgaaaatata aatcttgaaa 18660 ctccagatac ccatgtggtg tacaagccag gcaaagaaga tgatagctca gaaattaact 18720 tggtgcaaca gtccatgcc aacagaccca actacatcgg tttcagagac aactttgtgg 18780 gacttatgta ctacaacagc actggcaaca tgggcgtgct ggctggtcag gcctctcagt 18840 tgaatgctgt ggtcgacttg caagacagaa acaccgagct gtcttaccag ctcctgctag 18900 attctctggg tgacaggacc agatacttta gcatgtggaa ctctgcggtg gacagttatg 18960 atcccgatgt tagcatt gagaatcacg gtgtggaaga tgaacttccc aactattgct 19020 ttccattgga cggctcaggc actaattcag cttttcaagg tgtaaaaatt aagcaaaacc 19080 aagacggaga tgttaatgat gattgggaaa aggatgataa ggtatccact caaaaccaaa 19140 tttgcaaggg caacatttac gccatggaga tcaacctcca ggccaacctg tggaaaagtt 19200 ttctgtactc gaacgtggcc ctgtacctgc ccgactccta caagtacacg ccggccaacg 19260 tgacgctgcc cgccaacacc aacacctacg agtacatgaa cggccgcgtg gtagccccct 19320 cgctggtgga cgcctacatc aacatcggcg cccgctggtc gctagacccc atggacaatg 19380 tcaacccctt caaccaccac cgcaatgcgg gtctgcgcta ccgctccatg cttctgggca 19440 acggccgcta cgtgcccttc cacatccaag tgccccaaaaa gttctttgcc atcaagaacc 19500 tgctcctgct cccgggctcc tacacctacg agtggaactt ccgcaaggac gtcaacatga 19560 tcctgcagag ttcccttgga aacgacctgc gcgttgacgg cgcttcggtc cgcttgca 19620 gtgttaacct ctatgccacc ttcttcccca tggcgcacaa caccgcctcc accctggaag 19680 ccatgctgcg caacgacacc aacgaccagt ccttcaacga ctacctctcg gccgccaaca 19740 tgctctaccc tatcccggcc aaggccacca acgtgcccat ctccatcccc tcgcgcaact 19800 gggccgcctt ccgcggctgg agtttcaccc ggctcaagac caaggaaact ccctcccttg 19860 gctcgggttt tgacccctac tttgtctact cgggctccat ccctacctc gacgggacct 19920 tttacctgaa ccataccttc aagaaggtct ccatcatgtt cgactcctcg gtcagctggc 19980 ccggcaacga ccggctgctc acgccgaacg agttcgagat caagcgcagc gttgacgggg 20040 agggctacaa cgtggcccaa tgcaacatga ccaaggactg gttccttgtc cagatgctct 20100 cccactacaa catcggctac cagggcttcc acgtgcccga gggctacaag gaccgcatgt 20160 actcattct ccgcaacttc cagcccatga gcaggcaggt ggtcgatgag atcaactaca 20220 aggactacaa agccgtcacc ctgcccttc agcacaacaa ctcgggcttc accggctatc 20280 tcgcccccac catgcgccag gggcagccct accccgccaa cttcccatac ccgctcatcg 20340 gccagacagc cgtaccctcc gtcacccaga aaaagttcct ctgcgacagg gtcatgtggc 20400 gcatcccctt ttccagcaac ttcatgtcca tgggcgccct caccgacctg ggacagaaca 20460 tgctctatgc caactcggcc cacgcgctcg acataacttt tgaggtggac cccatggatg 20520 agcccaccct cctctatctt ccttcgaag tttcgacgt ggtcagagtg caccagccgc 20580 accgcggcgt catcgaggcc gtctacctgc gcacgccctt ctccgccggc aacgctacta 20640 cataagcatg agcggctcca gcgaacgaga gctcgcggcc atcgtgcgcg acctgggctg 20700 cgggccctac tttttgggaa cccacgacaa gcgcttccct ggcttccttg ccagcgacaa 20760 gctggcatgc gccatcgtca acacggccgg ccgcgagacc gggggcgtgc actggctcgc 20820 ctttggctgg aatccgcgct cgcgcacctg ctacatgttt gacccctttg ggttctcgga 20880 ccgccggctc aagcagattt acagcttcga gtacgaggcc atgctgcgcc gcagcgccct 20940 ggcctcctcg cccgaccgct gtctcagcct cgagcagtcc acccagaccg tgcaggggcc 21000 cgactccgcc gcctgcggac ttttttgttg catgtttttg catgcctttg tgcactggcc 21060 cgaccgaccc atggacggaa accccaccat gaacttgctg acgggggtgc ccaacggcat 21120 gctacaatcg ccacaggtgc tccccaccct caggcgcaac caggaggagc tctaccgctt 21180 cctcgcgcgc cactcccctt actttcgatc ccaccgcgcc gccattgaaa acgccaccgc 21240 ttttgataaa atgaaacaac tgcgtgtatc tcaataaaca gcactttatt ttacatgcac 21300 tggagtatat gcaagttatt taaaagtcga aggggttctc gcgctcgtcg ttgtgcgccg 21360 cgctggggag ggccacgttg cggtactgaa acttgggctg ccacttgaac tcggggatca 21420 ccagtttggg cactggggtc tcggggaagg tctcgctcca catgcgccgg ctcatctgca 21480 gggcgcccag tatgtcgggc gcggagatct tgaaatcgca gttgggacca gtgctctgcg 21540 cgcgcgagtt gcggtacacg gggttgcagc actggaacac catcaaactg gggtgcttca 21600 cgctggccag cacgctcttg tcgctgatct gatccttgtc caggtcctcg gcgttgctca 21660 ggccgaacgg ggtcatcttg cacagctggc ggcccaggaa gggcacgctg tgaggcttgt 21720 ggttacactc gcagtgcacg ggcatcagca tcatccccgc gccgcgctgc atattcgggt 21780 agagggcctt gacaaaggcc aagatctgct tgaaagcttg ctgggccttg gccccctcgc 21840 tgaagaacag gccgcagctc ttcccgctga actggttatt cccacacccg gcatcatgca 21900 cgcagcagcg cgcgtcatgg ctggtcagtt gcaccacgct tcggccccag cggttctggg 21960 tcaccttggc cttgctgggc tgctccttca acgcgcgctg gccgttctcg ctggtcacat 22020 ccatctccac cacgtggtcc ttgtggatca tcaccgtccc gtgcagacac ttgagctggc 22080 cctcgacatc ggtgcagccg tgttcccaca gggcgcagcc ggtgcactcc caattcttgt 22140 gcgcgatccc gctgtggctg aagatgtaac cttgcaacat gcggcccatc acggtgctaa 22200 atgatttact ggtgctgaag gtcagttgca ggccgcgggc ctcctcgttc atccatgtct 22260 gacacatttt ctggaagatc tcggtctgct cgggcatgag cttgtaggca tcgcgcaggc 22320 cgctgtcgac gcggtagcgt tccatcagca cgttcatggt atccatgccc ttctcccagg 22380 acgagaccag aggcagactc agggggttgc gcacgttcag gataccgggg gtcgcgggct 22440 cgacgatgcg ttttccgtcc tttccttcct tcaacagaac cggcggctgg ctgaatccca 22500 ctcccacgat cacggcgtct tcctggggca tctcttcgtc ggggtctacc ttggtcacat 22560 gcttggtctt tctggcttgc ttcttttttg gagggctgtc cacggggacc acgtcctcct 22620 cggaagaccc ggagcccacc cgctgatact ttcggcgctt ggtgggcaga ggaggtggcg 22680 gcggcgaggg gctcctctcc tgctccggcg gatagcgcgc cgacccgtgg ccccggggcg 22740 gagtggcctc tcggtccatg aaccggcgca cgtcctgact gccgccggcc attgtttcct 22800 aggggaagat ggaggagcag ccgcgtaagc aggagcagga ggaggactta accacccacg 22860 agcaacccaa aatcgagcag gacctgggct tcgaagagcc ggctcgtcta gaacccccac 22920 aggatgaaca ggagcacgag caagacgcag gccaggagga gaccgacgct gggctccagc 22980 atggctacct gggaggagag gaggatgtgc tgctgaaaca cctgcagcgc cagtccctca 23040 tcctccggga cgccctagcc gaccggagcg aaacccccct cagcgtcgag gagctgtgtc 23100 gggcctacga gctcaacctc ttctcgccgc gcatgccccc caaacgccag cccaatggca 23160 catgcgagcc caacccgcgt ctcaacttct atcccgtctt tgcggtcccc gaggcccttg 23220 ccacctatca catctttttc aagaaccaaa agatccccgt ctcctgccgc gccaaccgca 23280 cccgcgccga cgcgctcctt actctggggc ccggcgcgcg catacctgat atcgcttccc 23340 tggaagaggt gcccaagatc ttcgaagggc tcggtcggga cgagacgcgc gcggcgaacg 23400 ctctgaaaga aacagcagag gaagagggtc acactagcgc cctggtagag ttggaaggcg 23460 acaacgccag gctggccgtg ctcaagcgca gcgtcgagct cacccacttc gcctaccccg 23520 ccgtcaacct cccgcccaag gtcatgcgtc gcatcatgga tcagcttatc atgccccaca 23580 tcgaggccct cgatgaaagt caggagcagc gccccgagga cgcccggccc gtggtcagcg 23640 acgagatgct cgcgcgctgg ctcgggaccc gcgaccccca ggctttggaa cagcggcgca 23700 agctcatgct ggccgtggtc ctggtcaccc tcgagctgga atgcatgcgc cgcttcttca 23760 gcgaccccga gaccctgcgc aaggtcgagg agaccctgca ctacactttc aggcacggtt 23820 tcgtcaggca ggcctgcaag atctccaacg tggagctgac caacctggtc tcctgcctgg 23880 ggatcctgca cgagaaccgc ctgggccaga ccgtgctcca ctctaccctg aagggcgagg 23940 cgcggcggga ctacgtccgc gactgcgtct ttctctttct ctgccacaca tggcaagcgg 24000 ccatgggcgt gtggcagcag tgtctcgagg acgagaacct gaaggagctg gacaagcttc 24060 ttgctagaaa ccttaaaaag ctgtggacgg gcttcgacga gcgcaccgtc gcctcggacc 24120 tggccgagat cgtcttcccc gagcgcctga gacagacgct gaaaggcggg ctgcccgact 24180 tcatgagcca gagcatgttg caaaactacc gcactttcat tctcgagcga tctgggatgc 24240 tgcccgccac ctgcaacgcc ttcccctccg actttgtacc gctgagctac cgcgagtgtc 24300 ccccgccgct gtggagccac tgctacctct tgcagctggc caactacatc gcctaccact 24360 cggacgtgat cgaggacgtg agcggcgagg ggctgctcga gtgccactgt cgctgcaacc 24420 tgtgctcccc gcatcgctcc ctggtctgca acccccagct cctgagcgag acccaggtca 24480 tcggtacctt cgagctgcaa ggtccgcagg agtccaccgc tccgctgaaa ctcacgccgg 24540 ggttgtggac ttccgcgtac ctgcgcaaat ttgtacccga agactaccac gcccatgaga 24600 taaagttctt tgaggaccaa tcgcgcccgc agcacgcgga tctcacggcc tgcgtcatca 24660 cccagggcgc gatcctcgcc caattgcacg ccatccaaaa atcccgccaa gagtttcttc 24720 tgaaaaaggg tagaggggtc tacctggacc cccagacggg cgaggtgctc aacccggggtc 24780 tcccccagca tgccgagga gaagcaggag ccgctagtgg aggagatgga agaagaatgg gacagccagg cagaggag cgaatgggag gaggagag aggaggaga attggaagag gtggaagagc agcccgtcgc cgcaccatcc gcgccggcag ccccgccggt cacggataca 24960. acctccgctc cggtcaagcc tcctcgtaga tgggatcaaa tgaagggtga cggtaagcac gagcggcagg gctaccgatc atggagggcc cacaaagccg cgatcatcgc ctgcttgcaa gactgcgggg ggacatcgc ttttgcccgc cgctacctgc tcttccaccg cggggtgaac 25140 atcccccgca acgtgttgca ttactaccgt caccttcaca gctaagaaaa agcaagtaag aggagtcgcc gaggaggag gaggcctgag gatcgcggcg aacgagccct tgaccaccag 25260 25320. ggagctgagg aaccggatct tccccactct ttgccatt tttcagcaga gtcgaggtca 25380. gcagcaagag ctcaaagtaa aaaaccggtc tctgcgctcg ctcacccgca gttgcttgta ccacaaaaac gaagatcagc tgcagcgcac tctcgaagac gccgaggctc tgttccacaa 25440 gtactgcgcg ctcactctta aagactaagg cgcgcccacc cggaaaaaag gcgggaatta 25500 cctcatcgcc accatgagca aggagattcc caccccttac atgtggagct atcagcccca 25560 gatgggcctg gccgcgggcg cctcccagga ctactccacc cgcatgaact ggctcagtgc 25620 cggcccctcg atgatctcac aggtcaacgg ggtccgtaac catcgaaacc agatattgtt 25680 ggagcaggcg gcggtcacct ccacgcccag ggcaaagctc aacccgcgta attggccctc 25740 caccctggtg tatcaggaaa tccccgggcc gactaccgta ctacttccgc gtgacgcact 25800 ggccgaagtc cgcatgacta actcaggtgt ccagctggcc ggcggcgctt cccggtgccc 25860 gctccgccca caatcgggta taaaaccct ggtgatccga ggcagaggca cacagcttaa 25920 cgacgagttg gtgagctctt caatcggtct gcgaccggac ggagtgttcc aactagccgg 25980 agccgggaga tcctccttca ctccccacca ggcctacctg accttgcaga gcagctcttc 26040 ggagcctcgc tccggaggca tcggaaccct ccagttcgtg gaggagtttg tgccctcggt 26100 ctacttcaac cccttctcgg gatcgccagg cctctacccg gacgagttca taccgaactt 26160 cgatgcagtg agagaagcgg tggacggcta cgactgaatg tcccatggtg actcggctga 26220 gctcgctcgg ttgaggcatc tggaccactg ccgccgcctg cgctgcttcg cccgggagag 26280 ctgcggactc atctactttg agttcccga ggagcacccc aacggccctg cacacggagt 26340 gcggatcacc gtagagggca ccaccgagtc tcacctggtc aggttcttca cccagcaacc 26400 cttcctggtc gagcgggacc ggggcgccac cacctacacc gtctactgca tctgtcctac 26460 cccgaagttg catgagaatt tttgttgtac tctgtgtgct gagtttaata aaagctaaac 26520 tcctacaata ctctgggatc ccgtgtcgtc gcactcgcaa cgagatcttc aacctcacca 26580 accagactga ggtaaaactc aactgcagac cggggggcaa atacatcctc tggctctttg 26640 aaaacacttc cttcgcagtc tccaacgcct gcgccaacga cggtattgaa atacccaaca 26700 accttaccag tggactaact tacactacca gaaagactaa gctagtactc tacaatcctt 26760 ttgtagaggg aacctaccac tgccagagcg gaccttgctt ccacactttc actttggtga 26820 acgttaccga cagcagcaca gtcgctccag aaacatctaa cctttttgat actaacactc 26880 ctaaaaccgg aggtgagctc tgggttccct cttaacaga ggggggtaaa catattgaag 26940 cggttgggta tttgatttta ggggtggtcc tgggtgggtg catagcggtg ctgtattacc 27000 ttccttgctg gatcgaaatc aaaatcttta tctgctgggt catacattgt tgggaggaac 27060 catgaagggg ctcttgctga ttatcctttc cctggttggg ggtgtactgt catgccacga 27120 acagccacga tgtaacatca ccacaggcaa tgagaggagt gtgatatgca cagtagtcat 27180 caaatgcgag catgcatgtc ctctcaacat cacattcaag aataagacca tgggaaattc 27240 atgggtgggc gattgggaac caggagatga gcagaactac acggtcactg tccatggtag 27300 cgatgggaat cacactttcg gtttcaaatt catttttgaa gtcatgtgtg atattacact 27360 gcatgtggct agacttcatg gcttgtggcc ccctaccaag gagaacatgg ttgggttttc 27420 tttggcttt gtgatcatgg cctgctttat gtcaggtctg ctggtagggg ctttagtatg 27480 gttcctgaag cacaagccta ggtatggaaa tgaggagaag gaaaaattgc tataaatctt 27540 tttctcttcg cagaaccatg aatactttga ccagtgtcgt gctgctctct cttcttgtag 27600 cttttagtca ggcaggattt catactatca atgctacatg gtgggctaat gtaactttag 27660 tgggaccctc agatacgcca gtcacatggt atgataaaca gggaatgcag ttctgtgatg 27720 gaaatacagt taaaatcct caaataagac atgagtgtaa tgagcaaaac cttacactaa 27780 ttcatgtgaa caaaacccat gaaaggacat acatgggtta taatagacag agtactcata 27840 aggaagacta taaagtcata gttataccgc ctcctcctgc tactgtaaag ccacagtcag 27900 gtccagagta tgtatatgtt aatatgggag agaacaaac cttagttgga cctccaggaa 27960 taccagttac ttggtatgac ggagaaggaa ataaattctg cgatggagaa aaagttgaac 28020 atgcagaatt taatcataca tgtgacgagc aaaatcttac actgttgttt ataaatctta 28080 cacatgatgg ggcttatctt ggctataatc accagggaac taaaagaact tggtatgagg 28140 ttgtagtgac agatggtttt ccaaaatcag gggagatgaa aatcgaagat cagagtagac 28200 aaacagaaca aaaacaaaat gagcataaac agggtgggca gaaacaggag gggcaaaaag 28260 agacaagtca aaagaaagat aatgacaaac agaaggcgac acacaggagg ccatcaaaac 28320 taaagccgca cacacctgaa gcaaaactga ttacagtttc tagtgggtct aacttaacat 28380 tacttgggcc agatggaaag gtcacttggt atgatgatga tttaaaaaga ccatgcgagc 28440 ctgggtataa gttagggtgt aagtgtgaca atcaaaacct aacgctaatc aatgtaacta 28500 aactttatga gggagtttac tatggtacta atgacagagg caacagcaaa agatacagag 28560 taaaagtaaa cactactaat tctcaaagtg tgaaaattca gccatacaac aggtctacta 28620 ctcctgatca gaaacacaga tttgaattgc aaattgattc taatcaagac aatgacaaaa 28680 ttccatcaac cactgtggca atcgtggtgg gagtgattgc gggcttcata actataatca 28740 ttgtcattat gtgctacatc tgctgccgca agcgtcccag ggcatacaat catatggtag 28800 acccactact tagcttctct tactgagact cagtcacttt catttcagaa ccatgaaggc 28860 ttcacagct tgcgttctga ttagcatagt cacattagta tcagctgat aaaacaagt 28920 tcaagttagc agaggaggaa atattacatt agatggacca ttcgataata ctacatggac 28980 aagatatcat aatgatggac aaaaatgg ttggatgaaa atttgcacat ggactggagc 29040 aacatataaa tgtcacaata atggagcat tacttattt gctttcaca ttacatccgg 29100 agtttacaa gcagaagggt aaaaaaga ggttagaaca tttcatcta gaatcaaa 29160 acatacatt gagattctg gagattga acaaaaa attatctat atatctac 29220 aaattgaa ccgccaacta ctaaagcacc caccacagtt agacaacca cgcagacaac 29280 tactagggaa acacacatc acaccacac actcacacta acacacac 29340 cactacagtg cagaatacta ctttattgat tgggtttta ctaagaggaa atgaagtac 29400 tactgatcag acagaggcta cctcaagtgc cttcagcagc actgcaatt taacttcgct 29460 tgcttcggta atgaaacga tcgtgccgat gatgtatggc caacttact caggtttgga 29520 tattcaaatt acttttctgg ttgtctgtgg gatctttatt cttgtggttc ttttgtactt 29580 tgtctgctgc aaagccagag aaaaatctag gaggcccatc tacaggccag taatcgggga 29640 tcctcagcct ctccaagtgg aagggggtct aaggaatctt ctcttctctt tttcagtatg 29700 gtgattcagc catgattcct aggttcttcc tatttaacat cctcttctgt ctattcaacg 29760 tgtgcgctgc cttcgcggcc gtctcgcacg cctcgcccga ctgtcttggg cccttcccca 29820 cctaccttct ttttgccctg ctcacctgca cctgcgtctg cagcattgtc tgcctggtcg 29880 tcaccttcct gcagctcatc gactggtgct gcgcgcgcta caattatctc caccacagtc 29940 ccgaatacag ggacgagaac gtagccagaa tcttaaggct catttgacca tgcagactct 30000 gctcatactg ctatccctcc tctcccctgc cctcgctgat gatgattact ctaagtgcaa 30060 atttgtggag ctatggaatt tcttagactg ctatgatgct aaaatggata tgccatccta 30120 ttacttggtg attgtgggga tagtcatggt ctgctcctgc actttctttg ccatcatgat 30180 ctacccctgt tttgatctcg gctggaactc tgttgaggca ttcacataca cactagaaag 30240 cagttcacta gcttccacgc cgccacccac accgcctccc cgcagaaatc agtttcccat 30300 gattcagtac ttagaagagc cccctccccg gcccccttcc actgttagct actttcacat 30360 aaccggcggc gatgactgac aaccacctgg acctcgagat ggacggccag gcctccgagc 30420 agcgcatcct gcaactgcgc gtccgtcagc agcaggagcg ggccgccaag gagctcctcg 30480 atgccatcaa catccaccag tgcaagaagg gcatcttctg cctggtcaaa caggcaaaga 30540 tcacctacga gctcgtgtcc ggcggcaagc agcatcgcct cgcctatgag ctgccccagc 30600 agaagcagaa gttcacctgc atggtgggcg tcaaccccat agtcatcacc cagcagtcgg 30660 gcgagaccag cggctgcatc cactgctcct gcgaaagccc cgagtgcatc tactccctcc 30720 tcaagaccct ttgcggactt cgcgacctcc tccccatgaa ctgattgatt aaagcccaaa 30780 aaaccaatca aaccccttcc cattagcccc aaataaacaa tcattggaaa taatcattca 30840 ataaagatca cttacttgaa atctgaaagt atgtctctgg tgtagttgtt tagcagcacc 30900 tcggtaccct cctcccagct ctggtactcc agtccccggc gggcggcgaa ctttctccac 30960 accttgaaag ggatgtcaaa ttcctggtcc acaattttca ttgttttccc tctcagatgt 31020 caaagaggct tcgggtggaa gatgacttca accccgtcta cccctatgac tacgcgcgga 31080 atcagaatat ccccttcctc actcccccct ttgtctcctc cgatggattc caaaacttcc 31140 cccctggtgt cctgtcactc aaattggctg acccaatcgc tatcaccaat ggtgatgtct 31200 cactcaaggt gggaggggga cttactttac aagatggaac cggaaaacta actatcgaca 31260 ccaagactcc cttgcaagtt gcaaataata aattagaact tgcgtttgat gcaccattgt 31320 atgaaaaaaa tggaaaactt gctttaaaaa caggccatgg attagctgtt ttaaccaaag 31380 acattggcat accagaatta attggatccc ttgtgatctt aactggaaaa ggaattggga 31440 caggtactgt tgcaggagga ggaactatag atgtaagact gggtgatgat ggaggtttat 31500 catttgataa aaagggtgat ctagtagcct ggaataaaaa aaatgacagg cgcactttgt 31560 ggacaacacc tgatccatcc ccaaactgca gagtatcaga agataaagat tcaaaactaa 31620 ctttaattct tacaaaatgt ggaagtcaga tcctagcaag tttttcactg cttgtagtca 31680 aagggacgta tgcaactgtt gataaaaata caactaataa acaatttagc attaaactac 31740 tgtttgatgc aaatggaaag cttaaaagcg aatcaaacct tagtggttat tggaactata 31800 gaagtgataa tagtgttgtt agtactccct atgacaatgc agtgccttc atgccaaata 31860 ccacggctta tccaaaaatc attaatagta caactgatcc tgaaaataag aaaagttcgg 31920 ctaaaaaac tattgttggc aatgctacc tagaaggtaa tgcgggtcaa ccagtagccg 31980 ttgctattag tttcaataag gaaactactg ctgactattc aataacattt gactttgcgt 32040 ggagcaaagc tttgaaacc cctgtgcctt ttgacacctc ctccatgaca ttctcatata 32100 ttgcccagga aaatcaagac aaaggcgaat aaagtgtttt gaaatgaact tatgtatctt 32160 tattgattt tacaccagca cgggtagtca gtctcccacc accagcccat ttcacagtgt 32220 acacggttct ctcagcacgg gtggccttaa atagggaaat gttctgatta gtgcgggaac 32280 tggacttggg gtctataatc cacacagttt cctggcgagc caaacggggg tcggtgattg 32340 agatgaagcc gtcctctgaa aagtcatcca agcgggcctc acagtccaag gtcacagtct 32400 ggtgaaacga gaagaacgca cagattcata ctcggaaaac aggatgggtc tgtgcctctc 32460 catcagcgcc ctcaacagtc tctgccgccg gggctcggtg cggctgctgc agatgggatc 32520 gggatcgcaa gtctctctga ctatgatccc cacagccttc agcatcagtc tcctggtgcg 32580 tcgggcacag caccgcatcc tgatctctgc catgttctca cagtaagtgc agcacataat 32640 caccatgtta ttcagcagcc cataattcag ggtgctccag ccaaaactca tgttggggat 32700 gatggaaccc acgtgaccat cgtaccagat gcggcagtat atcagatgcc tgcccctcat 32760 gaacacactg cccatataca tgatctcttt gggcatgtct ctgttcacaa tctgacggta 32820 ccagggaaag cgctggttga acatgcaccc gtaaatgact ctcctgaacc acacggccag 32880 cagggtgcct cccgcccgac actgcaggga gcccggggat gaacagtggc aatgcaggat 32940 ccagcgctcg tacccgctca ccatctgagc tctcaccaag tccagggtag cggggcacag 33000 gcacactgac atacatcttt ttaaaatttt tattctctct ggagtcaaga tcatatccca 33060 ggggactgga aactcttgga gcagggtaaa gccagcagca catggtaatc cacggacaga 33120 acttacatta tgataatctg catgatcaca atcaggcac aggggatt gttcagtcag 33180 tgaagccctg gtttcctcat cagatcgtgg taaacggggcc ctgcgatatg gatgatggcg 33240 gagcgagctg gattgaatct cggtttgcat tgtagtgt tctcttgcgt accttgtcgt 33300 acttctgcca gcagaaatgg gccttgac agcagatacc cctcctgcgg ccgtccttc 33360 gctgctgccg ctcagtcatc caacgaagt acatccattc tcgagatc tggaagtt 33420 cctctgcatc tgatgaata aaaaacccgt ccatgcgaat tccctcatc acatcagcca 33480 ggactctgta ggccatcccc atccagttaa tgctgccttg tctatcattc aggggggcg 33540 gtggcaggat tggagaacc atttttc caaacggtct cgaaggacga taaagtgcaa 33600 gtcacgcagg tgacagcgtt cccctccgct gtgctggtgg aacagacag ccaggtcaa 33660 acccactcta ttttcaaggt gctcgaccgt ggcttcgagc agtggctcta cgcgtacatc 33720. cagcataaga atcacattaa aggctggccc tccatcgatt tcatcaatca tcaggttaca ttcctgcacc atccccaggt aattctcatt tttccagcct tggattatct ctacaaattg ttggtgtaag tccactccgc acatgtggaa aagctcccac agtgccccct ccactttcat aatcaggcag accttcataa tagaaacaga tcctgctgct ccaccacctg cagcgtgttc 34020. ggttctgccc tccgccctga gctcgcgcct caatgtcagc 34080. 34080. 34080. 34080. 34080. 34080. 34080. 34080. 34080 gtgggactgg caagcgtaag ggaaaacttt aatgctccga agctagcacc caaaaactgc atgctggaat aagctctctt tgtgtctccg gtgatgcctt ccaaaatgtg agtgataaag cgtggtagtt tttctttaat catttgcgta atagaaaagt cctgtaaata agtcactagg accccaggga ccacaatgtg gtagcttaca ccgcgtcgct gaagcatggt tagtagat 34320. gagagtctga aaaacagaaa gcatgcacta aactaaggtg gctattttca ctgaaggaaa aatcactctc tccagcaaca gggtacccac tgggtggccc ttgcggacat acaaaaatcg 34440 gtccgtgtga ttaaaaagca gcacagtaag ttcctgtctt cttccggcaa aaatcacatc 34500 ggactgggtt agtatgtccc tggcatggta gtcattcaag gccataaatc tgccctgata 34560 tccagtagga accagcacac tcacttttag gtgaagcaat accaccccat gcggaggaat 34620 gtggaaagat tcagggcaaa aaaaattata tctattgcta gtcccttcct ggacgggagc 34680 aatccctcca ggactatcta tgaaagcata cagagattca gccatagctc agcccgctta 34740 ccagtagaca gagagcacag cagtacaagc gccaacagca gcgactgact acccactgac 34800 ccagctccct atttaaaggc gccttacact gacgtaatga ccaaaggtct aaaaaccccg 34860 ccaaaaaaaa acacacacgc cctgggtgtt tttcgcgaaa acacttccgc gttctcactt 34920 cctcgtatcg atttcgtgac ttaacttccg ggttcccacg ttacgtcact tctgccctta 34980 catgtaactg agtcgtaggg cgccatcttg cccacgtcca aaatggcttc catgtccagc 35040 cacgcctccg cggcgaccgt tagccgtgcg tcgtgacgtc atttgcatcg tcttctctcg 35100 tccaatcagc gtcggctccg ccctaattc aaaagctcat ttgcatatta acttttgttt actttgtggg gtatattatt gatgatg <210> 2 <211> 6150 <212> DNA <213> The snowstorm <220> <223> pAdAptAd20-42-42.Empty <400> 2 ttaattaaca tcatcaata tataccccac aaagtaaaca aaagttaata tgcaaatgag cttttgaatt tagggcggag ccgacgctga ttggacgaga gaagacgatg caaatgacgt cacgacgcac ggctaacggt cgccgcggag gcgtggccta gcccggagc aagtcgcggg 180 gctgatgacg tataaaaaag cggactttag acccggaat ggccgatttt cccgcggcca cggccggata tgaggtaatt ctggggcggat gcaagtgaaa ttaggtcatt ttggcgcgaa aactgaatga ggaagtgaaa agcgaaaaat accgatcccg cccagggcgg aatatttacc gagggccgag agactttgac cgattacgtg ggggtttcga ttgcggtgtt tttttcgcga 420 atttccgcgt ccgtgtcaaa gtccggtgtt tatgtcacag atcagctgac ctaggtggtc 480 aatattggcc attagccata ttattcattg gttatatagc ataaatcaat attggctatt 540 ggccattgca tacgttgtat ccatatcata atatgtacat ttatattggc tcatgtccaa 600 cattaccgcc atgttgacat tgattattga ctagttatta atagtaatca attacggggt 660 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc 720 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag 780 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc 840 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg 900 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc 960 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca 1020 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca 1080 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg 1140 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc 1200 gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa 1260 gacaccggga ccgatccagc ctccgcggcc gggaacggtg cattggaagc ttggtaccgg 1320 tgaattcgct agcgttaacg gatcctctag acgagatccg aacttgttta ttgcagctta 1380 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 1440 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct cgatccaggt 1500 aggtttgagt agtgggcgtg gctaagatga ctataaaggt gggtgtctta cgagggtctt 1560 tttgcttttc tgcagacatc atgaacggga ccggcggggc cttcgaaggg gggcttttta 1620 gcccttattt gacaacccgc ctgccgggat gggccggagt tcgtcagaat gtgatgggat 1680 cgacggtgga cgggcgtcca gtgcttccag caaattcctc gaccatgacc tacgcgaccg 1740 tggggaactc gtcgctcgac agcaccgccg cagccgcggc agccgcagcc gccatgacag 1800 cgacgagact ggcttcgagc tacatgccca gcagcggtag cagcccctct gtgcccagtt 1860 ccatcatcgc cgaggagaaa ctgctggccc tcctggccga gctggaagcc ctgagccgcc agctggccgc cctgacccag caggtgtccg agctccgcga acagcagcag cagcaaaata aatgattcaa taaacacaga ttctgattca aacagcaaag catctttatt atttttttt tcgcgcgcgg tagccctgg tccacctctc ccgatcattg agagtgcggt ggattttttc caggacccgg tagagatggg attggatgtt gaggtacatg ggcatgagcc cgtcccgggg 2160 gtgaaggtag caccactgca tggcctcatg ctctggggtc gtgttgtag tgatccagtc atagcagggg cgctgggcgt ggtgctggat gatgtctttg aggaggac tgatggccac 2280 ggggagcccc ttggtgtagg tgttggcgaa gcggttgagc tgggagggat gcatgcgggg 2340 ggagatgatg tgcagtttgg cctggatctt gaggttggca atgttgccgc ccagatcccg ccggggggttc atgttgtgca ggaccaccag gacggtgtag cccgtgcact tggggaactt atcatgcaac ttggaaggga atgcgtggaa gaatttggag acgcccttgt gcccgcccag gttttccatg cactcatcca tgatgatggc gatgggcccg tgggctgcgg ctttggcaaa 2580 gacgtttctg gggtcagaga catcataatt atgctcctgg gtgagatcat cataagacat 2640 tttaatgaat ttggggcgga gggtgccaga ttgggggacg atcgttccct cgggccccgg 2700 ggcgaagttc ccctcacaga tctgcatctc ccaggctttc atctcggagg gggggatcat 2760 gtccacctgc ggggcgatga aaaaaacggt ttccggggcg ggggtgatga gctgcgagga 2820 gagcaggttt ctcaacagct gggacttgcc gcacccggtc gggccgtaga tgaccccgat 2880 gacgggttgc aggtggtagt tcaaggagat gcagctgccg tcttcccgga ggaggggggc 2940 cacctcgttg agcatgtctc tgacttggag gttttcccgg acgagctcgc cgaggaggcg 3000 gtccccgccc agcgagagca gctcttgcag ggaagcaaag tttttcaggg gcttgagccc 3060 gtcggccatg ggcatcttgg cgagggtctg cgagaggagt tccaagcggt cccagagctc 3120 ggtgacgtgc tctacggcat ctcgatccaa cagacttcct cgtttcgggg gttgggacga 3180 ctgcgactgt agggcacgag acgatgggcg tccagcgctg ccagcgtcat gtccttccag 3240 ggtctcaggg tccgcgtgag cgtggtctcc gtcacggtga aggggtgggc cccgggctgg 3300 gcgcttgcaa gggtgcgctt gagactcatc ctgctggtgc tgaaacgggc acggtcttcg 3360 ccctgcgcgt cggcgagata gcagttgacc atgagctcgt agttgagggc ctcggcggcg 3420 tggcccttgg cgcggagctt gcccttggaa gagcgcccgc aggcgggaca gaggagggat 3480 tgcagggcgt agagcttggg cgcgagaaag acggactcgg gggcgaaagc gtccgctccg 3540 cagtgggcgc agacggtctc gcactcgacg agccaggtga gctcgggatg ttcggggtca 3600 aaaaccagtt ttcccccgtt cttttgatg cgcttcttac ctcgcgtctc catgagtctg 3660 tgtccgcgct cggtgacaaa caggctgtct gtgtccccgt agacggactt gatgggcctg 3720 tcctgcaggg gcgtcccgcg gtcctcctcg tagagaaact cggaccactc tgagacgaag 3780 gcgcgcgtcc acgccaagac aaaggaggcc acgtgcgagg ggtagcggtc gttgtccacc 3840 aggggtcca ccttttccac cgtgtgcaga cacatgtccc cctcctcagc atccaagaag 3900 gtgattggct tgtaggtgta ggccacgtga ccgggggtcc ccgacgggg ggtataaaag 3960 ggggcgggtc tgtgctcgtc ctcactctct tccgcgtcgc tgtccacgag cgccagctgt 4020 tggggtaggt attccctctc gagattaatt aattcgaacc cataataccc ataatagctg 4080 tttgccatcg accaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt 4140 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg 4200 cggaacccct atttgttat ttttctaaat acattcaaat atgtatccgc tcatgagaca 4260 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt 4320 ccgtgtcgcc cttattccct ttttgcggc atttgcctt cctgtttttg ctcacccaga 4380 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga 4440 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat 4500 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca 4560 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt 4620 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac 4680 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct 4740 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga 4800 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac 4860 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat 4920 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg 4980 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc 5040 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc 5100 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg 5160 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta 5220 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg 5280 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga 5340 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt 5400 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag 5460 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa 5520 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag 5580 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca 5640 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac 5700 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa 5760 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc 5820 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg 5880 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc 5940 ctttttacgg ttcctggcct tttgctgggc tggtaagagc cgcgagcgat ccttgaagct 6000 gtccctgatg gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg ggcatcccga 6060 tgccgccgga agcgagaaga atcataatgg ggaaggccat ccagcctcgc gtcgatggca 6120 aacagctatt atgggtatta tgggttcgaa 6150 <210> 3 <211> 9350 <212> DNA <213> Artificial sequence <220> <223> pAdAptAd20-42-42.LacZ <400> 3 ttaattaaca tcatcaataa tataccccac aaagtaaaca aaagttaata tgcaaatgag 60 cttttgaatt tagggcggag ccgacgctga ttggacgaga gaagacgatg caaatgacgt 120 cacgacgcac ggctaacggt cgccgcggag gcgtggccta gcccggaagc aagtcgcggg 180 gctgatgacg tataaaaaag cggactttag acccggaaat ggccgatttt cccgcggcca 240 cggccggata tgaggtaatt ctgggcggat gcaagtgaaa ttaggtcatt ttggcgcgaa 300 aactgaatga ggaagtgaaa agcgaaaaat accgatcccg cccagggcgg aatatttacc 360 gagggccgag agactttgac cgattacgtg ggggtttcga ttgcggtgtt tttttcgcga 420 atttccgcgt ccgtgtcaaa gtccggtgtt tatgtcacag atcagctgac ctaggtggtc 480 aatattggcc attagccata ttattcattg gttatatagc ataaatcaat attggctatt 540 ggccattgca tacgttgtat ccatatcata atatgtacat ttatattggc tcatgtccaa 600 cattaccgcc atgttgacat tgattattga ctagttatta atagtaatca attacggggt 660 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc 720 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag 780 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc 840 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg 900 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc 960 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca 1020 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca 1080 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg 1140 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc 1200 gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa 1260 gacaccggga ccgatccagc ctccgcggcc gggaacggtg cattggaagc ttggtacctc 1320 taggcttttg caaaaagctt tgcaaagatg gataaagttt tccggaattc cgccaaaaag 1380 aagagaagg tagagaccc caaggacttt ccttcagaat tgctaagttt tttgagtcca 1440 agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa 1500 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc 1560 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt 1620 ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga ggccgatact 1680 gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta caccaacgta 1740 acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac gggttgttac 1800 tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg aattattttt 1860 gatggcgtta actcggcgtt tcatctgtgg tgcaacgggc gctgggtcgg ttacggccag 1920 gacagtcgtt tgccgtctga atttgacctg agcgcatttt tacgcgccgg agaaaaccgc 1980 ctcgcggtga tggtgctgcg ttggagtgac ggcagttatc tggagaatca ggatatgtgg 2040 cggatgagcg gcattttccg tgacgtctcg ttgctgcata aaccgactac acaaatcagc 2100 gatttccatg ttgccactcg ctttaatgat gatttcagcc gcgctgtact ggaggctgaa 2160 gttcagatgt gcggcgagtt gcgtgactac ctacgggtaa cagtttcttt atggcagggt 2220 gaaacgcagg tcgccagcgg caccgcgcct ttcggcggtg aaattatcga tgagcgtggt 2280 ggttatgccg atcgcgtcac actacgtctg aacgtcgaaa acccgaaact gtggagcgcc 2340 gaaatcccga atctctatcg tgcggtggtt gaactgcaca ccgccgacgg cacgctgatt 2400 gaagcagaag cctgcgatgt cggtttccgc gaggtgcgga ttgaaaatgg tctgctgctg 2460 ctgaacggca agccgttgct gattcgaggc gttaaccgtc acgagcatca tcctctgcat 2520 ggtcaggtca tggatgagca gacgatggtg caggatatcc tgctgatgaa gcagaacaac 2580 tttaacgccg tgcgctgttc gcattatccg aaccatccgc tgtggtacac gctgtgcgac 2640 cgctacggcc tgtatgtggt ggatgaagcc aatattgaaa cccacggcat ggtgccaatg 2700 aatcgtctga ccgatgatcc gcgctggcta ccggcgatga gcgaacgcgt aacgcgaatg 2760 gtgcagcgcg atcgtaatca cccgagtgtg atcatctggt cgctggggaa tgaatcaggc 2820 cacggcgcta atcacgacgc gctgtatcgc tggatcaaat ctgtcgatcc ttcccgcccg 2880 gtgcagtatg aaggcggcgg agccgacacc acggccaccg atattatttg cccgatgtac 2940 gcgcgcgtgg atgaagacca gcccttcccg gctgtgccga aatggtccat caaaaaatgg 3000 ctttcgctac ctggagagac gcgcccgctg atcctttgcg aatacgccca cgcgatgggt 3060 aacagtcttg gcggtttcgc taaatactgg caggcgtttc gtcagtatcc ccgtttacag 3120 ggcggcttcg tctgggactg ggtggatcag tcgctgatta aatatgatga aaacggcaac 3180 ccgtggtcgg cttacggcgg tgattttggc gatacgccga acgatcgcca gttctgtatg 3240 aacggtctgg tctttgccga ccgcacgccg catccagcgc tgacggaagc aaaacaccag 3300 cagcagtttt tccagttccg tttatccggg caaaccatcg aagtgaccag cgaatacctg 3360 ttccgtcata gcgataacga gctcctgcac tggatggtgg cgctggatgg taagccgctg 3420 gcaagcggtg aagtgcctct ggatgtcgct ccacaaggta aacagttgat tgaactgcct 3480 gaactaccgc agccggagag cgccgggcaa ctctggctca cagtacgcgt agtgcaaccg 3540 aacgcgaccg catggtcaga agccgggcac atcagcgcct ggcagcagtg gcgtctggcg 3600 gaaaacctca gtgtgacgct ccccgccgcg tcccacgcca tcccgcatct gaccaccagc 3660 gaaatggatt tttgcatcga gctgggtaat aagcgttggc aatttaaccg ccagtcaggc 3720 tttctttcac agatgtggat tggcgataaa aaacaactgc tgacgccgct gcgcgatcag 3780 ttcacccgtg caccgctgga taacgacatt ggcgtaagtg aagcgacccg cattgaccct 3840 aacgcctggg tcgaacgctg gaaggcggcg ggccattacc aggccgaagc agcgttgttg 3900 cagtgcacgg cagatacact tgctgatgcg gtgctgatta cgaccgctca cgcgtggcag 3960 catcagggga aaaccttatt tatcagccgg aaaacctacc ggattgatgg tagtggtcaa 4020 atggcgatta ccgttgatgt tgaagtggcg agcgatacac cgcatccggc gcggattggc 4080 ctgaactgcc agctggcgca ggtagcagag cgggtaaact ggctcggatt agggccgcaa 4140 gaaaactatc ccgaccgcct tactgccgcc tgttttgacc gctgggatct gccattgtca 4200 gacatgtata ccccgtacgt cttcccgagc gaaaacggtc tgcgctgcgg gacgcgcgaa 4260 ttgaattatg gcccacacca gtggcgcggc gacttccagt tcaacatcag ccgctacagt 4320 caacagcaac tgatggaaac cagccatcgc catctgctgc acgcggaaga aggcacatgg 4380 ctgaatatcg acggtttcca tatggggatt ggtggcgacg actcctggag cccgtcagta 4440 tcggcggaat tccagctgag cgccggtcgc taccattacc agttggtctg gtgtcaaaaa 4500 taataataac cgggcagggg ggattctgaa cttgtttatg gatcctctag acgagatccg 4560 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 4620 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 4680 tatcatgtct cgatccaggt aggtttgagt agtgggcgtg gctaagatga ctataaaggt 4740 gggtgtctta cgagggtctt tttgcttttc tgcagacatc atgaacggga ccggcggggc 4800 cttcgaaggg gggcttttta gcccttattt gacaacccgc ctgccgggat gggccggagt 4860 tcgtcagaat gtgatgggat cgacggtgga cgggcgtcca gtgcttccag caaattcctc 4920 gaccatgacc tacgcgaccg tggggaactc gtcgctcgac agcaccgccg cagccgcggc agccgcagcc gccatgacag cgacgagact ggcttcgagc tacatgccca gcagcggtag 5040. cagcccctct gtgcccagtt ccatcatcgc cgaggagaaa ctgctggccc tcctggccga 5100 gctggaagcc ctgagccgcc agctggccgc cctgacccag caggtgtccg agctccgcga 5160 acagcagcag cagcaaaata aatgattcaa taaacacaga ttctgattca aacagcaaag catctttatt atttattttt tcgcgcgcgg tagccctgg tccacctctc ccgatcattg 5280 agagtgcggt ggattttttc caggacccgg taggatggg attggatgtt gaggtacatg 5340 ggcatgagcc cgtcccgggg gtgaaggtag caccactgca tggcctcatg ctctggggtc 5400 gtgttgtag tgatccagtc atagcagggg cgctgggcgt ggtgctggat gatgtctttg 5460 aggaggac tgatggccac ggggagcccc ttggtgtagg tgttggcgaa gcggttgagc 5520 tgggagggat gcatgcgggg ggagatgatg tgcagtttgg cctggatctt gaggttggca 5580 atgttgccgc ccagatcccg ccggggggttc atgttgtgca ggaccaccag gacggtgtag 5640 cccgtgcact tggggaactt atcatgcaac ttggaaggga atgcgtggaa gaatttggag 5700 acgcccttgt gcccgcccag gttttccatg cactcatcca tgatgatggc gatgggcccg 5760 tgggctgcgg ctttggcaaa gacgtttctg gggtcagaga catcataatt atgctcctgg 5820 gtgagatcat cataagacat tttaatgaat ttggggcgga gggtgccaga ttgggggacg 5880 atcgttccct cgggccccgg ggcgaagttc ccctcacaga tctgcatctc ccaggctttc 5940 atctcggagg gggggatcat gtccacctgc ggggcgatga aaaaaacggt ttccggggcg 6000 ggggtgatga gctgcgagga gagcaggttt ctcaacagct gggacttgcc gcacccggtc 6060 gggccgtaga tgaccccgat gacgggttgc aggtggtagt tcaaggagat gcagctgccg 6120 tcttcccgga ggaggggggc cacctcgttg agcatgtctc tgacttggag gttttcccgg 6180 acgagctcgc cgaggaggcg gtccccgccc agcgagagca gctcttgcag ggaagcaaag 6240 tttttcaggg gcttgagccc gtcggccatg ggcatcttgg cgagggtctg cgagaggagt 6300 tccaagcggt cccagagctc ggtgacgtgc tctacggcat ctcgatccaa cagacttcct 6360 cgtttcgggg gttgggacga ctgcgactgt agggcacgag acgatgggcg tccagcgctg 6420 ccagcgtcat gtccttccag ggtctcaggg tccgcgtgag cgtggtctcc gtcacggtga 6480 aggggtggc cccgggctgg gcgcttgcaa gggtgcgctt gagactcatc ctgctggtgc 6540 tgaaacgggc acggtcttcg ccctgcgcgt cggcgagata gcagttgacc atgagctcgt 6600 agttgagggc ctcggcggcg tggcccttgg cgcggagctt gcccttggaa gagcgcccgc 6660 aggcgggaca gaggagggat tgcagggcgt agagcttggg cgcgagaaag acggactcgg 6720 gggcgaaagc gtccgctccg cagtgggcgc agacggtctc gcactcgacg agccaggtga 6780 gctcgggatg ttcggggtca aaaaccagtt ttcccccgtt cttttgatg cgcttcttac 6840 ctcgcgtctc catgagtctg tgtccgcgct cggtgacaaa caggctgtct gtgtccccgt 6900 agacggactt gatgggcctg tcctgcaggg gcgtcccgcg gtcctcctcg tagagaaact 6960 cggaccactc tgagacgaag gcgcgcgtcc acgccaagac aaaggaggcc acgtgcgagg 7020 ggtagcggtc gttgtccacc aggggtcca ccttttccac cgtgtgcaga cacatgtccc 7080 cctcctcagc atccaagaag gtgattggct tgtaggtgta ggccacgtga ccgggggtcc 7140 ccgacggggg ggtataaaag ggggcgggtc tgtgctcgtc ctcactctct tccgcgtcgc 7200 tgtccacgag cgccagctgt tggggtaggt attccctctc gagattaatt aattcgaacc 7260 cataataccc ataatagctg tttgccatcg accaattctt gaagacgaaa gggcctcgtg 7320 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc 7380 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat 7440 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag 7500 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt 7560 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt 7620 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc 7680 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta 7740 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac 7800 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa 7860 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg 7920 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc 7980 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg 8040 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta 8100 gcttccccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg 8160 cgctcggccc ttccggctgg ctggttatt gctgataaat ctggagccgg tgagcgtggg 8220 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc 8280 tacacgacgg ggagtcaggc aactatggat gacgaata gacagatcgc tgagataggt 8340 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt 8400 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc 8460 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag 8520 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa 8580 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg 8640 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag 8700 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg 8760 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga 8820 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc 8880 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc 8940 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga 9000 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt 9060 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg 9120 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctgggc tggtaagagc 9180 cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct acctgcctgg acagcatggc 9240 ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga atcataatgg ggaaggccat 9300 ccagcctcgc gtcgatggca aacagctatt atgggtatta tgggttcgaa 9350 <210> 4 <211> 7800 <212> DNA <213> Artificial Sequence <220> <223> pAdAptAd20-42-42.Luc+ <400> 4 ttaattaaca tcatcaataa tataccccac aaagtaaaca aaagttaata tgcaaatgag 60 cttttgaatt tagggcggag ccgacgctga ttggacgaga gaagacgatg caaatgacgt 120 cacgacgcac ggctaacggt cgccgcggag gcgtggccta gcccggaagc aagtcgcggg 180 gctgatgacg tataaaaaag cggactttag acccggaaat ggccgatttt cccgcggcca 240 cggccggata tgaggtaatt ctgggcggat gcaagtgaaa ttaggtcatt ttggcgcgaa 300 aactgaatga ggaagtgaaa agcgaaaaat accgatcccg cccagggcgg aatatttacc 360 gagggccgag agactttgac cgattacgtg ggggtttcga ttgcggtgtt tttttcgcga 420 atttccgcgt ccgtgtcaaa gtccggtgtt tatgtcacag atcagctgac ctaggtggtc 480 aatattggcc attagccata ttattcattg gttatatagc ataaatcaat attggctatt 540 ggccattgca tacgttgtat ccatatcata atatgtacat ttatattggc tcatgtccaa 600 cattaccgcc atgttgacat tgattattga ctagttatta atagtaatca attacggggt 660 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc 720 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag 780 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc 840 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg 900 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc 960 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca 1020 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca 1080 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg 1140 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc 1200 gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa 1260 gabaccggga ccgatccagc ctccgcggcc gggaacggtg cattggaagc ttggcattcc 1320 ggtactgttg gtaaagccac catggaagac gccaaaaaca taagaaagg cccggcgcca 1380 ttctatccgc tggagatgg aaccgctgga gagcactgc ataggctat gagagatac 1440 gccctggttc ctggaacaat tgctttaca gatgcacata tcgaggtgga catcacttac 1500 gctgagtact tcgaaatgtc cgttcggttg gcagaagcta tgaacgata tgggctgaat 1560 aaatcaca gatcgtcgt atgcagtga aactctctc aattcttat gccggtgttg 1620 ggcgcgttat tttacggagt tgcagttgcg cccgcgaacg acatttataa tgaacgtgaa 1680 ttgctcaaca gtatgggcat ttcgcagcct accgtggtgt tcgtttccaa aaaggggttg 1740 CAaaaattt tgaacgtgca aaaaaagctc ccaatcatcc aaaaaattt tatcatggat 1800 tctaaaacgg attaccagggg atttcagtcg atgtacacgt tcgtcacatc tcatcacct 1860 cccggtttta atgaatacga ttttgtgcca gagtccttcg ataggcacaa gatatgca 1920 ctgatcatga actcctctgg atctactggt ctgcctaaag gtgtcgctct gcctcataga 1980 actgcctgcg tgagattctc gcatgccaga gatcctattt ttggcaatca aatcattccg 2040 gatactgcga ttttaagtgt tgttccattc catcacggtt ttggaatgtt tactacactc 2100 ggatatttga tatgtggatt tcgagtcgtc ttaatgtata gatttgaaga agagctgttt 2160 ctgaggagcc ttcaggatta caagattcaa agtgcgctgc tggtgccaac cctattctcc 2220 ttcttcgcca aaagcactct gattgacaaa tacgatttat ctaatttaca cgaaattgct 2280 tctggtggcg ctcccctc taaggaagtc ggggaagcgg ttgccaagag gttccatctg 2340 ccaggtatca ggcaaggata tgggctcact gagactacat cagctattct gattacaccc 2400 gagggggatg ataaaccggg cgcggtcggt aaagttgttc cattttttga agcgaaggtt 2460 gtggatctgg ataccgggaa aacgctgggc gttaatcaaa gaggcgaact gtgtgtgaga 2520 ggtcctatga tttgtccgg ttatgtaaac aatccggaag cgaccaacgc cttgattgac 2580 aaggatggat ggctacattc tggagacata gcttactggg acgaagacga acacttcttc 2640 atcgttgacc gcctgaagtc tctgattaag tacaaaggct atcaggtggc tcccgctgaa 2700 ttggaatcca tcttgctcca acaccccaac atcttcgacg caggtgtcgc aggtcttccc 2760 gacgatgacg ccggtgaact tcccgccgcc gttgttgttt tggagcacgg aaagacgatg 2820 acggaaaaag agatcgtgga ttacgtcgcc agtcaagtaa caaccgcgaa aaagttgcgc 2880 ggaggagttg tgtttgtgga cgaagtaccg aaaggtctta ccggaaaact cgacgcaaga 2940 aaaatcagag agatcctcat aaaggccaag aagggcggaa agatcgccgt gtaattctag 3000 acgagatccg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 3060 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 3120 caatgtatct tatcatgtct cgatccaggt aggtttgagt agtgggcgtg gctaagatga 3180 ctataaaggt gggtgtctta cgagggtctt tttgcttttc tgcagacatc atgaacggga 3240 ccggcggggc cttcgaaggg gggcttttta gcccttattt gacaacccgc ctgccgggat 3300 gggccggagt tcgtcagaat gtgatgggat cgacggtgga cgggcgtcca gtgcttccag 3360 caaattcctc gaccatgacc tacgcgaccg tggggaactc gtcgctcgac agcaccgccg cagccgcggc agccgcagcc gccatgacag cgacgagact ggcttcgagc tacatgccca 3480. gcagcggtag cagcccctct gtgcccagtt ccatcatcgc cgaggagaaa ctgctggccc 3540 tcctggccga gctggaagcc ctgagccgcc agctggccgc cctgacccag caggtgtccg 3600 agctccgcga acagcagcag cagcaaaata aatgattcaa taaacacaga ttctgattca aacagcaaag catctttatt atttattttt tcgcgcgcgg tagccctgg tccacctctc ccgatcattg agagtgcggt ggattttttc caggacccgg tagagatggg attggatgtt 3780 3840. gaggtacatg ggcatgagcc cgtcccgggg gtgaaggtag caccactgca tggcctcatg ctctggggtc gtgttgtag tgatccagtc atagcagggg cgctgggcgt ggtgctggat 3900 gatgtctttg aggaggac tgatggccac ggggagcccc ttggtgtagg tgttggcgaa 3960 gcggttgagc tgggagggat gcatgcgggg ggagatgatg tgcagtttgg cctggatctt 4020 gaggttggca atgttgccgc ccagatcccg ccggggggttc atgttgtgca ggaccaccag 4080 gacggtgtag cccgtgcact tggggaactt atcatgcaac ttggaaggga atgcgtggaa 4140 gaatttggag acgcccttgt gcccgcccag gttttccatg cactcatcca tgatgatggc 4200 gatgggcccg tgggctgcgg ctttggcaaa gacgtttctg gggtcagaga catcataatt 4260 atgctcctgg gtgagatcat cataagacat tttaatgaat ttggggcgga gggtgccaga 4320 ttgggggacg atcgttccct cgggccccgg ggcgaagttc ccctcacaga tctgcatctc 4380 ccaggctttc atctcggagg gggggatcat gtccacctgc ggggcgatga aaaaaacggt 4440 ttccggggcg ggggtgatga gctgcgagga gagcaggttt ctcaacagct gggacttgcc 4500 gcacccggtc gggccgtaga tgaccccgat gacgggttgc aggtggtagt tcaaggagat 4560 gcagctgccg tcttcccgga ggaggggggc cacctcgttg agcatgtctc tgacttggag 4620 gttttcccgg acgagctcgc cgaggaggcg gtccccgccc agcgagagca gctcttgcag 4680 ggaagcaaag tttttcaggg gcttgagccc gtcggccatg ggcatcttgg cgagggtctg 4740 cgagaggagt tccaagcggt cccagagctc ggtgacgtgc tctacggcat ctcgatccaa 4800 cagacttcct cgtttcgggg gttgggacga ctgcgactgt agggcacgag acgatgggcg 4860 tccagcgctg ccagcgtcat gtccttccag ggtctcaggg tccgcgtgag cgtggtctcc 4920 gtcacggtga agggtgggc cccgggctgg gcgcttgcaa gggtgcgctt gagactcatc 4980 ctgctggtgc tgaaacgggc acggtcttcg ccctgcgcgt cggcgagata gcagttgacc 5040 atgagctcgt agttgagggc ctcggcggcg tggcccttgg cgcggagctt gcccttggaa 5100 gagcgcccgc aggcgggaca gaggagggat tgcagggcgt agagcttggg cgcgagaaag 5160 acggactcgg gggcgaaagc gtccgctccg cagtgggcgc agacggtctc gcactcgacg 5220 agccaggtga gctcgggatg ttcggggtca aaaaccagtt ttcccccgtt ctttttgatg 5280 cgcttcttac ctcgcgtctc catgagtctg tgtccgcgct cggtgacaaa caggctgtct 5340 gtgtccccgt agacggactt gatgggcctg tcctgcaggg gcgtcccgcg gtcctcctcg 5400 tagagaaact cggaccactc tgagacgaag gcgcgcgtcc acgccaagac aaaggaggcc 5460 acgtgcgagg ggtagcggtc gttgtccacc aggggtcca ccttttccac cgtgtgcaga 5520 cacatgtccc cctcctcagc atccaagaag gtgattggct tgtaggtgta ggccacgtga 5580 ccgggggtcc ccgacggggg ggtataaaag ggggcgggtc tgtgctcgtc ctcactctct 5640 tccgcgtcgc tgtccacgag cgccagctgt tggggtaggt attccctctc gagattaatt 5700 aattcgaacc cataataccc ataatagctg tttgccatcg accaattctt gaagacgaaa 5760 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac 5820 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 5880 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 5940 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc 6000 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga 6060 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga 6120 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg 6180 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc 6240 tcagaatgac ttggttgagt actcaccagt cagaaaag catcttacgg atggcatgac 6300 agtaagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact 6360 tctgacaacg atcgggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca 6420 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg 6480 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact 6540 acttactcta gcttccccggc aacaattaat agaactggatg gaggcggata aagttgcagg 6600 accacttctg cgctcggccc ttccggctgg ctggtttat gctgataaat ctggagccgg 6660 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat 6720 cgtagttatc tacacgacgg ggagtcaggc aactatggat gacgaata gacagatcgc 6780 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat 6840 actttagatt gatttaaaac ttcattttta atttaaaaagg atctaggtga agatccttt 6900 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc 6960 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 7020 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 7080 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt 7140 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 7200 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 7260 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 7320 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 7380 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 7440 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc 7500 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 7560 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctgggc 7620 tggtaagagc cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct acctgcctgg 7680 acagcatggc ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga atcataatgg 7740 ggaaggccat ccagcctcgc gtcgatggca aacagctatt atgggtatta tgggttcgaa 7800 <210> 5 <211> 6901 <212> DNA <213> Artificial sequence <220> <223> pAdAptAd20-42-42.eGFP <400> 5 ttaattaaca tcatcaataa tataccccac aaagtaaaca aaagttaata tgcaaatgag 60 cttttgaatt tagggcggag ccgacgctga ttggacgaga gaagacgatg caaatgacgt 120 cacgacgcac ggctaacggt cgccgcggag gcgtggccta gcccggaagc aagtcgcggg 180 gctgatgacg tataaaaaag cggactttag acccggaaat ggccgatttt cccgcggcca 240 cggccggata tgaggtaatt ctgggcggat gcaagtgaaa ttaggtcatt ttggcgcgaa 300 aactgaatga ggaagtgaaa agcgaaaaat accgatcccg cccagggcgg aatatttacc 360 gagggccgag agactttgac cgattacgtg ggggtttcga ttgcggtgtt tttttcgcga 420 atttccgcgt ccgtgtcaaa gtccggtgtt tatgtcacag atcagctgac ctaggtggtc 480 aatattggcc attagccata ttattcattg gttatatagc ataaatcaat attggctatt 540 ggccattgca tacgttgtat ccatatcata atatgtacat ttatattggc tcatgtccaa 600 cattaccgcc atgttgacat tgattattga ctagttatta atagtaatca attacggggt 660 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc 720 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag 780 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc 840 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg 900 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc 960 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca 1020 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca 1080 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg 1140 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc 1200 gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa 1260 gacaccggga ccgatccagc ctccgcggcc gggaacggtg cattggaagc ttggtaccgg 1320 tgaattcgct agcgttaacg gatccccggg taccggtcgc caccatggtg agcaagggcg 1380 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc 1440 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga 1500 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga 1560 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca 1620 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca 1680 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc 1740 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact 1800 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact 1860 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga 1920 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt 1980 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga 2040 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta 2100 gacgagatcc gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 2160 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 2220 tcaatgtatc ttatcatgtc tcgatccagg taggtttgag tagtgggcgt ggctaagatg 2280 actataaagg tgggtgtctt acgagggtct ttttgctttt ctgcagacat catgaacggg 2340 accggcgggg ccttcgaagg ggggcttttt agcccttatt tgacaacccg cctgccggga 2400 tgggccggag ttcgtcagaa tgtgatggga tcgacggtgg acgggcgtcc agtgcttcca 2460 gcaaattcct cgaccatgac ctacgcgacc gtggggaact cgtcgctcga cagcaccgcc 2520 gcagccgcgg cagccgcagc cgccatgaca gcgacgagac tggcttcgag ctacatgccc 2580 agcagcggta gcagcccctc tgtgcccagt tccatcatcg ccgaggagaa actgctggcc 2640 ctcctggccg agctggaagc cctgagccgc cagctggccg ccctgaccca gcaggtgtcc 2700 gagctccgcg aacagcagca gcagcaaaat aaatgattca ataaacacag attctgattc 2760 aaacagcaaa gcatctttat tatttatttt ttcgcgcgcg gtaggccctg gtccacctct 2820 cccgatcatt gagagtgcgg tggatttttt ccaggacccg gtagagatgg gattggatgt 2880 tgaggtacat gggcatgagc ccgtcccggg ggtgaaggta gcaccactgc atggcctcat 2940 gctctggggt cgtgttgtag atgatccagt catagcaggg gcgctgggcg tggtgctgga 3000 tgatgtcttt gaggaggaga ctgatggcca cggggagccc cttggtgtag gtgttggcga 3060 agcggttgag ctgggaggga tgcatgcggg gggagatgat gtgcagtttg gcctggatct 3120 tgaggttggc aatgttgccg cccagatccc gccgggggtt catgttgtgc aggaccacca 3180 ggacggtgta gcccgtgcac ttggggaact tatcatgcaa cttggaaggg aatgcgtgga 3240 agaatttgga gacgcccttg tgcccgccca ggttttccat gcactcatcc atgatgatgg 3300 cgatgggccc gtgggctgcg gctttggcaa agacgtttct ggggtcagag acatcataat 3360 3420. ttgctcctg ggtgagatca tcataagaca ttttaatgaa tttggggcgg agggtgccag attgggggac gatcgttccc tcggggccccg gggcgaagtt cccctcacag atctgcatct 3480. cccaggcttt catctcggag ggggggatca tgtccacctg cggggcgatg aaaaaaacgg 3540 tttccggggc gggggtgatg agctgcgagg agagcaggtt tctcaacagc tgggacttgc 3600 cgcacccggt cgggccgtag atgaccccga tgacgggttg caggtggtag ttcaaggaga 3660 tgcagctgcc gtcttcccgg aggagggggg ccacctcgtt gagcatgtct ctgacttgga 3720 ggttttcccg gacgagctcg ccgaggaggc ggtccccgcc cagcgagagc agctcttgca 3780 gggaagcaaa gtttttcagg ggcttgagcc cgtcggccat gggcatcttg gcgagggtct 3840. gcgagaggag ttccaagcgg tcccagagct cggtgacgtg ctctacggca tctcgatcca 3900 acagacttcc tcgtttcggg ggttgggacg actgcgactg tagggcacga gacgatgggc 3960. gtccagcgct gccagcgtca tgtccttcca gggtctcagg gtccgcgtga gcgtggtctc cgtcacggtg aaggggtggg ccccgggctg ggcgcttgca agggtgcgct tgagactcat 4080 cctgctggtg ctgaaacggg cacggtcttc gccctgcgcg tcggcgagat agcagttgac 4140 catgagctcg tagttgaggg cctcggcggc gtggcccttg gcgcggagct tgcccttgga 4200 agagcgcccg caggcgggac agaggaggga ttgcagggcg tagagcttgg gcgcgagaaa 4260 gacggactcg ggggcgaaag cgtccgctcc gcagtgggcg cagacggtct cgcactcgac 4320 gagccaggtg agctcgggat gttcggggtc aaaaaccagt tttcccccgt tctttttgat 4380 gcgcttctta cctcgcgtct ccatgagtct gtgtccgcgc tcggtgacaa acaggctgtc 4440 tgtgtccccg tagacggact tgatgggcct gtcctgcagg ggcgtcccgc ggtcctcctc 4500 gtagagaaac tcggaccact ctgagacgaa ggcgcgcgtc cacgccaaga caaaggaggc 4560 cacgtgcgag gggtagcggt cgttgtccac cagggggtcc accttttcca ccgtgtgcag 4620 acacatgtcc ccctcctcag catccaagaa ggtgattggc ttgtaggtgt aggccacgtg 4680 accgggggtc cccgacgggg gggtataaaa gggggcgggt ctgtgctcgt cctcactctc 4740 ttccgcgtcg ctgtccacga gcgccagctg ttggggtagg tattccctct cgagattaat 4800 taattcgaac ccataatacc cataatagct gtttgccatc gaccaattct tgaagacgaa 4860 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga 4920 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 4980 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 5040 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattcc ttttttgcgg 5100 cattttgcct tcctgttttt gctcacccag aaacgctggt gaagtaaa gatgctgaag 5160 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 5220 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg 5280 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt 5340 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga 5400 foot attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 5460 5520 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 5580 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac 5640 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 5700 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg 5760 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta 5820 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 5880 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata 5940 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 6000 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 6060 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 6120 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 6180 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag 6240 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 6300 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 6360 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 6420 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 6480 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 6540 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 6600 ctgcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 6660 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggg 6720 ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg 6780 gacagcatgg cctgcaacgc gggcatccg atgccgccgg aagcgagaag aatcataatg 6840 gggaaggcca tccagcctcg cgtcgatggc aaacagctat tatgggtatt atgggttcga 6900 at 6901 <210> 6 <211> 18499 <212> DNA <213> Artificial Sequence <220> <223> pBR.Ad20-42-42.SbfI final intermediate <400> 6 attaaaggta ggtttgagta gtgggcgtgg ctaagatgac tataaaggtg ggtgtcttac 60 gagggtcttt ttgcttttct gcagacatca tgaacgggac cggcggggcc ttcgaagggg 120 ggctttttag cccttatttg acaacccgcc tgccgggatg ggccggagtt cgtcagaatg 180 tgatgggatc gacggtggac gggcgtccag tgcttccagc aaattcctcg accatgacct 240 acgcgaccgt ggggaactcg tcgctcgaca gcaccgccgc agccgcggca gccgcagccg 300 ccatgacagc gacgagactg gcttcgagct acatgcccag cagcggtagc agcccctctg 360 tgcccagttc catcatcgcc gaggagaaac tgctggccct cctggccgag ctggaagccc 420 tgagccgcca gctggccgcc ctgacccagc aggtgtccga gctccgcgaa cagcagcagc 480 agcaaaataa atgattcaat aaacacagat tctgattcaa acagcaaagc atctttatta 540 tttatttttt cgcgcgcggt aggccctggt ccacctctcc cgatcattga gagtgcggtg 600 gattttttcc aggacccggt agagatggga ttggatgttg aggtacatgg gcatgagccc 660 gtcccggggg tgaaggtagc accactgcat ggcctcatgc tctggggtcg tgttgtagat 720 gatccagtca tagcaggggc gctgggcgtg gtgctggatg atgtctttga ggaggagact 780 gatggccacg gggagcccct tggtgtaggt gttggcgaag cggttgagct gggagggatg 840 catgcggggg gagatgatgt gcagtttggc ctggatcttg aggttggcaa tgttgccgcc 900 cagatcccgc cgggggttca tgttgtgcag gaccaccagg acggtgtagc ccgtgcactt 960 ggggaactta tcatgcaact tggaagggaa tgcgtggaag aatttggaga cgcccttgtg 1020 cccgcccagg ttttccatgc actcatccat gatgatggcg atgggcccgt gggctgcggc 1080 tttggcaaag acgtttctgg ggtcagagac atcataatta tgctcctggg tgagatcatc 1140 ataagacatt ttaatgaatt tggggcggag ggtgccagat tgggggacga tcgttccctc 1200 gggccccggg gcgaagttcc cctcacagat ctgcatctcc caggctttca tctcggaggg 1260 ggggatcatg tccacctgcg gggcgatgaa aaaaacggtt tccggggcgg gggtgatgag 1320 ctgcgaggag agcaggtttc tcaacagctg ggacttgccg cacccggtcg ggccgtagat 1380. gaccccgatg accggttgca ggtggtagtt caaggagatg cagctgccgt cttcccggag 1440 gaggggggcc acctcgttga gcatgtctct gacttggagg ttttcccgga cgagctcgcc gaggaggcgg tccccgccca gcgagagcag ctcttgcagg gaagcaaagt ttttcagggg 1560 cttgagcccg tcggccatgg gcatcttggc gagggtctgc gagaggagtt ccaagcggtc 1620 ccagagctcg gtgacgtgct ctacggcatc tcgatccaac agacttcctc gtttcggggg 1680. ttgggacgac tgcgactgta gggcacgaga cgatgggcgt ccagcgctgc cagcgtcatg 1740. tccttccagg gtctcagggt ccgcgtgagc gtggtctccg tcacggtgaa ggggtgggcc 1800. ccgggctggg cgcttgcaag ggtgcgcttg agactcatcc tgctggtgct gaaacgggca cggtcttcgc cctgcgcgtc ggcgagatag cagttgacca tgagctcgta gttgagggcc tcggcggcgt ggcccttggc gcggagcttg cccttggag agcgcccgca ggcgggacag 1980. aggagggatt gcagggcgta gagcttgggc gcgagaaaga cggactcggg ggcgaaagcg tccgctccgc agtgggcgca gacggtctcg cactcgacga gccaggtgag ctcgggatgt 2100 tcggggtcaa aaaccagttt tccccccgttc tttttgatgc gcttcttacc tcgcgtctcc 2160 atgagtctgt gtccgcgctc ggtgaaac aggctgtctg tgtccccgta gacggacttg 2220 atgggcctgt cctgcagggg cgtcccgcgg tccctctcgt agaaactc ggaccactct 2280 gagacgaagg cgcgcgtcca cgccaagaca aaagggcca cgtgcgaggg gtagcggtcg 2340 ttgtccacca gggggtccac cttttccacc gtgtgcagac acatgtcccc ctcctcagca 2400 tccaagaagg tgattggctt gtaggtgtag gccacgtgac cggggtccc cgacgggggg 2460 gtataaaagg gggcgggtct gtgctcgtcc tcactctctt ccgcgtcgct gtccacgagc 2520 gccagctgtt ggggtaggta ttccctctcg agagcgggca tgacctcggc actcaggttg 2580 tcagtttcta gaaggaga ggatttgatg ttggcctgcc ctgccgcaat gcttttgagt 2640 aaactctcat ccatctggtc agaaaagact atttttttat tgtcaagctt ggtggcgaag 2700 gagccataga gggcgttgga gagaagcttg gcgatggatc tcatggtctg attttgtca 2760 cggtcggcgc gctccttggc cgcgatgttt agctggacat attcgcgcgc gacacacttc 2820 cattcgggga agacggtggt gcgctcgtcg ggcacgatcc tgacgcgcca gccgcggtta 2880 tgcagggtga ccaggtccac gctggtggcc acctcgccgc gcaggggctc gttggtccag 2940 cagagtcgcc cgcctttgcg cgaacagaaa gggggcagca catcaagcag atgctcgtcg 3000 ggggggtccg catcgatggt gaagataccg ggacagagtt ccttgtcaaa atagtctatt 3060 tttgaggatg catcatccaa ggccatctgc cactcacggg cggccattgc tcgctcgtag 3120 gggttgaggg gcggacccca gggcatggga tgcgtgaggg cggaggcgta catgccgcag 3180 atgtcataga catagatggg ctccgagagg atgccgatgt aggtgggata acagcgcccc 3240 ccgcggatgc tggcgcgcac gtagtcatac aactcgtgcg agggggccaa gaaggcgggg 3300 ccgagattgg tgcgctgggg ctgctcggcg cggaagacga tctggcgaaa gatggcgtgc 3360 gagttggagg agatggtggg ccgttggaag atgttaaagt gggcgtgggg caagcggacc 3420 gagtcgcgga tgaagtgcgc gtaggagtct tgcagcttgg cgacgagctc ggcggtgacg 3480 aggacgtcca tggcgcagta gtccagcgtt tcgcggatga tgtcataacc cgtctctcct 3540 ttcttctccc acagctcgcg gttgagggcg tactctcgt catccttcca gtactcccgg 3600 agcgggaatc ctcgatcgtc cgcacggtaa gagcccagca tgtagaaatg gttcacggcc 3660 ttgtagggac agcagccctt ctccacgggg agggcgtaag cttgagcggc cttgcggagc 3720 gaggtgtgcg tcagggcgaa ggtgtccctg accatgactt tcaagaactg gtacttgaaa 3780 tccgagtcgt cgcagccgcc gtgctcccag agctcgaaat cggtgcgctt cttcgagagg 3840 gggttaggca gagcgaaagt gacgtcattg aagagaatct tgcctgcccg cggcatgaaa 3900 ttgcgggtga tgcggaaagg gcccgggacg gaggctcggt tgttgatgac ctgggcggcg 3960 aggacgatct cgtcgaagcc gttgatgttg tgcccgacga tgtagagttc catgaatcgc 4020 gggcggcctt tgatgtgcgg cagctttttg agctcctcgt aggtgaggtc ctcggggcat 4080 tgcaggccgt gctgctctag cgccactcc tggagatgtg ggttggcttg catgaaggaa 4140 gcccagagct cgcgggccat gagggtctgg agctcgtcac gaagaggcg gaactgctgg 4200 cccacggcca tctttcggg tgtgacgcag tagaaggtga gggggtcccg ctcccagcga 4260 tcccagcgta agcgcacggc gagatcgcga gcgagggcga ccagctctgg gtccccccgag 4320 aatttcatga ccagcatgaa ggggacgagc tgcttgccga aggaccccat ccaggtgtag 4380 gtttctacat cgtaggtgac aaagagccgc tccgtgcgag gatgagagcc gattgggaag 4440 aactggattt cctgccacca gttggacgag tggctgttga tgtgatgaaa gtagaaatcc 4500 cgccggcgaa ccgagcactc gtgctgatgc ttgtaaaagc gtccgcagta ctcgcagcgc 4560 tgcacgggct gtacctcatc cacgagatac acagcgcgtc ccttgaggag gaacttcagg 4620 agtggcggcc ctggctggtg gttttcatgt tcgcctgcgt gggactcacc ctggggctcc 4680 tcgaggacgg agaggctgac gagcccgcgt gggagccagg tccagatctc ggcgcggcgg 4740 gggcggagag cgaagacgag ggcgcgcagt tgggagctgt ccatggtgtc gcggagatcc 4800 aggtccgggg gcagggttct gaggttgacc tcgtagaggc gggtgagggc gtgcttgaga 4860 tgcagatggt acttgatttc tacgggtgag ttggtggccg tgtccacgca ttgcatgagc 4920 ccgtagctgc gcggggccac gaccgtgccg cggtgcgctt ttagaagcgg tgtcgcggac 4980 gcgctcccgg cggcagcggc ggttccggtc ccgcgggcag gggcggcaga ggcacgtcgg 5040 cgtggcgctc gggcaggtcc cggtgctgcg ccctgagagc gctggcgtgc gcgacgacgc 5100 ggcggttgac atcctggatc tgccgcctct gcgtgaagac cacgggcccc gtgactttga 5160 acctgaaaga cagttcaaca gaatcaatct cggcgtcatt gacggcggcc tgacgcagga 5220 tctcttgcac gtcgcccgag ttgtcctggt aggcgatctc ggacatgaac tgctcgatct 5280 cctcctcctg gagatcgccg cggcccgcgc gctcgacggt ggcggcgagg tcattcgaga 5340 tgcgacccat gagctgcgag aaggcgccca ggccgctctc gttccagacg cggctgtaga 5400 ccacgtcccc gtcggcgtcg cgcgcgcgca tgaccacctg cgcgaggttg agctccacgt 5460 gccgcgcgaa gacggcgtag ttgcgcaggc gctggaagag gtagttgagg gtggtggcga 5520 tgtgctcggt gacgaagaag tacatgatcc agcggcgcag aggcatctcg ctgatgtcgc 5580 cgatggcctc cagcctttcc atggcctcgt agaaatccac agcgaagttg aaaaactggg 5640 cgttgcgggc cgagaccgtg agctcgtctt ccaggagcct gatgagctcg gcgatggtgg 5700 cgcgcacctc gcgctcgaaa tccccggggg cctcctcctc ttcctcttct tccatgacga 5760 cctcttcttc tatttcttcc tctgggggcg gtggtggtgg cggggcccga cgacgacggc 5820 gacgcaccgg gagacggtcg acgaagcgct cgatcatctc cccgcggcgg cgacgcatgg 5880 tttcggtgac ggcgcgaccc cgttcgcgag gacgcagcgt gaagacgccg ccggtcatct 5940 cccggtaatg gggcgggtcc ccgttgggca gcgatagtgc gctgacgatg catcttatca 6000 attgcggtgt aggggacgtg agcgcgtcga gatcgaccgg atcggagaat ctttcgagga 6060 aagcgtctag ccaatcgcag tcgcaaggta agctcaaaca cgtagcagcc ctgtggacgc 6120 tgttagaatt gcggttgctg atgatgtaat tgaagtaggc gtttttgagg cggcggatgg 6180 tggcgaggag gaccaggtcc ttgggtcccg cttgctggat gcggagccgc tcggccatgc 6240 cccaggcctg gccctgacac cggctcaggt tcttgtagta gtcatgcatg agcctctcga 6300 tgtcatcact ggcggaggcg gagtcttcca tgcgggtgac cccaacgccc ctgagcggct 6360 gcacgagcgc caggtcggcg acgacgcgct cggcgaggat ggcctgttgc acgcgggtga 6420 gggtgtcctg gaagtcgtcc atgtcgacga agcggtggta ggcccctgtg ttgatggtgt 6480 aagtgcagtt ggccatgagc gaccagttga cggtctgcag gccgggctgc acgacctcgg 6540 agtacctgag ccgcgagaag gcgcgcgagt cgaagacgta gtcgttgcag gtgcgcacga 6600 ggtactggta tccgactagg aagtgcggcg gcggctggcg gtagagcggc cagcgctggg 6660 tggccggcgc gcccggggcc aggtcctcga gcatgaggcg gtggtagccg tagaggtagc 6720 gggacatcca ggtgatgccg gcggcggtgg tggaggcgcg cgggaactcg cggacgcggt 6780 tccagatgtt gcgcagcggc aggaaatagt ccatggtcgg cacggtctgg ccggtgagac 6840 gcgcgcagtc attgacgctc tagaggcaaa aacgaaagcg gttgagcggg ctattcctcc 6900 gtagcctggc ggaacgcaaa cgggttaggc cgcgtgtgta ccccggttcg agtcccctcg 6960 aatcaggctg gagccgcgac taacgtggta ttggcactcc cgtctcgacc cgagcccgat 7020 agctgccagg atacggcgga gagccctttt ggcggccgag gggggtcgct agacttgaaa 7080 gcggccgaaa accctgccgg gtagtggctc gcgcccgtag tctggagaag catcgccagg 7140 gttgagtcgc ggcagaaccc ggttcgcgga cggccgcgga gagcgggact tggtcacccc 7200 gccgatttaa aaacccacag ccagccgact tctccagtta cgggagcgag cccccttttt 7260 tctttttgcc agatgcatcc cgtcctgcgc caaatgcgtc ccaccccccc tccggcgacc 7320 accgcaaccg cggccgtagc aggcgccggc gctagccagc cacagccaca gacagagatg 7380 gacttggaag agggcgaagg gctggcgaga ttgggggcgc cgtctccgga gcgacacccc 7440 cgcgtgcagc tgcagaagga cgtgcgcccg gcgtacgtgc ctgcgcagaa cctgttcagg 7500 gaccgcagcg gggaggagcc cgaggagatg cgcgactgcc gctttcgggc gggcagggag 7560 ctgcgcgagg gtctggaccg ccagcgcgtg ctgcgcgatg aggatttcga gccgaacgag 7620 cagacgggga tcagccccgc gcgcgcgcac gtggcggcgg ccaacctggt gacggcctac 7680 gagcagacgg tgaagcagga gcgcaacttc caaaagagtt tcaacaacca cgtgcgcacg 7740 ctgatcgcgc gcgaggaggt ggccctgggc ctgatgcacc tgtgggacct ggcggaggcc 7800 atcgtgcaga acccggacag caagcctctg acggcgcagc tgttcctggt ggtgcagcac 7860 agtagggaca acgaggcgtt cagggaggcg ctgctgaaca tcgccgagcc cgagggtcgc 7920 tggctgctgg agctgatcaa catcttgcag agcatcgtag tgcaggagcg cagcctgagc 7980 ctggccgaga aagtggcggc gatcaactac tcggtgctga gcctgggcaa gttttacgcg 8040 cgcaagattt acaagacgcc atacgtgccc atagacaagg aggtgaagat agacagcttt 8100 tacatgcgca tggcgctcaa ggtgctgacg ctgagcgacg acctgggcgt gtaccgcaac 8160 gaccgcatcc acaaggccgt gagcgcgagc cggcggcgcg agctgagcga ccgcgagctg 8220 atgctgagcc tgcgccgggc gctggtaggg ggcgccgccg gcggcgagga gtcctacttc 8280 gacatggggg cggacctgca ttggcagccg agccggcgcg ccttggaagc cgcctacggt 8340 ccagaggact tggatgagga tgaggaagag gaggaggatg cacccgctgc ggggtactga 8400 cgcctccgtg atgtgttttt agatgtccca gcaagccccg gaccccgcca taagggcggc 8460 gctgcaaagc cagccgtccg gtctagcatc ggacgactgg gaggccgcga tgcaacgcat 8520 catggccctg acgacccgca accccgagtc ctttagacaa cagccgcagg ccaacagact 8580 ttcggccatt ctggaggcgg tggtcccctc tcgaaccaac cccacgcacg agaaggtgct 8640 ggcgatcgtg aacgcgctgg cggagaacaa ggccatccgt cccgacgagg ccggtctggt 8700 gtacaacgcc ctgctggagc gcgtgggccg ctacaacagc acaaacgtgc agtccaacct 8760 ggaccggctg gtgacggacg tgcgcgaggc cgtggcgcag cgcgagcggt tcaagaacga 8820 aggcctgggc tcgctggtgg cgctgaacgc cttcctggcg acgcagccgg cgaacgtgcc 8880 gcgcgggcag gacgattata ccaactttat aagcgcgctg cggctgatgg tgaccgaggt 8940 gccccagagc gaggtgtacc agtcgggccc ggattacttt ttccaaaacta gcacacaggg 9000 cctgcagacg gtgaacctga gccaggcctt taagaacctg cgcgggctgt ggggcgtgca 9060 ggcgccccgtg ggcgaccggt cgacggtgag cagcttgctg acgcccaact cgcggctgct 9120 gctgctgctg atcgcgccct tcaccgacag cggcagcgtg aaccgcaact cgtacctggg 9180 ccacctgctg acgctgtacc gcgaggccat aggccaggca caggtggacg agcagacctt 9240 ccaggagatc actagcgtga gccgcgcgct ggggcagaac gacaccgaca gtcttagggc 9300 caccctgaac ttcttgctga ccaatagaca gcaagaatc ccggcacagt acgcgctatc 9360 ggccgaggag gagcgcatcc tgagatatgt gcagcagagc gtagggctgt tcctgatgca 9420 ggagggggcc acccccagcg ccgcgctgga catgaccgcg cgcaacatgg aacctagcat 9480 gtacgctgcc aaccggccgt tcatcaataa gctgatggac tacctgcacc gcgcggcggc 9540 catgaactcg gactacttta caaacgccat cctgaacccg cactggctcc cgccgccggg 9600 gttctacacg ggcgagtacg atatgcccga ccccaacgac gggttcctgt gggacgacgt 9660 ggacagcgcg gtgttctccc cgaccttgca aaagcgccag gaggcggtgc gcacgcccgc 9720 gagcgagggt gcggtgggtc ggagcccctt tcctagctta gggagtttgc atagcttgcc 9780 gggctcggtg aacagcggca gggtgagccg accgcgcttg ctgggcgagg acgagtacct 9840 gaacgactcg ctgctgcagc cgccgcgggt caagaacgcc atggccaata acgggataga 9900 gagtctggtg gacaaactga accgctggaa gacctacgct caggaccata gggacgcgcc 9960 cgcgccgcgg cgacagcgcc acgaccggca gcggggcctg gtgtgggacg acgaggactc 10020 ggccgacgat agcagcgtgt tggacttggg cgggagcggt ggggccaacc cgttcgcgca 10080 tctgcagccc aaactggggc gacggatgtt ttgaaatgca aaataaact caccaaggcc 10140 atagcgtgcg ttctcttcct tgttagagat gaggcgtgcg gtggtgtctt cctctctccc 10200 tccctcgtac gagagcgtga tggcgcaggc gaccctggag gttccgtttg tgcctccgcg 10260 gtatatggct cctacggagg gcaagaacag cattcgttat tcggagctgg ctccgctgta 10320 cgacaccact cgcgtgtact tggtggacaa caagtcggcg gacatcgctt ccctgaacta 10380 ccaaaacgac cacagcaact tcctgaccac ggtggtgcag aacaacgatt tcacccccgc 10440 cgaggccagc acgcagacga taaattttga cgagcggtcg cggtggggcg gtgatctgaa 10500 gaccattctg cacacaaca tgcccaatgt gaacgagtac atgttcacca gcaagtttaa 10560 ggcgcgggtg atggtggcta gaaagcaccc agaaatgta gctaaagacg atttgagtca 10620 ggataagctt gaatatgagt ggtttgagtt taccctgccc gagggcaact tttccgagac 10680 catgaccata gacctgatga aaacgccat cttggaaaac tacttgcaag tggggcggca 10740 aaatggcgtg ctggagagcg atatcggagt caagtttgac agcagaaatt tcaagctggg 10800 ctgggacccg gtgaccaagt tggtgatgcc aggggtctac acctacgagg ccttccaccc 10860 ggacgtggtg ctgctgccgg gctgcggggt ggacttcacc gagagccgcc tgagcaacct 10920 cctgggcatt cgcaagaagc aacctttcca agaggggcttc agaatcatgt atgaagatct 10980 agtagggggc aacatcccag ccctcctgaa tgtcaaggag tatctgaagg ataaggaaga 11040 agctggcaca ccagatgcaa ataccattaa ggctcagaat gatgcagtcc caagaggaga 11100 taactatgca tcagcggcag aagccaaagc agcaggaaaa gaaattgagt tgaaggccat 11160 11220 ccgcagttgg tacctgtcct atacctacgg ggaccccgag aagggggtgc agtcgtggac 11280 gctgctcacc accccggacg tcacatgcgg cgcggagcaa gtctactggt cgctgccgga 11340 cctcatgcaa gaccccgtca ccttccgctc tacccagcaa gtcagcaact accccgtggt 11400 cggcgccgag ctcatgccct tccgcgccaa gagcttttac aacgacctcg ccgtctactc 11460 ccagctcatc cgcagctaca cctccctcac ccacgtcttc aaccgcttcc ccgacaacca 11520 gatcctctgc cgcccgcccg cgcccaccat caccaccgtc agtgaaaacg tgcctgctct 11580 cacagatcac gggacgctac cgctgcgcag foottccgc ggagtccagc gagtgaccgt 11640 cactgacgcc cgtcgccgca cctgtcccta cgtctacaag gccctgggca tagtcgcgcc 11700 gcgcgtgcta tccagtcgca ccttctaaaa aatgtctatt ctcatctcgc ccagcaataa 11760 caccggctgg ggtattacta ggcccagcac catgtacgga ggagccaaga agcgctccca 11820 gcagcacccc gtccgcgtcc gcggccactt ccgcgctccc tggggcgctt acaagcgcgg 11880 gcggacttct accgccgctg ccgtgcgtac caccgtcgac gacgtcatcg actcggtggt 11940 cgccgacgcg cgcaactaca cccccgcccc ctccaccgtg gacgcggtca ttgacagcgt 12000 ggtggccgac gcgcgcgact atgccagacg caagagccgg cggcgacgga tcgccaggcg 12060 ccaccggagc acgcccgcca tgcgcgccgc ccgggctctg ctgcgccgcg ccagacgcac 12120 aggccgccgg gccatgatgc gagccgcgcg ccgcgctgcc actgcaccca cccccgcagg 12180 caggactcgc agacgagcgg ccgccgccgc tgccgcggcc atctctagca tgaccagacc 12240 caggcgcgga aacgtgtact gggtgcgcga ctccgtcacg ggcgtgcgcg tgcccgtgcg 12300 cacccgtcct cctcgtccct gatctaatgc ttgtgtcctc ccccgcaagc gacgatgtca 12360 aagcgcaaaa tcaaggagga gatgctccag gtcgtcgccc cggagattta cggaccaacc 12420 caggcggacc agaaaccccg caaaatcaag cgggttaaaa aaaaggatga ggtggacgag 12480 ggggcagtag agtttgtgcg cgagttcgct ccgcggcggc gcgtaaattg gaaggggcgc 12540 agggtgcagc gcgtgttgcg gcccggcacg gcggtggtgt tcacgcctgg cgagcggtcc 12600 tcggtcagga gcaagcgtag ctatgacgag gtatacggcg acgacgacat cctggaccag 12660 gcggcggagc gggcgggcga gtttgcctac gggaagcggt cgcgcgaaga ggagctgatc 12720 tcgctgccgc tggacgaaag caaccccacg ccaagcctga agcccgtgac cctgcagcag 12780 gtgctgcccc aggcggtgct gctgccgagc cgcggggtca agcgcgaggg cgagaacatg 12840 tacccgacca tgcagatcat ggtgcccaag cgccggcgcg tggaggacgt gctggacacc 12900 gtgaaaatgg atgtggagcc cgaggtcaag gtgcgcccca tcaagcaggt ggcgccgggc 12960 ctgggcgtgc agaccgtgga cattcagatc cccaccgaca tggatgtcga caaaaaaccc 13020 tcgaccagca tcgaggtgca gaccgacccc tggctcccag cctccactgc taccgcctcc 13080 acttctaccg tcgccacggc caccgagcct cccaggaggc gaagatgggg ccctgccaac 13140 cggctgatgc ccaactacgt gttgcatcct tccatcatcc cgacgccggg ctaccgcggc 13200 actcggtact acgccagccg caggcgccca gccagcaaac gccgccgccg caccgccacc 13260 cgccgccgtc tggcccccgc ccgcgtgcgc cgcgtgacca cgcgccgggg ccgctcgctc 13320 gttctgccca ccgtgcgcta ccaccccagc atcctttaat ccgtgtgctg tgatactgtt 13380 gcagagagat ggctctcact tgccgcctgc gcatccccgt cccgaattac cgaggaagat 13440 cccgccgcag gagaggcatg gcaggcagtg gcctgaaccg ccgccggcgg cgggccatgc 13500 gcaggcgcct gagtggcggc tttctacccg cgctcatccc cataatcgcc gcggccatcg 13560 gcacgatccc gggcatagct tccgttgcgc tgcaggcgtc gcagcgccgt tgatgtgcga 13620 ataaagcctc tttagactct gacacacctg gtcctgtata tttttagaat ggaagacatc 13680 aattttgcgt ccctggctcc gcggcacggc acgcggccgt tcatgggcac ctggaacgag 13740 atcggcacca gccagctgaa cgggggcgcc ttcaattgga gcagtgtctg gagcgggctt 13800 aaaaatttcg gctcgacgct ccggacctat gggaacaagg cctggaatag tagcacgggg 13860 cagttgttaa gggaaaagct caaagaccag aacttccagc agaaggtggt ggacgggctg 13920 gcctcgggca ttaacggggt ggtggacatc gcgaaccagg ccgtgcagcg cgagataaac 13980 agccgcctgg acccgcgccc gcccacggtg gtggagatgg aagatgcaac tcttccgccg 14040 cccaaaggcg agaagcggcc gcggcccgac gcggaggaga cgatcctgca ggtggacgag 14100 ccgccctcgt acgaggaggc cgtgaaggcc ggcatgccca ccacgcgcat catcgcgccg 14160 ctggccacgg gtgtaatgaa acccgccacc cttgacctgc ctccaccacc cacgcccgct 14220 ccaccgaagg cagctccggt tgtgcagccc cctcctgtgg cgaccgccgt gcgccgcgtc 14280 cccgcccgcc gccaggccca gaactggcag agcacgctgc acagtatcgt gggcctggga 14340 gtgaaaagtc tgaagcgccg ccgatgctat tgagagagag gaaagaggac actaaaggga 14400 gagcttaact tgtatgtgcc ttaccgccag agaacgcgcg aagatggcca ccccctcgat 14460 gatgccgcag tgggcgtaca tgcacatcgc cgggcaggac gcctcggagt acctgagccc 14520 gggtctggtg cagtttgccc gcgccaccga cacgtacttc agcctgggca acaagtttag 14580 gaaccccacg gtggctccca cccacgatgt gaccacggac cggtcccagc gtctgacgct 14640 gcgcttcgtg cccgtggatc gcgaggacac cacgtactcg tacaaggcgc gcttcactct 14700 ggccgtgggc gacaaccggg tgctagacat ggccagcact tactttgaca tccgcggcgt 14760 cctggaccgc ggccccagct tcaaacccta ctcgggcacg gcctacaaca gtctggcccc 14820 caagggcgcc cccaactcca gccagtgggc tgacaaaga agggtgaatg gaggcggtaa caccaaagat gttacaaaga catttggtgt ggcagccatg ggtggagaag acattacgga aaagggtctg aagattgga ctgatacaac agaaatga ccaatttg cagacaaaaa ttttcagcct gaaccccaag tggagagga aaactggcag gaaacatttg ttttctatgg aggaagagct cttaaaaaag aaaccaaaat gaagccttgc tatggttcat ttgccagacc 15180. 15180. 15180. 15180. 15180. 15180. 15180. 15180. 15180 aaaccatgac father ctttctttga cactcctggt ggtacaatca caggaggtac tggtggtccc caught the cattgttatg caught the father tgaaactcca gatacccatg tggtgtacaa gccaggcaaa gaagatgata gctcagaaat 15420. 15420. 15420. 15420. 15420. 15420. 15420. 15420. 15420. 15420. 15420. 15420. 15420. 15420. 15420. 15420: 15480. tgtgggactt atgtactaca acagcactgg caacatgggc gtgctggctg gtcaggcctc tcagttgaat gctgtggtcg acttgcaaga cagaaacacc gagctgtctt accagctcct 15540 gctagattct ctgggtgaca ggaccagata ctttagcatg tggaactctg cggtggacag 15600 ttatgatccc gatgttagga tcattgagaa tcacggtgtg gaagatgaac ttcccaacta 15660 ttgctttcca ttggacggct caggcactaa ttcagctttt caaggtgtaa aaattaagca 15720 aaaccaagac ggagatgtta atgatgattg ggaaaaggat gataaggtat ccactcaaaa 15780 ccaaatttgc aagggcaaca tttacgccat ggagatcaac ctccaggcca acctgtggaa 15840 aagttttctg tactcgaacg tggccctgta cctgcccgac tcctacaagt acacgccggc 15900 caacgtgacg ctgcccgcca acaccaacac ctacgagtac atgaacggcc gcgtggtagc 15960 cccctcgctg gtggacgcct acatcaacat cggcgcccgc tggtcgctag accccatgga 16020 caatgtcaac cccttcaacc accaccgcaa tgcgggtctg cgctaccgct ccatgcttct 16080 gggcaacggc cgctacgtgc ccttccacat ccaagtgccc caaaagttct ttgccatcaa 16140 gaacctgctc ctgctcccgg gctcctacac ctacgagtgg aacttccgca aggacgtcaa 16200 catgatcctg cagagttccc ttggaaacga cctgcgcgtt gacggcgcct ccgtccgctt 16260 cgacagtgtt aacctctatg ccaccttctt ccccatggcg cacaacaccg cctccaccct 16320 ggaagccatg ctgcgcaacg acaccaacga ccagtccttc aacgactacc tctcggccgc 16380 caacatgctc taccctatcc cggccaaggc cattaattaa ttcgaaccca taatacccat 16440 aatagctgtt tgccatcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct 16500 tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac 16560 catcagggac agcttcaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 16620 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 16680 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 16740 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 16800 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 16860 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 16920 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 16980 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 17040 tggcctaact acggctcac tagaaggaca gtatttggta tctgcgctct gctgaagcca 17100 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 17160 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 17220 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 17280 ttggtcatga gattatcaaa aggatcttc acctagatcc ttttaaatta aaaatgaagt 17340 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 17400 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 17460 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 17520 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 17580 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 17640 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 17700 gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 17760 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 17820 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 17880 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 17940 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 18000 acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 18060 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 18120 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 18180 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 18240 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 18300 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 18360 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat 18420 aggcgtatca cgaggccctt tcgtcttcaa gaattggtcg atggcaaaca gctattatgg 18480 gtattatggg ttcgaatta 18499 <210> 7 <211> 17834 <212> DNA <213> Artificial Sequence <220> <223> pBrAd20-42-42 SrfI-rITR_dE3_5orf6 <400> 7 tgcaggtgga cgagccgccc tcgtacgagg aggccgtgaa ggccggcatg cccaccacgc 60 gcatcatcgc gccgctggcc acgggtgtaa tgaaacccgc cacccttgac ctgcctccac 120 cacccacgcc cgctccaccg aaggcagctc cggttgtgca gccccctcct gtggcgaccg 180 ccgtgcgccg cgtccccgcc cgccgccagg cccagaactg gcagagcacg ctgcacagta 240 tcgtgggcct gggagtgaaa agtctgaagc gccgccgatg ctattgagag agaggaaaga 300 ggacactaaa gggagagctt aacttgtatg tgccttaccg ccagagaacg cgcgaagatg 360 gccaccccct cgatgatgcc gcagtgggcg tacatgcaca tcgccgggca ggacgcctcg 420 gagtacctga gcccgggtct ggtgcagttt gcccgcgcca ccgacacgta cttcagcctg 480 ggcaacaagt ttaggaaccc cacggtggct cccacccacg atgtgaccac ggaccggtcc 540 cagcgtctga cgctgcgctt cgtgcccgtg gatcgcgagg acaccacgta ctcgtacaag 600 gcgcgcttca ctctggccgt gggcgacaac cgggtgctag acatggccag cacttacttt 660 gacatccgcg gcgtcctgga ccgcggcccc agcttcaaac cctactcggg cacggcctac 720 aacagtctgg cccccaaggg cgcccccaac tccagccagt gggctgacaa agaaagggtg 780 aatggaggcg gtaacaccaa agatgttaca aagacatttg gtgtggcagc catgggtgga 840 gaagacatta cggaaaaggg tctgaagatt ggaactgata caacagaaaa tgaaccaata 900 tttgcagaca aaaattttca gcctgaaccc caagtgggag aggaaaactg gcaggaaaca 960 tttgttttct atggaggaag agctcttaaa aaagaaacca aaatgaagcc ttgctatggt 1020 tcatttgcca gaccgactaa tgaaaaaggt ggccaggcta aatttataat tggagataat 1080 ggtcaaccca ctgaaaacca tgacataaca atggctttct ttgacactcc tggtggtaca 1140 atcacaggag gtactggtgg tccccaagat gaacttaaag ctgacattgt tatgtacact 1200 gaaaatataa atcttgaaac tccagatacc catgtggtgt acaagccagg caaagaagat 1260 gatagctcag aaattaactt ggtgcaacag tccatgccca acagacccaa ctacatcggt 1320 ttcagagaca actttgtggg acttatgtac tacaacagca ctggcaacat gggcgtgctg 1380 gctggtcagg cctctcagtt gaatgctgtg gtcgacttgc aagacagaaa caccgagctg 1440 tcttaccagc tcctgctaga ttctctgggt gacaggacca gatactttag catgtggaac 1500 tctgcggtgg acagttatga tcccgatgtt aggatcattg agaatcacgg tgtggaagat 1560 gaacttccca actattgctt tccattggac ggctcaggca ctaattcagc ttttcaaggt 1620 gtaaaaatta agcaaaacca agacggagat gttaatgatg attgggaaaa ggatgataag 1680 gtatccactc aaaaccaaat ttgcaagggc aacatttacg ccatggagat caacctccag 1740 gccaacctgt ggaaaagttt tctgtactcg aacgtggccc tgtacctgcc cgactcctac 1800 aagtacacgc cggccaacgt gacgctgccc gccaacacca acacctacga gtacatgaac 1860 ggccgcgtgg tagccccctc gctggtggac gcctacatca acatcggcgc ccgctggtcg 1920 ctagacccca tggacaatgt caaccccttc aaccaccacc gcaatgcggg tctgcgctac 1980 cgctccatgc ttctgggcaa cggccgctac gtgcccttcc acatccaagt gccccaaag 2040 ttctttgcca tcaagaacct gctcctgctc ccgggctcct acacctacga gtggaacttc 2100 cgcaaggacg tcaacatgat cctgcagagt tcccttggaa acgacctgcg cgttgacggc 2160 gcctccgtcc gcttgcacag tgttaacctc tatgccacct tcttccccat ggcgcacaac 2220 accgcctcca ccctggaagc catgctgcgc aacgacacca acgaccagtc cttcaacgac 2280 tacctctcgg ccgccaacat gctctaccct atcccggcca aggccaccaa cgtgcccatc 2340 tccatcccct cgcgcaactg ggccgccttc cgcggctgga gtttcacccg gctcaagacc 2400 areaactc cctcccttgg ctcgggtttt gacccctact ttgtctactc gggctccatc 2460 ccctacctcg acgggacctt ttacctgaac cataccttca agaaggtctc catcatgttc 2520 gactcctcgg tcagctggcc cggcaacgac cggctgctca cgccgaacga gttcgagatc 2580 aagcgcagcg ttgacggggga gggctacaac gtggcccaat gcaacatgac caaggactgg 2640 ttccttgtcc agatgctctc ccactacaac atcggctacc agggcttcca cgtgccccgag 2700 ggctacaagg accgcatgta ctcattcttc cgcaacttcc agcccatgag caggcaggtg 2760 gtcgatgaga tcaactacaa ggactacaaa gccgtcaccc tgcccttcca gcacaacaac 2820 tcgggcttca ccggctatct cgcccccacc atgcgccagg ggcagcccta ccccgccaac 2880 ttcccatacc cgctcatcgg ccagacagcc gtaccctccg tcacccagaa aaagttcctc 2940 tgcgacaggg tcatgtggcg catccccttt tccagcaact tcatgtccat gggcgccctc 3000 accgacctgg gacagaacat gctctatgcc aactcggccc acgcgctcga cataactttt 3060 gaggtggacc ccatggatga gcccaccctc ctctatcttc tcttcgaagt tttcgacgtg 3120 gtcagagtgc accagccgca ccgcggcgtc atcgaggccg tctacctgcg cacgcccttc 3180 tccgccggca acgctactac ataagcatga gcggctccag cgaacgagag ctcgcggcca 3300. tcgtgcgcga cctgggctgc gggccctact ttttgggac ccacgacaag cgcttccctg gcttccttgc cagcgacaag ctggcatgcg ccatcgtcaa cacggccggc cgcgagaccg 3360 ggggcgtgca ctggctcgcc tttggctgga atccgcgctc gcgcacctgc tacatgtttg 3420 acccctttgg gttctcggac cgccggctca bite cagcttcgag tacgaggcca 3480. tgctgcgccg cagcgccctg gcctcctcgc ccgaccgctg tctcagcctc gagcagtcca 3540 cccagaccgt gcaggggccc gactccgccg cctgcggact tttttgttgc atgtttttgc 3600 atgcctttgt gcactggccc gaccgaccca tggacggaa ccccaccatg aacttgctga 3660 cggggggtgcc caacggcatg ctacaatcgc cacaggtgct ccccaccctc aggcgcaacc 3720. aggaggagct ctaccgcttc ctcgcgcgcc actcccctta ctttcgatcc caccgcgccg 3780 ccattgaaaa cgccaccgct tttgataaaa tgaaacaact gcgtgtatct caataacag cactttattt tacatgcact ggagtatatg caagttattt aaaagtcgaa ggggttctcg cgctcgtcgt tgtgcgccgc gctggggagg gccacgttgc ggtactgaaa cttgggctgc 3960 cacttgaact cggggatcac cagtttgggc actggggtct cggggaaggt ctcgctccac 4020 atgcgccggc tcatctgcag ggcgcccagt atgtcgggcg cggagatctt gaaatcgcag 4080 ttgggaccag tgctctgcgc gcgcgagttg cggtacacgg ggttgcagca ctggaacacc 4140 atcaaactgg ggtgcttcac gctggccagc acgctcttgt cgctgatctg atccttgtcc 4200 aggtcctcgg cgttgctcag gccgaacggg gtcatcttgc acagctggcg gcccaggaag 4260 ggcacgctgt gaggcttgtg gttacactcg cagtgcacgg gcatcagcat catccccgcg 4320 ccgcgctgca tattcgggta gagggccttg acaaaggcca agatctgctt gaaagcttgc 4380 tgggccttgg ccccctcgct gaaacagg ccgcagctct tcccgctgaa ctggttattc 4440 ccacacccgg catcatgcac gcagcagcgc gcgtcatggc tggtcagttg caccacgctt 4500 cggccccagc ggttctgggt caccttggcc ttgctggggct gctccttcaa cgcgcgctgg 4560 ccgttctcgc tggtcacatc catctccacc acgtggtcct tgtggatcat caccgtcccg 4620 tgcagacact tgagctggcc ctcgacatcg gtgcagccgt gttcccacag ggcgcagccg 4680 gtgcactccc aattcttgtg cgcgatcccg ctgtggctga agatgtaacc ttgcaacatg 4740 cggcccatca cggtgctaaa tgatttactg gtgctgaagg tcagttgcag gccgcgggcc 4800 tcctcgttca tccatgtctg acacattttc tggaagatct cggtctgctc gggcatgagc 4860 ttgtaggcat cgcgcaggcc gctgtcgacg cggtagcgtt ccatcagcac gttcatggta 4920 tccatgccct tctcccagga cgagaccaga ggcagactca gggggttgcg cacgttcagg 4980 ataccggggg tcgcgggctc gacgatgcgt tttccgtcct ttccttcctt caacagaacc 5040 ggcggctggc tgaatcccac tcccacgatc acggcgtctt cctggggcat ctcttcgtcg 5100 gggtctacct tggtcacatg cttggtcttt ctggcttgct tcttttttgg agggctgtcc 5160 acggggacca cgtcctcctc ggaagacccg gagcccaccc gctgatactt tcggcgcttg 5220 gtgggcagag gaggtggcgg cggcgagggg ctcctctcct gctccggcgg atagcgcgcc 5280 gacccgtggc cccggggcgg agtggcctct cggtccatga accggcgcac gtcctgactg 5340 ccgccggcca ttgttccta ggggagatg gaggagcagc cgcgtaagca ggagcaggag 5400 gaggacttaa ccacccacga gcaaccaaa atcgagcagg acctggggctt cgaagagccg 5460 gctcgtctag aacccccaca ggatgaacag gagcacgagc aagacgcagg ccaggaggag 5520 accgacgctg ggctccacca tggtacctg ggaggagagg aggatgtgct gctgaacac 5580 ctgcagcgcc agtccctcat cctccgggac gcctagccg accggagcga aacccccctc 5640 agcgtcgagg agctgtgtcg gcctacgag ctcaacctct tctcgccgcg catgcccccc 5700 aaacgccagc ccaatggcac atgcgagccc aacccgcgtc tcaacttcta tcccgtctt 5760 gcggtccccg aggccttgc cacctatcac atctttttca agaaccaaa gatccccgtc 5820 tcctgccgcg ccaccgcac ccgcgccgac gcgctcctta ctctggggcc cggcgcgcgc 5880 atacctgata tcgcttccct ggaagaggtg cccagatct tcgaagggct cggtcgggac 5940 gagacgcgcg cggcgaacgc tctgaaagaa acagcagagg aagagggtca cactagcgcc 6000 ctggtagagt tggaggcga caacgccagg ctggccgtgc tcaagcgcag cgtcgagctc 6060 accaccttcg cctaccccgc cgtcaacctc ccgcccaagg tcatgcgtcg catcatggat 6120 cagcttatca tgccccacat cgaggccctc gatgaaagtc aggagcagcg ccccgaggac 6180 gcccggcccg tggtcagcga cgagatgctc gcgcgctggc tcgggacccg cgacccccag 6240 gctttggaac agcggcgcaa gctcatgctg gccgtggtcc tggtcaccct cgagctgggaa 6300 tgcatgcgcc gcttcttcag cgaccccgag accctgcgca aggtcgagga gaccctgcac 6360 tacactttca ggcacggttt cgtcaggcag gcctgcaaga tctccaacgt ggagctgacc 6420 aacctggtct cctgcctggg gatcctgcac gagaaccgcc tgggccagac cgtgctccac 6480 tctaccctga agggcgaggc gcggcgggac tacgtccgcg actgcgtctt tctctttctc 6540 tgccacacat ggcaagcggc catgggcgtg tggcagcagt gtctcgagga cgagaacctg 6600 aaggagctgg acaagcttct tgctagaaac cttaaaaagc tgtggacggg cttcgacgag 6660 cgcaccgtcg cctcggacct ggccgagatc gtttccccg agcgcctgag agagacgctg 6720 aaaggcgggc tgcccgactt catgagccag agcatgttgc aaaactaccg cactttcatt 6780 ctcgagcgat ctgggatgct gcccgccacc tgcacgcct tccctccga ctttgtaccg 6840 ctgagctacc gcgagtgtcc cccgccgctg tggagccact gctactct gcagctggcc 6900 aactacatcg cctaccactc ggacgtgatc gaggacgtga gcggcgaggg gctgctcgag 6960 tgccactgtc gctgcaacct gtgctccccg catcgctccc tgtctcca ccccagctc 7020 ctgagcgaga cccaggtcat cggtaccttc gagctgcaag gtccgcagga gtccaccgct 7080 ccgctgaaac tcacgccggg gttgtggact tccgcgtacc tgcgcaatt tgtacccgaa 7140 gactaccacg cccatgagat aaagttcttt gaggaccaat cgcgcccgca gcacgcggat 7200 ctcacggcct gcgtcatcac ccaggggcg atcctcgccc aattgcacgc catccaaaaa 7260 tcccgccaag agttctct gaaaaagggt agaggggtct acctggaccc ccagacggggc 7320 gaggtgctca acccggggtct cccccagcat gccgaggaag aagcaggagc cgctagtgga 7380 ggagatggaa gagaatggg acagccaggc agaggagg gatgggagg aggahaga 7440 ggaggaagaa tggaaggg tggagagca gcccgtcgcc gcaccatccg cgccggcagc 7500 cccgccggtc accggatacaa cctccgctcc ggtcaagcct cctcgtagat gggatcaaat gaagggtgac ggtaagcacg agcggcaggg ctaccgatca tggagggccc acaaagccgc 7620. gatcatcgcc tgcttgcaag actgcgggggg gacatcgct tttgcccgcc gctacctgct cttccaccgc ggggtgaaca tcccccgcaa cgtgttgcat tactaccgtc accttcacag 7740 ctaagaaaaa gcaagtaaga ggagtcgccg gaggagg aggcctgagg atcgcggcga 7800 acgagccctt gaccaccagg gagctgagga accggatctt ccccactctt tatgccattt ttcagcagag tcgaggtcag cagcaagagc tcaaagtaaa aaaccggtct ctgcgctcgc tcacccgcag ttgcttgtac cacaaaaaacg aagatcagct gcagcgcact ctcgaagacg ccgaggctct gttccacaag tactgcgcgc tcactcttaa agactaaggc gcgcccaccc ggaaaaaagg cgggaattac ctcatcgcca ccatgagcca ggagattccc accccttaca tgtggagcta tcagccccag atgggcctgg ccgcggggcgc ctcccaggac tactccaccc 8160. 8220. gcatgaactg gctcagtgcc ggcccctcga tgatctcaca ggtcaacggg gtccgtaacc atcgaaacca gatattgttg gagcaggcgg cggtcacctc cacgcccagg gcaaagctca 8280 acccgcgtaa ttggccctcc accctggtgt atcaggaaat ccccgggccg actaccgtac 8340 tacttccgcg tgacgcactg gccgaagtcc gcatgactaa ctcaggtgtc cagctggccg 8400 gcggcgcttc ccggtgcccg ctccgcccac aatcgggtat aaaaaccctg gtgatccgag 8460 gcagaggcac acagcttaac gacgagttgg tgagctcttc aatcggtctg cgaccggacg 8520 gagtgttcca actagccgga gccgggagat cctccttcac tccccaccag gcctacctga 8580 ccttgcagag cagctcttcg gagcctcgct ccggaggcat cggaaccctc cagttcgtgg 8640 aggagtttgt gccctcggtc tacttcaacc ccttctcggg atcgccaggc ctctacccgg 8700 acgagttcat accgaacttc gatgcagtga gagaagcggt ggacggctac gactgaatgt 8760 cccatggtga ctcggctgag ctcgctcggt tgaggcatct ggaccactgc cgccgcctgc 8820 gctgcttcgc ccgggagagc tgcggactca tctactttga gtttcccgag gagcacccca 8880 acggccctgc acacggagtg cggatcaccg tagagggcac caccgagtct cacctggtca 8940 ggttcttcac ccagcaaccc ttcctggtcg agcgggaccg gggcgccacc acctacaccg 9000 tctactgcat ctgtcctacc ccgaagttgc atgagaattt ttgttgtact ctgtgtgctg 9060 agtttaataa aagctaaact cctacaatac tctgggatcc cgtgtcgtcg cactcgcaac 9120 gagatcttca acctcaccaa ccagactgag gtaaaactca actgcagacc ggggggcaaa 9180 tacatcctct ggctctttga aaacacttcc ttcgcagtct ccaacgcctg ctgcactagt 9240 ccatgaactg attgattaaa gcccaaaaaa ccaatcaaac cccttcccat tagccccaaa 9300 taaacaatca ttggaaataa tcattcaata aagatcactt acttgaaatc tgaaagtatg 9360 tctctggtgt agttgtttag cagcacctcg gtaccctcct cccagctctg gtactccagt 9420 ccccggcggg cggcgaactt tctccacacc ttgaaaggga tgtcaaattc ctggtccaca 9480 attttcattg ttttccctct cagatgtcaa agaggcttcg ggtggaagat gacttcaacc 9540 ccgtctaccc ctatgactac gcgcggaatc agaatatccc cttcctcact cccccctttg 9600 tctcctccga tggattccaa aacttccccc ctggtgtcct gtcactcaaa ttggctgacc 9660 caatcgctat caccaatggt gatgtctcac tcaaggtggg agggggactt actttacaag atggaaccgg aaaactaact atcgacacca agactccctt gcaagttgca aataataat Tagaacttgc gtttgatgca ccattgtatg aaaaaaatgg aaaacttgct ttaaaaacag gccatggatt agctgtttta accaaagaca ttggcatacc agaattatt ggatcccttg tgatcttaac tggaaaagga attgggacag gtactgttgc aggagga actatagatg tagctggg tgatgatgga ggtttatcat ttgataaaaa gggtgatcta gtagcctgga ataaaaaaaa tgacaggcgc actttgtgga cacacctga tccatcccca aactgcagag 10140. 10140. 10140. 10140. 10140. 10140. 10140. 10140 tagcaagttt ttcactgctt gtagtcaaag ggacgtatgc aactgttgat aaaaatacaa ctaaaca atttagcatt aaactactgt ttgatgcaaa tggaagctt aaaagcgaat caaaccttag tggttattgg aactataga gtgataatag tgttgttag actccctatg acaatgcagt gcctttcatg ccaatacca cggcttatcc aaaaatcatt aatagtacaa ctgatcctga aaataagaaa agttcggcta aaaaaactat tgttggcaat gtctacctag 10440 aaggtaatgc gggtcaacca gtagccgttg ctattagttt aataaggaa actactgctg 10500 actattcaat aacatttgac tttgcgtgga gcaaagctta tgaaacccct gtgccttttg 10560 acacctcctc catgacattc tcatatattg cccaggaaaa tcaagaaaa ggcgaataaa 10620 gtgttttgaa atgaacttat gtatctttat tgatttttac accagcacgg gtagtcagtc 10680 tcccaccacc agcccatttc acagtgtaaa cagtcctttc tccccggctg gccttaaaaa 10740 gcatcatatc atgggtaaca gacatattct taggtgttat attccacacg gtttcctgtc 10800 gagccaaacg ctcatcagtg atattaataa actccccggg cagctcactt aagttcatgt 10860 cgctgtccag ctgctgagcc acaggctgct gtccaacttg cggttgctta acgggcggcg 10920 aaggaagt ccacgcctac atgggggtag agtcataatc gtgcatcagg atagggcggt 10980 ggtgctgcag cagcgcgcga ataaactgct gccgccgccg ctccgtcctg caggaataca 11040 acatggcagt ggtctcctca gcgatgattc gcaccgcccg cagcataagg cgccttgtcc 11100 tccgggcaca gcagcgcacc ctgatctcac ttaaatcagc acagtaactg cagcacagca 11160 ccacaatatt gttcaaaatc ccacagtgca aggcgctgta tccaaagctc atggcgggga 11220 ccacagaacc cacgtggcca tcataccaca agcgcaggta gattaagtgg cgacccctca 11280 taaacacgct ggacataaac attacctct ttggcatgtt gtaattcacc acctcccggt 11340 accatataaa cctctgatta aacatggcgc catccaccac catcctaaac cagctggcca 11400 aaacctgccc gccggctata cactgcaggg aaccgggact ggaacaatga cagtggagag 11460 cccaggactc gtaaccatgg atcatcatgc tcgtcatgat atcaatgttg gcacaacaca 11520 ggcacacgtg catacacttc ctcaggatta caagctcctc ccgcgttaga accatatccc 11580 agggaacaac ccattcctga atcagcgtaa atcccacact gcagggaaga cctcgcacgt 11640 aactcacgtt gtgcattgtc aaagtgttac attcgggcag cagcggatga tcctccagta 11700 tggtagcgcg ggtttctgtc tcaaaaggag gtagacgatc cctactgtac ggagtgcgcc 11760 gagacaaccg agatcgtgtt ggtcgtagtg tcatgccaaa tggaacgccg gacgtagtca 11820 tttcgctgct gccgctcagt catccaactg aagtacatcc attctcgaag attctggaga 11880 agttcctctg catctgatga aataaaaaac ccgtccatgc gaattcccct catcacatca 11940 gccaggactc tgtaggccat ccccatccag ttaatgctgc cttgtctatc attcagaggg 12000 ggcggtggca ggattggaag aaccattttt attccaaacg gtctcgaagg acgataaagt 12060 gcaagtcacg caggtgacag cgttcccctc cgctgtgctg gtggaaacag acagccaggt 12120 caaaacccac tctattttca aggtgctcga ccgtggcttc gagcagtggc tctacgcgta 12180 catccagcat aagaatcaca ttaaaggctg gccctccatc gatttcatca atcatcaggt 12240 tacattcctg caccatcccc aggtaattct catttttcca gccttggatt atctctacaa 12300 attgttggtg taagtccact ccgcacatgt ggaaaagctc ccacagtgcc ccctccactt 12360 12420 gttcaaaaca acaagattca ataaggttct gccctccgcc ctgagctcgc gcctcaatgt 12480 cagctgcaaa aagtcactta agtcctgggc cactacagct gacaattcag agccagggct 12540 aagcgtggga ctggcaagcg taagggaaaa cttaatgct ccgaagctag cacccaaaaa 12600 ctgcatgctg gaataagctc tctttgtgtc tccggtgatg ccttccaaaa tgtgagtgat 12660 aaagcgtggt agttttctt taatcatttg cgtaatagaa aagtcctgta aataagtcac 12720 taggacccca gggaccacaa tgtggtagct tacaccgcgt cgctgaagca tggttagtag 12780 agatgagagt ctgaaaaaca gaaagcatgc actaaactaa gttggctatt ttcactgaag 12840 gaaaaatcac tctctccagc aacagggtac ccactgggtg gcccttgcgg acatacaaaa 12900 atcggtccgt gtgattaaaa agcagcacag taagttcctg tcttcttccg gcaaaaatca 12960 catcggactg ggttagtatg tccctggcat ggtagtcatt caaggccata aatctgccct 13020 gatatccagt aggaaccagc acactcactt ttaggtgaag caataccacc ccatgcggag 13080 gaatgtggaa agattcaggg caaaaaaaat tatatctatt gctagtccct tcctggacgg 13140 gagcaatccc tccaggacta tctatgaaag catacagaga ttcagccata gctcagcccg 13200 cttaccagta gacagagagc acagcagtac aagcgccaac agcagcgact gactacccac 13260 tgacccagct ccctatttaa aggcgcctta cactgacgta atgaccaaag gtctaaaaac 13320 cccgccaaaa aaaaacacac acgccctggg tgtttttcgc gaaaacactt ccgcgttctc 13380 acttcctcgt atcgatttcg tgacttaact tccgggttcc cacgttacgt cacttctgcc 13440 cttacatgta actgagtcgt agggcgccat cttgcccacg tccaaaatgg cttccatgtc 13500 cagccacgcc tccgcggcga ccgttagccg tgcgtcgtga cgtcatttgc atcgtcttct 13560 ctcgtccaat cagcgtcggc tccgccctaa attcaaaagc tcatttgcat attaactttt 13620 gtttactttg tggggtatat tattgatgat gttaattaat tcgaacccat aatacccata 13680 atagctgttt gccatcgacg cgaggctgga tggccttccc cattatgatt cttctcgctt 13740 ccggcggcat cgggatgccc gcgttgcagg ccatgctgtc caggcaggta gatgacgacc 13800 atcagggaca gcttcaagga tcgctcgcgg ctcttaccag cccagcaaaa ggccaggaac 13860 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 13920 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 13980 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 14040 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat 14100 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 14160 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 14220 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 14280 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt 14340 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 14400 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 14460 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 14520 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 14580 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 14640 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 14700 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 14760 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 14820 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 14880 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 14940 cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct 15000 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 15060 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 15120 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 15180 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 15240 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa 15300 gtgctcatca ttggaaaacg ttcttcgggg cgaaaaactct caaggatctt accgctgttg 15360 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 15420 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 15480 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 15540 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taacaaata 15600 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc 15660 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca agaattggtc 15720 gatggcaaac agctattatg ggtattatgg gttcgaatta attaaactat gccagacgca 15780 agagccggcg gcgacggatc gccaggcgcc accggagcac gcccgccatg cgcgccgccc 15840 gggctctgct gcgccgcgcc agacgcacag gccgccgggc catgatgcga gccgcgcgcc 15900 gcgctgccac tgcacccacc cccgcaggca ggactcgcag acgagcggcc gccgccgctg 15960 ccgcggccat ctctagcatg accagaccca ggcgcggaaa cgtgtactgg gtgcgcgact 16020 ccgtcacggg cgtgcgcgtg cccgtgcgca cccgtcctcc tcgtccctga tctaatgctt 16080 gtgtcctccc ccgcaagcga cgatgtcaaa gcgcaaaatc aaggaggaga tgctccaggt 16140 cgtcgccccg gagatttacg gaccaaccca ggcggaccag aaaccccgca aaatcaagcg 16200 ggttaaaaaa areatgagg tggacgaggg ggcagtagag tttgtgcgcg agttcgctcc 16260 gcggcggcgc gtaaattgga aggggcgcag ggtgcagcgc gtgttgcggc ccggcacggc 16320 ggtggtgttc acgcctggcg agcggtcctc ggtcaggagc aagcgtagct atgacgaggt 16380 atacggcgac gacgacatcc tggaccaggc ggcggagcgg gcgggcgagt ttgcctacgg 16440 gaagcggtcg cgcgaagagg agctgatctc gctgccgctg gacgaaagca accccacgcc 16500 aagcctgaag cccgtgaccc tgcagcaggt gctgccccag gcggtgctgc tgccgagccg 16560 cggggtcaag cgcgagggcg agaacatgta cccgaccatg cagatcatgg tgcccaagcg 16620 ccggcgcgtg gaggacgtgc tggacaccgt gaaaatggat gtggagcccg aggtcaaggt 16680 gcgccccatc aagcaggtgg cgccgggcct gggcgtgcag accgtggaca ttcagatccc 16740 caccgacatg gatgtcgaca aaaaaccctc gaccagcatc gaggtgcaga ccgacccctg 16800 gctcccagcc tccactgcta ccgcctccac ttctaccgtc gccacggcca ccgagcctcc 16860 caggaggcga agatggggcc ctgccaaccg gctgatgccc aactacgtgt tgcatccttc 16920 catcatcccg acgccgggct accgcggcac tcggtactac gccagccgca ggcgcccagc 16980 cagcaaacgc cgccgccgca ccgccacccg ccgccgtctg gcccccgccc gcgtgcgccg 17040 cgtgaccacg cgccggggcc gctcgctcgt tctgcccacc gtgcgctacc accccagcat 17100 cctttaatcc gtgtgctgtg atactgttgc agagagatgg ctctcacttg ccgcctgcgc 17160 atccccgtcc cgaattaccg aggaagatcc cgccgcagga gaggcatggc aggcagtggc 17220 ctgaaccgcc gccggcggcg ggccatgcgc aggcgcctga gtggcggctt tctacccgcg 17280 ctcatcccca taatcgccgc ggccatcggc acgatcccgg gcatagcttc cgttgcgctg 17340 caggcgtcgc agcgccgttg atgtgcgaat aaagcctctt tagactctga cacacctggt 17400 cctgtatatt tttagaatgg aagacatcaa ttttgcgtcc ctggctccgc ggcacggcac 17460 gcggccgttc atgggcacct ggaacgagat cggcaccagc cagctgaacg ggggcgcctt 17520 caattggagc agtgtctgga gcgggcttaa aaatttcggc tcgacgctcc ggacctatgg 17580 gaacaaggcc tggaatagta gcacggggca gttgttaagg gaaaagctca aagaccagaa 17640 cttccagcag aaggtggtgg acgggctggc ctcgggcatt aacggggtgg tggacatcgc 17700 gaaccaggcc gtgcagcgcg agataaacag ccgcctggac ccgcgcccgc ccacggtggt 17760 ggagatggaa gatgcaactc ttccgccgcc caaaggcgag aagcggccgc ggcccgacgc 17820​​​​​​​​​​​​​​​​​attaacatca tcaataatat accccacaaa gtaaacaaaa gttaatatgc aaatgagctt 60 ttgaatttag ggcggagccg acgctgattg gacgagagaa gacgatgcaa atgacgtcac 120 gacgcacggc taacggtcgc cgcggaggcg tggcctagcc cggaagcaag tcgcggggct 180 gatgacgtat aaaaaagcgg actttagacc cggaaatggc cgattttccc gcggccacgg 240 ccggatatga ggtaattctg ggcggatgca agtgaaatta ggtcattttg gcgcgaaaac 300 tgaatgagga agtgaaaagc gaaaaatacc gatcccgccc agggcggaat atttaccgag 360 ggccgagaga ctttgaccga ttacgtgggg gtttcgattg cggtgttttt ttcgcgaatt 420 tccgcgtccg tgtcaaagtc cggtgtttat gtcacagatc agctgaccta ggtggtcaat 480 attggccatt agccatatta ttcattggtt atatagcata aatcaatatt ggctattggc 540 cattgcatac gttgtatcca tatcataata tgtacattta tattggctca tgtccaacat 600 taccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat 660 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg 720 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa 780 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact 840 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta 900 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt 960 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg 1020 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg 1080 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgcccc 1140 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt 1200 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac 1260 accgggaccg atccagcctc cgcggccggg aacggtgcat tggaagcttg gtacctctag 1320 gcttttgcaa aaagctttgc aaagatggat aaagttttcc ggaattccgc caaaaagaag 1380 agaaaggtag aagaccccaa ggactttcct tcagaattgc taagtttttt gagtccaagc 1440 ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt 1500 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc 1560 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg 1620 gcaccagaag cggtgccgga aagctggctg gagtgcgatc ttcctgaggc cgatactgtc 1680 gtcgtcccct caaactggca gatgcacggt tacgatgcgc ccatctacac caacgtaacc 1740 tatcccatta cggtcaatcc gccgtttgtt cccacggaga atccgacggg ttgttactcg 1800 ctcacattta atgttgatga aagctggcta caggaaggcc agacgcgaat tatttttgat 1860 ggcgttaact cggcgtttca tctgtggtgc aacgggcgct gggtcggtta cggccaggac 1920 agtcgtttgc cgtctgaatt tgacctgagc gcatttttac gcgccggaga aaaccgcctc 1980 gcggtgatgg tgctgcgttg gagtgacggc agttatctgg aagatcagga tatgtggcgg 2040 atgagcggca ttttccgtga cgtctcgttg ctgcataaac cgactacaca aatcagcgat 2100 ttccatgttg ccactcgctt taatgatgat ttcagccgcg ctgtactgga ggctgaagtt 2160 cagatgtgcg gcgagttgcg tgactaccta cgggtaacag tttctttatg gcagggtgaa 2220 acgcaggtcg ccagcggcac cgcgcctttc ggcggtgaaa ttatcgatga gcgtggtggt 2280 tatgccgatc gcgtcacact acgtctgaac gtcgaaaacc cgaaactgtg gagcgccgaa 2340 atcccgaatc tctatcgtgc ggtggttgaa ctgcacaccg ccgacggcac gctgattgaa 2400 gcagaagcct gcgatgtcgg tttccgcgag gtgcggattg aaaatggtct gctgctgctg 2460 aacggcaagc cgttgctgat tcgaggcgtt aaccgtcacg agcatcatcc tctgcatggt 2520 caggtcatgg atgagcagac gatggtgcag gatatcctgc tgatgaagca gaacaacttt 2580 aacgccgtgc gctgttcgca ttatccgaac catccgctgt ggtacacgct gtgcgaccgc 2640 tacggcctgt atgtggtgga tgaagccaat attgaaaccc acggcatggt gccaatgaat 2700 cgtctgaccg atgatccgcg ctggctaccg gcgatgagcg aacgcgtaac gcgaatggtg 2760 cagcgcgatc gtaatcaccc gagtgtgatc atctggtcgc tggggaatga atcaggccac 2820 ggcgctaatc acgacgcgct gtatcgctgg atcaaatctg tcgatccttc ccgcccggtg 2880 cagtatgaag gcggcggagc cgacaccacg gccaccgata ttatttgccc gatgtacgcg 2940 cgcgtggatg aagaccagcc cttcccggct gtgccgaaat ggtccatcaa aaaatggctt 3000 tcgctacctg gagagacgcg cccgctgatc ctttgcgaat acgcccacgc gatgggtaac 3060 agtcttggcg gtttcgctaa atactggcag gcgtttcgtc agtatccccg tttacagggc 3120 ggcttcgtct gggactgggt ggatcagtcg ctgattaaat atgatgaaaa cggcaacccg 3180 tggtcggctt acggcggtga ttttggcgat acgccgaacg atcgccagtt ctgtatgaac 3240 ggtctggtct ttgccgaccg cacgccgcat ccagcgctga cggaagcaaa acaccagcag 3300 cagtttttcc agttccgttt atccgggcaa accatcgaag tgaccagcga atacctgttc 3360 cgtcatagcg ataacgagct cctgcactgg atggtggcgc tggatggtaa gccgctggca 3420 agcggtgaag tgcctctgga tgtcgctcca caaggtaaac agttgattga actgcctgaa 3480 ctaccgcagc cggagagcgc cgggcaactc tggctcacag tacgcgtagt gcaaccgaac 3540 gcgaccgcat ggtcagaagc cgggcacatc agcgcctggc agcagtggcg tctggcggaa 3600 aacctcagtg tgacgctccc cgccgcgtcc cacgccatcc cgcatctgac caccagcgaa 3660 atggattttt gcatcgagct gggtaataag cgttggcaat ttaaccgcca gtcaggcttt 3720 ctttcacaga tgtggattgg cgataaaaaa caactgctga cgccgctgcg cgatcagttc 3780 acccgtgcac cgctggataa cgacattggc gtaagtgaag cgacccgcat tgaccctaac 3840 gcctgggtcg aacgctggaa ggcggcgggc cattaccagg ccgaagcagc gttgttgcag 3900 tgcacggcag atacacttgc tgatgcggtg ctgattacga ccgctcacgc gtggcagcat 3960 caggggaaaa ccttatttat cagccggaaa acctaccgga ttgatggtag tggtcaaatg 4020 gcgattaccg ttgatgttga agtggcgagc gatacaccgc atccggcgcg gattggcctg 4080 aactgccagc tggcgcaggt agcagagcgg gtaaactggc tcggattagg gccgcaagaa 4140 aactatcccg accgccttac tgccgcctgt tttgaccgct gggatctgcc attgtcagac 4200 atgtataccc cgtacgtctt cccgagcgaa aacggtctgc gctgcgggac gcgcgaattg 4260 aattatggcc cacaccagtg gcgcggcgac ttccagttca acatcagccg ctacagtcaa 4320 cagcaactga tggaaaccag ccatcgccat ctgctgcacg cggaagaagg cacatggctg 4380 aatatcgacg gtttccatat ggggattggt ggcgacgact cctggagccc gtcagtatcg 4440 gcggaattcc agctgagcgc cggtcgctac cattaccagt tggtctggtg tcaaaaataa 4500 taataaccgg gcagggggga ttctgaactt gtttatggat cctctagacg agatccgaac 4560 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 4620 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 4680 catgtctcga tccaggtagg tttgagtagt gggcgtggct aagatgacta taaaggtggg 4740 tgtcttacga gggtcttttt gcttttctgc agacatcatg aacgggaccg gcggggcctt 4800 cgaagggggg ctttttagcc cttatttgac aacccgcctg ccgggatggg ccggagttcg 4860 tcagaatgtg atgggatcga cggtggacgg gcgtccagtg cttccagcaa attcctcgac 4920 catgacctac gcgaccgtgg ggaactcgtc gctcgacagc accgccgcag ccgcggcagc 4980 cgcagccgcc atgacagcga cgagactggc ttcgagctac atgcccagca gcggtagcag 5040 cccctctgtg cccagttcca tcatcgccga ggagaaactg ctggccctcc tggccgagct ggagccctg agccgccagc tggccgccct gtgtccgagc tccgcgaaca 5160 gcagcagcag caaaataat gattcaata acacagattc tgattcaaac agcaaagcat ctttattatt tattttttcg cgcgcggtag gccctggtcc acctctcccg atcattgaga 5280 gtgcggtgga ttttttccag gacccggtag agatgggatt ggatgttgag gtacatgggc 5340 atgagcccgt cccggggggtg aaggtagcac cactgcatgg cctcatgctc tggggtcgtg 5400. ttgtagatga tccagtcata gcaggggcgc tggggcgtggt gctggatgat gtctttgagg 5460 aggagactga tggccacgggg gagccccttg gtgtaggtgt tggcgaagcg gttgagctgg 5520 gagggatgca tgcgggggga gatgatgtgc agtttggcct ggatcttgag gttggcaatg 5580 ttgccgccca gatcccgccg ggggttcatg ttgtgcagga ccaccaggac ggtgtagccc 5640. gtgcacttgg ggaacttatc atgcaacttg gaagggaatg cgtggaagaa tttggagacg cccttgtgcc cgcccaggtt ttccatgcac tcatccatga tgatggcgat gggcccgtgg 5760. gctgcggctt tggcaaagac gtttctgggg tcagagacat cataattatg ctcctgggtg agatcatcat aagacatttt aatgaatttg gggcggaggg tgccagattg ggggacgatc gttccctcgg gccccggggc gaagttcccc tcacagatct gcatctccca ggctttcatc 5940 tcggaggggg ggatcatgtc cacctgcggg gcgatgaaaa aaacggtttc cggggcgggg 6000 gtgatgagct gcgaggagag caggtttctc aacagctggg acttgccgca cccggtcggg 6060 ccgtagatga ccccgatgac gggttgcagg tggtagttca aggagatgca gctgccgtct tcccggagga ggggggccac ctcgttgagc atgtctctga cttggaggtt ttcccggacg 6180 agctcgccga ggaggcggtc cccgcccagc gagagcagct cttgcaggga agcaaagttt 6240 ttcaggggct tgagcccgtc ggccatgggc atcttggcga gggtctgcga gaggagttcc 6300. aagcggtccc agagctcggt gacgtgctct acggcatctc gatccaacag acttcctcgt 6360 ttcggggggtt gggacgactg cgactgtagg gcacgagacg atgggcgtcc agcgctgcca 6420. gcgtcatgtc cttccagggt ctcagggtcc gcgtgagcgt ggtctccgtc acggtgaagg 6480 ggtgggcccc gggctgggcg cttgcaaggg tgcgcttgag actcatcctg ctggtgctga 6540 aacgggcacg gtcttcgccc tgcgcgtcgg cgagatagca gttgaccatg agctcgtagt 6600 tgagggcctc ggcggcgtgg cccttggcgc ggagcttgcc cttggaagag cgcccgcagg 6660 cgggacagag gagggattgc agggcgtaga gcttgggcgc gagaaagacg gactcggggg 6720 cgaaagcgtc cgctccgcag tgggcgcaga cggtctcgca ctcgacgagc caggtgagct 6780 cgggatgttc ggggtcaaaa accagttttc ccccgttctt tttgatgcgc ttcttacctc 6840 gcgtctccat gagtctgtgt ccgcgctcgg tgacaaacag gctgtctgtg tccccgtaga 6900 cggacttgat gggcctgtcc tgcaggggcg tcccgcggtc ctcctcgtag agaaactcgg 6960 accactctga gacgaaggcg cgcgtccacg ccaagacaaa ggaggccacg tgcgaggggt 7020 agcggtcgtt gtccaccagg gggtccacct tttccaccgt gtgcagacac atgtccccct 7080 cctcagcatc caagaaggtg attggcttgt aggtgtaggc cacgtgaccg ggggtccccg 7140 acgggggggt ataaaagggg gcgggtctgt gctcgtcctc actctcttcc gcgtcgctgt 7200 ccacgagcgc cagctgttgg ggtaggtatt ccctctcgag agcgggcatg acctcggcac 7260 tcaggttgtc agtttctaga aacgaggagg atttgatgtt ggcctgccct gccgcaatgc 7320 ttttgagtaa actctcatcc atctggtcag aaaagactat ttttttattg tcaagcttgg 7380 tggcgaagga gccatagagg gcgttggaga gaagcttggc gatggatctc atggtctgat 7440 ttttgtcacg gtcggcgcgc tccttggccg cgatgtttag ctggacatat tcgcgcgcga 7500 cacacttcca ttcggggaag acggtggtgc gctcgtcggg cacgatcctg acgcgccagc 7560 cgcggttatg cagggtgacc aggtccacgc tggtggccac ctcgccgcgc aggggctcgt 7620 tggtccagca gagtcgcccg cctttgcgcg aacagaaagg gggcagcaca tcaagcagat 7680 gctcgtcggg ggggtccgca tcgatggtga agataccggg acagagttcc ttgtcaaaat 7740 agtctatttt tgaggatgca tcatccaagg ccatctgcca ctcacgggcg gccattgctc 7800 gctcgtaggg gttgaggggc ggaccccagg gcatgggatg cgtgagggcg gaggcgtaca 7860 tgccgcagat gtcatagaca tagatgggct ccgagaggat gccgatgtag gtgggataac 7920 agcgcccccc gcggatgctg gcgcgcacgt agtcatacaa ctcgtgcgag ggggccaaga 7980 aggcggggcc gagattggtg cgctggggct gctcggcgcg gaagacgatc tggcgaaaga 8040 tggcgtgcga gttggaggag atggtgggcc gttggaagat gttaaagtgg gcgtggggca 8100 agcggaccga gtcgcggatg aagtgcgcgt aggagtcttg cagcttggcg acgagctcgg 8160 cggtgacgag gacgtccatg gcgcagtagt ccagcgtttc gcggatgatg tcataacccg 8220 tctctccttt cttctcccac agctcgcggt tgagggcgta ctcctcgtca tccttccagt 8280 actcccggag cgggaatcct cgatcgtccg cacggtaaga gcccagcatg tagaaatggt 8340 tcacggcctt gtagggacag cagcccttct ccacggggag ggcgtaagct tgagcggcct 8400 tgcggagcga ggtgtgcgtc agggcgaagg tgtccctgac catgactttc aagaactggt 8460 acttgaaatc cgagtcgtcg cagccgccgt gctcccagag ctcgaaatcg gtgcgcttct 8520 tcgagagggg gttaggcaga gcgaaagtga cgtcattgaa gagaatcttg cctgcccgcg 8580 gcatgaaatt gcgggtgatg cggaaagggc ccgggacgga ggctcggttg ttgatgacct 8640 gggcggcgag gacgatctcg tcgaagccgt tgatgttgtg cccgacgatg tagagttcca 8700 tgaatcgcgg gcggcctttg atgtgcggca gctttttgag ctcctcgtag gtgaggtcct 8760 cggggcattg caggccgtgc tgctctagcg cccactcctg gagatgtggg ttggcttgca 8820 tgaaggaagc ccagagctcg cgggccatga gggtctggag ctcgtcacga aagaggcgga 8880 actgctggcc cacggccatc ttttcgggtg tgacgcagta gaaggtgagg gggtcccgct 8940 cccagcgatc ccagcgtaag cgcacggcga gatcgcgagc gagggcgacc agctctgggt 9000 cccccgagaa tttcatgacc agcatgaagg ggacgagctg cttgccgaag gaccccatcc 9060 aggtgtaggt ttctacatcg taggtgacaa agagccgctc cgtgcgagga tgagagccga 9120 ttgggaagaa ctggatttcc tgccaccagt tggacgagtg gctggtgatg tgatgaaagt 9180 agaaatcccg ccggcgaacc gagcactcgt gctgatgctt gtaaaagcgt ccgcagtact 9240 cgcagcgctg cacgggctgt acctcatcca cgagatacac agcgcgtccc ttgaggagga 9300 acttcaggag tggcggccct ggctggtggt tttcatgttc gcctgcgtgg gactcaccct 9360 ggggctcctc gaggacggag aggctgacga gcccgcgtgg gagccaggtc cagatctcgg 9420 cgcggcgggg gcggagagcg aagacgaggg cgcgcagttg ggagctgtcc atggtgtcgc 9480 ggagatccag gtccgggggc agggttctga ggttgacctc gtagaggcgg gtgagggcgt 9540 gcttgagatg cagatggtac ttgatttcta cgggtgagtt ggtggccgtg tccacgcatt 9600 gcatgagccc gtagctgcgc ggggccacga ccgtgccgcg gtgcgctttt agaagcggtg 9660 tcgcggacgc gctcccggcg gcagcggcgg ttccggtccc gcgggcaggg gcggcagagg 9720 cacgtcggcg tggcgctcgg gcaggtcccg gtgctgcgcc ctgagagcgc tggcgtgcgc 9780 gacgacgcgg cggttgacat cctggatctg ccgcctctgc gtgaagacca cgggccccgt 9840 gactttgaac ctgaaagaca gttcaacaga atcaatctcg gcgtcattga cggcggcctg 9900 acgcaggatc tcttgcacgt cgcccgagtt gtcctggtag gcgatctcgg acatgaactg 9960 ctcgatctcc tcctcctgga gatcgccgcg gcccgcgcgc tcgacggtgg cggcgaggtc 10020 attcgagatg cgacccatga gctgcgagaa ggcgcccagg ccgctctcgt tccagacgcg 10080 gctgtagacc acgtccccgt cggcgtcgcg cgcgcgcatg accacctgcg cgaggttgag 10140 ctccacgtgc cgcgcgaaga cggcgtagtt gcgcaggcgc tggaagaggt agttgagggt 10200 ggtggcgatg tgctcggtga cgaagaagta catgatccag cggcgcagag gcatctcgct 10260 gatgtcgccg atggcctcca gcctttccat ggcctcgtag aaatccacag cgaagttgaa 10320 aaactgggcg ttgcgggccg agaccgtgag ctcgtcttcc aggagcctga tgagctcggc 10380 gatggtggcg cgcacctcgc gctcgaaatc cccgggggcc tcctcctctt cctcttcttc 10440 catgacgacc tcttcttcta tttcttcctc tgggggcggt ggtggtggcg gggcccgacg 10500 acgacggcga cgcaccggga gacggtcgac gaagcgctcg atcatctccc cgcggcggcg 10560 acgcatggtt tcggtgacgg cgcgaccccg ttcgcgagga cgcagcgtga agacgccgcc 10620 ggtcatctcc cggtaatggg gcgggtcccc gttgggcagc gatagtgcgc tgacgatgca 10680 tcttatcaat tgcggtgtag gggacgtgag cgcgtcgaga tcgaccggat cggagaatct 10740 ttcgaggaaa gcgtctagcc aatcgcagtc gcaaggtaag ctcaaacacg tagcagccct 10800 gtggacgctg ttagaattgc ggttgctgat gatgtaattg aagtaggcgt ttttgaggcg 10860 gcggatggtg gcgaggagga ccaggtcctt gggtcccgct tgctggatgc ggagccgctc 10920 ggccatgccc caggcctggc cctgacaccg gctcaggttc ttgtagtagt catgcatgag 10980 cctctcgatg tcatcactgg cggaggcgga gtcttccatg cgggtgaccc caacgcccct 11040 gagcggctgc acgagcgcca ggtcggcgac gacgcgctcg gcgaggatgg cctgttgcac 11100 gcgggtgagg gtgtcctgga agtcgtccat gtcgacgaag cggtggtagg cccctgtgtt 11160 gatggtgtaa gtgcagttgg ccatgagcga ccagttgacg gtctgcaggc cgggctgcac 11220 gacctcggag tacctgagcc gcgagaaggc gcgcgagtcg aagacgtagt cgttgcaggt 11280 gcgcacgagg tactggtatc cgactaggaa gtgcggcggc ggctggcggt agagcggcca 11340 gcgctgggtg gccggcgcgc ccggggccag gtcctcgagc atgaggcggt ggtagccgta 11400 gaggtagcgg gacatccagg tgatgccggc ggcggtggtg gaggcgcgcg ggaactcgcg 11460 gacgcggttc cagatgttgc gcagcggcag gaaatagtcc atggtcggca cggtctggcc 11520 ggtgagacgc gcgcagtcat tgacgctcta gaggcaaaaa cgaaagcggt tgagcgggct 11580 attcctccgt agcctggcgg aacgcaaacg ggttaggccg cgtgtgtacc ccggttcgag 11640 tcccctcgaa tcaggctgga gccgcgacta acgtggtatt ggcactcccg tctcgacccg 11700 agcccgatag ctgccaggat acggcggaga gcccttttgg cggccgaggg gggtcgctag 11760 acttgaaagc ggccgaaaac cctgccgggt agtggctcgc gcccgtagtc tggagaagca 11820 tcgccagggt tgagtcgcgg cagaacccgg ttcgcggacg gccgcggaga gcgggacttg 11880 gtcaccccgc cgatttaaaa acccacagcc agccgacttc tccagttacg ggagcgagcc 11940 cccttttttc tttttgccag atgcatcccg tcctgcgcca aatgcgtccc accccccctc 12000 cggcgaccac cgcaaccgcg gccgtagcag gcgccggcgc tagccagcca cagccacaga 12060 cagagatgga cttggaagag ggcgaagggc tggcgagatt gggggcgccg tctccggagc 12120 gacacccccg cgtgcagctg cagaaggacg tgcgcccggc gtacgtgcct gcgcagaacc 12180 tgttcaggga ccgcagcggg gaggagccg aggagatgcg cgactgccgc tttcgggcgg 12240 gcagggagct gcgcgagggt ctggaccgcc agcgcgtgct gcgcgatgag gatttcgagc 12300 cgaacgagca gacggggatc agccccgcgc gcgcgcacgt ggcggcggcc aacctggtga 12360. cggcctacga gcagacggtg aagcaggagc gcaacttcca aaagagtttc aacaaccacg tgcgcacgct gatcgcgcgc gaggaggtgg ccctgggcct gatgcacctg tgggacctgg 12480 cggaggccat cgtgcagac ccggacagca agcctctgac ggcgcagctg ttcctggtgg 12540 tgcagcacag tagggacaac gaggcgttca gggaggcgct gctgaacatc gccgagcccg agggtcgctg gctgctggag ctgatcaaca tcttgcagag catcgtagtg caggagcgca gcctgagcct ggccgagaaa gtggcggcga tcaactactc ggtgctgagc ctgggcaagt tttacgcgcg caagatttac aagacgccat acgtgcccat agacaaggag gtgaagatag acagctttta catgcgcatg gcgctcaagg tgctgacgct gagcgacgac ctgggcgtgt accgcaacga ccgcatccac aaggccgtga gcgcgagccg gcggcgcgag ctgagcgacc 12900 gcgagctgat gctgagcctg cgccgggcgc tggtaggggg cgccgccggc ggcgaggagt 12960 cctacttcga catgggggcg gacctgcatt ggcagccgag ccggcgcgcc ttggaagccg 13020 cctacggtcc agaggacttg gatgaggatg aggaagagga ggaggatgca cccgctgcgg 13080 ggtactgacg cctccgtgat gtgtttttag atgtcccagc aagccccgga ccccgccata 13140 agggcggcgc tgcaaagcca gccgtccggt ctagcatcgg acgactggga ggccgcgatg 13200 caacgcatca tggccctgac gacccgcaac cccgagtcct ttagacaaca gccgcaggcc 13260 aacagacttt cggccattct ggaggcggtg gtcccctctc gaaccaaccc cacgcacgag 13320 aaggtgctgg cgatcgtgaa cgcgctggcg gagaacaagg ccatccgtcc cgacgaggcc 13380 ggtctggtgt acaacgccct gctggagcgc gtgggccgct acaacagcac aaacgtgcag 13440 tccaacctgg accggctggt gacggacgtg cgcgaggccg tggcgcagcg cgagcggttc 13500 aagaacgaag gcctgggctc gctggtggcg ctgaacgcct tcctggcgac gcagccggcg 13560 aacgtgccgc gcgggcagga cgattatacc aactttataa gcgcgctgcg gctgatggtg 13620 accgaggtgc cccagagcga ggtgtaccag tcgggcccgg attacttttt ccaaactagc 13680 agacagggcc tgcagacggt gaacctgagc caggccttta agaacctgcg cgggctgtgg 13740 ggcgtgcagg cgcccgtggg cgaccggtcg acggtgagca gcttgctgac gcccaactcg 13800 cggctgctgc tgctgctgat cgcgcccttc accgacagcg gcagcgtgaa cgcaactcg 13860 tacctgggcc acctgctgac gctgtaccgc gaggccatag gccaggcaca ggtggacgag 13920 cagaccttcc aggagatcac tagcgtgagc cgcgcgctgg ggcagaacga caccgacagt 13980 cttagggcca ccctgaactt cttgctgacc atagacagc agaagatccc ggcacagtac 14040 gcgctatcgg ccgaggagga gcgcatcctg agatatgtgc agcagagcgt agggctgttc 14100 ctgatgcagg agggggccac ccccagcgcc gcgctggaca tgaccgcgcg caacatggaa 14160 cctagcatgt acgctgccaa ccggccgttc atcaataagc tgatggacta cctgcaccgc 14220 gcggcggcca tgaactcgga ctactttaca aacgccatcc tgaacccgca ctggctcccg 14280 ccgccggggt tctacacggg cgagtacgat atgcccgacc ccaacgacgg gttcctgtgg 14340 gacgacgtgg acagcgcggt gttctccccg accttgcaaa agcgccagga ggcggtgcgc 14400 acgcccgcga gcgagggtgc ggtgggtcgg agcccctttc ctagcttagg gagtttgcat 14460 agcttgccgg gctcggtgaa cagcggcagg gtgagccgac cgcgcttgct gggcgaggac 14520 gagtacctga acgactcgct gctgcagccg ccgcgggtca agaacgccat ggccaataac 14580 gggatagaga gtctggtgga caaactgaac cgctggaaga cctacgctca ggaccatagg 14640 gacgcgcccg cgccgcggcg acagcgccac gaccggcagc ggggcctggt gtgggacgac 14700 gaggactcgg ccgacgatag cagcgtgttg gacttgggcg ggagcggtgg ggccaacccg 14760 ttcgcgcatc tgcagcccaa actggggcga cggatgtttt gaaatgcaaa ataaaactca 14820 ccaaggccat agcgtgcgtt ctcttccttg ttagagatga ggcgtgcggt ggtgtcttcc 14880 tctcctcctc cctcgtacga gagcgtgatg gcgcaggcga ccctggaggt tccgtttgtg 14940 cctccgcggt atatggctcc tacggagggc agaaacagca ttcgttattc ggagctggct 15000 ccgctgtacg acaccactcg cgtgtacttg gtggacaaca agtcggcgga catcgcttcc 15060 ctgaactacc aaaacgacca cagcaacttc ctgaccacgg tggtgcagaa caacgatttc 15120 acccccgccg aggccagcac gcagacgata aattttgacg agcggtcgcg gtggggcggt 15180 gatctgaaga ccattctgca caccaacatg cccaatgtga acgagtacat gttcaccagc 15240 aagtttaagg cgcgggtgat ggtggctaga aagcacccag agaatgtagc taaagacgat 15300 ttgagtcagg ataagcttga atatgagtgg tttgagttta ccctgcccga gggcaacttt 15360 tccgagacca tgaccataga cctgatgaac aacgccatct tggaaaacta cttgcaagtg 15420 gggcggcaaa atggcgtgct ggagagcgat atcggagtca agtttgacag cagaaatttc 15480 aagctgggct gggacccggt gaccaagttg gtgatgccag gggtctacac ctacgaggcc 15540 ttccacccgg acgtggtgct gctgccgggc tgcggggtgg acttcaccga gagccgcctg 15600 agcaacctcc tgggcattcg caagcaa cctttccaag agggcttcag atcatgtat 15660 gagatctag taggggca catcccagcc ctcctgaatg tcaggagta tctgaaggat 15720 aaggaagaag ctggcacagc agatgcaat accattaagg ctcagaatga tgcagtccca 15780 agaggagata actatgcatc agcggcagaa gccaaagcag caggaaaga aattgagttg 15840 aaggccattt tgaagatga ttcaacaga agctacaatg tgatcgaggg aaccacagac 15900 accctgtacc gcagttggta cctgtcctat acctacgggg acccgagaa gggggtgcag 15960 tcgtggacgc tgctcaccac cccggacgtc acatgcggcg cggagcaagt ctactgtcg 16020 ctgccggacc tcatgcaaga ccccgtcacc ttccgctta cccagcaagt cagcaacac 16080 cccgtgtcg gcgccgagct catgccctc cgcgccaaga gctttacaa cgacctcgcc 16140 gtctactccc agctcatccg cagctacacc tccctcaccc acgtcttca ccgcttcccc 16200 gawaccaga tcctctgccg cccgcccgcg cccaccatca cccaccgtcag tgaaaacgtg 16260 cctgctctca cagatcacgg gacgctaccg ctgcgcagca gtatccgcgg agtccagcga 16320 gtgaccgtca ctgacgcccg tcgccgcacc tgtccctacg tctacaaggc cctgggcata 16380 gtcgcgccgc gcgtgctatc cagtcgcacc ttctaaaaaa tgtctattct catctcgccc 16440 agcaataaca ccggctgggg tattactagg cccagcacca tgtacggagg agccaagaag 16500 cgctcccagc agcaccccgt ccgcgtccgc ggccacttcc gcgctccctg gggcgcttac 16560 aagcgcgggc ggacttctac cgccgctgcc gtgcgtacca ccgtcgacga cgtcatcgac 16620 tcggtggtcg ccgacgcgcg caactacacc cccgccccct ccaccgtgga cgcggtcatt 16680 gacagcgtgg tggccgacgc gcgcgactat gccagacgca agagccggcg gcgacggatc 16740 gccaggcgcc accggagcac gcccgccatg cgcgccgccc gggctctgct gcgccgcgcc 16800 agacgcacag gccgccgggc catgatgcga gccgcgcgcc gcgctgccac tgcacccacc 16860 cccgcaggca ggactcgcag acgagcggcc gccgccgctg ccgcggccat ctctagcatg 16920 accagaccca ggcgcggaaa cgtgtactgg gtgcgcgact ccgtcacggg cgtgcgcgtg 16980 cccgtgcgca cccgtcctcc tcgtccctga tctaatgctt gtgtcctccc ccgcaagcga 17040 cgatgtcaaa gcgcaaaatc aaggaggaga tgctccaggt cgtcgccccg gagatttacg 17100 gaccaaccca ggcggaccag aaaccccgca aaatcaagcg ggttaaaaaa aaggatgagg 17160 tggacgaggg ggcagtagag tttgtgcgcg agttcgctcc gcggcggcgc gtaaattgga 17220 aggggcgcag ggtgcagcgc gtgttgcggc ccggcacggc ggtggtgttc acgcctggcg 17280 agcggtcctc ggtcaggagc aagcgtagct atgacgaggt atacggcgac gacgacatcc 17340 tggaccaggc ggcggagcgg gcgggcgagt ttgcctacgg gaagcggtcg cgcgaagagg 17400 agctgatctc gctgccgctg gacgaaagca accccacgcc aagcctgaag cccgtgaccc 17460 tgcagcaggt gctgccccag gcggtgctgc tgccgagccg cggggtcaag cgcgagggcg 17520 agaacatgta cccgaccatg cagatcatgg tgcccaagcg ccggcgcgtg gaggacgtgc 17580 tggacaccgt gaaaatggat gtggagcccg aggtcaaggt gcgccccatc aagcaggtgg 17640 cgccgggcct gggcgtgcag accgtggaca ttcagatccc caccgacatg gatgtcgaca 17700 aaaaaccctc gaccagcatc gaggtgcaga ccgacccctg gctcccagcc tccactgcta 17760 ccgcctccac ttctaccgtc gccacggcca ccgagcctcc caggaggcga agatggggcc 17820 ctgccaaccg gctgatgccc aactacgtgt tgcatcctt catcatcccg acgccgggct 17880 accgcggcac tcggtactac gccagccgca ggcgcccagc cagcaaacgc cgccgccgca 17940 ccgccacccg ccgccgtctg gccccgccc gcgtgcgccg cgtgaccacg cgccggggcc 18000 gctcgctcgt tctgcccacc gtgcgctacc accccagcat cctttaatcc gtgtgctgtg 18060 atactgttgc agagagatgg ctctcacttg ccgcctgcgc atccccgtcc cgaattaccg 18120 aggaagatcc cgccgcagga gaggcatggc aggcagtggc ctgaaccgcc gccggcggcg 18180 ggccatgcgc aggcgcctga gtggcggctt tctacccgcg ctcatcccca taatcgccgc 18240 ggccatcggc acgatcccgg ccatagcttc cgttgcgctg caggcgtcgc agcgccgttg 18300 atgtgcgaat aaagcctctt tagactctga cacacctggt cctgtatatt tttagaatgg 18360 aagacatcaa ttgcgtcc ctggctccgc ggcacggcac gcggccgttc atgggcacct 18420 ggaacgagat cggcaccagc cagctgaacg ggggcgcctt cattggagc agtgtctgga 18480 gcggcttaa aaatttcggc tcgacgctcc ggacctatgg gacaaggcc tggaatagta 18540 gcacggggca gttgttaagg gaaagctca agaccagaa cttccagcag aaggtggtgg 18600 acggggctggc ctcgggcatt aacggggtgg tggacatcgc gaaccaggcc gtgcagcgcg 18660 agataacag ccgcctggac ccgcgccccgc ccacggtggt ggagatggaa gatgcaactc 18720 ttccgccgcc aaggcgag aagcggccgc ggccgacgc ggaggagacg atcctgcagg 18780 tggacgagcc gccctcgtac gaggaggccg tgaggccgg catgcccacc acgcgcatca 18840 tcgcgccgct gccacgggt gtaatgaaac ccgccaccct tgacctgcct ccaccaccca 18900 cgccccgctcc accgaggca gctccggttg tgcagccccc tcctgtggcg accgccgtgc 18960 gccgcgtccc cgcccgccgc caggcccaga actggcagag cacgctgcac agtatcgtgg 19020 gcctgggagt gaaagtctg aagcgccgcc gatgctttg aggagagga aagaggacac 19080 taaagggaga gcttaacttg tatgtgcctt accgccagag aacgcgcgaa gatggccacc 19140 ccctcgatga tgccgcagtg ggcgtacatg cacatcgccg ggcaggacgc ctcggagtac 19200 ctgagcccgg gtctggtgca gtttgcccgc gccaccgaca cgtacttcag cctgggcaac 19260 aagtttagga accccacggt ggctcccacc cacgatgtga ccacggaccg gtcccagcgt 19320 ctgacgctgc gcttcgtgcc cgtggatcgc gaggacacca cgtactcgta caaggcgcgc 19380 ttcactctgg ccgtgggcga caaccgggtg ctagacatgg ccagcactta ctttgacatc 19440 cgcggcgtcc tggaccgcgg ccccagcttc aaaccctact cgggcacggc ctacaacagt 19500 ctggccccca agggcgcccc caactccagc cagtgggctg acaaagaaag ggtgaatgga 19560 ggcggtaaca ccaaagatgt tacaaagaca tttggtgtgg cagccatggg tggagaagac 19620 attacggaaa agggtctgaa gattggaact gatacaacag aaaatgaacc aatatttgca 19680 gacaaaaatt ttcagcctga accccaagtg ggagaggaaa actggcagga aacatttgtt 19740 ttctatggag gagagctct taaaaaagaa accaaatga agccttgcta tggttcattt 19800 gccagaccga ctaatgaaaa aggtggccag gctaaattta taattggaga taatggtcaa 19860 cccactgaaa accatgacat aacaatggct ttcttgaca ctcctggtgg tacaatcaca 19920 ggaggtactg gtggtcccca agatgaactt aaagctgaca ttgttactgta cactgaaaat 19980 aaatcttg aaactccaga tacccatgtg gtgtacaagc caggcaag agatgatagc 20040 tcagaaatta acttggtgca acagtccatg cccaacagac ccactacat cggtttcaga 20100 gandaactttg tgggacttat gtactacaac agcactggca acatgggcgt gctggctggt 20160 caggccctc agttgaatgc tgtggtcgac ttgcagaca gaacaccga gctgtcttac 20220 cagctcctgc tagattct gggtgacagg accagatact ttagcatgtg gaactctgcg 20280 gtggacagtt atgatcccga tgttaggatc attgagaatc acggtgtgga atgatgaactt 20340 cccaactatt gctttccatt ggacggctca ggcactatt cagctttca aggtgtaaaa 20400 attaagcaa accaagacgg agatgttaat gatgatggg aaaaggatga taggtacc 20460 actcaaaacc aaatttgcaa gggcaacatt tacgccatgg agatcaacct ccaggccaac 20520 ctgtggaaaa gttttctgta ctcgaacgtg gccctgtacc tgcccgactc ctacaagtac 20580 acgccggcca acgtgacgct gcccgccaac accaacacct acgagtacat gaacggccgc 20640 gtggtagccc cctcgctggt ggacgcctac atcaacatcg gcgcccgctg gtcgctagac 20700 cccatggaca atgtcaaccc cttcaaccac caccgcaatg cgggtctgcg ctaccgctcc 20760 atgcttctgg gcaacggccg ctacgtgccc ttccacatcc aagtgcccca aaagttcttt 20820 gccatcaaga acctgctcct gctcccgggc tcctacacct acgagtggaa cttccgcaag 20880 gacgtcaaca tgatcctgca gagttccctt ggaaacgacc tgcgcgttga cggcgcctcc 20940 gtccgcttcg acagtgttaa cctctatgcc accttcttcc ccatggcgca caacaccgcc 21000 tccaccctgg aagccatgct gcgcaacgac accaacgacc agtccttcaa cgactacctc 21060 tcggccgcca acatgctcta ccctatcccg gccaaggcca ccaacgtgcc catctccatc 21120 ccctcgcgca actgggccgc cttccgcggc tggagtttca cccggctcaa gaccaaggaa 21180 actccctccc ttggctcggg ttttgacccc tactttgtct actcgggctc catcccctac 21240 ctcgacggga ccttttacct gaaccatacc ttcaagaagg tctccatcat gttcgactcc 21300 tcggtcagct ggcccggcaa cgaccggctg ctcacgccga acgagttcga gatcaagcgc 21360 agcgttgacg gggagggcta caacgtggcc caatgcaaca tgaccaagga ctggttcctt 21420 gtccagatgc tctcccacta caacatcggc taccagggct tccacgtgcc cgagggctac 21480 aaggaccgca tgtactcatt cttccgcaac ttccagccca tgagcaggca ggtggtcgat 21540 gagatcaact acaaggacta caaagccgtc accctgccct tccagcacaa caactcgggc 21600 ttcaccggct atctcgcccc caccatgcgc caggggcagc cctaccccgc caacttccca 21660 tacccgctca tcggccagac agccgtaccc tccgtcaccc agaaaaagtt cctctgcgac 21720 agggtcatgt ggcgcatccc cttttccagc aacttcatgt ccatgggcgc cctcaccgac 21780 ctgggacaga acatgctcta tgccaactcg gcccacgcgc tcgacataac ttttgaggtg 21840 gaccccatgg atgagcccac cctcctctat cttctcttcg aagttttcga cgtggtcaga 21900 gtgcaccagc cgcaccgcgg cgtcatcgag gccgtctacc tgcgcacgcc cttctccgcc 21960 ggcaacgcta ctacataagc atgagcggct ccagcgaacg agagctcgcg gccatcgtgc 22020 gcgacctggg ctgcgggccc tactttttgg gaacccacga caagcgcttc cctggcttcc 22080 ttgccagcga caagctggca tgcgccatcg tcaacacggc cggccgcgag accgggggcg 22140 tgcactggct cgcctttggc tggaatccgc gctcgcgcac ctgctacatg tttgacccct 22200 ttgggttctc ggaccgccgg ctcaagcaga tttacagctt cgagtacgag gccatgctgc 22260 gccgcagcgc cctggcctcc tcgcccgacc gctgtctcag cctcgagcag tccacccaga 22320 ccgtgcaggg gcccgactcc gccgcctgcg gacttttttg ttgcatgttt ttgcatgcct 22380 ttgtgcactg gcccgaccga cccatggacg gaaaccccac catgaacttg ctgacggggg 22440 tgcccaacgg catgctacaa tcgccacagg tgctccccac cctcaggcgc aaccaggagg 22500 agctctaccg cttcctcgcg cgccactccc cttactttcg atcccaccgc gccgccattg 22560 aaaacgccac cgcttttgat aaaatgaaac aactgcgtgt atctcaataa acagcacttt 22620 attttacatg cactggagta tatgcaagtt atttaaaagt cgaaggggtt ctcgcgctcg 22680 tcgttgtgcg ccgcgctggg gagggccacg ttgcggtact gaaacttggg ctgccacttg 22740 aactcgggga tcaccagttt gggcactggg gtctcgggga aggtctcgct ccacatgcgc 22800 cggctcatct gcagggcgcc cagtatgtcg ggcgcggaga tcttgaaatc gcagttggga 22860 ccagtgctct gcgcgcgcga gttgcggtac acggggttgc agcactggaa caccatcaaa 22920 ctggggtgct tcacgctggc cagcacgctc ttgtcgctga tctgatcctt gtccaggtcc 22980 tcggcgttgc tcaggccgaa cggggtcatc ttgcacagct ggcggcccag gaagggcacg 23040 ctgtgaggct tgtggttaca ctcgcagtgc acgggcatca gcatcatccc cgcgccgcgc 23100 tgcatattcg ggtagagggc cttgacaaag gccaagatct gcttgaaagc ttgctgggcc 23160 ttggccccct cgctgaagaa caggccgcag ctcttcccgc tgaactggtt attcccacac 23220 ccggcatcat gcacgcagca gcgcgcgtca tggctggtca gttgcaccac gcttcggccc 23280 cagcggttct gggtcacctt ggccttgctg ggctgctcct tcaacgcgcg ctggccgttc 23340 tcgctggtca catccatctc caccacgtgg tccttgtgga tcatcaccgt cccgtgcaga 23400 cacttgagct ggccctcgac atcggtgcag ccgtgttccc acagggcgca gccggtgcac 23460 tcccaattct tgtgcgcgat cccgctgtgg ctgaagatgt aaccttgcaa catgcggccc 23520 atcacggtgc taaatgattt actggtgctg aaggtcagtt gcaggccgcg ggcctcctcg 23580 ttcatccatg tctgacacat tttctggaag atctcggtct gctcgggcat gagcttgtag 23640 gcatcgcgca ggccgctgtc gacgcggtag cgttccatca gcacgttcat ggtatccatg 23700 cccttctccc aggacgagac cagaggcaga ctcagggggt tgcgcacgtt caggataccg 23760 ggggtcgcgg gctcgacgat gcgttttccg tcctttcctt ccttcaacag aaccggcggc 23820 tggctgaatc ccactcccac gatcacggcg tcttcctggg gcatctcttc gtcggggtct 23880 accttggtca catgcttggt ctttctggct tgcttctttt ttggagggct gtccacgggg 23940 accacgtcct cctcggaaga cccggagccc acccgctgat actttcggcg cttggtgggc 24000 agaggaggtg gcggcggcga ggggctcctc tcctgctccg gcggatagcg cgccgacccg 24060 tggccccggg gcggagtggc ctctcggtcc atgaaccggc gcacgtcctg actgccgccg 24120 gccattgttt cctaggggaa gatggaggag cagccgcgta agcaggagca ggaggaggac 24180 ttaaccaccc acgagcaacc caaaatcgag caggacctgg gcttcgaaga gccggctcgt 24240 ctagaacccc cacaggatga acaggagcac gagcaagacg caggccagga ggagaccgac 24300 gctgggctcc agcatggcta cctgggagga gaggaggatg tgctgctgaa acacctgcag 24360 cgccagtccc tcatcctccg ggacgcccta gccgaccgga gcgaaacccc cctcagcgtc 24420 gaggagctgt gtcgggccta cgagctcaac ctcttctcgc cgcgcatgcc ccccaaacgc 24480 cagcccaatg gcacatgcga gcccaacccg cgtctcaact tctatcccgt ctttgcggtc 24540 cccgaggccc ttgccaccta tcacatcttt ttcaagaacc aaaagatccc cgtctcctgc 24600 cgcgccaacc gcacccgcgc cgacgcgctc cttactctgg ggcccggcgc gcgcatacct 24660 gatatcgctt ccctggaaga ggtgcccaag atcttcgaag ggctcggtcg ggacgagacg 24720 cgcgcggcga acgctctgaa agaaacagca gaggaagagg gtcacactag cgccctggta 24780 gagttggaag gcgacaacgc caggctggcc gtgctcaagc gcagcgtcga gctcacccac 24840 ttcgcctacc ccgccgtcaa cctcccgccc aaggtcatgc gtcgcatcat ggatcagctt 24900 atcatgcccc acatcgaggc cctcgatgaa agtcaggagc agcgccccga ggacgcccgg 24960 cccgtggtca gcgacgagat gctcgcgcgc tggctcggga cccgcgaccc ccaggctttg 25020 gaacagcggc gcaagctcat gctggccgtg gtcctggtca ccctcgagct ggaatgcatg 25080 cgccgcttct tcagcgaccc cgagaccctg cgcaaggtcg aggagaccct gcactacact 25140 ttcaggcacg gtttcgtcag gcaggcctgc aagatctcca acgtggagct gaccaacctg 25200 gtctcctgcc tggggatcct gcacgagaac cgcctgggcc agaccgtgct ccactctacc 25260 ctgaagggcg aggcgcggcg ggactacgtc cgcgactgcg tctttctctt tctctgccac 25320 acatggcaag cggccatggg cgtgtggcag cagtgtctcg aggacgagaa cctgaaggag 25380 ctggacaagc ttcttgctag aaaccttaaa aagctgtgga cgggcttcga cgagcgcacc 25440 gtcgcctcgg acctggccga gatcgtcttc cccgagcgcc tgagacagac gctgaaaggc 25500 gggctgcccg acttcatgag ccagagcatg ttgcaaaact accgcacttt cattctcgag 25560 cgatctggga tgctgcccgc cacctgcaac gccttcccct ccgactttgt accgctgagc 25620 taccgcgagt gtcccccgcc gctgtggagc cactgctacc tcttgcagct ggccaactac 25680 atcgcctacc actcggacgt gatcgaggac gtgagcggcg aggggctgct cgagtgccac 25740 tgtcgctgca acctgtgctc cccgcatcgc tccctggtct gcaaccccca gctcctgagc 25800 gagacccagg tcatcggtac cttcgagctg caaggtccgc aggagtccac cgctccgctg 25860 aaactcacgc cggggttgtg gacttccgcg tacctgcgca aatttgtacc cgaagactac 25920 cacgcccatg agataaagtt ctttgaggac caatcgcgcc cgcagcacgc ggatctcacg 25980 gcctgcgtca tcacccaggg cgcgatcctc gcccaattgc acgccatcca aaaatcccgc 26040 caagagtttc ttctgaaaaa gggtagaggg gtctacctgg acccccagac gggcgaggtg 26100 ctcaacccgg gtctcccca gcatgccgag gaagaagcag gagccgctag tggaggagat 26160 gggaagaa tgggacagcc aggcagaga ggacgaatgg gagaggaga cagaggagga 26220 agaattgaa gaggtggaag agcagccccgt cgccgcacca tccgcgccgg cagccccgcc 26280 ggtcacggat acaacctccg ctccggtcaa gcctcctcgt agatgggatc aaatgaaggg 26340 tgacggtaag cacgagcggc agggctaccg atcatggagg gcccacaaag ccgcgatcat 26400 cgcctgcttg caagactgcg gggggaacat cgcttttgcc cgccgctacc tgctcttcca 26460 ccgcggggtg aacatcccc gcaacgtgtt gcattactac cgtcaccttc acagctaaga 26520 aaaagcaagt aagagagtc gccggaggag gagaggcct gagatcgcg gcgaacgagc 26580 ccttgaccac cagggagctg aggaaccgga tcttccccac tctttatgcc atttttcagc 26640 agagtcgagg tcagcagcaa gagctcaaag taaaaaaccg gtctctgcgc tcgctcaccc 26700 gcagttgctt gtaccacaaa aacgaagatc agctgcagcg cactctcgaa gacgccgagg 26760 ctctgttcca caagtactgc gcgctcactc ttaaagacta aggcgcgccc acccggaaaa 26820 aaggcgggaa ttacctcatc gccaccatga gcaaggagat tcccacccct tacatgtgga 26880 gctatcagcc ccagatgggc ctggccgcgg gcgctcccca ggactactcc acccgcatga 26940 actggctcag tgccggcccc tcgatgatct cacaggtcaa cggggtccgt aaccatcgaa 27000 accagatatt gttggagcag gcggcggtca cctccacgcc cagggcaaag ctcaacccgc 27060 gtaattggcc ctccaccctg gtgtatcagg aaatccccgg gccgactacc gtactacttc 27120 cgcgtgacgc actggccgaa gtccgcatga ctaactcagg tgtccagctg gccggcggcg 27180 cttcccggtg cccgctccgc ccacaatcgg gtataaaac cctggtgatc cgaggcagag 27240 gcacacagct taacgacgag ttggtgagct cttcaatcgg tctgcgaccg gacggagtgt 27300 tccaactagc cggagccggg agatcctcct tcactcccca ccaggcctac ctgaccttgc 27360 agagcagctc ttcggagcct cgctccggag gcatcggaac cctccagttc gtggaggagt 27420 ttgtgccctc ggtctacttc aaccccttct cgggatcgcc aggcctctac ccggacgagt 27480 tcataccgaa cttcgatgca gtgagagaag cggtggacgg ctacgactga atgtcccatg 27540 gtgactcggc tgagctcgct cggttgaggc atctggacca ctgccgccgc ctgcgctgct 27600 tcgcccggga gagctgcgga ctcatctact ttgagtttcc cgaggagcac cccaacggcc 27660 ctgcacacgg agtgcggatc accgtagagg gcaccaccga gtctcacctg gtcaggttct 27720 tcacccagca acccttcctg gtcgagcggg accggggcgc caccacctac accgtctact 27780 gcatctgtcc taccccgaag ttgcatgaga atttttgttg tactctgtgt gctgagttta 27840 ataaaagcta aactcctaca atactctggg atcccgtgtc gtcgcactcg caacgagatc 27900 ttcaacctca ccaaccagac tgaggtaaaa ctcaactgca gaccgggggg caaatacatc 27960 ctctggctct ttgaaaacac ttccttcgca gtctccaacg cctgctgcac tagtccatga 28020 actgattgat taaagcccaa aaaaccaatc aaaccccttc ccattagccc caaataaaca 28080 atcattggaa ataatcattc aataaagatc acttacttga aatctgaaag tatgtctctg 28140 gtgtagttgt ttagcagcac ctcggtaccc tcctcccagc tctggtactc cagtccccgg 28200 cgggcggcga actttctcca caccttgaaa gggatgtcaa attcctggtc cacaattttc 28260 attgttttcc ctctcagatg tcaaagaggc ttcgggtgga agatgacttc aaccccgtct 28320 acccctatga ctacgcgcgg aatcagaata tccccttcct cactcccccc tttgtctcct 28380 ccgatggatt ccaaaacttc ccccctggtg tcctgtcact caaattggct gacccaatcg 28440 ctatcaccaa tggtgatgtc tcactcaagg tgggaggggg acttacttta caagatggaa 28500 ccggaaaact aactatcgac accaagactc ccttgcaagt tgcaaataat aaattagaac 28560 ttgcgtttga tgcaccattg tatgaaaaaa atggaaaact tgctttaaaa acaggccatg 28620 gattagctgt tttaaccaaa gacattggca taccagaatt aattggatcc cttgtgatct 28680 taactggaaa aggaattggg acaggtactg ttgcaggagg aggaactata gatgtaagac 28740 tgggtgatga tggaggttta tcatttgata aaaagggtga tctagtagcc tggaataaaa 28800 aaaatgacag gcgcactttg tggacaacac ctgatccatc cccaaactgc agagtatcag 28860 aagataaaga ttcaaaacta actttaattc ttacaaaatg tggaagtcag atcctagcaa 28920 gtttttcact gcttgtagtc aaagggacgt atgcaactgt tgataaaaat acaactaata 28980 aacaatttag cattaacta ctgtttgatg caaatggaaa gcttaaaagc gaatcaaacc 29040 ttagtggtta ttggaactat agaagtgata atagtgttgt tagtactccc tatgacaatg 29100 cagtgccttt catgccaaat accacggctt atccaaaaat cattaatagt acaactgatc 29160 ctgaaaataa gaaaagttcg gctaaaaaaa ctattgttgg caatgtctac ctagaaggta 29220 atgcgggtca accagtagcc gttgctatta gtttcaataa ggaaactact gctgactatt 29280 caataacatt tgactttgcg tggagcaaag cttatgaaac ccctgtgcct tttgacacct 29340 cctccatgac attctcatat attgcccagg aaaatcaaga caaaggcgaa taaagtgttt 29400 tgaaatgaac ttatgtatct ttattgattt ttacaccagc acgggtagtc agtctcccac 29460 caccagccca tttcacagtg taaacagtcc tttctccccg gctggcctta aaagcatca 29520 tatcatgggt aacagacata ttcttaggtg ttatattcca cacggtttcc tgtcgagcca 29580 aacgctcatc agtgatatta ataaactccc cgggcagctc acttaagttc atgtcgctgt 29640 ccagctgctg agccacaggc tgctgtccaa cttgcggttg cttaacgggc ggcgaaggag 29700 aagtccacgc ctacatgggg gtagagtcat aatcgtgcat caggataggg cggtggtgct 29760 gcagcagcgc gcgaataaac tgctgccgcc gccgctccgt cctgcaggaa tacaacatgg 29820 cagtggtctc ctcagcgatg attcgcaccg cccgcagcat aaggcgcctt gtcctccggg 29880 cacagcagcg caccctgatc tcacttaaat cagcacagta actgcagcac agcaccacaa 29940 tattgttcaa aatcccacag tgcaaggcgc tgtatccaaa gctcatggcg gggaccacag 30000 aacccacgtg gccatcatac cacaagcgca ggtagattaa gtggcgaccc ctcataaaca 30060 cgctggacat aaacattacc tcttttggca tgttgtaatt caccacctcc cggtaccata 30120 taaacctctg attaaacatg gcgccatcca ccaccatcct aaaccagctg gccaaaacct 30180 gcccgccggc tatacactgc agggaaccgg gactggaaca atgacagtgg agagcccagg 30240 actcgtaacc atggatcatc atgctcgtca tgatatcaat gttggcacaa cacaggcaca 30300 cgtgcataca cttcctcagg attacaagct cctcccgcgt tagaaccata tcccagggaa 30360 caacccattc ctgaatcagc gtaaatccca cactgcaggg aagacctcgc acgtaactca 30420 cgttgtgcat tgtcaaagtg ttacattcgg gcagcagcgg atgatcctcc agtatggtag 30480 cgcgggtttc tgtctcaaaa ggaggtagac gatccctact gtacggagtg cgccgagaca 30540 accgagatcg tgttggtcgt agtgtcatgc caaatggaac gccggacgta gtcatttcgc 30600 tgctgccgct cagtcatcca actgaagtac atccattctc gaagattctg gagaagttcc 30660 tctgcatctg atgaaataaa aaacccgtcc atgcgaattc ccctcatcac atcagccagg 30720 actctgtagg ccatcccat ccagttaatg ctgccttgtc tatcattcag agggggcggt 30780 ggcaggattg gaagaaccat ttttattcca aacggtctcg aagtcgata aagtgcaagt 30840 cacgcaggtg acagcgttcc cctccgctgt gctgggtggaa acagacagcc aggtcaaaac 30900 ccactctatt ttcaaggtgc tcgaccgtgg cttcgagcag tggctctacg cgtacatcca 30960 gcatagaat cacattaaag gctggccctc catcgatttc atcaatcatc aggttacatt 31020 cctgcaccat ccccaggtaa ttctcatttt tccagccttg gattatctct acaaattgtt 31080 ggtgtaagtc cactccgcac atgtggaaaa gctcccacag tgccccctcc actttcataa 31140 tcaggcagac cttcataata gaacagatc ctgctgctcc accacctgca gcgtgttcaa 31200 aacaaaga ttcaataagg ttctgccctc cgccctgagc tcgcgcctca atgtcagctg 31260 caaaaagtca cttaagtcct gggccactac agctgacaat tcagagccag ggctaagcgt 31320 gggactggca agcgtaaggg aaaactttaa tgctccgaag ctagcaccca aaaactgcat 31380 gctgggaataa gctctctttg tgtctccggt gatgccttcc aaaatgtgag tgataaagcg 31440 tggtagtttt tctttaatca tttgcgtaat agaaaagtcc tgtaataag tcactaggac 31500 cccagggacc acatgtggt agcttacacc gcgtcgctga agcatggtta gtagagatga 31560 gagtctgaaaaagaaagc atgcactaaa ctaggtggc tattttcact gaaggaaaaa 31620 tcactctc cagcacagg gtaccactg gtggcctt gcggacatac aaaaatcggt 31680 ccgtgtgatt aaaaagcagc acagtaagtt cctgtctct tccggcaaaa atcacatcgg 31740 actgggttag tatgtccctg gcatgtagt cattcaggc cataaatctg ccctgatatc 31800 cagtaggaac cagcacactc acttttaggt gaagcaatac cacccatgc ggaggaatgt 31860 ggaaagattc agggcaaaa aaattatatc tattgctagt cccttccttgg acgggagcaa 31920 tccctccagg actatcttag aaagcataca gagattcagc catagctcag cccgcttacc 31980 agtagacaga gagacagca gtacaagcgc agcagcagc gactgactac ccactgaccc 32040 agctccctat ttaaaggcgc cttacactga cgtaatgacc aaaggtcta aaaccccgcc 32100 aaaaaaaaac acacacgccc tgggttttttcgcgaaaac acttccgcgt tccactcc 32160 tcgtatcgat ttcgtgactt aacttccggg ttcccacgtt acgtcacttc tgcccttaca 32220 tgtaactgag tcgtagggcg ccatcttgcc cacgtccaaa atggcttcca tgtccagcca 32280 cgcctccgcg gcgaccgtta gccgtgcgtc gtgacgtcat ttgcatcgtc ttctctcgtc 32340 caatcagcgt cggctccgcc ctaaattcaa aagctcattt gcatattaac ttttgtttac 32400 tttgtggggt atattattga tgatgttaat 32430 <210> 9 <211> 30880 <212> DNA <213> Artificial Sequence <220> <223> Ad20-42-42.Luc+.5ORF6 <400> 9 attaacatca tcaataatat accccacaaa gtaaacaaaa gttaatatgc aaatgagctt 60 ttgaatttag ggcggagccg acgctgattg gacgagagaa gacgatgcaa atgacgtcac 120 gacgcacggc taacggtcgc cgcggaggcg tggcctagcc cggaagcaag tcgcggggct 180 gatgacgtat aaaaaagcgg actttagacc cggaaatggc cgattttccc gcggccacgg 240 ccggatatga ggtaattctg ggcggatgca agtgaaatta ggtcattttg gcgcgaaaac 300 tgaatgagga agtgaaaagc gaaaaatacc gatcccgccc agggcggaat atttaccgag 360 ggccgagaga ctttgaccga ttacgtgggg gtttcgattg cggtgttttt ttcgcgaatt 420 tccgcgtccg tgtcaaagtc cggtgtttat gtcacagatc agctgaccta ggtggtcaat 480 attggccatt agccatatta ttcattggtt atatagcata aatcaatatt ggctattggc 540 cattgcatac gttgtatcca tatcataata tgtacattta tattggctca tgtccaacat 600 taccgccatg ttgacattga ttatgacta gttattaata gtaatcaatt acggggtcat 660 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg 720 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa 780 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact 840 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta 900 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt 960 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg 1020 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg 1080 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc 1140 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt 1200 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac 1260 accgggaccg atccagcctc cgcggccggg aacggtgcat tggaagcttg gcattccggt 1320 actgttggta aagccaccat ggaagacgcc aaaaacataa agaaaggccc ggcgccattc 1380 tatccgctgg aagatggaac cgctggagag caactgcata aggctatgaa gagatacgcc 1440 ctggttcctg gaacaattgc ttttacagat gcacatatcg aggtggacat cacttacgct 1500 gagtacttcg aaatgtccgt tcggttggca gaagctatga aacgatatgg gctgaataca 1560 aatcacagaa tcgtcgtatg cagtgaaaac tctcttcaat tctttatgcc ggtgttgggc 1620 gcgttattta tcggagttgc agttgcgccc gcgaacgaca tttataatga acgtgaattg 1680 ctcaacagta tgggcatttc gcagcctacc gtggtgttcg tttccaaaaa ggggttgcaa 1740 aaaattttga acgtgcaaaa aaagctccca atcatccaaa aaattattat catggattct 1800 aaaacggatt accagggatt tcagtcgatg tacacgttcg tcacatctca tctacctccc 1860 ggttttaatg aatacgattt tgtgccagag tccttcgata gggacaagac aattgcactg 1920 atcatgaact cctctggatc tactggtctg cctaaaggtg tcgctctgcc tcatagaact 1980 gcctgcgtga gattctcgca tgccagagat cctatttttg gcaatcaaat cattccggat 2040 actgcgattt taagtgttgt tccattccat cacggttttg gaatgtttac tacactcgga 2100 tatttgatat gtggatttcg agtcgtctta atgtatagat ttgaagaaga gctgtttctg 2160 aggagcctt aggattacaa gattcaaagt gcgctgctgg tgccaaccct attctccttc 2220 ttcgccaaaa gcactctgat tgacaaatac gatttatcta atttacacga aattgcttct 2280 ggtggcgctc ccctctctaa ggaagtcggg gaagcggttg ccaagaggtt ccatctgcca 2340 ggtatcaggc aaggatatgg gctcactgag actacatcag ctattctgat tacacccgag 2400 ggggatgata aaccgggcgc ggtcggtaaa gttgttccat tttttgaagc gaaggttgtg 2460 gatctggata ccgggaaac gctgggcgtt aatcaaagag gcgaactgtg tgtgagaggt 2520 cctatgatta tgtccggtta tgtaaacaat ccggaagcga ccaacgcctt gattgacaag 2580 gatggatggc tacattctgg agacatagct tactgggacg aagacgaaca cttcttcatc 2640 gttgaccgcc tgaagtctct gattaagtac aaaggctatc aggtggctcc cgctgaattg 2700 gaatccatct tgctccaaca ccccaacatc ttcgacgcag gtgtcgcagg tcttccccgac 2760 gatgacgccg gtgaacttcc cgccgccgtt gttgttttgg agcacggaaa gacgatgacg 2820 gaaaaagaga tcgtggatta cgtcgccagt caagtaacaa ccgcgaaaaa gttgcgcgga 2880 ggagttgtgt ttgtggacga agtaccgaaa ggtcttaccg gaaaactcga cgcaagaaaa 2940 atcagagaga tcctcataaa ggccaagaag ggcggaaaga tcgccgtgta attctagacg 3000 agatccgaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 3060 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 3120 tgtatcttat catgtctcga tccaggtagg tttgagtagt gggcgtggct aagatgacta 3180 taaaggtggg tgtcttacga gggtcttttt gcttttctgc agacatcatg aacgggaccg 3240 gcggggcctt cgaagggggg ctttttagcc cttatttgac aacccgcctg ccgggatggg 3300 ccggagttcg tcagaatgtg atgggatcga cggtggacgg gcgtccagtg cttccagcaa 3360 attcctcgac catgacctac gcgaccgtgg ggaactcgtc gctcgacagc accgccgcag 3420 ccgcggcagc cgcagccgcc atgacagcga cgagactggc ttcgagctac atgcccagca 3480 gcggtagcag cccctctgtg cccagttcca tcatcgccga ggagaaactg ctggccctcc 3540 tggccgagct ggaagccctg agccgccagc tggccgccct gacccagcag gtgtccgagc 3600 tccgcgaaca gcagcagcag caaaataaat gattcaataa acacagattc tgattcaaac 3660 agcaaagcat ctttattatt tattttttcg cgcgcggtag gccctggtcc acctctcccg 3720 atcattgaga gtgcggtgga ttttttccag gacccggtag agatgggatt ggatgttgag 3780 gtacatgggc atgagcccgt cccgggggtg aaggtagcac cactgcatgg cctcatgctc 3840 tggggtcgtg ttgtagatga tccagtcata gcaggggcgc tgggcgtggt gctggatgat 3900 gtctttgagg aggagactga tggccacggg gagccccttg gtgtaggtgt tggcgaagcg 3960 gttgagctgg gagggatgca tgcgggggga gatgatgtgc agtttggcct ggatcttgag 4020 gttggcaatg ttgccgccca gatccccgccg ggggttcatg ttgtgcagga ccaccaggac 4080 ggtgtagccc gtgcacttgg ggaacttatc atgcaacttg gaagggaatg cgtggaagaa tttggagacg cccttgtgcc cgcccaggtt ttccatgcac tcatccatga tgatggcgat 4200. gggcccgtgg gctgcggctt tggcaaagac gtttctgggg tcagagacat cataattatg 4260. ctcctgggtg agatcatcat aagacatttt aatgaatttg gggcggaggg tgccagattg ggggacgatc gttccctcgg gccccggggc gaagttcccc tcacagatct gcatctccca 4380 ggctttcatc tcggaggggg ggatcatgtc cacctgcggg gcgatgaaaa aaacggtttc 4440. cggggcgggg gtgatgagct gcgaggagag caggttctc aacagctggg acttgccgca 4500. cccggtcggg ccgtagtga ccccgatgac gggttgcagg tggtagttca aggagatgca 4560 gctgccgtct tcccggagga ggggggccac ctcgttgagc atgtctctga cttggaggtt 4620 ttcccggacg agctcgccga ggaggcggtc cccgcccagc gagagcagct cttgcaggga 4680 agcaaagttt ttcaggggct tgagcccgtc ggccatgggc atcttggcga gggtctgcga 4740 gaggagttcc aagcggtccc agagctcggt gacgtgctct acggcatctc gatccaacag 4800 acttcctcgt ttcgggggtt gggacgactg cgactgtagg gcacgagacg atgggcgtcc 4860 agcgctgcca gcgtcatgtc cttccagggt ctcagggtcc gcgtgagcgt ggtctccgtc 4920 acggtgaagg ggtgggcccc gggctgggcg cttgcaaggg tgcgcttgag actcatcctg 4980 ctggtgctga aacgggcacg gtcttcgccc tgcgcgtcgg cgagatagca gttgaccatg 5040 agctcgtagt tgagggcctc ggcggcgtgg cccttggcgc ggagcttgcc cttggaagag 5100 cgcccgcagg cgggacagag gagggattgc agggcgtaga gcttgggcgc gagaaagacg 5160 gactcggggg cgaaagcgtc cgctccgcag tgggcgcaga cggtctcgca ctcgacgagc 5220 caggtgagct cgggatgttc ggggtcaaaa accagttttc ccccgttctt tttgatgcgc 5280 ttcttacctc gcgtctccat gagtctgtgt ccgcgctcgg tgacaaacag gctgtctgtg 5340 tccccgtaga cggacttgat gggcctgtcc tgcaggggcg tcccgcggtc ctcctcgtag 5400 agaaactcgg accactctga gacgaaggcg cgcgtccacg ccaagacaaa ggaggccacg 5460 tgcgaggggt agcggtcgtt gtccaccagg gggtccacct tttccaccgt gtgcagacac 5520 atgtccccct cctcagcatc caagaaggtg attggcttgt aggtgtaggc cacgtgaccg 5580 ggggtccccg acgggggggt ataaaagggg gcgggtctgt gctcgtcctc actctcttcc 5640 gcgtcgctgt ccacgagcgc cagctgttgg ggtaggtatt ccctctcgag agcgggcatg 5700 acctcggcac tcaggttgtc agtttctaga aacgaggagg atttgatgtt ggcctgccct 5760 gccgcaatgc ttttgagtaa actctcatcc atctggtcag aaaagactat ttttttattg 5820 tcaagcttgg tggcgaagga gccatagagg gcgttggaga gaagcttggc gatggatctc 5880 atggtctgat ttttgtcacg gtcggcgcgc tccttggccg cgatgtttag ctggacatat 5940 tcgcgcgcga cacacttcca ttcggggaag acggtggtgc gctcgtcggg cacgatcctg 6000 acgcgccagc cgcggttatg cagggtgacc aggtccacgc tggtggccac ctcgccgcgc 6060 aggggctcgt tggtccagca gagtcgcccg cctttgcgcg aacagaaagg gggcagcaca 6120 tcaagcagat gctcgtcggg ggggtccgca tcgatggtga agataccggg acagagttcc 6180 ttgtcaaaat agtctatttt tgaggatgca tcatccaagg ccatctgcca ctcacgggcg 6240 gccattgctc gctcgtaggg gttgaggggc ggaccccagg gcatgggatg cgtgagggcg 6300 gaggcgtaca tgccgcagat gtcatagaca tagatgggct ccgagaggat gccgatgtag 6360 gtgggataac agcgcccccc gcggatgctg gcgcgcacgt agtcatacaa ctcgtgcgag 6420 ggggccaaga aggcggggcc gagattggtg cgctggggct gctcggcgcg gaagacgatc 6480 tggcgaaaga tggcgtgcga gttggaggag atggtgggcc gttggaagat gttaaagtgg 6540 gcgtggggca agcggaccga gtcgcggatg aagtgcgcgt aggagtcttg cagcttggcg 6600 acgagctcgg cggtgacgag gacgtccatg gcgcagtagt ccagcgtttc gcggatgatg 6660 tcataacccg tctctccttt cttctcccac agctcgcggt tgagggcgta ctcctcgtca 6720 tccttccagt actcccggag cgggaatcct cgatcgtccg cacggtaaga gcccagcatg 6780 tagaaatggt tcacggcctt gtagggacag cagcccttct ccacggggag ggcgtaagct 6840 tgagcggcct tgcggagcga ggtgtgcgtc agggcgaagg tgtccctgac catgactttc 6900 aagaactggt acttgaaatc cgagtcgtcg cagccgccgt gctcccagag ctcgaaatcg 6960 gtgcgcttct tcgagagggg gttaggcaga gcgaaagtga cgtcattgaa gagaatcttg 7020 cctgcccgcg gcatgaaatt gcgggtgatg cggaaagggc ccgggacgga ggctcggttg 7080 ttgatgacct gggcggcgag gacgatctcg tcgaagccgt tgatgttgtg cccgacgatg 7140 tagagttcca tgaatcgcgg gcggcctttg atgtgcggca gctttttgag ctcctcgtag 7200 gtgaggtcct cggggcattg caggccgtgc tgctctagcg cccactcctg gagatgtggg 7260 ttggcttgca tgaaggaagc ccagagctcg cgggccatga gggtctggag ctcgtcacga 7320 aagaggcgga actgctggcc cacggccatc ttttcgggtg tgacgcagta gaaggtgagg 7380 gggtcccgct cccagcgatc ccagcgtaag cgcacggcga gatcgcgagc gagggcgacc 7440 agctctgggt cccccgagaa tttcatgacc agcatgaagg ggacgagctg cttgccgaag 7500 gaccccatcc aggtgtaggt ttctacatcg taggtgacaa agagccgctc cgtgcgagga 7560 tgagagccga ttgggaagaa ctggatttcc tgccaccagt tggacgagtg gctgttgatg 7620 tgatgaaagt agaaatcccg ccggcgaacc gagcactcgt gctgatgctt gtaaaagcgt 7680 ccgcagtact cgcagcgctg cacgggctgt acctcatcca cgagatacac agcgcgtccc 7740 ttgaggagga acttcaggag tggcggccct ggctggtggt tttcatgttc gcctgcgtgg 7800 gactcaccct ggggctcctc gaggacggag aggctgacga gcccgcgtgg gagccaggtc 7860 cagatctcgg cgcggcgggg gcggagagcg aagacgaggg cgcgcagttg ggagctgtcc 7920 atggtgtcgc ggagatccag gtccgggggc agggttctga ggttgacctc gtagaggcgg 7980 gtgagggcgt gcttgagatg cagatggtac ttgatttcta cgggtgagtt ggtggccgtg 8040 tccacgcatt gcatgagccc gtagctgcgc ggggccacga ccgtgccgcg gtgcgctttt 8100 agaagcggtg tcgcggacgc gctcccggcg gcagcggcgg ttccggtccc gcgggcaggg 8160 gcggcagagg cacgtcggcg tggcgctcgg gcaggtcccg gtgctgcgcc ctgagagcgc 8220 tggcgtgcgc gacgacgcgg cggttgacat cctggatctg ccgcctctgc gtgaagacca 8280 cgggccccgt gactttgaac ctgaaagaca gttcaacaga atcaatctcg gcgtcattga 8340 cggcggcctg acgcaggatc tcttgcacgt cgcccgagtt gtcctggtag gcgatctcgg 8400 acatgaactg ctcgatctcc tcctcctgga gatcgccgcg gcccgcgcgc tcgacggtgg 8460 cggcgaggtc attcgagatg cgacccatga gctgcgagaa ggcgcccagg ccgctctcgt 8520 tccagacgcg gctgtagacc acgtccccgt cggcgtcgcg cgcgcgcatg accacctgcg 8580 cgaggttgag ctccacgtgc cgcgcgaaga cggcgtagtt gcgcaggcgc tggaagaggt 8640 agttgagggt ggtggcgatg tgctcggtga cgaagaagta catgatccag cggcgcagag 8700 gcatctcgct gatgtcgccg atggcctcca gcctttccat ggcctcgtag aaatccacag 8760 cgaagttgaa aaactgggcg ttgcgggccg agaccgtgag ctcgtcttcc aggagcctga 8820 tgagctcggc gatggtggcg cgcacctcgc gctcgaaatc cccgggggcc tcctcctctt 8880 cctcttcttc catgacgacc tcttcttcta tttcttcctc tgggggcggt ggtggtggcg 8940 gggcccgacg acgacggcga cgcaccggga gacggtcgac gaagcgctcg atcatctccc 9000 cgcggcggcg acgcatggtt tcggtgacgg cgcgaccccg ttcgcgagga cgcagcgtga 9060 agacgccgcc ggtcatctcc cggtaatggg gcgggtcccc gttgggcagc gatagtgcgc 9120 tgacgatgca tcttatcaat tgcggtgtag gggacgtgag cgcgtcgaga tcgaccggat 9180 cggagaatct ttcgaggaaa gcgtctagcc aatcgcagtc gcaaggtaag ctcaaacacg 9240 tagcagccct gtggacgctg ttagaattgc ggttgctgat gatgtaattg aagtaggcgt 9300 ttttgaggcg gcggatggtg gcgaggagga ccaggtcctt gggtcccgct tgctggatgc 9360 ggagccgctc ggccatgccc caggcctggc cctgacaccg gctcaggttc ttgtagtagt 9420 catgcatgag cctctcgatg tcatcactgg cggaggcgga gtcttccatg cgggtgaccc 9480 caacgcccct gagcggctgc acgagcgcca ggtcggcgac gacgcgctcg gcgaggatgg 9540 cctgttgcac gcgggtgagg gtgtcctgga agtcgtccat gtcgacgaag cggtggtagg 9600 cccctgtgtt gatggtgtaa gtgcagttgg ccatgagcga ccagttgacg gtctgcaggc 9660 cgggctgcac gacctcggag tacctgagcc gcgagaaggc gcgcgagtcg aagacgtagt 9720 cgttgcaggt gcgcacgagg tactggtatc cgactaggaa gtgcggcggc ggctggcggt 9780 agagcggcca gcgctgggtg gccggcgcgc ccggggccag gtcctcgagc atgaggcggt 9840 ggtagccgta gaggtagcgg gacatccagg tgatgccggc ggcggtggtg gaggcgcgcg 9900 ggaactcgcg gacgcggttc cagatgttgc gcagcggcag gaaatagtcc atggtcggca 9960 cggtctggcc ggtgagacgc gcgcagtcat tgacgctcta gaggcaaaaa cgaaagcggt 10020 tgagcgggct attcctccgt agcctggcgg aacgcaaacg ggttaggccg cgtgtgtacc 10080 ccggttcgag tcccctcgaa tcaggctgga gccgcgacta acgtggtatt ggcactcccg 10140 tctcgacccg agcccgatag ctgccaggat acggcggaga gcccttttgg cggccgaggg 10200 gggtcgctag acttgaaagc ggccgaaaac cctgccgggt agtggctcgc gcccgtagtc 10260 tggagaagca tcgccagggt tgagtcgcgg cagaacccgg ttcgcggacg gccgcggaga 10320 gcgggacttg gtcaccccgc cgatttaaaa acccacagcc agccgacttc tccagttacg 10380 ggagcgagcc cccttttttc tttttgccag atgcatcccg tcctgcgcca aatgcgtccc 10440 accccccctc cggcgaccac cgcaaccgcg gccgtagcag gcgccggcgc tagccagcca 10500 cagccacaga cagagatgga cttggaagag ggcgaagggc tggcgagatt gggggcgccg tctccggagc gacaccccg cgtgcagctg cagaaggacg tgcgcccggc gtacgtgcct 10620 gcgcagaacc tgttcaggga ccgcagcggg gaggagccg aggagatgcg cgactgccgc 10680 tttcgggcgg gcagggagct gcgcgagggt ctggaccgcc agcgcgtgct gcgcgatgag 10740 gatttcgagc cgaacgagca gacggggatc agccccgcgc gcgcgcacgt ggcggcggcc 10800. aacctggtga cggcctacga gcagacggtg aagcaggagc gcaacttcca aaagagtttc aacaaccacg tgcgcacgct gatcgcgcgc gaggaggtgg ccctgggcct gatgcacctg tgggacctgg cggaggccat cgtgcagaac ccggacagca agcctctgac ggcgcagctg ttcctggtgg tgcagcacag tagggacaac gaggcgttca gggaggcgct gctgaacatc gccgagcccg agggtcgctg gctgctggag ctgatcaaca tcttgcagag catcgtagtg caggagcgca gcctgagcct ggccgagaaa gtggcggcga tcaactactc ggtgctgagc 11160 ctgggcaagt tttacgcgcg caagatttac aagacgccat acgtgcccat agacaaggag 11220 gtgaagatag acagctttta catgcgcatg gcgctcaagg tgctgacgct gagcgacgac 11280 ctgggcgtgt accgcaacga ccgcatccac aaggccgtga gcgcgagccg gcggcgcgag 11340 ctgagcgacc gcgagctgat gctgagcctg cgccgggcgc tggtaggggg cgccgccggc 11400 ggcgaggagt cctacttcga catgggggcg gacctgcatt ggcagccgag ccggcgcgcc 11460 ttggaagccg cctacggtcc agaggacttg gatgaggatg aggaagagga ggaggatgca 11520 cccgctgcgg ggtactgacg cctccgtgat gtgtttttag atgtcccagc aagccccgga 11580 ccccgccata agggcggcgc tgcaaagcca gccgtccggt ctagcatcgg acgactggga 11640 ggccgcgatg caacgcatca tggccctgac gacccgcaac cccgagtcct ttagacaaca 11700 gccgcaggcc aacagacttt cggccattct ggaggcggtg gtcccctctc gaaccaaccc 11760 11820. aaggtgctgg cgatcgtgaa cgcgctggcg gagaacaagg ccatccgtcc cgacgaggcc ggtctggtgt acaacgccct gctggagcgc gtgggccgct acaacagcac 11880 aaacgtgcag tccaacctgg accggctggt gacggacgtg cgcgaggccg tggcgcagcg cgagcggttc aagaacgag gcctgggctc gctggtggcg ctgaacgcct tcctggcgac gcagccggcg aacgtgccgc gcgggcagga cgattatacc aactttataa gcgcgctgcg gctgatggtg accgaggtgc cccagagcga ggtgtaccag tcggggcccgg attacttttt 12120 ccaaactagc agacagggcc tgcagacggt gaacctgagc caggccttta agaacctgcg cgggctgtgg ggcgtgcagg cgcccgtggg cgaccggtcg acggtgagca gcttgctgac 12240 gcccaactcg cggctgctgc tgctgctgat cgcgcccttc accgacagcg gcagcgtgaa 12300 ccgcaactcg tacctgggcc acctgctgac gctgtaccgc gaggccatag gccaggcaca ggtggacgag cagaccttcc aggagatcac tagcgtgagc cgcgcgctgg ggcagaacga 12480. cccgacagt cttagggcca ccctgaactt cttgctgacc aatagacagc agaagatccc ggcacagtac gcgctatcgg ccgaggagga gcgcatcctg agatatgtgc agcagagcgt 12540 agggctgttc ctgatgcagg agggggccac ccccagcgcc gcgctggaca tgaccgcgcg 12600 caacatggaa cctagcatgt acgctgccaa ccggccgttc atcaataagc tgatggacta 12660 cctgcaccgc gcggcggcca tgaactcgga ctactttaca aacgccatcc tgaacccgca 12720 ctggctcccg ccgccggggt tctacacggg cgagtacgat atgcccgacc ccaacgacgg 12780 gttcctgtgg gacgacgtgg acagcgcggt gttctccccg accttgcaaa agcgccagga 12840 ggcggtgcgc acgcccgcga gcgagggtgc ggtgggtcgg agcccctttc ctagcttagg 12900 gagtttgcat agcttgccgg gctcggtgaa cagcggcagg gtgagccgac cgcgcttgct 12960 gggcgaggac gagtacctga acgactcgct gctgcagccg ccgcgggtca agaacgccat 13020 ggccaataac gggatagaga gtctggtgga caaactgaac cgctggaaga cctacgctca 13080 ggaccatagg gacgcgcccg cgccgcggcg acagcgccac gaccggcagc ggggcctggt 13140 gtgggacgac gaggactcgg ccgacgatag cagcgtgttg gacttgggcg ggagcggtgg 13200 ggccaacccg ttcgcgcatc tgcagcccaa actggggcga cggatgtttt gaaatgcaaa 13260 ataaaactca ccaaggccat agcgtgcgtt ctcttccttg ttagagatga ggcgtgcggt 13320 ggtgtcttcc tctcctcctc cctcgtacga gagcgtgatg gcgcaggcga ccctggaggt 13380 tccgtttgtg cctccgcggt atatggctcc tacggagggc agaaacagca ttcgttattc 13440 ggagctggct ccgctgtacg acaccactcg cgtgtacttg gtggacaaca agtcggcgga 13500 catcgcttcc ctgaactacc aaaacgacca cagcaacttc ctgaccacgg tggtgcagaa 13560 caacgatttc acccccgccg aggccagcac gcagacgata aattttgacg agcggtcgcg 13620 gtggggcggt gatctgaaga ccattctgca caccaacatg cccaatgtga acgagtacat 13680 gttcaccagc aagtttaagg cgcgggtgat ggtggctaga aagcacccag agaatgtagc 13740 taaagacgat ttgagtcagg ataagcttga atatgagtgg tttgagttta ccctgcccga 13800 gggcaacttt tccgagacca tgaccataga cctgatgaac aacgccatct tggaaaacta 13860 cttgcaagtg gggcggcaaa atggcgtgct ggagagcgat atcggagtca agtttgacag 13920 cagaaatttc aagctgggct gggacccggt gaccaagttg gtgatgccag gggtctacac 13980 ctacgaggcc ttccaccgg acgtggtgct gctgccggc tgcggggtgg acttcaccga 14040 gagccgcctg agcaacctcc tgggcattcg cagaagcaa cctttccaag agggcttcag 14100 aatcatgtat gaagatctag taggggca catcccagcc ctcctgaatg tcaggagta 14160 tctgaaggat aaggagaag ctggcacagc agatgcaat accattaagg ctcagaatga 14220 tgcagtccca agaggagata actatgcatc agcggcagaa gccaaagcag caggaaaaga 14280 aattgagttg aaggccattt tgaagatga ttcaacaga agctacaatg tgatcgaggg 14340 aaccacagac accctgtacc gcagttgta cctgtcctat acctacgggg acccgagaa 14400 gggggtgcag tcgtggacgc tgctcaccac cccggacgtc acatgcggcg cggagcaagt 14460 ctactgtcg ctgccggacc tcatgcaaga cccgtcacc tccgctcta cccagcaagt 14520 cagcaactac cccgtggtcg gcgccgagct catgcccttc cgcgccaaga gcttttacaa 14580 cgacctcgcc gtctactccc agctcatccg cagctacacc tccctcaccc acgtcttcaa 14640 ccgcttcccc gacaaccaga tcctctgccg cccgcccgcg cccaccatca ccaccgtcag 14700 tgaaaacgtg cctgctctca cagatcacgg gacgctaccg ctgcgcagca gtatccgcgg 14760 agtccagcga gtgaccgtca ctgacgcccg tcgccgcacc tgtccctacg tctacaaggc 14820 cctgggcata gtcgcgccgc gcgtgctatc cagtcgcacc ttctaaaaaa tgtctattct 14880 catctcgccc agcaataaca ccggctgggg tattactagg cccagcacca tgtacggagg 14940 agccaagaag cgctcccagc agcaccccgt ccgcgtccgc ggccacttcc gcgctccctg 15000 gggcgcttac aagcgcgggc ggacttctac cgccgctgcc gtgcgtacca ccgtcgacga 15060 cgtcatcgac tcggtggtcg ccgacgcgcg caactacacc cccgccccct ccaccgtgga 15120 cgcggtcatt gacagcgtgg tggccgacgc gcgcgactat gccagacgca agagccggcg 15180 gcgacggatc gccaggcgcc accggagcac gcccgccatg cgcgccgccc gggctctgct 15240 gcgccgcgcc agacgcacag gccgccgggc catgatgcga gccgcgcgcc gcgctgccac 15300 tgcacccacc cccgcaggca ggactcgcag acgagcggcc gccgccgctg ccgcggccat 15360 ctctagcatg accagaccca ggcgcggaaa cgtgtactgg gtgcgcgact ccgtcacggg 15420 cgtgcgcgtg cccgtgcgca cccgtcctcc tcgtccctga tctaatgctt gtgtcctccc 15480 ccgcaagcga cgatgtcaaa gcgcaaaatc aaggaggaga tgctccaggt cgtcgccccg 15540 gagatttacg gaccaaccca ggcggaccag aaaccccgca aaatcaagcg ggttaaaaaa 15600 zonetgagg tggacgagg ggcagtagag tttgtgcgcg agttcgctcc gcggcggcgc 15660 gtaaattgga aggggcgcag ggtgcagcgc gtgttgcggc ccggcacggc ggtggtgttc 15720 acgcctggcg agcggtcctc ggtcaggagc aagcgtagct atgacgaggt atacggcgac 15780 gacgacatcc tggaccaggc ggcggagcgg gcgggcgagt ttgcctacgg gaagcggtcg 15840 cgcgaagagg agctgatctc gctgccgctg gacgaaagca accccacgcc aagcctgaag 15900 cccgtgaccc tgcagcaggt gctgccccag gcggtgctgc tgccgagccg cggggtcaag 15960 cgcgagggcg agaacatgta cccgaccatg cagatcatgg tgcccaagcg ccggcgcgtg 16020 gaggacgtgc tggacaccgt gaaaatggat gtggagcccg aggtcaaggt gcgccccatc 16080 aagcaggtgg cgccgggcct gggcgtgcag accgtggaca ttcagatccc caccgacatg 16140 gatgtcgaca aaaaaccctc gaccagcatc gaggtgcaga ccgacccctg gctcccagcc 16200 tccactgcta ccgcctccac ttctaccgtc gccacggcca ccgagcctcc caggaggcga 16260 agatggggcc ctgccaaccg gctgatgccc aactacgtgt tgcatccttc catcatcccg 16320 acgccgggct accgcggcac tcggtactac gccagccgca ggcgcccagc cagcaaacgc 16380 cgccgccgca ccgccacccg ccgccgtctg gcccccgccc gcgtgcgccg cgtgaccacg 16440 cgccggggcc gctcgctcgt tctgcccacc gtgcgctacc accccagcat cctttaatcc 16500 gtgtgctgtg atactgttgc agagagatgg ctctcacttg ccgcctgcgc atccccgtcc 16560 cgaattaccg aggaagatcc cgccgcagga gaggcatggc aggcagtggc ctgaaccgcc 16620 gccggcggcg ggccatgcgc aggcgcctga gtggcggctt tctacccgcg ctcatcccca 16680 taatcgccgc ggccatcggc acgatcccgg ccatagcttc cgttgcgctg caggcgtcgc 16740 agcgccgttg atgtgcgaat aaagcctctt tagactctga cacacctggt cctgtatatt 16800 tttagaatgg aagacatcaa ttttgcgtcc ctggctccgc ggcacggcac gcggccgttc 16860 atgggcacct ggaacgagat cggcaccagc cagctgaacg ggggcgcctt caattggagc 16920 agtgtctgga gcgggcttaa aaatttcggc tcgacgctcc ggacctatgg gaacaaggcc 16980 tggaatagta cacggggca gttgttaagg gaaaagctca aagaccagaa cttccagcag 17040 aaggtggtgg acgggctggc ctcgggcatt aacggggtgg tggacatcgc gaaccaggcc 17100 gtgcagcgcg agataaacag ccgcctggac ccgcgcccgc ccacggtggt ggagatggaa 17160 gatgcaactc ttccgccgcc caaaggcgag aagcggccgc ggcccgacgc ggaggagacg 17220 atcctgcagg tggacgagcc gccctcgtac gaggaggccg tgaaggccgg catgcccacc 17280 acgcgcatca tcgcgccgct ggccacgggt gtaatgaaac ccgccaccct tgacctgcct 17340 ccaccaccca cgcccgctcc accgaaggca gctccggttg tgcagcccccc tcctgtggcg 17400 accgccgtgc gccgcgtccc cgcccgccgc caggcccaga actggcagag cacgctgcac 17460 agtatcgtgg gcctgggagt gaaaagtctg aagcgccgcc gatgctattg around 17520 aaggaccac taaagggaga gcttaacttg tatgtgcctt accgccagag aacgcgcgaa 17580 gatggccacc ccctcgatga tgccgcagtg ggcgtacatg cacatcgccg ggcaggacgc 17640 ctcggagtac ctgagcccgg gtctggtgca gtttgcccgc gccaccgaca cgtacttcag 17700 cctgggcaac aagtttagga accccacggt ggctcccacc cacgatgtga ccacggaccg 17760 gtcccagcgt ctgacgctgc gcttcgtgcc cgtggatcgc gaggacca cgtactcgta 17820 caaggcgcgc ttcactctgg ccgtgggcga caaccgggtg ctagacatgg ccagcactta 17880 ctttgacatc cgcggcgtcc tggaccgcgg ccccagcttc aaaccctact cgggcacggc 17940 ctacaacagt ctggccccca agggcgcccc caactccagc cagtgggctg acaaagaaag 18000 ggtgaatgga ggcggtaaca ccaaagatgt tacaaagaca tttggtgtgg cagccatggg 18060 tggagaagac attacggaaa agggtctgaa gattggaact gatacaacag aaaatgaacc 18120 aatatttgca gacaaaaatt ttcagcctga accccaagtg ggagaggaaa actggcagga 18180 aacatttgtt ttctatggag gaagagctct taaaaaagaa accaaaatga agccttgcta 18240 tggttcattt gccagaccga ctaatgaaaa aggtggccag gctaaattta taattggaga 18300 taatggtcaa cccactgaaa accatgacat aacaatggct ttctttgaca ctcctggtgg 18360 tacaatcaca ggaggtactg gtggtcccca agatgaactt aaagctgaca ttgttatgta 18420 cactgaaaat ataaatcttg aaactccaga tacccatgtg gtgtacaagc caggcaaaga 18480 agatgatagc tcagaaatta acttggtgca acagtccatg cccaacagac ccaactacat 18540 cggtttcaga gacaactttg tgggacttat gtactacaac agcactggca acatgggcgt 18600 gctggctggt caggcctctc agttgaatgc tgtggtcgac ttgcaagaca gaaacaccga 18660 gctgtcttac cagctcctgc tagattctct gggtgacagg accagatact ttagcatgtg 18720 gaactctgcg gtggacagtt atgatcccga tgttaggatc attgagaatc acggtgtgga 18780 agatgaactt cccaactatt gctttccatt ggacggctca ggcactaatt cagcttttca 18840 aggtgtaaaa attaagcaaa accaagacgg agatgttaat gatgattggg aaaaggatga 18900 taaggtatcc actcaaaacc aaatttgcaa gggcaacatt tacgccatgg agatcaacct 18960 ccaggccaac ctgtggaaaa gttttctgta ctcgaacgtg gccctgtacc tgcccgactc 19020 ctacaagtac acgccggcca acgtgacgct gcccgccaac accaacacct acgagtacat 19080 gaacggccgc gtggtagccc cctcgctggt ggacgcctac atcaacatcg gcgcccgctg 19140 gtcgctagac cccatggaca atgtcaaccc cttcaaccac caccgcaatg cgggtctgcg 19200 ctaccgctcc atgcttctgg gcaacggccg ctacgtgccc ttccacatcc aagtgcccca 19260 aaagttcttt gccatcaaga acctgctcct gctcccgggc tcctacacct acgagtggaa 19320 cttccgcaag gacgtcaaca tgatcctgca gagttccctt ggaaacgacc tgcgcgttga 19380 cggcgcctcc gtccgcttcg acagtgttaa cctctatgcc accttcttcc ccatggcgca 19440 caacaccgcc tccaccctgg aagccatgct gcgcaacgac accaacgacc agtccttcaa 19500 cgactacctc tcggccgcca acatgctcta ccctatcccg gccaaggcca ccaacgtgcc 19560 catctccatc ccctcgcgca actgggccgc cttccgcggc tggagtttca cccggctcaa 19620 gaccaaggaa actccctccc ttggctcggg ttttgacccc tactttgtct actcgggctc 19680 catcccctac ctcgacggga ccttttacct gaaccatacc ttcaagaagg tctccatcat 19740 gttcgactcc tcggtcagct ggcccggcaa cgaccggctg ctcacgccga acgagttcga 19800 gatcaagcgc agcgttgacg gggagggcta caacgtggcc caatgcaaca tgaccaagga 19860 ctggttcctt gtccagatgc tctcccacta caacatcggc taccagggct tccacgtgcc 19920 cgagggctac aaggaccgca tgtactcatt cttccgcaac ttccagccca tgagcaggca 19980 ggtggtcgat gagatcaact acaaggacta caaagccgtc accctgccct tccagcacaa 20040 caactcgggc ttcaccggct atctcgcccc caccatgcgc caggggcagc cctaccccgc 20100 caacttccca tacccgctca tcggccagac agccgtaccc tccgtcaccc agaaaaagtt 20160 cctctgcgac agggtcatgt ggcgcatccc cttttccagc aacttcatgt ccatgggcgc 20220 cctcaccgac ctgggacaga acatgctcta tgccaactcg gcccacgcgc tcgacataac 20280 ttttgaggtg gaccccatgg atgagcccac cctcctctat cttctcttcg aagttttcga 20340 cgtggtcaga gtgcaccagc cgcaccgcgg cgtcatcgag gccgtctacc tgcgcacgcc 20400 cttctccgcc ggcaacgcta ctacataagc atgagcggct ccagcgaacg agagctcgcg 20460 gccatcgtgc gcgacctggg ctgcgggccc tactttttgg gaacccacga caagcgcttc 20520 cctggcttcc ttgccagcga caagctggca tgcgccatcg tcaacacggc cggccgcgag 20580 accgggggcg tgcactggct cgcctttggc tggaatccgc gctcgcgcac ctgctacatg 20640 tttgacccct ttgggttctc ggaccgccgg ctcaagcaga tttacagctt cgagtacgag 20700 gccatgctgc gccgcagcgc cctggcctcc tcgcccgacc gctgtctcag cctcgagcag 20760 tccacccaga ccgtgcaggg gcccgactcc gccgcctgcg gacttttttg ttgcatgttt 20820 ttgcatgcct ttgtgcactg gcccgaccga cccatggacg gaaaccccac catgaacttg 20880 ctgacggggg tgcccaacgg catgctacaa tcgccacagg tgctccccac cctcaggcgc 20940 aaccaggagg agctctaccg cttcctcgcg cgccactccc cttactttcg atcccaccgc 21000 gccgccattg aaaacgccac cgcttttgat aaaatgaaac aactgcgtgt atctcaataa 21060 acagcacttt attttacatg cactggagta tatgcaagtt atttaaaagt cgaaggggtt 21120 ctcgcgctcg tcgttgtgcg ccgcgctggg gagggccacg ttgcggtact gaaacttggg 21180 ctgccacttg aactcgggga tcaccagttt gggcactggg gtctcgggga aggtctcgct 21240 ccacatgcgc cggctcatct gcagggcgcc cagtatgtcg ggcgcggaga tcttgaaatc 21300 gcagttggga ccagtgctct gcgcgcgcga gttgcggtac acggggttgc agcactggaa 21360 caccatcaaa ctggggtgct tcacgctggc cagcacgctc ttgtcgctga tctgatcctt 21420 gtccaggtcc tcggcgttgc tcaggccgaa cggggtcatc ttgcacagct ggcggcccag 21480 gaagggcacg ctgtgaggct tgtggttaca ctcgcagtgc acgggcatca gcatcatccc 21540 cgcgccgcgc tgcatattcg ggtagagggc cttgacaaag gccaagatct gcttgaaagc 21600 ttgctgggcc ttggccccct cgctgaagaa caggccgcag ctcttcccgc tgaactggtt 21660 attcccacac ccggcatcat gcacgcagca gcgcgcgtca tggctggtca gttgcaccac 21720 gcttcggccc cagcggttct gggtcacctt ggccttgctg ggctgctcct tcaacgcgcg 21780 ctggccgttc tcgctggtca catccatctc caccacgtgg tccttgtgga tcatcaccgt 21840 cccgtgcaga cacttgagct ggccctcgac atcggtgcag ccgtgttccc acagggcgca 21900 gccggtgcac tcccaattct tgtgcgcgat cccgctgtgg ctgaagatgt aaccttgcaa 21960 catgcggccc atcacggtgc taaatgattt actggtgctg aaggtcagtt gcaggccgcg 22020 ggcctcctcg ttcatccatg tctgacacat tttctggaag atctcggtct gctcgggcat 22080 gagcttgtag gcatcgcgca ggccgctgtc gacgcggtag cgttccatca gcacgttcat 22140 ggtatccatg cccttctccc aggacgagac cagaggcaga ctcagggggt tgcgcacgtt 22200 caggataccg ggggtcgcgg gctcgacgat gcgttttccg tcctttcctt ccttcaacag 22260 aaccggcggc tggctgaatc ccactcccac gatcacggcg tcttcctggg gcatctcttc 22320 gtcggggtct accttggtca catgcttggt ctttctggct tgcttctttt ttggagggct 22380 gtccacgggg accacgtcct cctcggaaga cccggagccc acccgctgat actttcggcg 22440 cttggtgggc agaggaggtg gcggcggcga ggggctcctc tcctgctccg gcggatagcg 22500 cgccgacccg tggccccggg gcggagtggc ctctcggtcc atgaaccggc gcacgtcctg 22560 actgccgccg gccattgttt cctaggggaa gatggaggag cagccgcgta agcaggagca 22620 ggaggaggac ttaaccaccc acgagcaacc caaaatcgag caggacctgg gcttcgaaga 22680 gccggctcgt ctagaacccc cacaggatga acaggagcac gagcaagacg caggccagga 22740 ggagaccgac gctgggctcc agcatggcta cctgggagga gaggaggatg tgctgctgaa 22800 acacctgcag cgccagtccc tcatcctccg ggacgcccta gccgaccgga gcgaaacccc 22860 cctcagcgtc gaggagctgt gtcgggccta cgagctcaac ctcttctcgc cgcgcatgcc 22920 ccccaaacgc cagcccaatg gcacatgcga gcccaacccg cgtctcaact tctatcccgt 22980 ctttgcggtc cccgaggccc ttgccaccta tcacatcttt ttcaagaacc aaaagatccc 23040 cgtctcctgc cgcgccaacc gcacccgcgc cgacgcgctc cttactctgg ggcccggcgc 23100 gcgcatacct gatatcgctt ccctggaaga ggtgcccaag atcttcgaag ggctcggtcg 23160 ggacgagacg cgcgcggcga acgctctgaa agaaacagca gaggaagagg gtcacactag 23220 cgccctggta gagttggaag gcgacaacgc caggctggcc gtgctcaagc gcagcgtcga 23280 gctcacccac ttcgcctacc ccgccgtcaa cctcccgccc aaggtcatgc gtcgcatcat 23340 ggatcagctt atcatgcccc acatcgaggc cctcgatgaa agtcaggagc agcgccccga 23400 ggacgcccgg cccgtggtca gcgacgagat gctcgcgcgc tggctcggga cccgcgaccc 23460 ccaggctttg gaacagcggc gcaagctcat gctggccgtg gtcctggtca ccctcgagct 23520 ggaatgcatg cgccgcttct tcagcgaccc cgagaccctg cgcaaggtcg aggagaccct 23580 gcactacact ttcaggcacg gtttcgtcag gcaggcctgc aagatctcca acgtggagct 23640 gaccaacctg gtctcctgcc tggggatcct gcacgagaac cgcctgggcc agaccgtgct 23700 ccactctacc ctgaagggcg aggcgcggcg ggactacgtc cgcgactgcg tctttctctt 23760 tctctgccac acatggcaag cggccatggg cgtgtggcag cagtgtctcg aggacgagaa 23820 cctgaaggag ctggacaagc ttcttgctag aaaccttaaa aagctgtgga cgggcttcga 23880 cgagcgcacc gtcgcctcgg acctggccga gatcgtcttc cccgagcgcc tgagacagac 23940 gctgaaaggc gggctgcccg acttcatgag ccagagcatg ttgcaaaact accgcacttt 24000 cattctcgag cgatctggga tgctgcccgc cacctgcaac gccttcccct ccgactttgt 24060 accgctgagc taccgcgagt gtcccccgcc gctgtggagc cactgctacc tcttgcagct 24120 ggccaactac atcgcctacc actcggacgt gatcgaggac gtgagcggcg aggggctgct 24180 cgagtgccac tgtcgctgca acctgtgctc cccgcatcgc tccctggtct gcaaccccca 24240 gctcctgagc gagacccagg tcatcggtac cttcgagctg caaggtccgc aggagtccac 24300 cgctccgctg aaactcacgc cggggttgtg gacttccgcg tacctgcgca aatttgtacc 24360 cgaagactac cacgcccatg agataaagtt ctttgaggac caatcgcgcc cgcagcacgc 24420 ggatctcacg gcctgcgtca tcacccaggg cgcgatcctc gcccaattgc acgccatcca 24480 aaaatcccgc caagagtttc ttctgaaaaa gggtagaggg gtctacctgg acccccagac 24540 gggcgaggtg ctcaacccgg gtctcccca gcatgccgag gaagaagcag gagccgctag 24600 tggagagat ggaagaagaa tgggacagcc aggcagagga ggacgaatgg gaggagaga 24660 agaggga agaattggga gaggtgggaag agcagccccgt cgccgcacca tccgcgccgg 24720 cagccccgcc ggtcacggat acaacctccg ctccggtcaa gcctcctcgt agatgggatc 24780 aaatgaaggg tgacggtaag cacgagcggc agggctaccg atcatggagg gcccacaaag 24840 ccgcgatcat cgcctgcttg caagactgcg ggggaacat cgcttttgcc cgccgctacc 24900 tgctcttcca ccgcggggtg aacatcccc gcaacgtgtt gcattactac cgtcaccttc 24960 acagctaaga aaaagcaagt aagagagtc gccggaggag gagaggcct gagatcgcg 25020 gcgaacgagc ccttgaccac cagggagctg aggaaccgga tcttccccac tctttatgcc 25080 atttttcagc agagtcgagg tcagcagcaa gagctcaaag taaaaaaccg gtctctgcgc 25140 tcgctcaccc gcagttgctt gtaccacaaa aacgaagaatc agctgcagcg cactctcgaa 25200 gacgccgagg ctctgttcca caagtactgc gcgctcactc ttaaaagacta aggcgcgccc 25260 acccggaaaa aaggcgggaa ttacctcatc gccaccatga gcaaggagat tcccacccct 25320 tacatgtgga gctatcagcc ccagatgggc ctggccgcgg gcgctcccca ggactactcc 25380 acccgcatga actggctcag tgccggcccc tcgatgatct cacaggtcaa cggggtccgt 25440 aaccatcgaa accagatatt gttggagcag gcggcggtca cctccacgcc cagggcaaag 25500 ctcaacccgc gtaattggcc ctccaccctg gtgtatcagg aaatccccgg gccgactacc 25560 gtactacttc cgcgtgacgc actggccgaa gtccgcatga ctaactcagg tgtccagctg 25620 gccggcggcg cttcccggtg cccgctccgc ccacaatcgg gtataaaaac cctggtgatc 25680 cgaggcagag gcacacagct taacgacgag ttggtgagct cttcaatcgg tctgcgaccg 25740 gacggagtgt tccaactagc cggagccggg agatcctcct tcactcccca ccaggcctac 25800 ctgaccttgc agagcagctc ttcggagcct cgctccggag gcatcggaac cctccagttc 25860 gtggaggagt ttgtgccctc ggtctacttc aaccccttct cgggatcgcc aggcctctac 25920 ccggacgagt tcataccgaa cttcgatgca gtgagagaag cggtggacgg ctacgactga 25980 atgtcccatg gtgactcggc tgagctcgct cggttgaggc atctggacca ctgccgccgc 26040 ctgcgctgct tcgcccggga gagctgcgga ctcatctact ttgagtttcc cgaggagcac 26100 cccaacggcc ctgcacacgg agtgcggatc accgtagagg gcaccaccga gtctcacctg 26160 gtcaggttct tcacccagca acccttcctg gtcgagcggg accggggcgc caccacctac 26220 accgtctact gcatctgtcc taccccgaag ttgcatgaga atttttgttg tactctgtgt 26280 gctgagttta ataaaagcta aactcctaca atactctggg atcccgtgtc gtcgcactcg 26340 caacgagatc ttcaacctca ccaaccagac tgaggtaaaa ctcaactgca gaccgggggg 26400 caaatacatc ctctggctct ttgaaaacac ttccttcgca gtctccaacg cctgctgcac 26460 tagtccatga actgattgat taaagcccaa aaaaccaatc aaaccccttc ccattagccc 26520 caaataaaca atcattggaa ataatcattc aataaagatc acttacttga aatctgaaag 26580 tatgtctctg gtgtagttgt ttagcagcac ctcggtaccc tcctcccagc tctggtactc 26640 cagtccccgg cgggcggcga actttctcca caccttgaaa gggatgtcaa attcctggtc 26700 cacaattttc attgttttcc ctctcagatg tcaaagaggc ttcgggtgga agatgacttc 26760 aaccccgtct acccctatga ctacgcgcgg aatcagaata tccccttcct cactcccccc 26820 tttgtctcct ccgatggatt ccaaaacttc ccccctggtg tcctgtcact caaattggct 26880 gacccaatcg ctatcaccaa tggtgatgtc tcactcaagg tgggaggggg acttacttta 26940 caagatggaa ccggaaaact aactatcgac accaagactc ccttgcaagt tgcaaataat 27000 aaattagaac ttgcgtttga tgcaccattg tatgaaaaaa atggaaaact tgctttaaaa 27060 acaggccatg gattagctgt tttaaccaaa gacattggca taccagaatt aattggatcc 27120 cttgtgatct taactggaaa aggaattggg acaggtactg ttgcaggagg aggaactata 27180 gatgtaagac tgggtgatga tggaggttta tcatttgata aaaagggtga tctagtagcc 27240 tggaataaaa aaaatgacag gcgcactttg tggacaacac ctgatccatc cccaaactgc 27300 agagtatcag aagataaaga ttcaaaacta actttaattc ttacaaaatg tggaagtcag 27360 atcctagcaa gtttttcact gcttgtagtc aaagggacgt atgcaactgt tgataaaaat 27420 acaactaata aacaatttag cattaacta ctgtttgatg caaatggaaa gcttaaaagc 27480 gaatcaaacc ttagtggtta ttggaactat agaagtgata atagtgttgt tagtactccc 27540 tatgacaatg cagtgccttt catgccaaat accacggctt atccaaaaat cattaatagt 27600 acaactgatc ctgaaaataa gaaaagttcg gctaaaaaaa ctattgttgg caatgtctac 27660 ctagaaggta atgcgggtca accagtagcc gttgctatta gtttcaataa ggaaactact 27720 gctgactatt caataacatt tgactttgcg tggagcaaag cttatgaaac ccctgtgcct 27780 tttgacacct cctccatgac attctcatat attgcccagg aaaatcaaga caaaaggcgaa 27840 taaagtgttt tgaaatgaac ttatgtatct ttattgattt ttacaccagc acgggtagtc 27900 agtctcccac caccagccca tttcacagtg taaacagtcc tttctccccg gctggcctta 27960 aaaagcatca tatcatgggt aacagacata ttcttaggtg ttatattcca cacggtttcc 28020 tgtcgagcca aacgctcatc agtgatatta ataaactccc cgggcagctc acttaagttc 28080 atgtcgctgt ccagctgctg agccacaggc tgctgtccaa cttgcggttg cttaacgggc 28140 ggcgaaggag aagtccacgc ctacatgggg gtagagtcat aatcgtgcat caggataggg 28200 cggtggtgct gcagcagcgc gcgaataaac tgctgccgcc gccgctccgt cctgcaggaa 28260 tacaacatgg cagtggtctc ctcagcgatg attcgcaccg cccgcagcat aaggcgcctt 28320 gtcctccggg cacagcagcg caccctgatc tcacttaaat cagcacagta actgcagcac 28380 agcaccacaa tattgttcaa aatcccacag tgcaaggcgc tgtatccaaa gctcatggcg 28440 gggaccacag aacccacgtg gccatcatac cacaagcgca ggtagattaa gtggcgaccc 28500 ctcataaaca cgctggacat aaacattacc tcttttggca tgttgtaatt caccacctcc 28560 cggtaccata taaacctctg attaaacatg gcgccatcca ccaccatcct aaaccagctg 28620 gccaaaacct gcccgccggc tatacactgc agggaaccgg gactggaaca...

Claims

1. An isolated nucleic acid encoding a chimeric adenoviral capsid, wherein the chimeric adenoviral capsid comprises: a fiber polypeptide consisting of the amino acid sequence of SEQ ID NO: 11; a hexon polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; and a penton polypeptide consisting of the amino acid sequence of SEQ ID NO:

13.

2. A vector comprising the isolated nucleic acid of claim 1.

3. The vector of claim 2, wherein the vector is an adenoviral vector.

4. The vector of claim 3, wherein the adenoviral vector further comprises a transgene. The vector of claim 4 , wherein the adenoviral vector further comprises an E1 deletion.

6. The vector of claim 4, wherein the adenoviral vector further comprises an E3 deletion.

7. The vector of claim 4, wherein the adenoviral vector is a chimeric adenoviral vector comprising one or more adenoviral nucleic acid sequences from at least one of human adenovirus-4, human adenovirus-5, human adenovirus-26, or human adenovirus-35.

8. The vector of claim 7, wherein the adenoviral vector comprises human adenovirus-5 (HAdV-5) E4 orf6.

9. The vector of claim 4, wherein the adenoviral vector comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 9, and SEQ ID NO:

10.

10. The vector of claim 4, wherein the transgene is located at the E1 deletion, the E3 deletion, and / or adjacent to the right inverted terminal repeat (rITR).

11. A recombinant cell comprising the vector according to any one of claims 2 to 10.

12. A method for producing a vector, comprising: (a) growing the recombinant cell of claim 11 under conditions for producing an adenoviral vector; and (b) isolating the vector from the recombinant cell.

13. An immunogenic composition comprising the adenoviral vector according to any one of claims 3 to 10 and a pharmaceutically acceptable carrier.

14. Use of the immunogenic composition of claim 13 in the preparation of a vaccine for inducing an immune response in a subject in need thereof.

15. A method of producing a vaccine, the method comprising combining the adenoviral vector of any one of claims 3 to 10 with a pharmaceutically acceptable carrier.

16. Use of the vector of any one of claims 4 to 10 in the preparation of a medicament for treating or preventing a disease or condition in a subject in need thereof, wherein expression of the transgene in the subject in need thereof treats or prevents the disease or condition.

17. The use of claim 16, wherein the subject is contacted with the vector, and wherein contacting the subject with the vector comprises isolating cells from the subject, and contacting the cells with the vector.

18. The use of claim 16 or 17, wherein the subject in need thereof is a human subject.

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