Abstract:
Disclosed are replicatable viral DNA vectors encoding a site-specific DNA-altering enzyme and a DNA target recognized by the enzyme, the enzyme selectively converting, in a cell expressing the enzyme, the DNA vector to a rearranged form. The invention further relates to methods for assembling recombinant adenoviral DNAs. These methods include the steps of: (a) providing a first linearized DNA vector including a restriction site and a cos site and a second linearized DNA vector including the restriction site, an adenoviral nucleic acid molecule, and a cos site; and (b) ligating the first and second linearized DNA vectors, the ligation assembling a recombinant adenoviral DNA.

Description:
CROSS REFERENCE TO RELATED APPLICATIONS  
       [0001]    This application is a continuation of International Application No. PCT/US01/27682, filed Sep. 7, 2001, which claims benefit of U.S. provisional application Nos. 60/246,904, and 60/231,053, filed Nov. 8, 2000, and Sep. 8, 2000, respectively, all of which are hereby incorporated by reference in their entirety. 
     
    
     
       BACKGROUND OF THE INVENTION  
         [0002]    The invention relates to DNA vectors.  
           [0003]    Mammalian cell expression vectors based on DNA viruses have been widely discussed as gene delivery vehicles for genetic therapy. Among the different DNA viruses proposed for this purpose have been adenoviruses, baculovirus, Epstein Barr virus, and herpes simplex virus. In addition other smaller viruses that have an intranuclear phase in which the viral genome is present as a double stranded DNA, such as retroviruses and parvoviruses, have been proposed as gene delivery vehicles.  
           [0004]    Adenoviral vectors (AdV), for example, have a recognized potential for gene delivery, founded in their broad host range, robust growth in culture, and capacity to infect mitotically quiescent cells (Graham and Prevec, Manipulation of adenovirus vectors, p. 109-128, In E. J. Murray (ed.), Methods in Molecular Biology, vol. 7, Humana, Clifton, N.J., 1991; Trapnell and Gorziglia, Curr. Opin. Biotechnol. 5:617-625, 1994). AdV can be propagated in a helper cell line, 293, a human embryonic kidney cell line transformed by adenovirus type 5 (Graham et al., J. Gen. Virol. 36:59-72, 1994). 293 cells express the viral E1 gene products (E1a and E1b) that are the master regulatory proteins for subsequent viral gene expression. E1 deleted viruses can propagate in 293 cells, but not in other cells. Although it would be expected that E1 deleted viruses lack the machinery to express viral genes, several studies have demonstrated that cellular E1-like components can stimulate viral gene expression (Imperiale et al., Mol. Cell. Biol. 4:867-74, 1984; Onclercq et al., J. Virol. 62:4533-7,1988; Spergel et al., J. Virol. 66:1021-30, 1992). The expression of these viral genes results in the relatively rapid elimination of transduced cells in vivo as a result of cytotoxic T cell responses (Yang et al., Immunity 1:433-42, 1994; Yang et al., Gene Ther. 3:137-44, 1996; Yang et al., J. Virol. 69:2004-15, 1995).  
           [0005]    Thus attention has been focused on eliminating the remaining vestiges of viral expression. Viral genes that have been deleted for this purpose include the gene for E4 proteins (Armentano et al., Hum. Gene Ther. 6:1343-53, 1995; Kochanek et al., Proc. Natl. Acad. Sci. USA 93:5731-6, 1996; and Yeh et al., J. Virol. 70:559-565, 1996), DNA binding protein (Engelhardt et al., Proc. Natl. Acad. Sci. USA 21:6196-6200, 1994; and Gorziglia et al., J. Virol. 70:4173-8, 1996), DNA polymerase (Amalfitano et al., J. Virol. 72:926-33, 1998), and the preterminal protein (Schaack et al., Proc. Natl. Acad. Sci. USA 93:14686-91, 1996). The most aggressive approach has been the creation of helper virus-dependent vectors that lack all viral genes (Hardy et al., J. Virol. 71:1842-9, 1997; Kochaneketal., Proc. Natl. Acad. Sci. USA 93:5731-6, 1996; Lieber et al., J. Virol. 70:8944-60, 1996; Mitani et al., Proc. Natl. Acad. Sci. USA 92:3854-8, 1995; and Parks et al., Proc. Natl. Acad. Sci. USA 93:13565-13570, 1996). These vectors have high capacity, evoke reduced cellular immune responses and show prolonged expression in vivo (Morsy et al., Proc. Natl. Acad. Sci. USA 95:7866-71, 1998). However to deploy these viruses on the scale required for human clinical application presents major challenges because a cesium chloride (CsCl) gradient is needed to remove the helper virus.  
         SUMMARY OF THE INVENTION  
         [0006]    In one aspect, the invention features a replicatable viral DNA vector encoding a site-specific DNA-altering enzyme and a DNA target recognized by the enzyme, the enzyme selectively converting, in a cell expressing the enzyme, the DNA vector to a rearranged form.  
           [0007]    In preferred embodiments, the rearranged form includes an autonomously replicating episome and a linear DNA product. In other preferred embodiments, the vector comprises adenoviral DNA.  
           [0008]    In yet other preferred embodiments, the vector includes a genetically-engineered recombination site (such as a target of Cre or FLP). Preferably, such a recombination site includes a recognition sequence of a site-specific DNA altering enzyme.  
           [0009]    In another preferred embodiment, the site-specific DNA altering enzyme is a recombinase (such as Cre or FLP) or an integrase. Preferably, such an enzyme is functional in a mammalian cell. Preferred embodiments of the vector also include an origin of replication that functions in a mammalian cell (such as an Epstein Barr Virus replicon). Moreover, the vector typically includes a gene of interest (such as a therapeutic gene that encodes a protein or polypeptide or an RNA product).  
           [0010]    In another aspect, the invention features a method for assembling a recombinant adenoviral DNA. The method, in general, includes the steps of: (a) providing a first linearized DNA vector comprising a restriction site and a cos site and a second linearized DNA vector comprising the restriction site, an adenoviral nucleic acid molecule, and a cos site; and (b) ligating the first and second linearized DNA vectors, the ligation assembling a recombinant adenoviral DNA.  
           [0011]    In preferred embodiments, the first linearized DNA vector comprises a selectable marker (such as a gene encoding a polypeptide that confers, on a host cell expressing such a polypeptide, resistance to an antibiotic). In other preferred embodiments, the first linearized DNA vector includes an adenoviral left-end inverted terminal repeat, a gene of interest, or both. In still other preferred embodiments, the second linearized DNA vector includes a selectable marker. Preferably, the second linearized DNA vector includes an adenoviral right-end inverted terminal repeat.  
           [0012]    The method further includes packaging the assembled adenoviral DNA into a phage and infecting a host cell. Typically the first and second linearized DNAs include cosmid vector DNA. In addition, such adenoviral DNA is typically flanked by cleavage sites (such as intron endonuclease cleavage sites).  
           [0013]    In another aspect, the invention features an adenovirus producer cell having a nucleic acid molecule that expresses a dominant negative site-specific DNA-altering enzyme. In preferred embodiments, the site-specific DNA altering enzyme is a dominant negative recombinase (for example, a Cre recombinase such as CreY324C or a Flp recombinase). Exemplary adenovirus producer cells include, without limitation, 293 human embryonic kidney cells, per.C6 cells, and N52 cells.  
           [0014]    In yet another aspect, the invention features a vector comprising, in the 5′ to 3′ direction, a first genetically engineered cis-acting target recognized by a site-specific DNA altering enzyme; a gene of interest; a lineage-specific gene promoter; a second genetically engineered cis-acting target recognized by a site-specific DNA altering enzyme; and a nucleic acid molecule encoding a site-specific DNA altering enzyme.  
           [0015]    In still another aspect, the invention features a vector including, in the 5′ to 3′ direction, a first genetically engineered cis-acting target recognized by a site-specific DNA altering enzyme; a gene of interest; a bi-directional promoter, comprising a second genetically engineered cis-acting target recognized by a site-specific DNA altering enzyme; and a nucleic acid molecule encoding a site-specific DNA altering enzyme.  
           [0016]    In related aspects, the invention features a method of gene therapy including the administration to a patient in need of gene therapy a therapeutically effective amount of the vector of the invention, which is expressed in the patient. The invention further relates to a population of cells transfected with the vector of the invention.  
           [0017]    Accordingly, the invention further relates to the use of a recombinant viral vector or use of a recombinant viral particle for gene therapy. Such vectors and viral particles may be introduced either in vitro into a host cell removed from the patient, or directly in vivo, into the body to be treated, according to standard methods known in the art.  
           [0018]    The invention also relates to a pharmaceutical composition that includes a therapeutically effective amount of a recombinant viral vector or viral particle prepared according to the methods disclosed herein, in combination with a vehicle that is acceptable from a pharmaceutical standpoint. Such a pharmaceutical composition may be prepared according to the techniques commonly employed and administered by any known administration route, for example systemically (in particular, by intravenous, intratracheal, intraperitoneal, intramuscular, subcutaneous, intratumoral, or intracranial routes) or by aerosolization or intrapulmonary administration.  
           [0019]    One skilled in the art will appreciate that suitable methods of administering a vector (particularly an adenoviral vector) of the present invention to an animal for purposes of gene therapy, chemotherapy, and vaccination are available, and, although more than one route can be used for administration, one particular route may provide a more immediate and more effective reaction than another. Pharmaceutically acceptable excipients also are well known to those who are skilled in the art, and are readily available. The choice of excipient will be determined, in part, by the particular method used to administer the recombinant vector or particle. Accordingly, there are a wide variety of suitable formulations for use in the context of the present invention.  
           [0020]    By “recombinant DNA vector” is meant a DNA sequence containing a desired sequence (such as a gene of interest) and an appropriate regulatory element(s) necessary for the expression of the operably linked sequence in a particular host organism (such as a mammal).  
           [0021]    By “operably linked” is meant that a gene and a regulatory element(s) are connected to permit gene expression when the appropriate molecules (for example, transcriptional activator proteins) are bound to the regulatory sequence(s).  
           [0022]    By “regulatory element” is meant a genetic element that controls some aspect of the expression of a nucleic acid sequence. For example, a promoter is a regulatory element that facilitates the initiation of transcription of an operably linked coding region. Other genetic regulatory elements include, without limitation, splicing signals, polyadenylation signals, and termination signals. For example, transcriptional regulatory elements in eukaryotes include promoter and enhancer elements. Promoters and enhancers include arrays of DNA sequences that interact directly or indirectly with cellular proteins involved in transcription. Promoter and enhancer elements have been isolated from a variety of eukaryotic sources including genes in mammalian cells and viruses.  
           [0023]    By “transfection” is meant the introduction of foreign DNA into eukaryotic cells. Transfection is typically accomplished by a variety of means known in the art including, without limitation, calcium phosphate-DNA co-precipitation, DEAE-dextran-mediated transfection, electroporation, microinjection, liposome fusion, lipofection, protoplast fusion, and biolistics.  
           [0024]    By “stably transfected” is meant the introduction of foreign DNA into the genome of the transfected cell. In general, transfer and expression of transgenes in mammalian cells are now routine practices to those skilled in the art, and have become major tools to carry out gene expression studies and to generate vectors useful in gene therapy.  
           [0025]    By “gene of interest” is meant a gene inserted into a vector whose expression is desired in a host cell. Genes of interest include, without limitation, genes having therapeutic value, as well as reporter genes. A variety of such genes are useful in the invention, including genes of interest encoding a protein, which provides a therapeutic function. In addition, the gene of interest, if a therapeutic gene, can render its effect at the level of RNA, for instance, by encoding an antisense message or ribozyme, a protein which affects splicing or 3′ processing (e.g., polyadenylation), or it can encode a protein which acts by affecting the level of expression of another gene within the cell (i.e., where gene expression is broadly considered to include all steps from initiation of transcription through production of a processed protein), for example, by mediating an altered rate of mRNA accumulation, an alteration of mRNA transport, and/or a change in post-transcriptional regulation.  
           [0026]    By “reporter gene” is meant a gene sequence that encodes a reporter molecule (including an enzyme). A “reporter molecule” is detectable in any detection system, including, but not limited to, enzyme (e.g., ELISA, as well as enzyme-based histochemical assays), fluorescent, radioactive, and luminescent systems. Exemplary reporter gene systems include the  E. coli  beta-galactosidase or glucuronidase genes, green fluorescent protein (GFP), blue fluorescent protein (BFP), the human placental alkaline phosphatase gene, and the chloramphenicol acetyltransferase (CAT) gene; other reporter genes are known in the art and may be employed as desired.  
           [0027]    By “transgene” is meant any piece of DNA that is inserted by artifice into a cell, and becomes part of the genome of the organism that develops from that cell. Such a transgene may include a gene that is partly or entirely heterologous (i.e., foreign) to the transgenic organism, or may represent a gene homologous to an endogenous gene of the organism.  
           [0028]    By “transgenic” is meant any cell that includes a DNA sequence that is inserted by artifice into a cell, and becomes part of the genome of the organism that develops from that cell.  
           [0029]    By “polypeptide” is meant any chain of amino acids, regardless of length or post-translational modification (for example, glycosylation or phosphorylation).  
           [0030]    By “derived from” is meant isolated from or having the sequence of a naturally occurring sequence (e.g., a cDNA, genomic DNA, synthetic, or combination thereof).  
           [0031]    By “nucleic acid” is meant a polynucleotide (DNA or RNA).  
           [0032]    By “gene” is meant any nucleic acid sequence coding for a protein or an RNA molecule.  
           [0033]    By “gene product” is meant either an untranslated RNA molecule transcribed from a given gene or coding sequence (such as mRNA or antisense RNA) or the polypeptide chain translated from the mRNA molecule transcribed from the given gene or coding sequence. Nucleic acids according to the invention can be wholly or partially synthetically made, can comprise genomic or complementary DNA (cDNA) sequences, or can be provided in the form of either DNA or RNA.  
           [0034]    The presently claimed invention affords a number of advantages. For example, applicants&#39; gene therapy vehicles, particularly those based on recombinant adenoviruses, minimize the propensity of the vectors to activate host immune surveillance, and thereby maximize the persistence for the DNA transduced. The invention therefore facilitates the development of gene delivery vectors designed to enhance persistence of virally delivered genes and evade the cellular immune response by severing the connection between the sole adenoviral enhancer and the sequences encoding potentially antigenic viral proteins.  
           [0035]    As described in more detail below, the mechanism by which this is accomplished differs significantly from any other previous approaches. For example, to reduce the immunogenicity of vectors it is widely acknowledged that some intervention, such as the removal of key genes, or the prevention of their expression in the cells targeted for therapy, is important; however, many related approaches are directed at the host and have generally focused on the selective induction of tolerance to adenoviral antigens, or similar strategies directed at inducing a temporally restricted or antigen-specific compromise of the immune system.  
           [0036]    In addition, the poor persistence of transduced DNA appears to be due in part to immunological rejection of transduced cells and to the inability of the viral DNA to replicate, a feature generally inherent in the design of adenoviral vectors, but one which is not associated with applicants&#39; claimed gene therapy vehicles.  
           [0037]    Moreover, some contemporary adenoviral vectors are designed to propagate in specific host cells which provide essential replication factors in trans. These vectors are typically based on cell lines which express the master regulatory proteins of the E1 complex, which are required for induction of adenoviral DNA replication. In cells expressing E1 genes, the best studied of which is a human embryonic kidney cell line transformed by DNA from human adenovirus 5 (called HEK293, or simply 293), viruses lacking E1 genes propagate well. Such viruses do not propagate on cell lines which do not express E1, and do not generally propagate well in the target cells to which the therapeutic gene is to be delivered. Cells transduced with E1-deleted adenovirus vectors also do not express high levels of viral genes in the absence of E1. However, the weak residual expression that remains in such vectors appears to be sufficient to induce cellular immune responses that contribute to the destruction of the transduced cells.  
           [0038]    In addition, the gene therapy vectors claimed herein are hybrid vectors capable of self-rearrangement to form circular and linear DNA products. The linear DNA has a compromised ability to express adenoviral genes, and therefore has a lower immunological profile. And the circular DNA behaves like a mammalian plasmid, encoding the gene of interest and persisting by autonomous replication in the nucleus.  
           [0039]    For example, the circularization of an adenoviral vector via the action of Cre recombinase beneficially places a gene of interest (for example, a therapeutic gene) on a self-replicating episome. Vector circularization occurs in a tissue-targeted manner, for example, as a result of the activation of a synthetic liver-specific promoter upstream of the recombinase Cre. Once circularized, the EBV replicon in the episome confers improved persistence on the therapeutic gene as detected by reporter gene expression and direct assay for the presence of vector DNA sequences.  
           [0040]    Furthermore, the invention eliminates the requirement for a helper virus, thus avoiding two potential limitations of that system. First, the continuous expression of Cre recombinase may lead to toxicity in host cells, either as a direct consequence of the protein&#39;s activity or via its immunogenicity. Second, the Cre helper virus may itself produce antigenic viral proteins that contribute to the immunologic elimination of infected host cells. In contrast, the self-resolving adenovirus/EBV vector system disclosed herein advantageously provides no alternative source of viral proteins, and Cre expression is terminated upon rearrangement.  
           [0041]    In addition, the invention described herein provides tools for analyzing the roles of the enhancer in viral gene regulation and virus growth.  
           [0042]    The invention also provides a convenient general system for creating recombinant adenoviruses, which increase their attractiveness as gene transduction tools for basic research. The system, for example, employs two conventional plasmid vectors and a γ phage packaging step. The entire recombinant AdV genome is assembled into a single cosmid that is easily amplified in  E. coli . The use of intron endonuclease recognition sequences flanking the ITRs enhances virus production while simplifying insertion of therapeutic gene sequences into the pLEP shuttle plasmid. The convenience of this vector system has facilitated the construction of over two hundred recombinant viruses to date.  
           [0043]    Other embodiments and advantages of the invention will be apparent from the detailed description thereof, and from the claims. 
       
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
       [0044]    The application file contains drawings executed in color (FIGS. 7A, 7B,  7 C,  10 C,  11 ,  15 A,  17 , and  21 ). Copies of this patent or patent application with color drawings will be provided by the U.S. Patent and Trademark Office upon request and payment of the necessary fee.  
         [0045]    [0045]FIG. 1A is a schematic diagram of the structure of an adenoviral type A vector and its fate in a target cell. “enh” refers to the Ad2 enhancer; “GFP” refers to the marker gene green fluorescent protein; “EBV” refers to the Epstein Barr Virus replicon; “TetO 7 ” refers to a heptamer of Tet operator; “TetR” refers to the Tet repressor; “VP 16” refers to the viral protein 16 of Herpes simplex virus, SD refers to the splice donor site; and SA refers to the splice acceptor site.  
         [0046]    [0046]FIG. 1B is a schematic diagram of the structure of an adenoviral type B vector and its fate in a target cell. enh refers to the Ad2 enhancer; GFP refers to the marker gene green fluorescent protein; EBV refers to the Epstein Barr Virus replicon; SD refers to the splice donor site; and SA refers to the splice acceptor site.  
         [0047]    [0047]FIG. 2A shows a schematic diagram of the pLEP cosmid polylinker region and its position relative to the adenoviral left ITR. The adenovirus enhancer/packaging sequence (ψ) is boxed.  
         [0048]    [0048]FIG. 2B is a schematic diagram showing the generation of a single cosmid encoding the AdV genome by the direct ligation of two smaller plasmids. A gene expression unit, CMVGFP, was inserted into the pLEP cosmid at the polylinker region. pLEP and pREP cosmids were digested with an intron endonuclease (PI-PspI), ligated, and packaged in vitro to generate pAd2CMVGFP. This DNA was then digested with another intron endonuclease (1-CeuI) to expose the ITRs at both ends of the viral genome. Finally, cosmid digestion mixtures were transfected into 293 cells. Plaques generated by recombinant viruses are detected in 7-10 days.  
         [0049]    [0049]FIG. 3A shows the restriction analysis of cosmids carrying the full length AdV DNA showing uniform generation of the desired vector DNA. 2 μg DNA samples from four pAd2-7CMVGFP colonies were digested with Bgl II, resolved on a 1% agarose gel and stained with ethidium bromide. The predicted sizes of the DNA fragments are: 13261, 7684, 5228, 5088, 2284, 1757, 1549, 1270, 351, and 275 base pairs (bp). The 5228 and 5088 fragments appear as a doublet, and the 351 and 275 bp fragments are too small to be seen on the gel.  
         [0050]    [0050]FIG. 3B shows the release of the recombinant Ad DNA from cosmids by I-CeuI digestion. 2 μg of pAd2-7CMV DNA from two clones was digested with I-CeuI. The position of the released recombinant AdV DNA and the vector fragments of approximately 35 kb and 5 kb, respectively, are indicated.  
         [0051]    [0051]FIG. 4A shows the appearance of plaques in 293 cells transfected with 10 μg of pIAdGFPB with no ITR exposed (undigested), one ITR exposed (BsaBI or I-CeuI), or both ITRs exposed (BsaBI plus I-CeuI). Values represent the mean plaque counts per dish and the time required for plaque development in 293 cells from three separate experiments. “I” designates I-CeuI; and “B” designates Bsa BI.  
         [0052]    [0052]FIG. 4B shows the viral titers obtained from plaques that were allowed to grow over 10 days after transfection. Viruses were harvested and the titer of each virus stock was determined by a GFP based semi-quantitative titration procedure described herein. Values represent the mean±SE of three independent determinations.  
         [0053]    [0053]FIG. 5 is a schematic diagram showing a linear AdV that resolves into a circular episome. The elements involved in the self-directed rearrangement of the vector are shown schematically in pLEP1BHCRGFP/EBV and in the corresponding AdV. Starting from the left ITR, the elements are shown as following sequence: left ITR, 147 bp; first 34 bp loxP site; 185 bp enhancer/packaging signal; 64 bp splicing acceptor (SA) from EF1α gene first intron; 720 bp GFP cDNA; 230 bp SV40 poly(A); 1.7 kb TK-EBNA-1/OriP; 970 bp HCR12 promoter; 1 kb EF1α gene first intron containing splicing donor (SD) and acceptor (SA) sites with the second loxP site inserted at 64 bp upstream of the 3′end; 1.2 kb Cre gene tagged with AU1 and a nuclear localization signal; ˜120 bp poly(A) signal and PI-PspI site. After infection of liver cells, the HCR12 promoter drives the expression of Cre which results in the cleavage of the two loxP sites. This results in the circularization of the fragment containing the EBV replicon. The excision severs the connection between the enhancer/packaging signals and the remainder of the AdV genome. The Cre gene becomes promoterless and is left on the AdV genome fragment. After excision, the HCR12 promoter drives the expression of the GFP reporter gene. The EBV replicon maintains the excised circle as an episome in host cells.  
         [0054]    [0054]FIG. 6A is a schematic representation of the loxP sites and EBNA-1 locations in the AdV genome. The relevant Bgl II site is also shown.  
         [0055]    [0055]FIG. 6B shows the time course of rearrangement in HepG2 and Hela cells at an equal multiplicity of infection (moi) of 1,000 particles per cell. Cells were infected with Ad2HCRGFP/EBV viruses for 2 hours at 37° C. Hirt DNA samples were extracted from the cells. ˜5 μg of Hirt DNA samples were digested with Bgl II, fractionated on a 1% agarose gel, and analyzed by Southern blot techniques using a  32 P-labeled EBNA-1 fragment as the hybridization probe.  
         [0056]    [0056]FIG. 6C shows the DNA blot results obtained from Hela cells infected at a moi of 10,000; and HepG2 at 1,000. The upper bands (4915 bp) represent the circularized DNA fragments whereas the lower bands (3162 bp) represent the non-circularized AdV.  
         [0057]    [0057]FIG. 7A shows green fluorescent protein (GFP) expression in liver and non-liver cells infected with the Ad2HCRGFP/EBV viruses. Cells were cultured in 35 mm dishes and infected with the Ad2HCRGFP/EBV virus at desired moi. HepG2 cells were infected with 1,000 particles per cell, whereas Hela cells were infected with a moi of 10,000. GFP expression was examined at the indicated time points after infection. Fluorescent cells were photographed using an Olympus SC35 mm camera mounted on an Olympus IX70 fluorescent microscope, at 200× magnification, using a filter with peak excitation and emission wavelengths of 450 nm and 510 nm, respectively.  
         [0058]    [0058]FIG. 7B shows the expression of GFP in HepG2, Hela, A431, and HT29 cells. Cells were seeded in 35 mm dishes and infected with the Ad2HCRGFP/EBV virus at a moi of 10,000 particles per cell. GFP expression was examined at 72 hours after infection.  
         [0059]    [0059]FIG. 7C shows the expression of GFP in human primary hepatocytes. These cells were photographed under bright field (left) and fluorescent conditions (right).  
         [0060]    [0060]FIG. 8A shows the results of RT-PCR that was performed to detect the tripartite leader sequence (upper panel) for virus late gene expression; and PCR was performed in the DNA samples for detection of the AdV genomes. The specific target sequences are described in detail infra. PCR analyses of adenovirus late gene expression in cells infected with the first generation AdVs or the self-resolving Ad2HCRGFP/EBV was analyzed. HepG2 cells were cultured in 35 mm dishes and infected with increasing moi (0, 10, 100, 1000, 10,000, and 100,000) of adenoviral vectors. RNA and DNA were isolated in parallel from the cells at 72 hours after infection.  
         [0061]    [0061]FIG. 8B shows a summary of quantitative RT-PCR and PCR results. Each determinant was the average of three experiments.  
         [0062]    [0062]FIG. 9A is a schematic diagram depicting the deletion analysis of the OriP and EBNA-1 regions of the EBV replicon. Structures of the deletions in EBNA-1 and OriP are schematically represented. Elements considered important for episomal maintenance are indicated. “FR” refers to the family of repeats; “DS” designates the region of dyad symmetry; “LR1” refers to the so-called linker region 1; “GA” refers to gly-ala repeats; “LR2” refers to linker region 2; and “Dimerization” designates the dimerization domain.  
         [0063]    [0063]FIG. 9B is a graph depicting fractions of GFP positive cells carrying the EBV replicons represented in FIG. 9A.  
         [0064]    [0064]FIG. 10A shows the positions and identities of Cre mutants tested for their dominant negative Cre activities.  
         [0065]    [0065]FIG. 10B is a schematic diagram of the substrate Cre plasmid (ad2239) used to test dominant negative functions of Cre mutants.  
         [0066]    [0066]FIG. 10C shows GFP expression in cells cotransfected with the substrate Cre plasmid (ad2239) and the indicated Cre mutants.  
         [0067]    [0067]FIGS. 10D and 10E show Cre mutants tested for their ability to inhibit rearrangement. Only those showing the strongest inhibitory activities were retested in FIG. 10E. GFP intensity was normalized to that of cells in the absence of inhibition.  
         [0068]    [0068]FIG. 11 shows GFP expression in 293 TetON cells and #17 cells transfected with ad2239. The ability of #17 cells to inhibit Cre activity is demonstrated by the weak GFP signal in cells treated with 2 μM doxycycline.  
         [0069]    [0069]FIG. 12 is a schematic diagram depicting the tetracycline mediated auto-regulatory circuit.  
         [0070]    [0070]FIGS. 13A and 13B show the effects of different basal elements on synthetic TetO promoter activity. FIG. 13A shows a schematic diagram of the components of various auto-regulatory synthetic TetO promoters. FIG. 13B shows a comparison of the strength of auto-regulatory synthetic TetO promoters bearing different basal elements, in the presence and absence of tetracycline, using GFP as a marker in HepG2 cells.  
         [0071]    [0071]FIG. 14 shows the structure of a Cre substrate plasmid (ad2265). The promoter, Ef1α, and the gene, BFP, are interrupted by two loxP sites, which can be joined by Cre-mediated recombination. “PA” stands for poly A; “BFP” for blue fluorescent protein.  
         [0072]    [0072]FIGS. 15A and 15B show the estrogen regulation of Cre recombinase activity. 293 cells infected with type B virus, AD121.5, in which the Cre enzyme is fused with estrogen ligand binding domain at both the N- and C-termini were cultured in the presence or absence of 1 μM estrogen. Cre-mediated rearrangement in the presence of estrogen is shown in FIG. 15A, whereas blot analysis of extrachromosomal DNA from the same cells is shown in FIG. 15B. “L” represents the position corresponding to the unrearranged adenoviral DNA; and “C” represents the position corresponding to the circular form of DNA.  
         [0073]    [0073]FIG. 16 shows the rearrangement of adenoviral sequences in vivo. Extrachromosomal DNA from the livers of Rag-2 mice sacrificed 2.5 hours post injection of type A adenovirus, AD102.7, was analyzed by DNA blot. “L” represents the size corresponding to linear adenoviral DNA; and “C” represents the size corresponding to rearranged circular DNA.  
         [0074]    [0074]FIG. 17 is a photomicrograph depicting high level GFP expression in Rag2 mouse hepatic tissues 48 hours post type A adenovirus (AD102.7) injection.  
         [0075]    [0075]FIGS. 18A and 18B show schematic diagrams of the structures of adenoviral vectors and their fates in target cells. “enh” refers to Ad2 enhancer; “GFP” refers to green fluorescent protein; “EBV” refers to Epstein Barr Virus replicon; “TetO 7 ” refers to heptamer of Tet operator; “TetR” refers to Tet repressor; “VP16” refers to transcriptional activator domain from HSV protein 16; “SD” refers to splice donor site; and “SA” refers to splice acceptor site.  
         [0076]    [0076]FIG. 19A shows the structure of a FLP substrate plasmid, ad2879. The promoter, Ef1α, and the gene, GFP, are interrupted by 2 FRT sites, which can be joined by the FLP-mediated recombination. “PA” stands for poly A; “BFP” for blue fluorescent protein.  
         [0077]    [0077]FIG. 19B shows the structure of a cre substrate plasmid, ad2204.  
         [0078]    [0078]FIG. 20 shows the structures of several FLPe anti-sense plasmids.  
         [0079]    [0079]FIG. 21 is a panel of photomicrographs showing inhibition of FLP enzyme activity by anti-sense FLP. 293 cells were transfected with FLP substrate (FIG. 12) and plasmids indicated in each photo. High GFP intensity indicate the higher expression of FLP and less inhibition by the anti-sense expressed.  
         [0080]    [0080]FIG. 22 shows a schematic diagram of FRT/Cre and loxP/FLP adenovirus. 
     
    
     DETAILED DESCRIPTION  
       [0081]    Described herein are systems for the regulated self-rearrangement of DNA vectors, for example, gene therapy vectors. Such regulated self-rearrangement has the potential to prevent unwanted expression of vector genes not required for a therapeutic effect, and to allow the stable association of the therapeutic gene with the target cell.  
         [0082]    The essential elements of the regulated DNA rearrangement system are a gene that encodes one or more proteins that induce DNA rearrangement, a method for regulating the activity of those proteins or their abundance, and a target DNA sequence on which those proteins act. Particularly desirable are methods for regulating the activity of the proteins or their abundance which can be easily carried out on an intact organism, such as administration or withdrawal of a drug, hormone, or environmental stimulus such as heat or irradiation, which induces the activity or abundance of the proteins which cause DNA rearrangement.  
         [0083]    Especially desirable are regulated DNA rearrangement systems in which all of the components can be delivered in a single vector. An example of this is a virus that bears both the cis-acting sequences for DNA rearrangement as well as the protein or proteins that act on those sequences, and the regulatory apparatus which controls the activity or abundance of those proteins. However, it is not necessary that the different elements be encoded in a single nucleic acid.  
         [0084]    The important elements of this strategy are: the compromise of vector gene function by regulated rearrangement of DNA topology, the generation of plasmid circles from vector DNA in a regulated manner, and the removal of enhancer or promoter elements from the vector DNA by regulated excision. It is also important that the circular DNA generated by site-specific recombination possesses a mechanism for stable association with the host genome in some form, here conferred by the EBV replicon. In other embodiments, the circular DNA might possess the ability to direct its integration into the host chromosomes by a site-specific integration. Site-specific integration into the host chromosomes may also be generated by the action of a regulated site-specific recombinase on a linear template without passing through a circular intermediate.  
         [0085]    Also described herein is one particular self-rearranging vector that begins as a hybrid adenovirus vector which is capable of converting itself into two unlinked molecules, a circular and a linear DNA. After this event the linear DNA product is deleted for two important cis-acting sequences: the packaging signals, which are required for insertion of the viral DNA into the viral capsid, and the enhancer, which increases the expression of other promoters encoded in the viral DNA. The remaining linear DNA is thereby compromised in its ability to express adenoviral genes, endowing the vector with a lower immunological profile. The circular DNA generated by the excision event is a mammalian cell plasmid which has the capacity to persist by autonomous replication in the nucleus. This capacity is encoded in genetic elements derived from the Epstein Barr virus (EBV). A schematic diagram of such a vector is illustrated in FIG. 5.  
         [0086]    Epstein Barr virus is a human herpes virus which is the etiologic agent of infectious mononucleosis and which has been implicated in the genesis of Burkitt&#39;s lymphoma, a B cell neoplasm, and is thought to be a predisposing factor for some forms of nasopharyngeal carcinoma. Approximately 85% of the adult Western population has a persistent population of B cells which contain a circular latent form of the viral-genome, maintained in cells by the action of Epstein Barr Nuclear Antigen 1 (EBNA1), a DNA replication protein that acts on the viral latent phase origin of replication, OriP. EBNA1 in and of itself is not thought to promote neoplasia; current thinking places greater weight on the actions of the EBNA2 proteins and LMP, latent membrane protein, in the inception of EBV-associated neoplasm.  
         [0087]    Mammalian cell plasmids have been created which bear the EBNA1 gene and OriP. In nonrodent cells, these plasmids persist by replication with each transit of the cell cycle. Multiple transcription units can be borne by these plasmids, allowing regulated expression of diverse gene products.  
         [0088]    Preferred adenoviral vectors, shown in FIGS. 1A and 1B, are linear forms of an EBV plasmid flanked by loxP sites, cis-acting sequences required for site-specific recombination directed by the bacteriophage P1 cre protein. To prepare an adenovirus bearing both the cre protein and loxP sites, it is necessary to insure that the cre protein is not expressed while the vector is being propagated in 293 cells. To lower the immunological profile of the vector, it is also desirable that the cre protein not be expressed after the vector delivered its payload to the target cell and the cre protein performed its function.  
         [0089]    To accomplish these objectives, two general approaches have been developed for the production of adenoviral chromosomes that circularize following the regulated expression of site-specific recombinases. In each case, the vector is engineered to allow for the production of viruses in 293 cells, and to provide transitory expression of recombinase that induces rearrangement in target tissues. The major difference between the two strategies lies in the way the deinduction of recombinase is achieved.  
         [0090]    In the first approach, adenoviral vectors are engineered to turn an activating transcription factor into a repressor upon chromosomal rearrangement. Vectors employing this approach are referred to herein as type A vectors (FIG. 1A). In the second approach, the recombinase promoter is redirected following chromosomal rearrangement. Vectors utilizing the second approach are referred to as type B vectors (FIG. 1B). In both cases a linear chromosome is converted to its circular episomal form and a resulting deleted linear form. The circular DNA contains an Epstein Barr virus (EBV) replicon, which allows synchronous replication of the episome with the host mitotic cycle (Reisman et al., Mol. Cell Biol. 8: 1822-32, 1985; Yates et al., Nature 313: 812-15, 1985). The linear DNA is deleted for the enhancer and E1 genes.  
         [0091]    One self-regulated gene switch, employing the type A vector strategy, was designed based on the bacterial transposon Tn10 tetracycline repressor (tetR) gene. In its natural context, the tetR protein binds to specific sequences (tet operator sequences) upstream of a tetracycline resistance gene, preventing transcription of the gene unless tetracycline is present. To adapt this protein for eukaryotic gene regulation, a gene fusion is created between tetR and an active portion of a strong eukaryotic transcriptional activator, the herpes simplex virus VP16 protein. The fusion protein exerts its action on a synthetic promoter created by the insertion of multiple tet operator sequences upstream of a basal promoter element. This configuration allows high-level gene expression whenever the tetR-VP16 fusion protein binds to its cognate operator sequences. Because the tetR protein normally does not bind to its operator in the presence of tetracycline, the activity of this synthetic promoter is high in the absence of tetracycline and low in its presence.  
         [0092]    One example of a type A vector is shown in FIG. 1A. This self-regulated gene expression cassette, present in a hybrid adenovirus, consists of a bi-directional promoter element in which central tetR binding sites are flanked by divergently oriented basal promoter elements. In one direction the promoter directs the formation of a transcript encoding the cre protein; in the other direction, the promoter directs the formation of a tetR-VP16 fusion protein. The latter differs from the conventional version in bearing a loxP site between the tetR component and the VP16 component. When tetracycline is present this gene switch is silent. As shown in FIG. 1A, upon introduction into a target cell in the absence of tetracycline, the tetR-loxP-VP16 fusion protein is produced, stimulating further production of the fusion protein, and the cre protein. The cre protein then acts to promote site specific recombination between the loxP site in the tetR-loxP-VP16 coding sequences, and a distant loxP site. As a result of this recombination, the fusion protein coding sequence is disrupted so that the promoter no longer directs the formation of a tetR-loxP-VP16 fusion protein, but gives rise to an inert tetR-loxP-VP16 fusion protein for binding to the promoter upstream elements, thereby extinguishing promoter activity.  
         [0093]    As shown in FIG. 1A, the excised circular DNA element contains at least two transcription units. In addition, other transcription units or internal ribosome entry site elements may be used to allow the coexpression of gene products which are useful for extending the persistence of the delivered DNA, regulating expression of the gene of interest, or providing for ablation of the transduced cells once their presence is no longer desirable. In addition, the linear DNA remaining after excision of the circular gene expression plasmid lacks both viral packaging sequences and the cis-acting enhancer. Within this linear DNA, additional loxP sites may be placed to provide for the rearrangement of the remaining vector DNA in the target cell, disrupting the normal topology of the genes, and further thwarting expression.  
         [0094]    Using the type B vector design strategy, described in greater detail below, a recombinant adenoviral gene delivery system that is capable of undergoing growth phase-dependent site-specific recombination has also been constructed.  
         [0095]    The following examples are presented for the purpose of illustrating, not limiting, the invention.  
       Type B Vectors—Experimental Results  
       [0096]    Several experimental examples for constructing type B vectors and for carrying out the general approaches of the invention are now described below.  
         [0097]    Two-Cosmid System for Efficient Construction of Recombinant AdV  
         [0098]    To simplify and facilitate the generation of recombinant AdV, a system was established to assemble the desired AdV genome in a single plasmid by ligation (shown in FIGS. 2A and 2B). The system consists of two component vectors, a left end plasmid, pLEP, and a right end plasmid, pREP. The left end Ad sequences (nt 1-376) in pLEP include the viral inverted terminal repeat, the cis-acting packaging sequences, and the viral enhancer. Nucleotide (nt) positions described herein refer to the wild type Ad2 sequence in GenBank (J019017). The Ad sequences are followed by the gene expression unit intended for delivery and an intron endonuclease (PI-PspI) cleavage site. The right end plasmid contains a PI-PspI site followed by the Ad2 genome from the end of the E1 locus rightward (nt 3527-35937).  
         [0099]    pLEP is a small tractable vector for cloning, whereas pREP is much larger and contains less frequently manipulated genes. Both pLEP and pREP contain a bacteriophage γ cos site, oriented to generate a single cosmid of appropriate length for in vitro packaging following ligation of the two plasmids at the PI-PspI cleavage site. pLEP is tetracycline resistant (Tet r ) and pREP is ampicillin (Amp r ) resistant, allowing the recombinants to be selectively isolated by co-selection for both markers. In the resulting assembled cosmid, the adenoviral sequences are closely flanked by cleavage sites for the intron endonuclease I-CeuI. Digestion with I-CeuI liberates the entire recombinant AdV genome from the parent cosmid (see FIG. 2B).  
         [0100]    Three classes of pREP have been constructed to allow the preparation of AdVs bearing E1 (pREP7; SEQ ID NO.: 2), E1 and E3 (pREP8; SEQ ID NO.: 3), or E1, E3, and E4 (pREP12; SEQ ID NO.: 4) deletions. pREP7 (SEQ ID NO.: 2) contains nt 3527-35937 of the Ad2 genome, and pREP8 (SEQ ID NO.: 3) carries an additional deletion in the E3 region (Δ nt 27901-30841). pREP12 (SEQ ID NO.: 4) has deleted open reading frames (ORF) 1-4 of the E4 region (Δ nt 34121-35469, 1348 bp). AdV generated with these cosmids should be able to accommodate 5, 8, and 10 kb inserts, respectively.  
         [0101]    These aforementioned vectors were constructed as follows. The EcoRI to BsaI fragment that spans the ampicillin resistance gene in pBR322 was deleted and replaced by a synthetic adapter, and the bacteriophage γ cos site was inserted between the unique StyI and BsmI sites. A PCR amplified Ad2 fragment containing the left end ITR (L.ITR), enhancer elements, and the encapsidation signal (nt 1-376) was created and inserted into the adapter (FIGS. 2A, 2B) to yield the tetracycline-resistant left-end plasmid pLEP. The right end of Ad2 from the AflII site to the right end (nt 3527-35937) was assembled into an ampicillin resistant cosmid vector, pACKrr3 (SEQ ID NO.: 1), by multiple steps of PCR amplification and fragment interchange. The resultant cosmid was termed pREP7 (SEQ ID NO.: 2). To expand vector capacity, two deletions were incorporated into the pREP7 (SEQ ID NO.: 2) cosmid, an E3 gene deletion (nt 27901-30841, 2840 bp); cosmid pREP8 (SEQ ID NO.: 3) and a 1.3 kb deletion (nt 34121-35469) in the E4 region of the Ad2 region; pREP12 (SEQ ID NO.: 4) An example of the construction of an AdV carrying a CMV-GFP expression unit is outlined in FIG. 2. pLEPCMVGFP (Tet r ) was digested with PI-PspI and ligated to the pREP7 (SEQ ID NO.: 2; ΔE 1, Amp r ) digested with the same enzyme. The ligation mixture was packaged with γ phage extracts (MaxPlax lambda packaging extracts, Epicentre Technologies) and a fraction of the packaged phage was used to infect a recombination-deficient  E. coli  host, with selection for the assembled plasmid on Amp/Tet plates. Transductants containing pLEP fused to pREP were selected on agar containing 25 μg/ml ampicillin and 12.5 μg/ml tetracycline (Amp/Tet). Colonies were selected and DNA isolated (Qiagen). DNA was used either for restriction analysis or for tranfection of 293 cells as described herein.  
         [0102]    [0102]FIG. 3A shows typical results for the Bgl II digestion pattern of a pLEP3CMVGFP/pREP7 hybrid cosmid, pAd2-7CMVGFP DNA. Because of the size minimum (˜40 kbp) for γ phage in vitro packaging and the double antibiotic selection, most of the colonies growing on Amp/Tet plates were the desired hybrid cosmids, and undesired rearrangements were rarely seen. In the present example, all four pAd2-7CMVGFP clones exhibited the digestion pattern predicted from the inferred sequence. The entire recombinant AdV genome was then released from the cosmid by I-CeuI digestion (FIG. 3B). I-CeuI digestion leaves ten nucleotides to the left of the left ITR and eight nucleotides to the right of the right ITR. Short flanking sequences have been reported to be eliminated during replication of recombinant viruses after transfecting the DNA into 293 (human embryonic kidney) cells (Hanahan et al., Mol. Cell. Biol. 4:302-309, 1984).  
         [0103]    The digestion reaction can be transfected into 293 cells without purification as follows. 293 cells, obtained from Microbix Bisosystems (Ontario, Canada), were cultured in 10 cm dishes in complete Dulbecco&#39;s Modified Eagle&#39;s Medium (DMEM) supplemented with 10% FBS, 2 mM glutamine and penicillin/streptomycin (Gibco BRL), and maintained at 37° C. and 5% CO 2  atmosphere in an incubator. Cells were grown to ˜50% confluence on the day of transfection. Ten fig of cosmid DNA were digested with I-CeuI in a volume of 50 μl. The reaction mixture was transfected into 293 cells by calcium phosphate precipitation (Graham and Prevec, Manipulation of adenovirus vectors, p. 109-128, In E. J. Murray (ed.), Methods in Molecular Biology, vol. 7, Humana, Clifton, N.J., 1991) without purification. After transfection, cells were cultured and examined daily for the appearance of cytopathic effects (CPE). Virus propagation, purification, plaque assay, and viral DNA isolation were performed using established protocols (Graham and Prevec, supra). At day six post-transfection, 5-30 viral plaques/10 cm dish/10 ug DNA were usually apparent, which compared favorably with the 30-50 plaques/10 cm dish/10 μg DNA found for 293 cells transfected with purified wild type Ad2 DNA.  
         [0104]    To compare the efficiency of recombinant virus production, similar viruses were also generated by homologous recombination. 20 μg of pREP7 (SEQ ID NO.: 2) was co-transfected into 293 cells with 10 μg of a plasmid encoding the left end of the adenoviral genome and a green fluorescent reporter gene (pLITREF1αGFP). pLITREF1αGFP contained the Ad2 left end nt 1-376, an EF1α promoter/GFP expression unit and Ad2 sequence (from 3525-8120) that overlaps with the same sequence in pREP7 (SEQ ID NO.: 2). This overlap fragment served as the region for homologous recombination. Each co-transfection was performed in duplicate. Initial plaques took longer to appear (14 days post transfection) and were less abundant (0-3 plaques per plate).  
         [0105]    Data in the literature suggest that exposed ITR ends favor efficient virus production (Hanahan et al., supra). To assess the importance of this effect, an AdV cosmid, pIAdEF1αGFPB, in which the AdV ITRs were flanked with a different restriction site at each end was constructed. pIAdEF1αGFPB DNA was digested with BsaBI to expose the right ITR, I-CeuI to expose the left ITR, or the two enzymes were used together to expose both ends. Digested cosmid DNA samples were transfected into 293 cells and plaques were allowed to develop. Virus propagation, purification, plaque assay, and viral DNA isolation were performed using established protocols described in Graham and Prevec. (Manipulation of adenovirus vectors, In E. J. Murray (ed.), Methods in Molecular Biology, vol. 7. Humana, Clifton, N.J., pp. 109-128, 1991).  
         [0106]    Ten days after transfection the viruses were harvested and viral titers were determined. The average titer for the viral stocks (FIGS. 4A and 4B) was 1.3×10 4  pfu (plaque forming unit)/ml from transfection with undigested DNA; 2.4×10 5  pfu/ml from BsaBI linearized DNA (free right ITR); 1.1×10 5  pfu/ml from I-CeuI linearized DNA (free left ITR); and 2.7×10 6  pfu/ml for the BsaBI/I-CeuI double digested DNA (both ITRs free). Thus liberation of each end resulted in an approximate increase in the efficiency of generating virus by a factor of ten (FIGS. 4A and 4B).  
         [0107]    Construction of an AdV Capable of Self-Rearrangement  
         [0108]    One approach to attenuating adenoviral gene expression and improving transgene persistence is the creation of viruses capable of undergoing internal, self-directed rearrangement upon delivery to the target tissue. In principle, this objective can be achieved through the regulated expression of site-specific recombinases in vectors that contain the cis-acting target of recombinase action. To allow such vectors to be created, the recombinase activity must be suppressed during propagation in the packaging cell line. As described in more detail below, the use of a lineage-specific promoter to control recombinase expression has been successfully employed to achieve this end.  
         [0109]    An example of this is shown in FIG. 5. The expression of Cre recombinase was controlled by a liver-specific promoter constructed as follows. The human hepatic control region 1 and 2 (HCR1 and 2) of the ApoE/C gene locus (Allan et al., J. Biol. Chem. 270:26278-81, 1995; and Dang et al., J. Biol. Chem. 270:22577-85, 1995) were amplified by PCR using 293 cell genomic DNA as the template. The following primers were used to amplify both HCR1 and HCR2 fragment: HCRtop-5′gcggaattcggcttggtgacttagagaacagag 3′ (SEQ ID NO.:5); HCRbot-5′ gcgggatccttgaacccggaccctctcacacta 3′ (SEQ ID NO.:6). The amplified PCR fragments (˜0.39 kb) were cloned into pUC19. The HCR1 and HCR2 sequences were confirmed by dideoxy DNA sequencing. The two fragments were assembled in a head to tail orientation, fused with a synthetic basal TATA element and cloned in a parental pLEP vector containing a GFP reporter gene. The resultant plasmid was named pLEPHCR12GFP. The synthetic liver-specific, as demonstrated below, provided a means to control Cre recombinase expression during propagation of the vector in 293 cells, and allowed for testing the consequences of abstracting the enhancer from the linear vector DNA upon delivery of the DNA to the target cells.  
         [0110]    In 293 cells, this promoter is silent, allowing the viral chromosome to be propagated with minimal rearrangement. Any rearranged viruses that are formed lack packaging signals and so disappear from the pool of propagating vectors. In liver cells the Cre recombinase is induced by the action of the tissue-specific promoter. The resulting Cre-induced recombination excises a circular episome and redirects the transcriptional output of the liver-specific promoter so that it directs the synthesis of the transgene of interest. The remaining linear fragment consists of an adenoviral genome lacking the enhancer and packaging signals and a Cre expression unit devoid of promoter sequences.  
         [0111]    In the form discussed here, one loxP site is located at nucleotide 147 of the Ad2 genome, between the left ITR and the enhancer/packaging sequences, and the second loxP site is placed inside an intron a few bases upstream of the splice acceptor sequence. Hence the loxP site does not appear in the resulting mature transcript. The Cre coding sequence that remains on the right end linear fragment after rearrangement lies downstream from a splice acceptor that lacks a splice donor or upstream promoter sequences. This effectively terminates the expression of Cre following excision.  
         [0112]    Prior to recombination, the Cre recombinase gene is under the control of a synthetic promoter (referred to as HCR12), consisting of hepatic locus control elements from the human ApoE/C locus fused to the first intron of the human EF1α gene. After cyclization the HCR12 promoter lies upstream of the transgene (in this case GFP) and the distal segment of the intron (beyond the loxP site) contains the adenoviral enhancer. To facilitate manipulation of the plasmids in  E. coli , the human IgG 1 hinge-CH2 intron (118 bp) was inserted in the Cre coding sequence at nucleotide 237, suppressing Cre expression in bacteria. The circularized episome contains the latent origin of replication (OriP) and trans-acting DNA replication protein (EBNA-1) of Epstein Barr virus, and hence is capable of autonomous replication in synchrony with the host mitotic cycle (Yates et al., Nature 313:812-815, 1985).  
         [0113]    Using the two cosmid system described above, the pLEP plasmid containing the self-resolving components, pLEP1BHCR12, was ligated with pREP8 (SEQ ID NO.: 3; ΔE1ΔE3) to create pAdVHCRGFP/EBV. The latter was digested with I-CeuI and transfected into 293 cells. Appearance of plaques from AdVHCRGFP/EBV was retarded (by 8 days) compared to non-rearranging viruses, perhaps as a result of basal expression of the liver-specific promoter in 293 cells. However high titer viral stocks of 10 12  nominal (absorbance-determined) particles/ml was achieved.  
         [0114]    Rearrangement in Target and Nontarget Cells  
         [0115]    To test excision efficiency, HepG2 (hepatocellular carcinoma) and Hela (cervical carcinoma) cells, obtained from ATCC, were infected with virus at a multiplicity of infection (moi) of 1,000 nominal particles/cell. This titer corresponds to approximately 10 plaque forming units per cell. For these experiments, HepG2 and Hela cells were seeded in 35 mm dishes and cultured to approximately 80% confluence in DMEM/FBS as described herein. Cells were infected with the desired multiplicity of virus in a volume of 1 ml at 37° C. for 2 hours. At the end of the incubation, cells were washed with PBS twice and cultured in 2 ml of medium. Cells were collected in parallel at desired points for low molecular weight DNA and RNA extraction. Cells were examined for GFP expression by fluorescence microscopy (Olympus, IX70) or microtiter plate reader (PerSeptive Biosystem, CytoFluor II) before extraction of DNA for analysis of chromosomal rearrangement.  
         [0116]    DNA analysis of chromosomal rearrangement was performed as follows. 5 μg of Hirt DNA was digested with Bgl II and analyzed by DNA blot techniques using a labeled EBNA-1 gene fragment as probe (FIG. 6). The Bgl II fragment from the non-circularized AdV is 3162 bp, generated from the 5 ′end of the AdV to the first BglII site in the AdV. The circularized fragment created from the two loxP sites has a size of 4915 bp (FIG. 6A). Densitometry revealed that at 72 hours post infection, 95% or more of the input genomes had undergone circularization in HepG2 cells. In contrast, low but detectable levels of circularized fragment was visualized in Hela cells infected at the same time and at the same multiplicity of infection used for the HepG2 cells (FIG. 6B).  
         [0117]    At the time of infection (t=0, FIG. 6B), the amount of input viral DNA detected by DNA blot was higher for HepG2 cells than for Hela cells when similar virus multiplicities were applied (moi of 1,000). This may reflect differences in AdV adsorption or infection efficiency between the two cell types, possibly as a result of the lower levels of coxsackievirus-adenovirus receptor on the Hela cells surface. To achieve similar viral genome input into HepG2 and Hela cells, Hela cells were infected with ten-fold more virus (moi of ˜10,000) than HepG2 cells (moi of 1,000). Episomal DNA samples were extracted and analyzed by blotting. The results (FIG. 6C) indicated that when comparable amounts of viral genome are present in the nucleus, the cyclization rate in both cell types was similar. Because the level of subsequent GFP expression is much higher in HepG2 cells than in HeLa cells (FIG. 7A), it is likely that very small amounts of Cre recombinase suffice to promote rearrangement, and that recombinase expression is not limiting for rearrangement in either HepG2 or HeLa cells.  
         [0118]    GFP expression cannot be detected until rearrangement has taken place, so the measurement of the fraction of GFP positive cells provided a simple alternate method for assessing the degree of productive rearrangement. FIG. 7A shows that GFP expression developed quickly in transduced HepG2 cells, but that only a few GFP positive cells can be detected in Hela cells infected with a ten fold higher moi, conditions that allow circularization to a comparable extent to that seen in HepG2 cells (FIG. 6C).  
         [0119]    The HCR12 promoter specificity was also tested by infecting two additional non-hepatic cell lines, A431 (human epidermoid carcinoma) and HT29 (human colon adenocarcinoma), with the Ad2HCRGFP/EBV vector. Both cell lines were obtained from ATCC and cultured using DMEM/FBS as described herein. A few cells, with weak GFP signal, were detected at 72 hours after infection in these cells (FIG. 7B). In contrast, these non-hepatic cells could be infected efficiently with a first generation AdV, Ad2CMVGFP virus (data not shown), indicating that the low GFP signal was not due to the low infectivity of these cells by AdV.  
         [0120]    To further assess the utility of the AdV genome rearrangement, primary human hepatocytes were infected with the Ad2HCRGFP/EBV vector. For these experiments, primary human hepatocytes, generously provided by Dr. Albert Edge (Diacrin, Inc., Charlestown, Mass.) were isolated and cultured as described by Gunsalus et al. (Nat. Med. 3:48-53, 1997), infected with adenovirus, and GFP expression was analyzed. As shown in FIG. 7C, GFP expression was readily detected 72 hours after infection.  
         [0121]    Diminished Viral Gene Expression in Rearranged AdV  
         [0122]    After excision, the adenovirus major enhancer/packaging signal segregates with the episomal DNA, yielding a linear fragment containing the remainder of the AdV genome without this important cis-element (FIG. 5). To assess the impact of enhancer deletion, PCR amplification and quantitative RT-PCR measurement of late viral gene expression was performed as follows.  
         [0123]    Four μg of total RNA was reverse transcribed into cDNA using M-MLV RT by a standard protocol (Promega). 1 μl of the cDNA from each sample was used in subsequent PCR reactions. PCR primers were designed to amplify the tripartite leader sequence of the adenovirus late genes: TPL1-5′ act ctc ttc cgc atc gct gt 3′ (SEQ ID NO.: 7) and TPL2-5′ ctt gcg act gtg act ggt tag 3′ (SEQ ID NO.:8). For detection of the AdV genome in the Hirt DNA samples, 1 μg DNA was employed in the PCR amplification using the following primers which are specific for the adenovirus DNA in the fiber gene: Fiber1-5′ ccg cac cca cta tct tca ta 3′ (SEQ ID NO.: 9) and Fiber2-5′ ggt gtc caa agg ttc gga ga 3′ (SEQ ID NO.: 10). PCR reactions were performed as 95° C. 30 seconds; 54° C. 30 seconds; 72° C. 30 seconds for 30 cycles. All amplified products were analyzed on a 2% agarose gel.  
         [0124]    For quantitative PCR, a molecular beacon based universal amplification and detection system was used (Intergen). A common leading sequence (Z sequence, 5′ act gaa cct gac cgt aca 3′) was added to the TPL1 and Fiber1 primers. The TPL2 and Fiber2 primers, described above, were used in the quantitative PCR reactions. 1 μl of the cDNA and one μg of Hirt DNA from each sample were used in the assay. The PCR were carried out in a 96-well spectrofluorometric thermal cycler (Applied Biosystems Prism 7700). The number of template molecules in the PCR reaction was calculated from the standard curve using linearized plasmid as templates.  
         [0125]    As most late adenoviral genes transcripts share a common ˜200 bp tripartite leader sequence (TPL) (Akusjarvi and Persson, Nature 292:420-6, 1981), the TPL sequence was chosen as a marker of viral gene expression. HepG2 cells were infected with the first generation vectors Ad2CMVGFP and Ad2HCRGFP, or the self-resolving vector, Ad2HCRGFP/EBV, using increasing multiplicities of infection. Total cellular RNA and low molecular weight DNA were isolated in parallel as described by Hirt (J. Mol. Biol. 26:365-9, 1967) and total RNA was prepared using RNAzol solution (Te1-Test. Inc.). RT-PCR was performed to quantitate the amount of RNA encoding the TPL in the cDNA samples. PCR amplification of a 201 bp fiber gene fragment from the AdV genome was used to detect the amount of viral genome in the DNA samples. A representative result of three experiments is shown in FIG. 8A. TPL sequences were detected, 72 hours post-infection, with either 100 or 1000 viruses infected per cell, using both of the first generation adenoviruses (upper panel).  
         [0126]    In contrast, no TPL signal was detected in the self-resolving Ad2HCRGFP/EBV infected cells, even at a moi of 100,000/cell. PCR amplification of the AdV fiber gene revealed comparable levels of AdV genomic DNA in cells infected at comparable moi&#39;s. (FIG. 8A, lower panel). The cDNA samples in which the TPL signals were detected were further analyzed by real-time fluorescence PCR. The corresponding genomic DNA samples were also analyzed to determine the number of AdV genomes present in each sample. The results are summarized in FIG. 8B. There were approximately 1×10 4  TPL per 1×10 6  AdV genomes detected in the Ad2HCRGFP infected cells, but no detectable TPL in the self-resolving Ad2HCRGFP/EBV infected cells. These results indicate that adenoviral gene expression was dramatically reduced by the separation of the viral enhancer sequences occasioned by the re-arrangement of the self-resolving vector.  
       Type A And Type B Vectors—Experimental Results  
       [0127]    Additional experimental examples now follow that further illustrate the general approaches of the invention relating to using and constructing type A and type B vectors. For generating such adenoviral vectors, DNA sequences important for gene expression in the target tissue were placed between two loxP sites. The first loxP site was inserted between the Ad2 left-end inverted terminal repeat (ITR) and the enhancer sequence, replacing a BspLU11I and BstZ17 fragment of Ad2. A target gene expression cassette, comprising a promoter, a gene of interest, polyadenylation signals, the EBV replicon, and site specific recombinase expression unit were inserted in place of the E1 locus.  
         [0128]    In type A adenoviral vectors, the second loxP site is placed between TetR and VP16, preserving the coding frame of both (FIG. 1A). A bidirectional promoter in which a central heptamer of tetracycline operator sites (TetO) (Gossen and Bujard, Proc. Natl. Acad. Sci. USA 89:5547-5551, 1992) was flanked by two divergently oriented basal elements, directs the expression of TetR loxP VP 16 from a synthetic TATA element, whereas Cre recombinase is controlled by the same heptamer of operator upstream of the HIV LTR basal element.  
         [0129]    In the case of type B viruses (FIG. 1B), the second loxP site was inserted in the first intron of the Ef1α gene, which contains the transcription stimulating sequences described herein. In addition, a splice acceptor sequence was added to the 5′ end of the coding sequence of the gene of interest. To avoid rearrangement during plasmid construction in bacteria, the Cre recombinase coding sequence was interrupted by the addition of the human IgG1 hinge-CH2 intron (between amino acids Q78 and A79), as described herein.  
         [0130]    Designing a Compact EBV Replicon  
         [0131]    Most plasmids employing the EBV latent origin of replication exceed 10 kb in length. To provide a means for increasing the capacity of the recombinant adenoviral type A or type B vectors to accommodate a therapeutic gene, a compact EBV replicon having episomal stability was designed. To this end, deletions were generated in both the cis-acting origin of replication, OriP, and the sequences encoding the trans-acting replication protein, Epstein Barr virus nuclear antigen-1 (EBNA-1) (FIG. 9A). Episomal persistence was assessed with a green fluorescent protein (GFP)-bearing test plasmid by determining the fraction of cells retaining green fluorescence as a function of time, assuming that the half-life of GFP, in daughter cells that have not received an episome as a result of segregation failure, is approximately 1.4 days (Fukumura et al., Cell 94:715-725, 1998).  
         [0132]    EBNA-1 contains a central repeated structure that consists entirely of Gly and Ala residues, termed the GA repeats (FIG. 9A). Although deletion of this structure has been reported to have little consequence, a deletion mutant consisting of both a short OriP and a short EBNA-1 (SoriP +SEBNA1) was generated and found not to support plasmid maintenance effectively (approximately 40% loss per cell division). A version of this mutant, reconstructed with 40 GA repeats, in which the short OriP was paired with a short EBNA-1 provided significantly better plasmid stability (20% loss per cell division vs. 10% per cell division for the wild type) (FIG. 9B). Since most target tissues are relatively quiescent mitotically, this level of segregation fidelity provides reasonable stability in a compact replicon.  
         [0133]    Producing Cell Lines that Express Cre- or FLP-Dominant Negative Mutants  
         [0134]    As discussed herein, one obstacle to creating adenovirus carrying both recombinase and target sites has been the difficulty of controlling recombinase activity during virus propagation. Since efficient recombinase activity is needed in target cells, recombinase activity is best tempered in the production cell line.  
         [0135]    Vector-independent methods to suppress recombinase activity during the production phase are attractive because they allow vector design objectives to be pursued with fewer constraints. In principle, dominant negative recombinase mutants provide the desired antagonism of recombinase activity. Cell lines expressing such recombinase dominant negative mutants were produced as follows.  
         [0136]    Dominant negative Cre mutants were selected from known point mutants (Wierzbicki et al., J. Mol. Biol. 195:785-794, 1987) that are defective in recombination function but are likely to retain dimerization function FIGS.  10 A-E. Several mutants were screened for their abilities to inhibit Cre activity of a type B vector construct (ad2239 in FIG. 10B) in a transient cotransfection assay. Under these conditions, Cre activity is detected by the expression of GFP that occurs upon rearrangement. FIGS. 10D and 10E show the point mutants that were assessed and their relative activities in the transient cotransfection assay. Dilution studies, in which increasing amounts of substrate/Cre plasmid were cotransfected with the mutant forms, were conducted and based on its favorable profile, one mutant recombinase, designated CreY324C, was chosen for further development (FIG. 10D).  
         [0137]    Strong constitutive expression of CreY324C, under control of the Ef1α promoter failed to yield stable cell lines. Stable clones were obtained when the Ef1α promoter was replaced with a tetracycline regulated promoter (Gossen et al., Science 268:1766-1769, 1995). Clones were then tested for the ability to inhibit Cre enzymatic activities, and one clone, designated cell line #17, was selected for additional experiments. When a plasmid bearing Cre and capable of undergoing Cre-directed rearrangement to create a GFP transcription unit (ad2239) was transfected into #17 cells or parental 2930N cells, GFP expression in the #17 cells in the presence of 2 μM doxycycline was significantly lower than those of controls (FIG. 11), showing that Cre enzyme activity can be inhibited in #17 cells.  
         [0138]    In addition to dominant negative Cre mutants, dominant negative FLP mutants may also be identified. FLP belongs to the same family of site-specific recombinases as Cre recombinase. A number of FLP mutations that show defects in either cleavage or ligation of FRT sites have been identified. Mutant FLP defective in cleaving FRT site (for example, H309L, L315P, G328R, G28E, N329D, S336Y, S336F, A339D, Y343F, and H345L) are generated using standard methods. Mutants that inhibit the wild type enzyme are then identified for generating stable cell lines according to the methods described above. These and the other cell lines (described herein) are then used for producing FRT/FLP containing virus.  
         [0139]    As mentioned above, difficulties creating stable cell lines expressing Cre dominant negative mutants were occasionally encountered. This difficulty was not limited to Cre mutants, but was also observed with the wild-type Cre enzyme. In contrast, 293 cell lines stably expressing a thermostable FLP, referred to as FLPe (Buckholz et al., Nat. Biotechnol. 16:657-662, 1998), were created, suggesting that FLPe might not be as cytostatic as Cre protein. To demonstrate this, 293 cells were transfected with plasmids expressing either Cre or FLPe, and puromycin resistant colonies were selected. To generate stable cell lines expressing Cre or FLP mutants, 293 TetON cells were transfected with linearized plasmid expressing Cre or FLP mutants and puromycin acetyltransferase and selected with 1 μg/ml of puromycin. Puromycin resistant colonies were characterized further for their ability to inhibit Cre recombinase using the cre (ad2239) or flp (ad2879) substrate plasmids. Table 1 shows that there are more puromycin resistant colonies selected from FLPe transfected cells than from Cre transfected cells. From this result, it is expected that stable cell lines expressing a reasonably high level of dominant negative FLP may be readily created.  
                                           TABLE 1                           Puromycin Resistant Colonies Formed When Cre       Expressing or FLPe Expressing Plasmid was       Used to Transfect 293 Cells                Expression Plasmid   Number of colonies (2 μg/ml puromycin)                            Control   236           Cre    92           FLPe   127                      
 
         [0140]    Cre or Cre dominant negative mutants were also found to inhibit FLP activity (Table 2). Accordingly, cell lines, such as cell line #17, that stably express a Cre dominant negative mutant (for example, CreY324C), are useful for producing FLP/FRT carrying adenovirus.  
                                   TABLE 2                           Cre Inhibition of FLP Activity in trans            Plasmids   Arbitrary GFP intensity                    Ef1α FLP + FLP substrate + vector control   4.9       Ef1α FLP + FLP substrate + Ef1α Cre   0.78       Ef1α FLP + FLP substrate + Ef1α Cre R173C   2.23                          
 
         [0141]    Transcriptional Regulation of Cre or FLP Recombinases  
         [0142]    It has been relatively difficult to achieve high-level promoter inducibility in a replicating adenovirus. The challenge is similar to that of achieving faithful control of transcription in a transient expression setting. One approach to increase the induction ratio in a transient setting is the use of auto-regulatory (feed-forward) circuits. One such system, based on tetracycline dependent activation, is shown in FIG. 12. A central heptamer of tetracycline promoter operator sites (TetO sites) was placed between two divergently oriented basal TATA elements. The leftward TATA controls the expression of the TetR-VP16 fusion protein, in which a loxP (or FRT) site has been placed between the TetR DNA binding domain and the VP16 transcriptional activator. The rightward TATA box directs the synthesis of recombinase, either Cre or the yeast FLP enzyme. In the presence of tetracycline, the promoter has reduced activity in both directions. Upon removal of tetracycline, the synthesis of both TetR-VP16 and recombinase are induced (FIG. 1A). The induced recombinase then disjoins the TetR DNA binding element from the transcriptional activation contributed by VP16. Any existing TetR-VP16 fusions thereafter promote transcription of TetR, which competes with TetR-VP16 for TetO, resulting in deinduction of recombinase transcription.  
         [0143]    When a model target cell line, HepG2, was tested with this type of adenovirus, the efficiency of circularization was low relative to that seen in 293 cells (data not shown), indicating a cell dependence of the bidirectional TetO promoter. To correct this, the TATA element of the TetO synthetic promoter (derived from the CMV immediate early promoter) was replaced with that of the HIV LTR. Constructs bearing differing components of the HIV basal promoter were analyzed for strength and regulation in 293 and HepG2 cells (FIG. 13A). Among the constructs tested, one version bearing the HIV LTR TATA and Sp1 elements (D in FIG. 13A) showed the least basal expression in 293 cells (data not shown) and the greatest induction in HepG2 cells (FIG. 13B).  
         [0144]    Using this promoter, a construct (ad3400) containing the auto-regulatory structure D as shown in FIG. 13A was engineered, and Cre activities in the presence and in the absence of tetracycline were assayed. Plasmid ad2265 (FIG. 14) in which a blue fluorescent protein (BFP) expression unit is interrupted by two loxP sites and transcription termination sequences was used as a substrate for Cre. Cre-mediated recombination joins BFP to the promoter resulting in BFP expression. As shown in Table 3, no difference was found in the intensity of BFP expression, either in the presence or absence of tetracycline. One possible explanation for this is that very little Cre protein is required for activity. Consistent with this idea, standard imunohistochemical techniques failed to reveal the presence of Cre enzyme in cells that were fully induced (data not shown).  
                                         TABLE 3                           Cre Recombinase Activity Regulation in Type A Constructs            Construct   Cre Form   +tet −tam   −tet −tam   +tet +tam   −tet +tam               ad3400   Cre   2.58    3.39   2.71    6.20       ad4394   Cre-LBD   0.081   0.22   0.056   1.02       ad4705   LBD-Cre-   ND   ND   ND   ND           LBD                                                  
 
         [0145]    Deletion of the PolyA Consensus Sequence from Cre or FLP Transcription Units  
         [0146]    To reduce the expression of FLP or Cre recombinase further, the consensus polyA addition signals from the Cre or FLP transcript unit were deleted from vector constructs, leaving polyadenylation dependent on distal downstream sequences, for example, in gene IX. The activity of Crc using type B proviral constructs with or without the polyA signal was measured. As shown Table 4, the construct without polyA signals (AD229.3) showed a significant reduction of GFP intensity compared to a construct bearing the polyA signal (AD230.5). When FLPe constructs of similar structure were evaluated, similar results were found (data not shown). These data show that Cre and FLPe enzyme activity levels can be modulated by attenuating polyadenylation.  
                                                   TABLE 4                           Effect of Deleting polyA Addition Signal From the Cre       Expression Unit on Cre Enzyme Activity Level                    polyA   Relative GFP Intensity                            AD229.3   −   0.25           AD230.5   +   1                      
 
         [0147]    Post-Transcriptional Regulation of Cre Recombinase Activity  
         [0148]    Post-transcriptional control mechanisms of Cre recombinase activity were also evaluated. Translational fusions between Cre and the ligand binding domain (LBD) of estrogen receptor have been reported to be regulated by estrogen (Feil et al., Proc. Natl. Acad. Sci., U.S.A 93:10887-10890, 1996; Gossen et al., Proc. Natl. Acad. Sci., U.S.A. 89:5547-5551, 1994), or, in the case of mutant estrogen receptors (Metzger et al., Proc. Natl. Acad. Sci. U.S.A. 92:6991-6995, 1995), by the partial antagonist tamoxifen.  
         [0149]    Use of a ligand-dependent recombinase (ad4394 in Table 3), in combination with the HIV LTR-based autoregulated Tet system, allowed for a small degree of regulation by tetracycline, but not by ligand, as assayed using the ad2265 rearrangement assay (Table 3). One interpretation of this finding is that fusion of the estrogen receptor LBD to Cre provides only modest control of recombinase activity, but attenuates enzyme potency to a level so that transcriptional regulation can be measured.  
         [0150]    To increase control of recombinase activity, the LBD was fused both to the N-terminus and C-terminus of Cre (LBD-Cre-LBD) and inserted into the coding sequence of both type A and type B vectors. When the LBD-Cre-LBD construct of type A was transfected into 293 cells, it showed no significant Cre enzyme activity even in the presence of ligand (Table 3). This result confirmed that the Cre recombinase activity is attenuated by N-terminal or C-terminal extension.  
         [0151]    When the LBD fusion Cre enzymes were assayed in the type B vector context, only LBD-Cre-LBD fusions (pk8-ad4626) showed ligand-dependent regulation of Cre enzyme activities (Table 5). It appears that attenuated Cre activity in LBD-Cre-LBD, in the absence of ligand, is low enough to fall below the upper limit of the Cre assay.  
                                     TABLE 5                           Cre Enzyme Activities of Type B Provirus                Cre Form   Provirus   −tam   +tam                       Cre   pk8-ad2239   ND   ND           Cre-LBD   pk8-ad4332   4.1   6           LBD-Cre-LBD   pk8-ad4626   0.05   4.2                                              
 
         [0152]    Consistent with this notion, only the construct carrying two LBDs, pk8-ad4626, was able to produce virus (AD121.5) by transfection and propagate in 293 cells, while pk8-ad4332, which carried one LBD, produced virus (AD100.9) initially (following transfection of the cognate DNA), but was unable to propagate in 293 cells (Table 6). In the case of wild type Cre, no virus was produced in 293 cells by transfection.  
                                     TABLE 6                           Production and Propagation of Type B Adenovirus                        Viral   Viral               Viral   Prop-   Prop-           Type B   Production in   agation in   agation in       Cre Form   adenovirus   293 cells   293 cells   #17 cells               Cre   Pack8-2239   −   −   −       Cre-LBD   AD100.9   +   −   +           (Pack8-4332)       LBD-Cre-LBD   AD121.5   +   +   +           (Pack8-4626)                  
 
         [0153]    The AD100.9 virus was able to propagate in #17 cells expressing the dominant negative Cre Y324C, demonstrating that modulation of Cre activity is important for viral production. Thus, adenovirus carrying both two loxP sites and Cre in two different configurations were generated by controlling Cre activity.  
         [0154]    Viral Rearrangement in Culture  
         [0155]    Cre/loxP mediated rearrangement of the adenovirus in tissue culture cells has also been analyzed. As shown in FIG. 15A, the AD121.5 virus showed a significant increase in GFP expression in the presence of the ligand, estrogen, suggesting a successful rearrangement of the virus by Cre recombinases. When non-chromosomal DNA (Hirt, J. Mol. Biol. 40:141-144, 1969) was made from the cells and analyzed by DNA blot analysis, the viral DNA from estrogen treated cells was identified mostly in circular form (C in FIG. 15B), while the DNA from cells not treated with estrogen was found mainly in linear form (L in FIG. 15B).  
         [0156]    To evaluate the efficiency of the self-rearranging viruses in vivo, high titer stocks of AD102.7 (a type A virus carrying LBD-Cre, pk8-ad4394) in #17 cells was prepared and purified by CsCl gradient ultracentrifugation. The titer of AD102.7 (4−6×10 12 /ml by OD) is comparable to or slightly exceeds that of control viruses (2−4×10 12 /ml by OD), which carry neither Cre nor a loxP site. To determine the efficiency of viral rearrangement in vivo and whether such rearrangement is dependent on the presence of ligand, AD102.7 virus (4×10 11  pfu/mouse as determined by optical density) were injected via tail vein into Rag-2 mice that were pretreated with vehicle alone or 110 μg/day of tamoxifen for 7 days as follows.  
         [0157]    Rag-2 mice were injected with either PBS (mock) or 4×10 11  adenovirus particles (as determined by OD 260 ) of type A virus, AD102.7, via the tail vein. At various times after injection, animals were sacrificed and the liver tissues were removed and frozen rapidly on dry ice. To visualize GFP expression in animal tissues, mice were anaesthetized and perfused with 4% paraformaldehyde containing 0.2% glutaraldehyde intracardially (Kafri et al., Natl Genet. 17:314-317, 1997), and the liver tissues were removed and fixed overnight at room temperature in the perfusion buffer containing 30% sucrose. The fixed tissues were sectioned serially and observed under confocal scanning laser microscopy. In experiments evaluating the responses of ligand-regulated recombinase, mice were injected either with vehicle (vegetable oil) alone or with 110 μg/day of tamoxifen for 7 days prior to adenoviral injection.  
         [0158]    Liver tissues from these animals were harvested at 2.5 hours post injection (the earliest time point taken after injection) and Hirt DNA from approximately 250 mg of frozen hepatic tissue was prepared and analyzed by blot analysis. As shown in FIG. 16, the majority of adenoviral DNA was found in circular form in tissues from untreated mice, as well as tamoxifen-treated mice. It can be concluded from these data that the Cre enzyme activity present in the tissue, even in the absence of ligand was sufficient for efficient self rearrangement of virus. As expected, the hepatic tissues from the Rag2 mice injected with AD102.7 showed strong expression of GFP (FIG. 17).  
         [0159]    Demonstrating that AD102.7 virus, produced efficiently in 293 cells at high titres by the conventional means, can self rearrange efficiently in vivo provides the proof of the concept that potentially safer adenoviral gene therapy vectors can be produced.  
         [0160]    Adenoviruses Carrying Both FRT and FLP Recombinase  
         [0161]    Type A and type B proviral constructs carrying both FRT (FLP recombinase recognition site) and FLP recombinase were also generated. Structures of these viruses are analogous to those of loxP/Cre carrying viruses except that loxP sites are replaced by FRT sites and Cre coding sequence is replaced by FLP coding sequence (FIGS. 18A, 18B).  
         [0162]    Virus Production at Reduced Temperature  
         [0163]    Temperature dependence of the Ef1α promoter using GFP expression as a marker was also examined. As shown in Table 7, Ef1α promoter activity is strongly reduced at 32° C. in comparison to 37° C. or 39° C. The temperature sensitive nature of the Ef1α promoter was used to propagate type B adenovirus carrying FLP at 32° C. following initial production of the virus by DNA transfection (pk8-ad3302) at 37° C. HepG2 cells infected with these viruses (AD41.4) showed strong GFP expression, but with an approximately 12 hour delay compared to GFP expressing viruses, suggesting that FLP recombinase activity may be impaired at 37° C. To improve the activity of FLP recombinase, viral constructs were created using a thermostable FLP (referred to as “FLPe”) described by Buchholz et al. (Nat. Biotechnol. 16:657-662, 1998).  
                                                                             TABLE 7                           Effects of Temperature on Ef1α Promoter Strength       as Shown by GFP Intensities                Arbitrary GFP Intensities                Tester plasmids       32° C.   37° C.   39° C.                            Ef1α GFP   16 hrs   1475   7886   11409               41 hrs   6472   36699   50787               86 hrs   16256   53370   54424           Ef1α Cre + ad2204   16 hrs   243   1141   2132               41 hrs   1094   9119   9784               86 hrs   695   3219   8144                                              
 
         [0164]    The activities of FLP and FLPe using a FLP substrate plasmid (ad2879, FIG. 19A) in 293 cells were compared. As shown in Table 8, FLPe is significantly more active than FLP under these conditions.  
                                                   TABLE 8                           FLPe is Significantly More Active than FLP Recombinase                Plasmid       Mean GFP intensity                            ad4821   Ef1α FLPe   2.39           ad2949   Ef1α FLP   0.01                                  
 
         [0165]    In addition, a tamoxifen-regulated FLPe was created by fusing the ligand-binding domain from a mutant form of estrogen receptor to the FLPe coding sequence at its C-terminus (FLPe-LBD). The FLPe-LBD was found to be regulated by the ligand, tamoxifen (Table 9). Although FLP activity was retained by C-terminal fusion (FLP-LBD), addition of a short oligopeptide tag to the N-terminus of FLP abolished its activity (data not shown).  
                                     TABLE 9                           Tamoxifen Regulation of FLPe as       Determined by GFP Intensities                Plasmid   FLPe   −tam   +tam                       ad4821 + ad2879   FLPe   ++++   ++++           ad5022 + ad2879   FLPe-LBD(tam)   +   +++                                              
 
         [0166]    Inhibition of FLPe Activities by Anti-sense FLPe  
         [0167]    An anti-sense approach to inhibit FLP enzyme activity was also employed. This approach tested the notion that incorporation of an open reading frame into an antisense transcript would stabilize the transcript and potentiate antisense activity. Two approaches were utilized. In one approach, the BFP coding sequence was placed upstream of anti-FLPe. In the second approach, an anti-FLPe was placed upstream of an internal ribosome entry sequence (IRES) and the BFP coding sequence (FIG. 20). The ability of these constructs to inhibit FLPe was assayed using a FLP substrate plasmid, ad2879 (FIG. 19A) and the result is shown in FIG. 21. These data show that anti-sense FLPe is more effective in inhibiting FLPe function when it is fused to BFP, which can presumably be replaced with any other stable protein.  
       Other Self-Rearranging Adenoviruses  
       [0168]    Mixed Infection With Adenoviruses Carrying loxP/FLP and FRT/Cre  
         [0169]    One of the ways to produce adenoviruses that can be rearranged in target cells, but not producer cells, is to engineer two separate viruses, each carrying one recombinase and the target sequence for the other. To test this system, type B adenoviral constructs carrying Cre recombinase and FRT sites and FLPe recombinase and loxP sites were created (FIG. 22). In target cells infected with both viruses, Cre catalyzes recombination between the two loxP sites in the FLP virus, and FLP carries out FRT mediated recombination in the Cre virus, resulting in two circular plasmids. The loxP virus contained BFP, whereas the FRT virus contained GFP. Measurement of the fluorescent intensities of GFP and BFP, after cotransfecting the two constructs, revealed that BFP expression (mediated by Cre enzyme) was greater than GFP expression (mediated by FLP enzyme), suggesting that the Cre enzyme functions more efficiently than FLP.  
         [0170]    Accordingly, these two recombinase activities in the target cells need to be balanced for complete circularization of both viral vectors. Exemplary methods for modulating Cre/FLP activity include the use of transcriptional regulation (such as by varying promoter strength and/or with or without poly A addition signal sequence) and translational and/or post-translational regulation (such as by changing FLP to FLPe and making LBD fusion proteins), and post viral production control (such as by changing the ratio of two viruses).  
         [0171]    In one approach, Cre was replaced by Cre-LBD and FLP was replaced by FLPe. To improve identification of the rearrangement products, BFP was replaced with RFP as a marker for Cre recombination. As shown in Table 10, in the presence of estrogen, expression of GFP (FLPe mediated) and RFP (Cre mediated) were similar.  
                                                                         TABLE 10                           RFP and GFP Expression of Cells Cotransfected       With Type B Proviral Constructs       Carrying Cre-LBD/FRT(GFP) or FLPe/loxP (RFP)                    GFP   RFP       Plasmids   Genotypes   intensity   intensity                    pk8-ads120 +   Cre-LBD/FRT (GFP) +   Estrogen            pk8-ads113   FLPe/loxP (RFP)   −   +   −   +               3222   3183   46   1954                  
 
         [0172]    To insure that both Cre and FLP carrying viruses with an optimal ratio infect each target cell, these viruses can be cross-linked prior to infection. For example, Cre carrying virus is labeled by biotin while FLP carrying virus is labeled by avidin. Mixing two types of modified viruses generates virus complexes of desired proportions as well. Biotinylation or avidinylation can be carried out using commercially available reagents such as EZ-Link TFP-PEO biotin (Pierce) and EZ-Link maleimide activated NeutrAvidin (Pierce). The extent of the biotin/virus and avidin/virus will be empirically determined to ensure the viability of the virus and to obtain an optimal ratio of two viruses in the complex. Optimal ratios will be those resulting in 1:1 Cre and FLP recombinase activities in target cells. The modifications will be done following manufacture&#39;s instructions.  
         [0173]    This approach not only increases the effective capacity of adenoviral vector but also opens new avenue of applications involving multiple proteins, some of which cannot be coexpressed in production cell line as a result of combination toxicity.  
         [0174]    All references mentioned herein are hereby incorporated by reference.  
         [0175]    Other embodiments are within the claims.  
     
       
       
         1 
         
           
             10  
           
           
             1  
             2341  
             DNA  
             Artificial Sequence  
             
               derived from Adenovirus  
             
           
            1 

gaatcggcca gcgcgaattc gattatcatc atcataatat accttatttt ggattgaagc     60 

caatatgata atgagggggt ggagtttgtg acgtggcgcg gggcgtggga acggggcggg    120 

tgacgtaggt tttagggcgg agtaacttgc atgtattggg aattgtagtt tttttaaaat    180 

gggaagttac gtacgcggca tcgatgcgcg ggatatcgcg gcggctagcg acatgaggtt    240 

gccccgtatt cagtgtcgct gatttgtatt gtctgaagtt gtttttacgt taagttgatg    300 

cagatcaatt aatacgatac ctgcgtcata attgattatt tgacgtggtt tgatggcctc    360 

cacgcacgtt gtgatatgta gatgataatc attatcactt tacgggtcct ttccggtgat    420 

ccgacaggtt acggggcggc gacctcgcgg gttttcgcta tttatgaaaa ttttccggtt    480 

taaggcgttt ccgttcttct tcgtcataac ttaatgtttt tatttaaaat accctctgaa    540 

aagaaaggaa acgacaggtg ctgaaagcga ggctttttgg cctctgtcgt ttcctttctc    600 

tgtttttgtc cgtggaatga acaatggaag ttaacggatc caggccgcga gcaaaaggcc    660 

agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc    720 

cccctgacga gcatcacaaa aatcaacgct caagtcagag gtggcgaaac ccgacaggac    780 

tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc    840 

tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata    900 

gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc    960 

acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca   1020 

acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag   1080 

cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta   1140 

gaagaacagt atttggtatc tgcgctctgc caaagccagt taccttcgga aaaagagttg   1200 

gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc   1260 

agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt   1320 

ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatcaga ttatcaaaaa   1380 

ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat   1440 

atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga   1500 

tctgtctatt tcgttcatcc atagttgcct gactccccgt agtgtagata actacgatac   1560 

gggagggctt accatccggc cccagtgctg caatgatacc gcgtgaccca cgctcaccgg   1620 

ctcctgattt atcagcaata aaccagccag ccggaagtgc cgagcgcaga agtggtcctg   1680 

caactttatc cgcctccatc cagtctatta gttgttgccg ggaagctaga gtaagtagtt   1740 

cgccagttaa tagttttcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct   1800 

cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat   1860 

cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatagtt gtcagaagta   1920 

agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca   1980 

tgccatccgt aagatgcttt tctgtgactg gtgagtattc aaccaagaat acgggataat   2040 

accgcgccac atagcagaac tttaaaagtg ctcatcattg ggaaacgttc ttcggggcga   2100 

aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgcgcaccc   2160 

aagtgatctt ctgcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg   2220 

caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catacttttc   2280 

ctttttcaat attattgaag catttatcag ggttattgtc tcatcagcgg atacatattt   2340 

g                                                                   2341 

 
           
             2  
             34616  
             DNA  
             Artificial Sequence  
             
               derived from Adenovirus  
             
           
            2 

gaatcggcca gcgcgaatta actataacgg tcctaaggta gcgtcatcat cataatatac     60 

cttattttgg attgaagcca atatgataat gagggggtgg agtttgtgac gtggcgcggg    120 

gcgtgggaac ggggcgggtg acgtaggttt tagggcggag taacttgcat gtattgggaa    180 

ttgtagtttt tttaaaatgg gaagttacgt atcgtgggaa aacggaagtg aagatttgag    240 

gaagttgtgg gttttttggc tttcgtttct gggcgtaggt tcgcgtgcgg ttttctgggt    300 

gttttttgtg gactttaacc gttacgtcat tttttagtcc tatatatact cgctctgtac    360 

ttggcccttt ttacactgtg actgattgag ctggtgccgt gtcgagtggt gttttttaat    420 

aggttttttt actggtaagg ctgactgtta tggctgccgc tgtggaagcg ctgtatgttg    480 

ttctggagcg ggagggtgct attttgccta ggcaggaggg tttttcaggt gtttatgtgt    540 

ttttctctcc tattaatttt gttatacctc ctatgggggc tgtaatgttg tctctacgcc    600 

tgcgggtatg tattcccccg ggctatttcg gtcgcttttt agcactgacc gatgttaacc    660 

aacctgatgt gtttaccgag tcttacatta tgactccgga catgaccgag gaactgtcgg    720 

tggtgctttt taatcacggt gaccagtttt tttacggtca cgccggcatg gccgtagtcc    780 

gtcttatgct tataagggtt gtttttcctg ttgtaagaca ggcttctaat gtttaaatgt    840 

ttttttttgt tattttattt tgtgtttaat gcaggaaccc gcagacatgt ttgagagaaa    900 

aatggtgtct ttttctgtgg tggttccgga acttacctgc ctttatctgc atgagcatga    960 

ctacgatgtg cttgcttttt tgcgcgaggc tttgcctgat tttttgagca gcaccttgca   1020 

ttttatatcg ccgcccatgc aacaagctta cataggggct acgctggtta gcatagctcc   1080 

gagtatgcgt gtcataatca gtgtgggttc ttttgtcatg gttcctggcg gggaagtggc   1140 

cgcgctggtc cgtgcagacc tgcacgatta tgttcagctg gccctgcgaa gggacctacg   1200 

ggatcgcggt atttttgtta atgttccgct tttgaatctt atacaggtct gtgaggaacc   1260 

tgaatttttg caatcatgat tcgctgcttg aggctgaagg tggagggcgc tctggagcag   1320 

atttttacaa tggccggact taatattcgg gatttgctta gagacatatt gataaggtgg   1380 

cgagatgaaa attatttggg catggttgaa ggtgctggaa tgtttataga ggagattcac   1440 

cctgaagggt ttagccttta cgtccacttg gacgtgaggg cagtttgcct tttggaagcc   1500 

attgtgcaac atcttacaaa tgccattatc tgttctttgg ctgtagagtt tgaccacgcc   1560 

accggagggg agcgcgttca cttaatagat cttcattttg aggttttgga taatcttttg   1620 

gaataaaaaa aaaaaaaaca tggttcttcc agctcttccc gctcctcccg tgtgtgactc   1680 

gcagaacgaa tgtgtaggtt ggctgggtgt ggcttattct gcggtggtgg atgttatcag   1740 

ggcagcggcg catgaaggag tttacataga acccgaagcc agggggcgcc tggatgcttt   1800 

gagagagtgg atatactaca actactacac agagcgagct aagcgacgag accggagacg   1860 

cagatctgtt tgtcacgccc gcacctggtt ttgcttcagg aaatatgact acgtccggcg   1920 

ttccatttgg catgacacta cgaccaacac gatctcggtt gtctcggcgc actccgtaca   1980 

gtagggatcg cctacctcct tttgagacag agacccgcgc taccatactg gaggatcatc   2040 

cgctgctgcc cgaatgtaac actttgacaa tgcacaacgt gagttacgtg cgaggtcttc   2100 

cctgcagtgt gggatttacg ctgattcagg aatgggttgt tccctgggat atggttctga   2160 

cgcgggagga gcttgtaatc ctgaggaagt gtatgcacgt gtgcctgtgt tgtgccaaca   2220 

ttgatatcat gacgagcatg atgatccatg gttacgagtc ctgggctctc cactgtcatt   2280 

gttccagtcc cggttccctg cagtgcatag ccggcgggca ggttttggcc agctggttta   2340 

ggatggtggt ggatggcgcc atgtttaatc agaggtttat atggtaccgg gaggtggtga   2400 

attacaacat gccaaaagag gtaatgttta tgtccagcgt gtttatgagg ggtcgccact   2460 

taatctacct gcgcttgtgg tatgatggcc acgtgggttc tgtggtcccc gccatgagct   2520 

ttggatacag cgccttgcac tgtgggattt tgaacaatat tgtggtgctg tgctgcagtt   2580 

actgtgctga tttaagtgag atcagggtgc gctgctgtgc ccggaggaca aggcgtctca   2640 

tgctgcgggc ggtgcgaatc atcgctgagg agaccactgc catgttgtat tcctgcagga   2700 

cggagcggcg gcggcagcag tttattcgcg cgctgctgca gcaccaccgc cctatcctga   2760 

tgcacgatta tgactctacc cccatgtagg cgtggacttc cccttcgccg cccgttgagc   2820 

aaccgcaagt tggacagcag cctgtggctc agcagctgga cagcgacatg aacttaagcg   2880 

agctgcccgg ggagtttatt aatatcactg atgagcgttt ggctcgacag gaaaccgtgt   2940 

ggaatataac acctaagaat atgtctgtta cccatgatat gatgcttttt aaggccagcc   3000 

ggggagaaag gactgtgtac tctgtgtgtt gggagggagg tggcaggttg aatactaggg   3060 

ttctgtgagt ttgattaagg tacggtgatc aatataagct atgtggtggt ggggctatac   3120 

tactgaatga aaaatgactt gaaattttct gcaattgaaa aataaacacg ttgaaacata   3180 

acatgcaaca ggttcacgat tctttattcc tgggcaatgt aggagaaggt gtaagagttg   3240 

gtagcaaaag tttcagtggt gtattttcca ctttcccagg accatgtaaa agacatagag   3300 

taagtgctta cctcgctagt ttctgtggat tcactagtgc cattaagtgt aatggtaagt   3360 

atcataggtt tagttttatc accatgcaag taaacttgac tgacaatgtt atttttagca   3420 

gtttgacttt gggtttttgg ataggctaga aggttaggca taaatccaac tgcatttgtg   3480 

tatggatttg cattagttga gttcccattt ctaaagttcc agtaatgttt tttaagtgag   3540 

gagttctcca ttagaacacc gttttggtca aatctaagga atatactaac acttgcaacg   3600 

gtgcctgtca tggatgaaag atctccagat acagccaaag cagctacagt agctagtact   3660 

tgactcccac attttgtaag aaccaaagta aatttgcagt cattatctga atgaattctg   3720 

cagttaggag atgggtctgg ggttgtccac agggtaagtt tgtcatcatt tttgtttcct   3780 

attgtaatgg cccctgagtt gtcaaagctt aaacccgctc caagtttagt aatcatggca   3840 

ccgttttcat tgtaatcaat gccagagcca attttagttt ttattgggtt gatatctgga   3900 

gactcagatg tgtttgtatc aaactccaga ccctttcctg catttatagc tatggcagta   3960 

ttatcaaagt ttagtccact ggattttttt atgctaactt ccagtttttt agtattgttt   4020 

gatgcattaa aaaggtatag gcctctgtta tagtttatgt ccaagttatg agatgcatta   4080 

atatacaggg gtccctgccc cagtttaaga cgtagttttg tttgagcatc aaatgggtaa   4140 

tccacatcta gaattaacaa gttgttattt atacgcatgc caccgcccgt tttaatttcc   4200 

atgttgtttg atgaatcata accaatagct cctgcaactt tggttctaag ggagttttgt   4260 

tcaacggtga cacctggtcc agtaactact gttagtgtat cggagttttg tgctacttgc   4320 

aaaggaccgc ttattttaat tcctattttt ccattattta cataaatagg atcttccatg   4380 

ttaatgccca agctacccgt ggcagtagtt agcgggggtg atgcagttac agtaagggtg   4440 

tcgctgtcac tgccagagag gggggctgat gtttgcaggg ctagctttcc atctgacact   4500 

gtaatgggcc ctttagtagc aatgcttagt ttggagtctt gcacggtcag tggggcttgt   4560 

gactgtacgc taagagcgcc gctagtaact atcagaggag cggtggttgc cactgttagg   4620 

gcgcctgagg taattgtaag tggtgcggag gtgtccaaac ttatgtttga ctttgttttt   4680 

ttaagtggct gagtaacagt ggttacattt tgggaggtga ggtttccggc cttgtctagg   4740 

gtaagaccgc tgcccatttt aagcgcaagc atgccgtggg aggtgtccaa aggttcggag   4800 

acgcgtagag agagaactcc agggggactt tcttggaaac cattgggtga aacaaatgga   4860 

ggggtaagaa agggcacagt tggaggcccg gtttctgtgt catatggata cacggggttg   4920 

aaggtgtctt cagacggtct ggcgcgtttc atctgcaaca atatgaagat agtgggtgcg   4980 

gagggacaag aacatgagga atttgacatc ccatttaaac tttggagaaa gtttgcagct   5040 

aaaaggcggc tgagatacca gagttgggag gaaggaaagg aggtgatgct gaataagctg   5100 

gacaaagatt tgctgactga ttttaagtaa gtaatttatt gtgtgtttat gttagttgaa   5160 

tggaataaga tctctaatac cacacatggt tttaataaga gtgcagaggt cctctggacc   5220 

ctgatagggg aagtgcaggc agccctctgt ttctgccgag tgctgggtga cggtgatagg   5280 

tttttctccc accataagca ccagtttttg gcgctgggtg ggtagcttgt agctgaggcg   5340 

gttgccggta gtggtttttt cgtaggtaag tttggcctgc ttgaccacac aaaagatacc   5400 

tcttttacac tggtgtaggt taaccatgtc ttcaacttct tgttttaggc gttctcgctc   5460 

ggacgccgcc ttgcgccttt ctagtaggcg ctgttcggtg ttaattccat ccaattctag   5520 

atctagagat tcagtcatct ccacctgtca aattaaagta gctaatctca gtgggggtgg   5580 

gagaaggggg gcgaggctga ttgattgggg caataacctg ttgcagtggt atgacagcgg   5640 

gcactgggaa agtagggtgg ttcatggcat ctatggcatt ccagccaatg tcaaggtatg   5700 

gatatatggc tagggcaaaa atggtactgc aaaaaaccat gacagagatg atggcgtata   5760 

accaggcttc tgacaaatcg ctctgtttgt tgtagcagct gggaatgttc catatttgag   5820 

tgaatctgca ggaaatatgt cttttgggag gcgctgaggt ttgggagcaa agcacaggta   5880 

gggcgcaaaa aatcagcaaa acaaaaatga cactccgttt cataattaaa gaattctgag   5940 

aagatcagct atagtcctgt ctctgtattg cggatggtgc ctgaggtacg caatgcgcac   6000 

acaaacccag tcaatgaact gaatgaaggc gatgactaca gtgacgaggc tgcagatgag   6060 

gataagggtg acaaatccgt aaagcaggta aactgtgaaa ggtgggatgc aatctacttc   6120 

gatgtgagcg accgcggcca atgtagagca cgcacagaaa agcgcaacaa gggtcaataa   6180 

tataagaact cgaggaatca tgtctcattt aatcatactg taaaagaaga gaacatggtt   6240 

tcagaccgtc caatctatga attttttcat tgtgtgggtt gagcacaatg ataggcctat   6300 

agatgggggg tctggcgcgt ctgcgcttta ggcaacaaat aagccacata ataataaggc   6360 

aaacaaacat aagcgctatg gaaaaccacc acatgtccaa gctcgcccag tcattgacaa   6420 

aggcatgaac ttggggtaaa tttagggcag atgttagtcc ggtagcagtg gtgttgcgat   6480 

agtccgttgt gggcgcgatg gttgagccgg tcatctctgg agcaggcaag ctgaagctgg   6540 

gtttgatcaa atttgcagtg caggcgctgg cagaaatcag gcgctaacgt ccaggaaagt   6600 

ttgatttgaa ggttgtgggt ataatcttgc ccgcctggag catatcccac atagagtaaa   6660 

ttgtccaggg gaatacaagc aagcggaaaa tcaaggcatt ttcttttcat caataaaact   6720 

gcgtctgctt ttgtatttga gataaagtaa ggtacatacc aaagcaagcg ctgtaataag   6780 

cagagcggtg gaacaaaagg tgccagtgtt ctctaaacac ttttgtgggg gccacaactt   6840 

gtactgtttg ctcatgtaca tggtaatatc gcacatttca taaaatggaa atttatacat   6900 

aaaagtttta cgattttcac cttggaagac tgtgacatta tagtcgttag tgtcacctgg   6960 

ctgccaaata gcatatacag catacttgcc aattttgtct ttgtggcgaa taataagctt   7020 

ttcatgttct gtggtgcatt ttataagagt agtgcattca ttagcttctg atttaaatgt   7080 

aacattgcaa gctggttcct taaactcaac ctttttggca gcgctgcaga ctgccgcaag   7140 

ggcgagcaag cctaaaatca tgtacctcat cttggatgtt gcccccagcg tttaaaaagc   7200 

tgacaatagg tacaaacgtg cgtgcagcag gcggcaaccc taaggcacag aagtgctagt   7260 

ataagaataa acagaattac aagagtaagg ataaccccga ccccaattcc agaaaaatta   7320 

gacaagcttg tagagttact tgaattgctc atatacttaa ttaaaaaatc ccagcacccc   7380 

gcaaaatgct tttttgacct gagttccggg agttgagctc acctcctgtt ttggaaaaat   7440 

gggagtaatg tctggttacg ctcaggctgt aggtgtgggc gcagcaaccg gtgacgcact   7500 

cgtacgttcc cggcaggtga ggagggtggt ggtggtggtg tttttcttga cggtgtagtt   7560 

gaagccgaga aggttgtgtg gcaaacttac ttcgtctcgc tggaaactgt tgtaaattac   7620 

aaatgaagag ccgttaaagt accaggtaag gtacttattg gcccgcttgt gcaaaccgga   7680 

ggtgaggttt gctttggtct gctttgggtg ggtaaaaacg gtggcgttca caggatggcg   7740 

acaggagccc cagtagattc taatttctgt atttattata ctcagcacag agatgacaac   7800 

aaagatcttg atgtaatcca gggttaggac agttgcaaac cacggtcaga acacagggac   7860 

cccgctcccg ctccactagc agggggcgct tggtaaactc ccgaatcagg ctacgtgtaa   7920 

gctctacctg ggtggtgagc cggacgccgt gcgccgggcc ctcgatatgc tcttcgggca   7980 

attcaaagta acaaaactca ccggagccgc gggcaaagca cttgtggcgg cggcagtggt   8040 

cgaggtgtgt caggcgcagt cgctctgcct ctccactggt cattcagtcg tagccgtccg   8100 

ccgagtcttt caccgcgtca aagttgggaa taaactggtc cgggtagtgg ccgggaggtc   8160 

cagaaaaggg gttgaagtaa accgaaggca cgaactcctc aataaattgt agagttccaa   8220 

tgcctccgga gcgcggctcc gaggacgagg tctgcagagt taggatcgcc tgacggggcg   8280 

taaatgaaga gcggccagcg ccgccgatct gaaatgtccc gtccggacgg agaccaagag   8340 

aggagctcac cgactcgtcg ttgagctgaa tacctcgccc tctgattttc aggtgagtta   8400 

taccctgccc gggcgaccgc accctgtgac gaaagccgcc cgcaagctgc gcccctgagt   8460 

tagtcatctg aacttcggcc tgggcgtctc tgggaagtac cacagtggtg ggagcgggac   8520 

tttcctggta caccagggca gcgggccaac tacggggatt aaggttatta cgaggtgtgg   8580 

tggtaatagc cgcctgttcg aggagaattc ggtttcggtg ggcgcggatt ccgttgaccc   8640 

gggatatcat gtggggtccc gcgctcatgt agtttattcg ggttgagtag tcttgggcag   8700 

ctccagccgc aagtcccatt tgtggctggt aactccacat gtagggcgtg ggaatttcct   8760 

tgctcataat ggcgctgacg acaggtgctg gcgccgggtg tggccgctgg agatgacgta   8820 

gttttcgcgc ttaaatttga gaaagggcgc gaaactagtc cttaagagtc agcgcgcagt   8880 

atttgctgaa gagagcctcc gcgtcttcca gcgtgcgccg aagctgatct tcgcttttgt   8940 

gatacaggca gctgcgggtg agggagcgca gagacctgtt ttttattttc agctcttgtt   9000 

cttggcccct gctttgttga aatatagcat acagagtggg aaaaatccta tttctaagct   9060 

cgcgggtcga tacgggttcg ttgggcgcca gacgcagcgc tcctcctcct gctgctgccg   9120 

ccgctgtgga tttcttgggc tttgtcagag tcttgctatc cggtcgcctt tgcttctgtg   9180 

tgaccgctgc tgttgctgcc gctgccgctg ccgccggtgc agtaggggct gtagagatga   9240 

cggtagtaat gcaggatgtt acgggggaag gccacgccgt gatggtagag aagaaagcgg   9300 

cgggcgaagg agatgttgcc cccacagtct tgcaagcaag caactatggc gttcttgtgc   9360 

ccgcgccacg agcggtagcc ttggcgctgt tgttgctctt gggctaacgg cggcggctgc   9420 

ttagacttac cggccctggt tccagtggtg tcccatctac ggttgggtcg gcgaacaggc   9480 

agtgccggcg gcgcctgagg agcggaggtt gtagcgatgc tgggaacggt tgccaatttc   9540 

tggggcgccg gcgaggggaa tgcgaccgag ggtgacggtg tttcgtctga cacctcttcg   9600 

gcctcggaag cttcgtctag gctgtcccag tcttccatca tctcctcctc ctcgtccaaa   9660 

acctcctctg cctgactgtc ccagtattcc tcctcgtccg tgggtggcgg cggcggcagc   9720 

tgcagcttct ttttgggtgc catcctggga agcaagggcc cgcggctgct gatagggctg   9780 

cggcggcggg gggattgggt tgagctcctc gccggactgg gggtccaggt aaaccccccg   9840 

tccctttcgt agcagaaact cttggcgggc tttgttgatg gcttgcaatt ggccaaggat   9900 

gtggccctgg gtaatgacgc aggcggtaag ctccgcattt ggcgggcggg attggtcttc   9960 

gtagaaccta atctcgtggg cgtggtagtc ctcaggtaca aatttgcgaa ggtaagccga  10020 

cgtccacagc cccggagtga gtttcaaccc cggagccgcg gacttttcgt caggcgaggg  10080 

accctgcagc tcaaaggtac cgataatttg actttcgcta agcagttgcg aattgcagac  10140 

cagggagcgg tgcggggtgc ataggttgca gcgacagtga cactccagta ggccgtcacc  10200 

gctcacgtct tccatgatgt cggagtggta ggcaaggtag ttggctagct gcagaaggta  10260 

gcagtgaccc caaagcggcg gagggcattc acggtactta atgggcacaa agtcgctagg  10320 

aagcgcacag caggtggcgg gcagaattcc tgaacgctct aggataaagt tcctaaagtt  10380 

ttgcaacatg ctttgactgg tgaagtctgg cagaccctgt tgcagggttt taagcaggcg  10440 

ttcggggaag ataatgtccg ccaggtgcgc ggccacggag cgctcgttga aggccgtcca  10500 

taggtccttc aagttttgct ttagcagctt ctgcagctcc tttaggttgc gctcctccag  10560 

gcattgctgc cacacgccca tggccgtttg ccaggtgtag cacagaaata agtaaacgca  10620 

gtcgcggacg tagtcgcggc gcgcctcgcc cttgagcgtg gaatgaagca cgttttgccc  10680 

gaggcggttt tcgtgcaaaa ttccaaggta ggagaccagg ttgcagagct ccacgttgga  10740 

aattttgcag gcctggcgca cgtagccctg gcgaaaggtg tagtgcaacg tttcctctag  10800 

cttgcgctgc atctccgggt cagcaaagaa ccgctgcatg cactcaagct ccacggtaac  10860 

aagcactgcg gccatcatta gcttgcgtcg ctcctccaag tcggcaggct cgcgcgtctc  10920 

aagccagcgc gccagctgct catcgccaac tgcgggtagg ccctcctcgg tttgttcttg  10980 

caagtttgca tccctctcca ggggtcgtgc acggcgcacg atcagctcgc tcatgactgt  11040 

gctcataacc ttggggggta ggttaagtgc cgggtaggca aagtgggtga cctcgatgct  11100 

gcgtttcagc acggctaggc gcgcgttgtc accctcaagt tccaccagca ctccacagtg  11160 

actttcattt tcgctgtttt cttgttgcag agcgtttgcc gcgcgtttct cgtcgcgtcc  11220 

aagaccctca aagatttttg gcacttcgtc gagcgaggcg atatcaggta tgacagcgcc  11280 

ctgccgcaag gccagctgct tgtccgctcg gctgcggttg gcacggcagg ataggggtat  11340 

cttgcagttt tggaaaaaga tgtgataggt ggcaagcacc tctggcacgg caaatacggg  11400 

gtagaagttg aggcgcgggt tgggctcgca tgtgccgttt tcttggcgtt tggggggtac  11460 

gcgcggtgag aacaggtggc gttcgtaggc aaggctgaca tccgctatgg cgaggggcac  11520 

atcgctgcgc tcttgcaacg cgtcgcagat aatggcgcac tggcgctgca gatgcttcaa  11580 

cagcacgtcg tctcccacat ctaggtagtc gccatgcctt tggtcccccc gcccgacttg  11640 

ttcctcgttt gcctctgcgt cgtcctggtc ttgcttttta tcctctgttg gtactgagcg  11700 

atcctcgtcg tcttcgctta caaaacctgg gtcctgctcg ataatcactt cctcctcctc  11760 

aagcgggggt gcctcgacgg ggaaggtggt aggcgcgttg gcggcatcgg tggaggcggt  11820 

ggtggcgaac tcaaaggggg cggttaggct gtcctccttc tcgactgact ccatgatctt  11880 

tttctgccta taggagaagg aaatggccag tcgggaagag gagcagcgcg aaaccacccc  11940 

cgagcgcgga cgcggtgcgg cgcgacgtcc accaaccatg gaggacgtgt cgtccccgtc  12000 

gccgtcgccg ccgcctcccc gcgcgccccc aaaaaagcgg ctgaggcggc gtctcgagtc  12060 

cgaggacgaa gaagactcgt cacaagatgc gctggtgccg cgcacaccca gcccgcggcc  12120 

atcgacctcg acggcggatt tggccattgc gtccaaaaag aaaaagaagc gcccctctcc  12180 

caagcccgag cgcccgccat ccccagaggt gatcgtggac agcgaggaag aaagagaaga  12240 

tgtggcgcta caaatggtgg gtttcagcaa cccaccggtg ctaatcaagc acggcaaggg  12300 

aggtaagcgc acggtgcggc ggctgaatga agacgaccca gtggcgcggg gtatgcggac  12360 

gcaagaggaa aaggaagagt ccagtgaagc ggaaagtgaa agcacggtga taaacccgct  12420 

gagcctgccg atcgtgtctg cgtgggagaa gggcatggag gctgcgcgcg cgttgatgga  12480 

caagtaccac gtggataacg atctaaaggc aaacttcaag ctactgcctg accaagtgga  12540 

agctctggcg gccgtatgca agacctggct aaacgaggag caccgcgggt tgcagctgac  12600 

cttcaccagc aacaagacct ttgtgacgat gatggggcga ttcctgcagg cgtacctgca  12660 

gtcgtttgca gaggtaacct acaagcacca cgagcccacg ggctgcgcgt tgtggctgca  12720 

ccgctgcgct gagatcgaag gcgagcttaa gtgtctacac gggagcatta tgataaataa  12780 

ggagcacgtg attgaaatgg atgtgacgag cgaaaacggg cagcgcgcgc tgaaggagca  12840 

gtctagcaag gccaagatcg tgaagaaccg gtggggccga aatgtggtgc agatctccaa  12900 

caccgacgca aggtgctgcg tgcatgacgc ggcctgtccg gccaatcagt tttccggcaa  12960 

gtcttgcggc atgttcttct ctgaaggcgc aaaggctcag gtggctttta agcagatcaa  13020 

ggctttcatg caggcgctgt atcctaacgc ccagaccggg cacggtcacc ttctgatgcc  13080 

actacggtgc gagtgcaact caaagcctgg gcatgcaccc tttttgggaa ggcagctacc  13140 

aaagttgact ccgttcgccc tgagcaacgc ggaggacctg gacgcggatc tgatctccga  13200 

caagagcgtg ctggccagcg tgcaccaccc ggcgctgata gtgttccagt gctgcaaccc  13260 

tgtgtatcgc aactcgcgcg cgcagggcgg aggccccaac tgcgacttca agatatcggc  13320 

gcccgacctg ctaaacgcgt tggtgatggt gcgcagcctg tggagtgaaa acttcaccga  13380 

gctgccgcgg atggttgtgc ctgagtttaa gtggagcact aaacaccagt atcgcaacgt  13440 

gtccctgcca gtggcgcata gcgatgcgcg gcagaacccc tttgattttt aaacggcgca  13500 

gacggcaagg gtggggggta aataatcacc cgagagtgta caaataaaaa catttgcctt  13560 

tattgaaagt gtctcctagt acattatttt tacatgtttt tcaagtgaca aaaagaagtg  13620 

gcgctcctaa tctgcgcact gtggctgcgg aagtagggcg agtggcgctc caggaagctg  13680 

tagagctgtt cctggttgcg acgcagggtg ggctgtacct ggggactgtt aagcatggag  13740 

ttgggtaccc cggtaataag gttcatggtg gggttgtgat ccatgggagt ttggggccag  13800 

ttggcaaagg cgtggagaaa catgcagcag aatagtccac aggcggccga gttgggcccc  13860 

tgcacgcttt gggtggactt ttccagcgtt atacagcggt cgggggaaga agcaatggcg  13920 

ctacggcgca ggagtgactc gtactcaaac tggtaaacct gcttgagtcg ttggtcagaa  13980 

aagccaaagg gctcaaagag gtagcatgtt tttgagcgcg ggttccaggc aaaggccatc  14040 

cagtgtacgc ccccagtctc gcgaccggcc gtattgacta tggcgcaggc gagcttgtgt  14100 

ggagaaacaa agcctggaaa gcgcttgtca taggtgccca aaaaatatgg cccacaacca  14160 

agatctttga caatggcttt cagttcctgc tcactggagc ccatggcggc agctgttgtt  14220 

gatgttgctt gcttctttta tgttgtggcg ttgccggccg agaagggcgt gcgcaggtac  14280 

acggtctcga tgacgccgcg gtgcggctgg tgcacacgga ccacgtcaaa gacttcaaac  14340 

aaaacataaa gaagggtggg ctcgtccatg ggatccacct caaaagtcat gtctagcgcg  14400 

tgggcggagt tggcgtagag aaggttttgg cccaggtctg tgagtgcgcc catggacata  14460 

aagttactgg agaatgggat gcgccaaagg gtgcgatcgc aaagaaactt tttctgggta  14520 

atactgtcaa ccgcggtttt gcctattagt gggtagggca cgttggcggg gtaagcctgt  14580 

ccctcgcgca tggtgggagc gaggtagcct acgaatcctg agttgttatg ctggtgaaga  14640 

attccaacct gctgatactc cttgtattta gtatcgtcaa ccacttgccg gctcatgggc  14700 

tggaagtttc tgaagaacga gtacatgcgg tccttgtagc tttctggaat gtagaagccc  14760 

tggtagccaa tattgtagtt ggccaacatc tgcaccagga accagtcctt ggtcatgttg  14820 

cactgagcta cgttgtagcc ctccccgtca actgagcgtt taatctcaaa ctcattggga  14880 

gtaagcaggc ggtcgttgcc cggccagcta acagaagagt caaaggtaat ggccaccttc  14940 

ttaaaggtgt gattaagata gaaggttccg tcaaggtatg gtatggagcc agagtaggtg  15000 

tagtaagggt cgtagcctga tcccagggaa ggggtttcct ttgtcttcaa gcgtgtgaag  15060 

gcccaaccgc gaaatgctgc ccagttgcgc gatgggatgg agatgggcac gttggtggcg  15120 

ttggcgggta tggggtatag catgttggcg gcggaaaggt agtcattaaa ggactggtcg  15180 

ttggtgtcat ttctgagcat ggcttccagc gtggaggccg tgttgtgggc catggggaag  15240 

aaggtggcgt aaagacaaat gctgtcaaac ttaatgctag ccccgtcaac tctaagatcg  15300 

tttcccagag agctctgcag aaccatgtta acatccttcc tgaagttcca ttcatatgta  15360 

tatgagcctg gcaggaggag gaggttttta atggcaaaaa acttttgggg cacctgaatg  15420 

tgaaagggca cgtagcggcc gtttcccaac aacatggagc gataacggag gcccgcattg  15480 

cggtggtggt taaagggatt aacgttgtcc atgtagtcca gagaccagcg cgccccaagg  15540 

ttaatgtagc agtctacaag cccgggagcc accactcgct tgttcatgta gtcgtaggtg  15600 

ttggggttgt cagatatttc cacattggtg gggttgtatt ttagcttgtc tggcaggtac  15660 

agcgcaatat tggagtaaag gaaatttctc cataggttgg catttaggtt aatttccatg  15720 

gcaaagttgt tacccactcc tatttcatta cgtgttgcaa aagtttcatc ttttgtccat  15780 

gtagtatctc cattatcgcc tgagccattg ccattagcct taatagcttg ataggtgtca  15840 

gttaccccaa tacccccaag aggaaaacaa taatttggca attcatcctc agttccatgg  15900 

ttttcaatga ttctaacatc tggatcatag ctgtctacag cctgattcca catagaaaaa  15960 

tatctggttc tatcacctat ggaatcaagc aagagttgat aggacagctc tgtgtttctg  16020 

tcttgcaaat ctaccacggc atttagctgc gatgcctgac cagcaagaac acccatgttg  16080 

ccagtgctgt tataatacat taggccaata aaattgtccc tgaaagcaat gtaattgggt  16140 

ctgtttggca tagattgttg acccaacata gctttagaat tttcatcacc ttttccaggt  16200 

ttgtaagaca gatgtgtgtc tggggtttcc atatttacat cttcactgta caaaaccact  16260 

tttggtttag tagcattgcc ttgccggtcg ttcaaagagg tagtatttga gaagaattgc  16320 

aagtcaacct ttggaagagg cacccctttt tcatccggaa ccagaacgga ttgaccacca  16380 

aaaggatttg taggcctggc ataagatcca tagcatggtt tcatgggagt tgttttttta  16440 

agcactctcc ctcctgccgc attagcatca gcttcgttcc actgagattc gccaatttga  16500 

ggttctggtt gataggaagg atctgcgtat acaggtttag cttgtgtttc tgcattgtct  16560 

gatcctattt gtagcccgct ttttgtaatt gtttctccag acaaaggagc ctgggcatag  16620 

acatgtgttt tcttagtagc ctgatctcga gcgttttgct cttcttcttc ctcttcttca  16680 

tcttcatctt cctcttcttc atcctcggca actgcccggc cgctatcttc ggtttgttcc  16740 

cactcacagg agttaggagc gcccttggga gctagagcgt tgtaggcagt gccggagtag  16800 

ggcttaaaag taggccccct gtccagcacg ccgcggatgt caaagtacgt ggaagccata  16860 

tcaagcacac ggttgtcacc cacagccagg gtgaaccgcg ctttgtacga gtacgcggta  16920 

tcctcgcggt ccacagggat gaaccgcagc gtcaaacgct gggaccggtc tgtggttacg  16980 

tcgtgcgtag gtgccaccgt ggggtttcta aacttgttat tcaggctgaa gtacgtctcg  17040 

gtggcgcggg caaactgcac cagcccgggg ctcaggtact ccgaggcgtc ctggcccgag  17100 

atgtgcatgt aagaccactg cggcatcatc gaaggggtag ccatcttgga aagcgggcgc  17160 

acggcggctc agcagctcct ctggcggcga catggacgca tacatgacac atacgacacg  17220 

ttagctattt agaagcatcg tcggcgcttc agggattgca cccccagacc cacgatgctg  17280 

ttcagtgtgc tttgccagtt gccactggct acgggccgca tcgatcgcgg accgctggcg  17340 

gcacggcgca gggacgcgcg gctagggcgg gttacaacaa cggcggacgg ccctggcagc  17400 

acaggtttct gctgggtgtc agcgggggga ggcaggtcca gcgttacagg tgtgtgctgg  17460 

cccagcactc cggtagccat gggcgcgatg ggacgggtgg tgggcaggcc ttgctttagt  17520 

gcctcctcgt acgagggagg ctcatctatt tgcgtcacca gagtttcttc cctgtcgggc  17580 

cgcggacgct tttcgccacg cccctctgga gacactgtct ccacggccgg tggaggctcc  17640 

tctacgggag ggcggggatc aagcttactg ttaatcttat tttgcactgc ctggttggcc  17700 

aggtccacca ccccgctaat gccagaggcc aggccatcta ccaccttttg ttggaaattt  17760 

tgctctttca acttgtccct cagcatctgg cctgtgctgc tgttccaggc cttgctgcca  17820 

tagttcttaa tggtggaacc gaaattttta atgccgctcc acagcgagcc ccagctgaag  17880 

gcgccaccgc tcatattgct ggtgccgata tcttgccagt ttcccatgaa cgggcgcgag  17940 

ccgtgtcgcg gggccagaga cgcaaagttg atgtcttcca ttctacaaaa tagttacagg  18000 

accaagcgag cgtgagactc cagacttttt attttgattt ttccacatgc aacttgtttt  18060 

taatcagtgt ctctgcgcct gcaaggccac ggatgcaatt ccgggcacgg cgccaatcgc  18120 

cgcggcgatc agtggaataa ggaggggcag gataccgccg cgcatgcgac ggtgcgacgc  18180 

gcgccgccgc cggtggtgcg cacgacgcat gccgcccgtc aggccgtggc cggccatgcc  18240 

cctcctacgg tgcattcttc ctcggaatcc cggcaccggg aaacggaggc ggcaggtgag  18300 

ggccatatct gcaagaacca caaagaccgg cttttaaacg atgctggggt ggtagcgcgc  18360 

tgttggcagc accagggtcc tgcctccttc gcgagccacc ctgcgcacgg aaatcggggc  18420 

cagcacgggc tggcgacggc gacggcggcg gcgggttcca gtggtggttc ggcgtcgggt  18480 

agtcgctcgt cttctggggc ggtaggtgta gccacgatag ccgggggtag gcgcgatgga  18540 

aggatgtagg gcatattcgg gcagtagtgc gctggcggtg ccgtacttcc tggaacggcg  18600 

cgggcgccgg ggggctgaaa cgcgaaacat ccacgggtcc gtttgcacct ccgtagaggt  18660 

tttggacgcg gccgcagcgg ccgcctgcac cgcggcatct gccaccgccg aggcaaccgg  18720 

ggacgtttgt gtctccatgc cctctgtggc agtggcaata ctagtgctac tggtggtggg  18780 

tatctgaacg tccacggtct gcacgcccag tcccggtgcc acctgcttga ttggccgcac  18840 

gcggacctcg ggctccagcc caggctccac ggtcattttt tccaagacat cttccagtcg  18900 

ctggcgcttg ggtaccatca gctgcacggt gggtgccaag tcaccagact cgcgctttag  18960 

gccgcgcttt tcttcggacg gtgcaagcgt gggcagcacc tgctgcagtg tcacgggctt  19020 

taggctaggt gttgggttgc cctcgtccag cggcaacgcc aacatgtcct tatgccgctt  19080 

tccgtaggca aactccccga ggcgctcgtt ggcctgctca agcaggtcct cgtcgccgta  19140 

cacctcatca tacacgcgct tgtaggtgcg ggtggagcgc tcaccgggcg taaaaactac  19200 

ggtggtgccg ggtcgcaaaa cacgtcttac gcgtcgacct ttccactgta cccgccgcct  19260 

gggcgcggtt gcgtgcagca gttccacctc gtcgtcaagt tcatcatcat catcatcttt  19320 

ctttttcttt ttgacccgct ttagctttcg gggcttgtaa tcctgctctt ccttcttcgg  19380 

ggggccatag atctccggcg cgatgacctg gagcatctct tctttgattt tgcgcttgga  19440 

catagcttcg ttgcgcgccg ccgccgctgg atacatacaa cagtacgagt ctaagtagtt  19500 

ttttcttgca atctagttgc gcggggggcg ggtgcgcacg ggcacgcgca ggccgctaac  19560 

cgagtcgcgc acccagtaca cgttgcccct gcgaccctga gtcatagcac taatggccgc  19620 

ggctgctgcg gcggccgctc gtcgcctgga cctggggggc acagtgacaa tacccgcggc  19680 

cagccttcga gcggcccgca tggccgcccg tcggccggtg cgacgtgcgc ggttaagcag  19740 

ggccgccgcc gcgcgttggg cggcagtgcc gggtcggcgg cggtggcgac gtgctacgcg  19800 

cctccgccgt ctcttcattt tagcataacg ccgggctccg cgcaccacgg tctgaatggc  19860 

cgcgtccact gtggacactg gtggcggcgt gggcgtgtag ttgcgcgcct cctccaccac  19920 

cgcgtcaatg gcgtcatcga cggtggtgcg cccagtgcgg ccgcgtttgt gcgcgcccca  19980 

gggcgcgcgg tagtgcccgc gcacgcgcac tgggtgttgg tcggagcgct tctttgcccc  20040 

gccaaacatc ttgcttggga agcgcaggcc ccagcctgtg ttattgctgg gcgatataag  20100 

gatggacatg tttgctcaaa aagtgcggct cgataggacg cgcggcgaga ctatgcccag  20160 

ggccttgtaa acgtaggggc aggtgcggcg tctggcgtca gtaatggtca ctcgctggac  20220 

tcctccgatg ctgttgcgca gcggtagcgt cccgtgatct gtgagagcag gaacgttttc  20280 

actgacggtg gtgatggtgg gggctggcgg gcgcgccaaa atctggttct cgggaaagcg  20340 

attgaacacg tgggtcagag aggtaaactg gcggatgagc tgggagtaga cggcctggtc  20400 

gttgtagaag ctcttggagt gcacgggcaa cagctcggcg cccaccaccg gaaagttgct  20460 

gatctggctc gtggagcgga aggtcacggg gtcttgcatc atgtctggca acgaccagta  20520 

gacctgctcc gagccgcagg ttacgtcagg agtgcaaagg agggtccatg agcggatccc  20580 

ggtctgaggg tcgccgtagt tgtatgcaag gtaccagctg cggtactggg tgaaggtgct  20640 

gtcattgctt attaggttgt aactgcgttt cttgctgtcc tctgtcaggg gtttgatcac  20700 

cggtttcttc tgaggcttct cgacctcggg ttgcgcagcg ggggcggcag cttctgccgc  20760 

tgcctcggcc tcagcgcgct tctcctccgc ccgtgtggca aaggtgtcgc cgcgaatggc  20820 

atgatcgttc atgtcctcca ccggctgcat tgccgcggct gccgcgttgg agttctcttc  20880 

cgcgccgctg ccactgttgt tgccgccgcc tgcgccatcc ccgccctgtt cggtgtcatc  20940 

ttttaagctt gcctggtagg cgtccacatc caacagtgcg ggaatgttac caccctccag  21000 

gtcatcgtag gtgatcctaa agccctcctg gaagggttgc cgcttgcgga tgcccaacaa  21060 

gttgctcagg cggctgtggg tgaagtccac cccgcatcct ggcagcaaaa tgatgtctgg  21120 

atggaaggct tcgtttgtat ataccccagg catgacaaga ccagtgactg ggtcaaaccc  21180 

cagtctgaag ttgcgggtgt caaactttac cccgatgtcg ctttccagaa ccccgttctg  21240 

cctgcccact ttcaagtagt gctccacgat cgcgttgttc ataaggtcta tggtcatggt  21300 

ctcggagtag ttgccctcgg gcagcgtgaa ctccacccac tcatatttca gctccacctg  21360 

tttgtcctta gtaagcgagc gcgacaccat cacccgcgcc ttaaacttat tggtaaacat  21420 

gaactcgttc acatttggca tgttggtatg caggatggtt ttcaggtcgc cgccccagtg  21480 

cgaacggtcg tcaagattga tggtctgtgt gcttgcctcc cccgggctgt agtcattgtt  21540 

ttgaatgacc gtggttagaa agttgctgtg gtcgttctgg tagttcaggg atgccacatc  21600 

cgttgacttg ttgtccacaa ggtacacacg ggtggtgtcg aataggggtg ccaactcaga  21660 

gtaacggatg ctgtttctcc ccccggtagg ccgcaggtac cgcggaggca caaacggcgg  21720 

gtccagggga gcatcgaagg gggaacccag cgccgccgcc actggcgccg cgctcaccac  21780 

gctctcgtag gagggaggag gaccttcctc atacatcgcc gcgcgctgca tactaagggg  21840 

aatacaagaa aaccaacgct cggtgccatg gccttggtga gttttttatt ttgcatcatg  21900 

cttttttttt ttttttttaa aacattctcc ccagcctggg gcgaaggtgc gcaaacgggt  21960 

tgccactccc tcccaaatcc aggacgctgc tgtcgtctgc cgagtcatcg tcctcccaca  22020 

ccagaccccg ctgacggtcg tgcctttgac gacgggtggg cgggcgcggg ccgggcacat  22080 

ccctgtgctc ctgcgcatac gtcttccatc tactcatctt gtccactagg ctctctatcc  22140 

cgttgttggg aaatgccgga ggcaggttct tttcgcgctg cggctgcagc agcgagttgt  22200 

ttaggtactc ctcctcgccc agcaggcgcg ggcgggtggt gcgagtgctg gtaaaagacc  22260 

ctatcaagct tggaaatggg ctactcgcat ctgaccgcgg ggccgcagcg cctagatcgg  22320 

acaagctgct tggcctgcgg aagctttcct ttcgcagcgc cgcctctgcc tgctcgcgct  22380 

gttgcaactc tagcagggtc tgcggttgcg gggaaaacac gctgtcgtct atgtcgtccc  22440 

agaggaatcc atcgttaccc tcgggcacct caaatccccc ggtgtagaaa ccagggggcg  22500 

gtagccagtg cgggttcaag atggcattgg tgaaatactc ggggttcacg gcggccgcgc  22560 

gatgcaagta gtccattagg cgattgataa acggccggtt tgaggcatac atgcccggtt  22620 

ccatgttgcg cgcggtcatg tccagcgcca cgctgggcgt taccccgtcg cgcatcaggt  22680 

taaggctcac gctctgctgc acatagcgca agatgcgctc ctcctcgctg tttaaactgt  22740 

gcaacgaggg gatcttctgc cgccggttgg tcagcaggta gttcagggtt gcctccaggc  22800 

tgcccgtgtc ctcctgcccc agcgcgcggc tgacacttgt aatctcctgg aaagtatgct  22860 

cgtccacatg cgcctgacct atggcctcgc ggtacagtgt cagcaagtga cctaggtatg  22920 

tgtcccggga cacgctgcca ctgtccgtga agggcgctat tagcagcagc aacaggcgcg  22980 

agttgggcgt cagcaagcta gacacggtcg cgcggtcgcc tgtgggagcc cgcacccccc  23040 

acagcccctg caagttcttg aaagcctggc tcaggtttac ggtctgcagg ccttgtctac  23100 

tggtctggaa aaaatagtct ggcccggact ggtacacctc actttgcggt gtctcagtca  23160 

ccattagccg cagtgcgctc acaaagttgg tgtagtcctc ctgtccccgc ggcacgttgg  23220 

cgggctgtgt actcaggaag gcgtttagtg caaccatgga gcccaggttg ccctgctgct  23280 

gcgcgcgctc acgctgcgcc acggcctcgc gcacatcccc caccagccgg tccaggttgg  23340 

tctgcacgtt gccgctgttg taacgagcca cgcgctgaag cagcgcgtcg tagaccaggc  23400 

cggcctcatc gggccggatg gccctgtttt cggccagcgc gtttacgatc gccagcacct  23460 

tctcgtgcgt ggggtttgcg cgcgccggga ccaccgcttc cagaattgcg gagagccggt  23520 

tggcctgcgg ctgctgccgg aacgcgtcag ggttacgcgc agtcagcgac atgatgcggt  23580 

ccatgacctg gcgccagtcg tccgtggagt taaggccgga cggctggctc tgcagcgccg  23640 

cccgcaccgc cgggtccgtt gcgtcttgca tcatctgatc agaaacatca ccgcttagta  23700 

ctcgccgtcc tctggctcgt actcatcgtc ctcgtcatat tcctccacgc cgccgacgtt  23760 

gccagcgcgc gcgggtgcca ccgccagccc aggtccggcc ccagctgcct ccagggcgcg  23820 

tcggcttggg gcccagcgca ggtcagcgcc cgcgtcaaag taggactcgg cctctctatc  23880 

gccgctgccc gtgccagcca gggccctttg caggctgtgc atcagctcgc ggtcgctgag  23940 

ctcgcgccgc cggctcacgc tcacggcctt gtggatgcgc tcgttgcgat aaacgcccag  24000 

gtcgtcgctc aaggtaagca ccttcaacgc catgcgcatg tagaacccct cgatctttac  24060 

ctccttgtct atgggaacgt aaggggtatg gtatatcttg cgggcgtaaa acttgcccag  24120 

actgagcatg gaatagttaa tggcggccac cttgtcagcc aggctcaagc tgcgctcctg  24180 

caccactatg ctctgcagaa tgtttatcaa atcgagcagc cagcggccct cgggctctac  24240 

tatgtttagc agcgcatccc tgaatgcctc gttgtccctg ctgtgctgca ctataaggaa  24300 

cagctgcgcc atgagcggct tgctatttgg gttttgctcc agcgcgctta caaagtccca  24360 

cagatgcatc agtcctatag ccacctcctc gcgcgccaca agcgtgcgca cgtggttgtt  24420 

aaagcttttt tgaaagttaa tctcctggtt caccgtctgc tcgtacgcgg ttaccaggtc  24480 

ggcggccgcc acgtgtgcgc gcgcgggact aatcccggtc cgcgcgtcgg gctcaaagtc  24540 

ctcctcgcgc agcaaccgct cgcggttcag gccatgccgc aactcgcgcc ctgcgtggaa  24600 

ctttcgatcc cgcatctcct cgggctcctc tccctcgcgg tcgcgaaaca ggttctgccg  24660 

cggcacgtac gcctcgcgcg tgtcacgctt cagctgcacc cttgggtgtc gctcaggaga  24720 

gggcgctcct agccgcgcca ggccctcgcc ctcctccaag tccaggtagt gccgggcccg  24780 

gcgccgcggg ggttcgtaat caccatctgc cgccgcgtca gccgcggatg ttgcccctcc  24840 

tgacgcggta ggagaagggg agggtgccct gcatgtctgc cgctgctctt gctcttgccg  24900 

ctgctgagga ggggggcgca tctgccgcag caccggatgc atctgggaaa agcaaaaaag  24960 

gggctcgtcc ctgtttccgg aggaatttgc aagcggggtc ttgcatgacg gggaggcaaa  25020 

cccccgttcg ccgcagtccg gccggcccga gactcgaacc gggggtcctg cgactcaacc  25080 

cttggaaaat aaccctccgg ctacagggag cgagccactt aatgctttcg ctttccagcc  25140 

taaccgctta cgccgcgcgc ggccagtggc caaaaaagct agcgcagcag ccgccgcgcc  25200 

tggaaggaag ccaaaaggag cgctcccccg ttgtctgacg tcgcacacct gggttcgaca  25260 

cgcgggcggt aaccgcatgg atcacggcgg acggccggat ccggggttcg aaccccggtc  25320 

gtccgccatg atacccttgc gaatttatcc accagaccac ggaagagtgc ccgcttacag  25380 

gctctccttt tgcacggtct agagcgtcaa cgactgcgca cgcctcaccg gccagagcgt  25440 

cccgaccatg gagcactttt tgccgctgcg caacatctgg aaccgcgtcc gcgactttcc  25500 

gcgcgcctcc accaccgccg ccggcatcac ctggatgtcc aggtacatct acggatatca  25560 

tcgccttatg ttggaagacc tcgcccccgg agccccggcc accctacgct ggcccctcta  25620 

ccgccagccg ccgccgcact ttttggtggg atatcagtac ctggtgcgga cttgcaacga  25680 

ctacgtcttt gactcaaggg cttactcgcg tctcaggtac accgagctct cgcagccggg  25740 

tcaccagacc gttaactggt ccgttatggc caactgcact tacaccatca acacgggcgc  25800 

ataccaccgc tttgtggaca tggatgactt ccagtctacc ctcacgcagg tgcagcaggc  25860 

catattagcc gagcgcgttg tcgccgacct ggccctgctt cagccgatga ggggcttcgg  25920 

ggtcacacgc atgggaggaa gagggcgcca cctacggcca aactccgccg ccgccgtagc  25980 

gatagatgca agagatgcag gacaagagga aggagaagaa gaagtgccgg tagaaaggct  26040 

catgcaagac tactacaaag acctgcgccg atgtcaaaac gaagcctggg gcatggccga  26100 

ccgcctgcgc attcagcagg ccggacccaa ggacatggtg cttctgtcga ccatccgccg  26160 

tctcaagacc gcctacttta attacatcat cagcagcacc tccgccagaa acaaccccga  26220 

ccgccacccg ctgccgcccg ccacggtgct cagcctacct tgcgactgtg actggttaga  26280 

cgcctttctc gagaggtttt ccgatccggt cgatgcggac tcgctcaggt ccctcggtgg  26340 

cggagtacct acacaacaat tgttgagatg catcgttagc gccgtatccc tgccgcacgg  26400 

cagccccccg ccaacccata accgggacat gacgggcggc gtcttccaac tgcgcccccg  26460 

cgagaacggc cgcgccgtca ccgagaccat gcgccgtcgc cgcggggaga tgatcgagcg  26520 

ctttgtcgac cgcctcccgg tgcgccgtcg tcgccgccgt gtcccccctc ccccaccgcc  26580 

gccagaagaa gaagaagaag gggaggccct tatggaagag gagattgaag aagaagaggc  26640 

ccctgtagcc tttgagcgcg aggtgcgcga cactgtcgcc gagctcatcc gtcttctgga  26700 

ggaggagtta accgtgtcgg cgcgcaactc ccagtttttc aacttcgccg tggacttcta  26760 

cgaggccatg gagcgccttg aggccttggg ggatatcaac gaatccacgt tgcgacgctg  26820 

ggttatgtac ttcttcgtgg cagaacacac cgccaccacc ctcaactacc tctttcagcg  26880 

cctgcgaaac tacgccgtct tcgcccggca cgtggagctc aatctcgcgc aggtggtcat  26940 

gcgcgcccgc gatgccgaag ggggcgtggt ctacagccgc gtctggaacg agggaggcct  27000 

caacgccttc tcgcagctca tggcccgcat ctccaacgac ctcgccgcca ccgtggagcg  27060 

agccggacgc ggagatctcc aggaggaaga gatcgagcag ttcatggccg aaatcgccta  27120 

tcaagacaac tcaggagacg tgcaggagat tttgcgccag gccgccgtca acgacaccga  27180 

aattgattct gtcgaactct ctttcaggtt caagctcacc gggcccgtcg tcttcacgca  27240 

gaggcgccag attcaggaga tcaaccgccg cgtcgtcgcg ttcgccagca acctccgcgc  27300 

gcagcaccag ctcctgcccg cgcgcggcgc cgacgtgccc ctgccccctc tcccggcggg  27360 

tcccgagccc cccctacctc cgggggcccg cccgcgtcac cgcttttaga tgcatcatcc  27420 

aaggacaccc ccgcggccca ccgcccgccg cgcggtaccg tagtcgcgcc gcggggatgc  27480 

ggcctcttgc aagtcatcga cgccgccacc aaccagcccc tggaaatcag gtatcacctg  27540 

gacctagccc gcgccctgac ccggctatgc gaggtaaacc tgcaggagct cccgcctgac  27600 

ctgtcgccgc gggagctcca gaccatggac agctcccatc tgcgcgatgt tgtcatcaag  27660 

ctccgaccgc cgcgcgcgga catctggact ttgggctcgc gcggcgtggt ggtccgatcc  27720 

accataactc ccctcgagca gccagacggt caaggacaag cagccgaagt agaagaccac  27780 

cagccaaacc cgccaggcga ggggctcaaa ttcccactct gcttccttgt gcgcggtcgt  27840 

caggtcaacc tcgtgcagga tgtacagccc gtgcaccgct gccagtactg cgcacgtttt  27900 

tacaaaagcc agcacgagtg ttcggcccgt cgcagggact tctactttca ccacatcaac  27960 

agccactcct ccaactggtg gcgggagatc cagttcttcc cgatcggctc gcatcctcgc  28020 

accgagcgtc tctttgtcac ctacgatgta gagacctata cttggatggg ggcctttggg  28080 

aagcagctcg tgcccttcat gctggttatg aagttcggcg gagatgagcc tctggtgacc  28140 

gccgcgcgag acctagccgt ggaccttgga tgggaccgct gggaacaaga cccgcttacc  28200 

ttctactgca tcaccccaga aaaaatggcc ataggtcgcc agtttaggac ctttcgcgac  28260 

cacctgcaaa tgctaatggc ccgtgacctg tggagctcat tcgtcgcttc caaccctcat  28320 

cttgcagact gggccctgtc agaacacggg ctcagctccc ctgaggagct cacctacgag  28380 

gaacttaaaa aattgccctc catcaagggc accccgcgct tcttggaact ttacatcgtg  28440 

ggccacaaca tcaacggctt cgacgagatc gtgctcgccg cccaggtaat taacaaccgt  28500 

tccgaggtgc cgggaccctt ccgcatcaca cgcaacttta tgcctcgcgc gggaaagata  28560 

cttttcaacg atgtcacctt cgccctgcca aacccgcgtt ccaaaaagcg cacggacttt  28620 

ttgctctggg agcagggcgg atgcgacgac actgacttca aataccagta cctcaaagtc  28680 

atggttaggg acacctttgc gctcacccac acctcgctcc ggaaggccgc gcaggcatac  28740 

gcgctacccg tagaaaaggg atgctgcgcc taccaggccg tcaaccagtt ctacatgcta  28800 

ggctcttacc gttcggaggc cgacgggttt ccgatccaag agtactggaa agaccgcgaa  28860 

gagtttgtcc tcaaccgcga gctgtggaaa aaaaagggac aggataagta tgacatcatc  28920 

aaggaaaccc tggactactg cgccctagac gtgcaggtca ccgccgagct ggtcaacaag  28980 

ctgcgcgact cctacgcctc cttcgtgcgt gacgcggtag gtctcacaga cgccagcttc  29040 

aacgtcttcc agcgtccaac catatcatcc aactcacatg ccatcttcag gcagatagtc  29100 

ttccgagcag agcagcccgc ccgtagcaac ctcggtcccg acctcctcgc tccctcgcac  29160 

gaactatacg attacgtgcg cgccagcatc cgcggtggaa gatgctaccc tacatatctt  29220 

ggaatactca gagagcccct ctacgtttac gacatttgcg gcatgtacgc ctccgcgctc  29280 

acccacccca tgccatgggg tcccccactc aacccatacg agcgcgcgct tgccgcccgc  29340 

gcatggcagc aggcgctaga cttgcaagga tgcaagatag actacttcga cgcgcgcctg  29400 

ctgcccgggg tctttaccgt ggacgcagac cccccggacg agacgcagct agacccacta  29460 

ccgccattct gttcgcgcaa gggcggccgc ctctgctgga ccaacgagcg cctacgcgga  29520 

gaggtagcca ccagcgttga ccttgtcacc ctgcacaacc gcggttggcg cgtgcacctg  29580 

gtgcccgacg agcgcaccac cgtctttccc gaatggcggt gcgttgcgcg cgaatacgtg  29640 

cagctaaaca tcgcggccaa ggagcgcgcc gatcgcgaca aaaaccaaac cctgcgctcc  29700 

atcgccaagt tgctgtccaa cgccctctac gggtcgtttg ccaccaagct tgacaacaaa  29760 

aagattgtct tttctgacca gatggacgcg gccaccctca aaggcatcac cgcgggccag  29820 

gtgaatatca aatcctcctc gtttttggaa actgacaatc ttagcgcaga agtcatgccc  29880 

gcttttgaga gggagtactc accccaacag ctggccctcg cagacagcga tgcggaagag  29940 

agtgaggacg aacgcgcccc cacccccttt tatagccccc cttcaggaac acccggtcac  30000 

gtggcctaca cctataaacc aatcaccttc cttgatgccg aagagggcga catgtgtctt  30060 

cacaccctgg agcgagtgga ccccctagtg gacaacgacc gctacccctc ccacttagcc  30120 

tccttcgtgc tggcctggac gcgagccttc gtctcagagt ggtccgagtt tctatacgag  30180 

gaggaccgcg gaacaccgct cgaggacagg cctctcaagt ctgtatacgg ggacacggac  30240 

agccttttcg tcaccgagcg tggacaccgg ctcatggaaa ccagaggtaa gaaacgcatc  30300 

aaaaagcatg ggggaaacct ggtttttgac cccgaacggc cagagctcac ctggctcgtg  30360 

gaatgcgaga ccgtctgcgg ggcctgcggc gcggatgcct actccccgga atcggtattt  30420 

ctcgcgccca agctctacgc ccttaaaagt ctgcactgcc cctcgtgcgg cgcctcctcc  30480 

aagggcaagc tgcgcgccaa gggccacgcc gcggaggggc tggactatga caccatggtc  30540 

aaatgctacc tggccgacgc gcagggcgaa gaccggcagc gcttcagcac cagcaggacc  30600 

agcctcaagc gcaccctggc cagcgcgcag cccggagcgc accccttcac cgtgacccag  30660 

actacgctga cgaggaccct gcgcccgtgg aaagacatga ccctggcccg tctggacgag  30720 

caccgactac tgccgtacag cgaaagccgc cccaacccgc gaaacgagga gatatgctgg  30780 

atcgagatgc cgtagagcac gtgaccgagc tgtgggaccg cctggaactg cttggtcaaa  30840 

cgctcaaaag catgcctacg gcggacggcc tcaaaccgtt gaaaaacttt gcttccttgc  30900 

aagaactgct atcgctgggc ggcgagcgcc ttctggcgca tttggtcagg gaaaacatgc  30960 

aagtcaggga catgcttaac gaagtggccc ccctgctcag ggatgacggc agctgcagct  31020 

ctcttaacta ccagttgcag ccggtaatag gtgtgattta cgggcccacc ggctgcggta  31080 

agtcgcagct gctcaggaac ctgctttctt cccagctgat ctcccctacc ccggaaacgg  31140 

ttttcttcat cgccccgcag gtagacatga tccccccatc tgaactcaaa gcgtgggaaa  31200 

tgcaaatctg tgagggtaac tacgcccctg ggccggatgg aaccattata ccgcagtctg  31260 

gcaccctccg cccgcgcttt gtaaaaatgg cctatgacga tctcatcctg gaacacaact  31320 

atgacgttag tgatcccaga aatatcttcg cccaggccgc cgcccgtggg cccattgcca  31380 

tcattatgga cgaatgcatg gaaaatctcg gaggtcacaa gggcgtctcc aagttcttcc  31440 

acgcatttcc ttctaagcta catgacaaat ttcccaagtg caccggatac actgtgctgg  31500 

tggttctgca caacatgaat ccccggaggg atatggctgg gaacatagcc aacctaaaaa  31560 

tacagtccaa gatgcatctc atatccccac gtatgcaccc atcccagctt aaccgctttg  31620 

taaacactta caccaagggc ctgcccctgg caatcagctt gctactgaaa gacattttta  31680 

ggcaccacgc ccagcgctcc tgctacgact ggatcatcta caacaccacc ccgcagcatg  31740 

aagctctgca gtggtgctac ctccacccca gagacgggct tatgcccatg tatctgaaca  31800 

tccagagtca cctttaccac gtcctggaaa aaatacacag gaccctcaac gaccgagacc  31860 

gctggtcccg ggcctaccgc gcgcgcaaaa cccctaaata aagacagcaa gacacttgct  31920 

tgatcaaaat ccaaacagag tctggttttt atttatgttt taaaccgcat tgggagggga  31980 

ggaagccttc agggcagaaa cctgctggcg cagatccaac agctgctgag aaacgacatt  32040 

aagttcccgg gtcaaagaat ccaattgtgc caaaagagcc gtcaacttgt catcgcgggc  32100 

ggatgaacgg gaagctgcac tgcttgcaag cgggctcagg aaagcaaagt cagtcacaat  32160 

cccgcgggcg gtggctgcag cggctgaagc ggcggcggag gctgcagtct ccaacggcgt  32220 

tccagacacg gtctcgtagg tcaaggtagt agagtttgcg ggcaggacgg ggcgaccatc  32280 

aatgctggag cccatcacat tctgacgcac cccggcccat gggggcatgc gcgttgtcaa  32340 

atatgagctc acaatgcttc catcaaacga gttggcgctc atggcggcgg ctgctgcaaa  32400 

acagatacaa aactacatga gacccccacc ttatatattc tttcccaccc ttaagccccg  32460 

cccatcgatg gcaaacagct attatgggta ttatgggtgc tagcgacatg aggttgcccc  32520 

gtattcagtg tcgctgattt gtattgtctg aagttgtttt tacgttaagt tgatgcagat  32580 

caattaatac gatacctgcg tcataattga ttatttgacg tggtttgatg gcctccacgc  32640 

acgttgtgat atgtagatga taatcattat cactttacgg gtcctttccg gtgatccgac  32700 

aggttacggg gcggcgacct cgcgggtttt cgctatttat gaaaattttc cggtttaagg  32760 

cgtttccgtt cttcttcgtc ataacttaat gtttttattt aaaataccct ctgaaaagaa  32820 

aggaaacgac aggtgctgaa agcgaggctt tttggcctct gtcgtttcct ttctctgttt  32880 

ttgtccgtgg aatgaacaat ggaagttaac ggatccaggc cgcgagcaaa aggccagcaa  32940 

aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct  33000 

gacgagcatc acaaaaatca acgctcaagt cagaggtggc gaaacccgac aggactataa  33060 

agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg  33120 

cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca  33180 

cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa  33240 

ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg  33300 

gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg  33360 

tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga  33420 

acagtatttg gtatctgcgc tctgccaaag ccagttacct tcggaaaaag agttggtagc  33480 

tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag  33540 

attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac  33600 

gctcagtgga acgaaaactc acgttaaggg attttggtca tcagattatc aaaaaggatc  33660 

ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag  33720 

taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt  33780 

ctatttcgtt catccatagt tgcctgactc cccgtagtgt agataactac gatacgggag  33840 

ggcttaccat ccggccccag tgctgcaatg ataccgcgtg acccacgctc accggctcct  33900 

gatttatcag caataaacca gccagccgga agtgccgagc gcagaagtgg tcctgcaact  33960 

ttatccgcct ccatccagtc tattagttgt tgccgggaag ctagagtaag tagttcgcca  34020 

gttaatagtt ttcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg  34080 

tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc  34140 

atgttgtgca aaaaagcggt tagctccttc ggtcctccga tagttgtcag aagtaagttg  34200 

gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca  34260 

tccgtaagat gcttttctgt gactggtgag tattcaacca agaatacggg ataataccgc  34320 

gccacatagc agaactttaa aagtgctcat cattgggaaa cgttcttcgg ggcgaaaact  34380 

ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgcg cacccaagtg  34440 

atcttctgca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa  34500 

tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac ttttcctttt  34560 

tcaatattat tgaagcattt atcagggtta ttgtctcatc agcggataca tatttg      34616 

 
           
             3  
             31672  
             DNA  
             Artificial Sequence  
             
               derived from Adenovirus  
             
           
            3 

gaatcggcca gcgcgaatta actataacgg tcctaaggta gcgtcatcat cataatatac     60 

cttattttgg attgaagcca atatgataat gagggggtgg agtttgtgac gtggcgcggg    120 

gcgtgggaac ggggcgggtg acgtaggttt tagggcggag taacttgcat gtattgggaa    180 

ttgtagtttt tttaaaatgg gaagttacgt atcgtgggaa aacggaagtg aagatttgag    240 

gaagttgtgg gttttttggc tttcgtttct gggcgtaggt tcgcgtgcgg ttttctgggt    300 

gttttttgtg gactttaacc gttacgtcat tttttagtcc tatatatact cgctctgtac    360 

ttggcccttt ttacactgtg actgattgag ctggtgccgt gtcgagtggt gttttttaat    420 

aggttttttt actggtaagg ctgactgtta tggctgccgc tgtggaagcg ctgtatgttg    480 

ttctggagcg ggagggtgct attttgccta ggcaggaggg tttttcaggt gtttatgtgt    540 

ttttctctcc tattaatttt gttatacctc ctatgggggc tgtaatgttg tctctacgcc    600 

tgcgggtatg tattcccccg ggctatttcg gtcgcttttt agcactgacc gatgttaacc    660 

aacctgatgt gtttaccgag tcttacatta tgactccgga catgaccgag gaactgtcgg    720 

tggtgctttt taatcacggt gaccagtttt tttacggtca cgccggcatg gccgtagtcc    780 

gtcttatgct tataagggtt gtttttcctg ttgtaagaca ggcttctaat gtttaaatgt    840 

ttttttttgt tattttattt tgtgtttaat gcaggaaccc gcagacatgt ttgagagaaa    900 

aatggtgtct ttttctgtgg tggttccgga acttacctgc ctttatctgc atgagcatga    960 

ctacgatgtg cttgcttttt tgcgcgaggc tttgcctgat tttttgagca gcaccttgca   1020 

ttttatatcg ccgcccatgc aacaagctta cataggggct acgctggtta gcatagctcc   1080 

gagtatgcgt gtcataatca gtgtgggttc ttttgtcatg gttcctggcg gggaagtggc   1140 

cgcgctggtc cgtgcagacc tgcacgatta tgttcagctg gccctgcgaa gggacctacg   1200 

ggatcgcggt atttttgtta atgttccgct tttgaatctt atacaggtct gtgaggaacc   1260 

tgaatttttg caatcatgat tcgctgcttg aggctgaagg tggagggcgc tctggagcag   1320 

atttttacaa tggccggact taatattcgg gatttgctta gagacatatt gataaggtgg   1380 

cgagatgaaa attatttggg catggttgaa ggtgctggaa tgtttataga ggagattcac   1440 

cctgaagggt ttagccttta cgtccacttg gacgtgaggg cagtttgcct tttggaagcc   1500 

attgtgcaac atcttacaaa tgccattatc tgttctttgg ctgtagagtt tgaccacgcc   1560 

accggagggg agcgcgttca cttaatagat cttcattttg aggttttgga taatcttttg   1620 

gaataaaaaa aaaaaaaaca tggttcttcc agctcttccc gctcctcccg tgtgtgactc   1680 

gcagaacgaa tgtgtaggtt ggctgggtgt ggcttattct gcggtggtgg atgttatcag   1740 

ggcagcggcg catgaaggag tttacataga acccgaagcc agggggcgcc tggatgcttt   1800 

gagagagtgg atatactaca actactacac agagcgagct aagcgacgag accggagacg   1860 

cagatctgtt tgtcacgccc gcacctggtt ttgcttcagg aaatatgact acgtccggcg   1920 

ttccatttgg catgacacta cgaccaacac gatctcggtt gtctcggcgc actccgtaca   1980 

gtagggatcg cctacctcct tttgagacag agacccgcgc taccatactg gaggatcatc   2040 

cgctgctgcc cgaatgtaac actttgacaa tgcacaacgt gagttacgtg cgaggtcttc   2100 

cctgcagtgt gggatttacg ctgattcagg aatgggttgt tccctgggat atggttctga   2160 

cgcgggagga gcttgtaatc ctgaggaagt gtatgcacgt gtgcctgtgt tgtgccaaca   2220 

ttgatatcat gacgagcatg atgatccatg gttacgagtc ctgggctctc cactgtcatt   2280 

gttccagtcc cggttccctg cagtgcatag ccggcgggca ggttttggcc agctggttta   2340 

ggatggtggt ggatggcgcc atgtttaatc agaggtttat atggtaccgg gaggtggtga   2400 

attacaacat gccaaaagag gtaatgttta tgtccagcgt gtttatgagg ggtcgccact   2460 

taatctacct gcgcttgtgg tatgatggcc acgtgggttc tgtggtcccc gccatgagct   2520 

ttggatacag cgccttgcac tgtgggattt tgaacaatat tgtggtgctg tgctgcagtt   2580 

actgtgctga tttaagtgag atcagggtgc gctgctgtgc ccggaggaca aggcgtctca   2640 

tgctgcgggc ggtgcgaatc atcgctgagg agaccactgc catgttgtat tcctgcagga   2700 

cggagcggcg gcggcagcag tttattcgcg cgctgctgca gcaccaccgc cctatcctga   2760 

tgcacgatta tgactctacc cccatgtagg cgtggacttc cccttcgccg cccgttgagc   2820 

aaccgcaagt tggacagcag cctgtggctc agcagctgga cagcgacatg aacttaagcg   2880 

agctgcccgg ggagtttatt aatatcactg atgagcgttt ggctcgacag gaaaccgtgt   2940 

ggaatataac acctaagaat atgtctgtta cccatgatat gatgcttttt aaggccagcc   3000 

ggggagaaag gactgtgtac tctgtgtgtt gggagggagg tggcaggttg aatactaggg   3060 

ttctgtgagt ttgattaagg tacggtgatc aatataagct atgtggtggt ggggctatac   3120 

tactgaatga aaaatgactt gaaattttct gcaattgaaa aataaacacg ttgaaacata   3180 

acatgcaaca ggttcacgat tctttattcc tgggcaatgt aggagaaggt gtaagagttg   3240 

gtagcaaaag tttcagtggt gtattttcca ctttcccagg accatgtaaa agacatagag   3300 

taagtgctta cctcgctagt ttctgtggat tcactagtgc cattaagtgt aatggtaagt   3360 

atcataggtt tagttttatc accatgcaag taaacttgac tgacaatgtt atttttagca   3420 

gtttgacttt gggtttttgg ataggctaga aggttaggca taaatccaac tgcatttgtg   3480 

tatggatttg cattagttga gttcccattt ctaaagttcc agtaatgttt tttaagtgag   3540 

gagttctcca ttagaacacc gttttggtca aatctaagga atatactaac acttgcaacg   3600 

gtgcctgtca tggatgaaag atctccagat acagccaaag cagctacagt agctagtact   3660 

tgactcccac attttgtaag aaccaaagta aatttgcagt cattatctga atgaattctg   3720 

cagttaggag atgggtctgg ggttgtccac agggtaagtt tgtcatcatt tttgtttcct   3780 

attgtaatgg cccctgagtt gtcaaagctt aaacccgctc caagtttagt aatcatggca   3840 

ccgttttcat tgtaatcaat gccagagcca attttagttt ttattgggtt gatatctgga   3900 

gactcagatg tgtttgtatc aaactccaga ccctttcctg catttatagc tatggcagta   3960 

ttatcaaagt ttagtccact ggattttttt atgctaactt ccagtttttt agtattgttt   4020 

gatgcattaa aaaggtatag gcctctgtta tagtttatgt ccaagttatg agatgcatta   4080 

atatacaggg gtccctgccc cagtttaaga cgtagttttg tttgagcatc aaatgggtaa   4140 

tccacatcta gaattaacaa gttgttattt atacgcatgc caccgcccgt tttaatttcc   4200 

atgttgtttg atgaatcata accaatagct cctgcaactt tggttctaag ggagttttgt   4260 

tcaacggtga cacctggtcc agtaactact gttagtgtat cggagttttg tgctacttgc   4320 

aaaggaccgc ttattttaat tcctattttt ccattattta cataaatagg atcttccatg   4380 

ttaatgccca agctacccgt ggcagtagtt agcgggggtg atgcagttac agtaagggtg   4440 

tcgctgtcac tgccagagag gggggctgat gtttgcaggg ctagctttcc atctgacact   4500 

gtaatgggcc ctttagtagc aatgcttagt ttggagtctt gcacggtcag tggggcttgt   4560 

gactgtacgc taagagcgcc gctagtaact atcagaggag cggtggttgc cactgttagg   4620 

gcgcctgagg taattgtaag tggtgcggag gtgtccaaac ttatgtttga ctttgttttt   4680 

ttaagtggct gagtaacagt ggttacattt tgggaggtga ggtttccggc cttgtctagg   4740 

gtaagaccgc tgcccatttt aagcgcaagc atgccgtggg aggtgtccaa aggttcggag   4800 

acgcgtagag agagaactcc agggggactt tcttggaaac cattgggtga aacaaatgga   4860 

ggggtaagaa agggcacagt tggaggcccg gtttctgtgt catatggata cacggggttg   4920 

aaggtgtctt cagacggtct ggcgcgtttc atctgcaaca atatgaagat agtgggtgcg   4980 

gagggacaag aacatgagga atttgacatc ccatttaaac tttggagaaa gtttgcagct   5040 

aaaaggcggc tgagatacca gagttgggag gaaggaaagg aggtgatgct gaataagctg   5100 

gacaaagatt tgctgactga ttttaagtaa gtaatttatt cagtcgtagc cgtccgccga   5160 

gtctttcacc gcgtcaaagt tgggaataaa ctggtccggg tagtggccgg gaggtccaga   5220 

aaaggggttg aagtaaaccg aaggcacgaa ctcctcaata aattgtagag ttccaatgcc   5280 

tccggagcgc ggctccgagg acgaggtctg cagagttagg atcgcctgac ggggcgtaaa   5340 

tgaagagcgg ccagcgccgc cgatctgaaa tgtcccgtcc ggacggagac caagagagga   5400 

gctcaccgac tcgtcgttga gctgaatacc tcgccctctg attttcaggt gagttatacc   5460 

ctgcccgggc gaccgcaccc tgtgacgaaa gccgcccgca agctgcgccc ctgagttagt   5520 

catctgaact tcggcctggg cgtctctggg aagtaccaca gtggtgggag cgggactttc   5580 

ctggtacacc agggcagcgg gccaactacg gggattaagg ttattacgag gtgtggtggt   5640 

aatagccgcc tgttcgagga gaattcggtt tcggtgggcg cggattccgt tgacccggga   5700 

tatcatgtgg ggtcccgcgc tcatgtagtt tattcgggtt gagtagtctt gggcagctcc   5760 

agccgcaagt cccatttgtg gctggtaact ccacatgtag ggcgtgggaa tttccttgct   5820 

cataatggcg ctgacgacag gtgctggcgc cgggtgtggc cgctggagat gacgtagttt   5880 

tcgcgcttaa atttgagaaa gggcgcgaaa ctagtcctta agagtcagcg cgcagtattt   5940 

gctgaagaga gcctccgcgt cttccagcgt gcgccgaagc tgatcttcgc ttttgtgata   6000 

caggcagctg cgggtgaggg agcgcagaga cctgtttttt attttcagct cttgttcttg   6060 

gcccctgctt tgttgaaata tagcatacag agtgggaaaa atcctatttc taagctcgcg   6120 

ggtcgatacg ggttcgttgg gcgccagacg cagcgctcct cctcctgctg ctgccgccgc   6180 

tgtggatttc ttgggctttg tcagagtctt gctatccggt cgcctttgct tctgtgtgac   6240 

cgctgctgtt gctgccgctg ccgctgccgc cggtgcagta ggggctgtag agatgacggt   6300 

agtaatgcag gatgttacgg gggaaggcca cgccgtgatg gtagagaaga aagcggcggg   6360 

cgaaggagat gttgccccca cagtcttgca agcaagcaac tatggcgttc ttgtgcccgc   6420 

gccacgagcg gtagccttgg cgctgttgtt gctcttgggc taacggcggc ggctgcttag   6480 

acttaccggc cctggttcca gtggtgtccc atctacggtt gggtcggcga acaggcagtg   6540 

ccggcggcgc ctgaggagcg gaggttgtag cgatgctggg aacggttgcc aatttctggg   6600 

gcgccggcga ggggaatgcg accgagggtg acggtgtttc gtctgacacc tcttcggcct   6660 

cggaagcttc gtctaggctg tcccagtctt ccatcatctc ctcctcctcg tccaaaacct   6720 

cctctgcctg actgtcccag tattcctcct cgtccgtggg tggcggcggc ggcagctgca   6780 

gcttcttttt gggtgccatc ctgggaagca agggcccgcg gctgctgata gggctgcggc   6840 

ggcgggggga ttgggttgag ctcctcgccg gactgggggt ccaggtaaac cccccgtccc   6900 

tttcgtagca gaaactcttg gcgggctttg ttgatggctt gcaattggcc aaggatgtgg   6960 

ccctgggtaa tgacgcaggc ggtaagctcc gcatttggcg ggcgggattg gtcttcgtag   7020 

aacctaatct cgtgggcgtg gtagtcctca ggtacaaatt tgcgaaggta agccgacgtc   7080 

cacagccccg gagtgagttt caaccccgga gccgcggact tttcgtcagg cgagggaccc   7140 

tgcagctcaa aggtaccgat aatttgactt tcgctaagca gttgcgaatt gcagaccagg   7200 

gagcggtgcg gggtgcatag gttgcagcga cagtgacact ccagtaggcc gtcaccgctc   7260 

acgtcttcca tgatgtcgga gtggtaggca aggtagttgg ctagctgcag aaggtagcag   7320 

tgaccccaaa gcggcggagg gcattcacgg tacttaatgg gcacaaagtc gctaggaagc   7380 

gcacagcagg tggcgggcag aattcctgaa cgctctagga taaagttcct aaagttttgc   7440 

aacatgcttt gactggtgaa gtctggcaga ccctgttgca gggttttaag caggcgttcg   7500 

gggaagataa tgtccgccag gtgcgcggcc acggagcgct cgttgaaggc cgtccatagg   7560 

tccttcaagt tttgctttag cagcttctgc agctccttta ggttgcgctc ctccaggcat   7620 

tgctgccaca cgcccatggc cgtttgccag gtgtagcaca gaaataagta aacgcagtcg   7680 

cggacgtagt cgcggcgcgc ctcgcccttg agcgtggaat gaagcacgtt ttgcccgagg   7740 

cggttttcgt gcaaaattcc aaggtaggag accaggttgc agagctccac gttggaaatt   7800 

ttgcaggcct ggcgcacgta gccctggcga aaggtgtagt gcaacgtttc ctctagcttg   7860 

cgctgcatct ccgggtcagc aaagaaccgc tgcatgcact caagctccac ggtaacaagc   7920 

actgcggcca tcattagctt gcgtcgctcc tccaagtcgg caggctcgcg cgtctcaagc   7980 

cagcgcgcca gctgctcatc gccaactgcg ggtaggccct cctcggtttg ttcttgcaag   8040 

tttgcatccc tctccagggg tcgtgcacgg cgcacgatca gctcgctcat gactgtgctc   8100 

ataaccttgg ggggtaggtt aagtgccggg taggcaaagt gggtgacctc gatgctgcgt   8160 

ttcagcacgg ctaggcgcgc gttgtcaccc tcaagttcca ccagcactcc acagtgactt   8220 

tcattttcgc tgttttcttg ttgcagagcg tttgccgcgc gtttctcgtc gcgtccaaga   8280 

ccctcaaaga tttttggcac ttcgtcgagc gaggcgatat caggtatgac agcgccctgc   8340 

cgcaaggcca gctgcttgtc cgctcggctg cggttggcac ggcaggatag gggtatcttg   8400 

cagttttgga aaaagatgtg ataggtggca agcacctctg gcacggcaaa tacggggtag   8460 

aagttgaggc gcgggttggg ctcgcatgtg ccgttttctt ggcgtttggg gggtacgcgc   8520 

ggtgagaaca ggtggcgttc gtaggcaagg ctgacatccg ctatggcgag gggcacatcg   8580 

ctgcgctctt gcaacgcgtc gcagataatg gcgcactggc gctgcagatg cttcaacagc   8640 

acgtcgtctc ccacatctag gtagtcgcca tgcctttggt ccccccgccc gacttgttcc   8700 

tcgtttgcct ctgcgtcgtc ctggtcttgc tttttatcct ctgttggtac tgagcgatcc   8760 

tcgtcgtctt cgcttacaaa acctgggtcc tgctcgataa tcacttcctc ctcctcaagc   8820 

gggggtgcct cgacggggaa ggtggtaggc gcgttggcgg catcggtgga ggcggtggtg   8880 

gcgaactcaa agggggcggt taggctgtcc tccttctcga ctgactccat gatctttttc   8940 

tgcctatagg agaaggaaat ggccagtcgg gaagaggagc agcgcgaaac cacccccgag   9000 

cgcggacgcg gtgcggcgcg acgtccacca accatggagg acgtgtcgtc cccgtcgccg   9060 

tcgccgccgc ctccccgcgc gcccccaaaa aagcggctga ggcggcgtct cgagtccgag   9120 

gacgaagaag actcgtcaca agatgcgctg gtgccgcgca cacccagccc gcggccatcg   9180 

acctcgacgg cggatttggc cattgcgtcc aaaaagaaaa agaagcgccc ctctcccaag   9240 

cccgagcgcc cgccatcccc agaggtgatc gtggacagcg aggaagaaag agaagatgtg   9300 

gcgctacaaa tggtgggttt cagcaaccca ccggtgctaa tcaagcacgg caagggaggt   9360 

aagcgcacgg tgcggcggct gaatgaagac gacccagtgg cgcggggtat gcggacgcaa   9420 

gaggaaaagg aagagtccag tgaagcggaa agtgaaagca cggtgataaa cccgctgagc   9480 

ctgccgatcg tgtctgcgtg ggagaagggc atggaggctg cgcgcgcgtt gatggacaag   9540 

taccacgtgg ataacgatct aaaggcaaac ttcaagctac tgcctgacca agtggaagct   9600 

ctggcggccg tatgcaagac ctggctaaac gaggagcacc gcgggttgca gctgaccttc   9660 

accagcaaca agacctttgt gacgatgatg gggcgattcc tgcaggcgta cctgcagtcg   9720 

tttgcagagg taacctacaa gcaccacgag cccacgggct gcgcgttgtg gctgcaccgc   9780 

tgcgctgaga tcgaaggcga gcttaagtgt ctacacggga gcattatgat aaataaggag   9840 

cacgtgattg aaatggatgt gacgagcgaa aacgggcagc gcgcgctgaa ggagcagtct   9900 

agcaaggcca agatcgtgaa gaaccggtgg ggccgaaatg tggtgcagat ctccaacacc   9960 

gacgcaaggt gctgcgtgca tgacgcggcc tgtccggcca atcagttttc cggcaagtct  10020 

tgcggcatgt tcttctctga aggcgcaaag gctcaggtgg cttttaagca gatcaaggct  10080 

ttcatgcagg cgctgtatcc taacgcccag accgggcacg gtcaccttct gatgccacta  10140 

cggtgcgagt gcaactcaaa gcctgggcat gcaccctttt tgggaaggca gctaccaaag  10200 

ttgactccgt tcgccctgag caacgcggag gacctggacg cggatctgat ctccgacaag  10260 

agcgtgctgg ccagcgtgca ccacccggcg ctgatagtgt tccagtgctg caaccctgtg  10320 

tatcgcaact cgcgcgcgca gggcggaggc cccaactgcg acttcaagat atcggcgccc  10380 

gacctgctaa acgcgttggt gatggtgcgc agcctgtgga gtgaaaactt caccgagctg  10440 

ccgcggatgg ttgtgcctga gtttaagtgg agcactaaac accagtatcg caacgtgtcc  10500 

ctgccagtgg cgcatagcga tgcgcggcag aacccctttg atttttaaac ggcgcagacg  10560 

gcaagggtgg ggggtaaata atcacccgag agtgtacaaa taaaaacatt tgcctttatt  10620 

gaaagtgtct cctagtacat tatttttaca tgtttttcaa gtgacaaaaa gaagtggcgc  10680 

tcctaatctg cgcactgtgg ctgcggaagt agggcgagtg gcgctccagg aagctgtaga  10740 

gctgttcctg gttgcgacgc agggtgggct gtacctgggg actgttaagc atggagttgg  10800 

gtaccccggt aataaggttc atggtggggt tgtgatccat gggagtttgg ggccagttgg  10860 

caaaggcgtg gagaaacatg cagcagaata gtccacaggc ggccgagttg ggcccctgca  10920 

cgctttgggt ggacttttcc agcgttatac agcggtcggg ggaagaagca atggcgctac  10980 

ggcgcaggag tgactcgtac tcaaactggt aaacctgctt gagtcgttgg tcagaaaagc  11040 

caaagggctc aaagaggtag catgtttttg agcgcgggtt ccaggcaaag gccatccagt  11100 

gtacgccccc agtctcgcga ccggccgtat tgactatggc gcaggcgagc ttgtgtggag  11160 

aaacaaagcc tggaaagcgc ttgtcatagg tgcccaaaaa atatggccca caaccaagat  11220 

ctttgacaat ggctttcagt tcctgctcac tggagcccat ggcggcagct gttgttgatg  11280 

ttgcttgctt cttttatgtt gtggcgttgc cggccgagaa gggcgtgcgc aggtacacgg  11340 

tctcgatgac gccgcggtgc ggctggtgca cacggaccac gtcaaagact tcaaacaaaa  11400 

cataaagaag ggtgggctcg tccatgggat ccacctcaaa agtcatgtct agcgcgtggg  11460 

cggagttggc gtagagaagg ttttggccca ggtctgtgag tgcgcccatg gacataaagt  11520 

tactggagaa tgggatgcgc caaagggtgc gatcgcaaag aaactttttc tgggtaatac  11580 

tgtcaaccgc ggttttgcct attagtgggt agggcacgtt ggcggggtaa gcctgtccct  11640 

cgcgcatggt gggagcgagg tagcctacga atcctgagtt gttatgctgg tgaagaattc  11700 

caacctgctg atactccttg tatttagtat cgtcaaccac ttgccggctc atgggctgga  11760 

agtttctgaa gaacgagtac atgcggtcct tgtagctttc tggaatgtag aagccctggt  11820 

agccaatatt gtagttggcc aacatctgca ccaggaacca gtccttggtc atgttgcact  11880 

gagctacgtt gtagccctcc ccgtcaactg agcgtttaat ctcaaactca ttgggagtaa  11940 

gcaggcggtc gttgcccggc cagctaacag aagagtcaaa ggtaatggcc accttcttaa  12000 

aggtgtgatt aagatagaag gttccgtcaa ggtatggtat ggagccagag taggtgtagt  12060 

aagggtcgta gcctgatccc agggaagggg tttcctttgt cttcaagcgt gtgaaggccc  12120 

aaccgcgaaa tgctgcccag ttgcgcgatg ggatggagat gggcacgttg gtggcgttgg  12180 

cgggtatggg gtatagcatg ttggcggcgg aaaggtagtc attaaaggac tggtcgttgg  12240 

tgtcatttct gagcatggct tccagcgtgg aggccgtgtt gtgggccatg gggaagaagg  12300 

tggcgtaaag acaaatgctg tcaaacttaa tgctagcccc gtcaactcta agatcgtttc  12360 

ccagagagct ctgcagaacc atgttaacat ccttcctgaa gttccattca tatgtatatg  12420 

agcctggcag gaggaggagg tttttaatgg caaaaaactt ttggggcacc tgaatgtgaa  12480 

agggcacgta gcggccgttt cccaacaaca tggagcgata acggaggccc gcattgcggt  12540 

ggtggttaaa gggattaacg ttgtccatgt agtccagaga ccagcgcgcc ccaaggttaa  12600 

tgtagcagtc tacaagcccg ggagccacca ctcgcttgtt catgtagtcg taggtgttgg  12660 

ggttgtcaga tatttccaca ttggtggggt tgtattttag cttgtctggc aggtacagcg  12720 

caatattgga gtaaaggaaa tttctccata ggttggcatt taggttaatt tccatggcaa  12780 

agttgttacc cactcctatt tcattacgtg ttgcaaaagt ttcatctttt gtccatgtag  12840 

tatctccatt atcgcctgag ccattgccat tagccttaat agcttgatag gtgtcagtta  12900 

ccccaatacc cccaagagga aaacaataat ttggcaattc atcctcagtt ccatggtttt  12960 

caatgattct aacatctgga tcatagctgt ctacagcctg attccacata gaaaaatatc  13020 

tggttctatc acctatggaa tcaagcaaga gttgatagga cagctctgtg tttctgtctt  13080 

gcaaatctac cacggcattt agctgcgatg cctgaccagc aagaacaccc atgttgccag  13140 

tgctgttata atacattagg ccaataaaat tgtccctgaa agcaatgtaa ttgggtctgt  13200 

ttggcataga ttgttgaccc aacatagctt tagaattttc atcacctttt ccaggtttgt  13260 

aagacagatg tgtgtctggg gtttccatat ttacatcttc actgtacaaa accacttttg  13320 

gtttagtagc attgccttgc cggtcgttca aagaggtagt atttgagaag aattgcaagt  13380 

caacctttgg aagaggcacc cctttttcat ccggaaccag aacggattga ccaccaaaag  13440 

gatttgtagg cctggcataa gatccatagc atggtttcat gggagttgtt tttttaagca  13500 

ctctccctcc tgccgcatta gcatcagctt cgttccactg agattcgcca atttgaggtt  13560 

ctggttgata ggaaggatct gcgtatacag gtttagcttg tgtttctgca ttgtctgatc  13620 

ctatttgtag cccgcttttt gtaattgttt ctccagacaa aggagcctgg gcatagacat  13680 

gtgttttctt agtagcctga tctcgagcgt tttgctcttc ttcttcctct tcttcatctt  13740 

catcttcctc ttcttcatcc tcggcaactg cccggccgct atcttcggtt tgttcccact  13800 

cacaggagtt aggagcgccc ttgggagcta gagcgttgta ggcagtgccg gagtagggct  13860 

taaaagtagg ccccctgtcc agcacgccgc ggatgtcaaa gtacgtggaa gccatatcaa  13920 

gcacacggtt gtcacccaca gccagggtga accgcgcttt gtacgagtac gcggtatcct  13980 

cgcggtccac agggatgaac cgcagcgtca aacgctggga ccggtctgtg gttacgtcgt  14040 

gcgtaggtgc caccgtgggg tttctaaact tgttattcag gctgaagtac gtctcggtgg  14100 

cgcgggcaaa ctgcaccagc ccggggctca ggtactccga ggcgtcctgg cccgagatgt  14160 

gcatgtaaga ccactgcggc atcatcgaag gggtagccat cttggaaagc gggcgcacgg  14220 

cggctcagca gctcctctgg cggcgacatg gacgcataca tgacacatac gacacgttag  14280 

ctatttagaa gcatcgtcgg cgcttcaggg attgcacccc cagacccacg atgctgttca  14340 

gtgtgctttg ccagttgcca ctggctacgg gccgcatcga tcgcggaccg ctggcggcac  14400 

ggcgcaggga cgcgcggcta gggcgggtta caacaacggc ggacggccct ggcagcacag  14460 

gtttctgctg ggtgtcagcg gggggaggca ggtccagcgt tacaggtgtg tgctggccca  14520 

gcactccggt agccatgggc gcgatgggac gggtggtggg caggccttgc tttagtgcct  14580 

cctcgtacga gggaggctca tctatttgcg tcaccagagt ttcttccctg tcgggccgcg  14640 

gacgcttttc gccacgcccc tctggagaca ctgtctccac ggccggtgga ggctcctcta  14700 

cgggagggcg gggatcaagc ttactgttaa tcttattttg cactgcctgg ttggccaggt  14760 

ccaccacccc gctaatgcca gaggccaggc catctaccac cttttgttgg aaattttgct  14820 

ctttcaactt gtccctcagc atctggcctg tgctgctgtt ccaggccttg ctgccatagt  14880 

tcttaatggt ggaaccgaaa tttttaatgc cgctccacag cgagccccag ctgaaggcgc  14940 

caccgctcat attgctggtg ccgatatctt gccagtttcc catgaacggg cgcgagccgt  15000 

gtcgcggggc cagagacgca aagttgatgt cttccattct acaaaatagt tacaggacca  15060 

agcgagcgtg agactccaga ctttttattt tgatttttcc acatgcaact tgtttttaat  15120 

cagtgtctct gcgcctgcaa ggccacggat gcaattccgg gcacggcgcc aatcgccgcg  15180 

gcgatcagtg gaataaggag gggcaggata ccgccgcgca tgcgacggtg cgacgcgcgc  15240 

cgccgccggt ggtgcgcacg acgcatgccg cccgtcaggc cgtggccggc catgcccctc  15300 

ctacggtgca ttcttcctcg gaatcccggc accgggaaac ggaggcggca ggtgagggcc  15360 

atatctgcaa gaaccacaaa gaccggcttt taaacgatgc tggggtggta gcgcgctgtt  15420 

ggcagcacca gggtcctgcc tccttcgcga gccaccctgc gcacggaaat cggggccagc  15480 

acgggctggc gacggcgacg gcggcggcgg gttccagtgg tggttcggcg tcgggtagtc  15540 

gctcgtcttc tggggcggta ggtgtagcca cgatagccgg gggtaggcgc gatggaagga  15600 

tgtagggcat attcgggcag tagtgcgctg gcggtgccgt acttcctgga acggcgcggg  15660 

cgccgggggg ctgaaacgcg aaacatccac gggtccgttt gcacctccgt agaggttttg  15720 

gacgcggccg cagcggccgc ctgcaccgcg gcatctgcca ccgccgaggc aaccggggac  15780 

gtttgtgtct ccatgccctc tgtggcagtg gcaatactag tgctactggt ggtgggtatc  15840 

tgaacgtcca cggtctgcac gcccagtccc ggtgccacct gcttgattgg ccgcacgcgg  15900 

acctcgggct ccagcccagg ctccacggtc attttttcca agacatcttc cagtcgctgg  15960 

cgcttgggta ccatcagctg cacggtgggt gccaagtcac cagactcgcg ctttaggccg  16020 

cgcttttctt cggacggtgc aagcgtgggc agcacctgct gcagtgtcac gggctttagg  16080 

ctaggtgttg ggttgccctc gtccagcggc aacgccaaca tgtccttatg ccgctttccg  16140 

taggcaaact ccccgaggcg ctcgttggcc tgctcaagca ggtcctcgtc gccgtacacc  16200 

tcatcataca cgcgcttgta ggtgcgggtg gagcgctcac cgggcgtaaa aactacggtg  16260 

gtgccgggtc gcaaaacacg tcttacgcgt cgacctttcc actgtacccg ccgcctgggc  16320 

gcggttgcgt gcagcagttc cacctcgtcg tcaagttcat catcatcatc atctttcttt  16380 

ttctttttga cccgctttag ctttcggggc ttgtaatcct gctcttcctt cttcgggggg  16440 

ccatagatct ccggcgcgat gacctggagc atctcttctt tgattttgcg cttggacata  16500 

gcttcgttgc gcgccgccgc cgctggatac atacaacagt acgagtctaa gtagtttttt  16560 

cttgcaatct agttgcgcgg ggggcgggtg cgcacgggca cgcgcaggcc gctaaccgag  16620 

tcgcgcaccc agtacacgtt gcccctgcga ccctgagtca tagcactaat ggccgcggct  16680 

gctgcggcgg ccgctcgtcg cctggacctg gggggcacag tgacaatacc cgcggccagc  16740 

cttcgagcgg cccgcatggc cgcccgtcgg ccggtgcgac gtgcgcggtt aagcagggcc  16800 

gccgccgcgc gttgggcggc agtgccgggt cggcggcggt ggcgacgtgc tacgcgcctc  16860 

cgccgtctct tcattttagc ataacgccgg gctccgcgca ccacggtctg aatggccgcg  16920 

tccactgtgg acactggtgg cggcgtgggc gtgtagttgc gcgcctcctc caccaccgcg  16980 

tcaatggcgt catcgacggt ggtgcgccca gtgcggccgc gtttgtgcgc gccccagggc  17040 

gcgcggtagt gcccgcgcac gcgcactggg tgttggtcgg agcgcttctt tgccccgcca  17100 

aacatcttgc ttgggaagcg caggccccag cctgtgttat tgctgggcga tataaggatg  17160 

gacatgtttg ctcaaaaagt gcggctcgat aggacgcgcg gcgagactat gcccagggcc  17220 

ttgtaaacgt aggggcaggt gcggcgtctg gcgtcagtaa tggtcactcg ctggactcct  17280 

ccgatgctgt tgcgcagcgg tagcgtcccg tgatctgtga gagcaggaac gttttcactg  17340 

acggtggtga tggtgggggc tggcgggcgc gccaaaatct ggttctcggg aaagcgattg  17400 

aacacgtggg tcagagaggt aaactggcgg atgagctggg agtagacggc ctggtcgttg  17460 

tagaagctct tggagtgcac gggcaacagc tcggcgccca ccaccggaaa gttgctgatc  17520 

tggctcgtgg agcggaaggt cacggggtct tgcatcatgt ctggcaacga ccagtagacc  17580 

tgctccgagc cgcaggttac gtcaggagtg caaaggaggg tccatgagcg gatcccggtc  17640 

tgagggtcgc cgtagttgta tgcaaggtac cagctgcggt actgggtgaa ggtgctgtca  17700 

ttgcttatta ggttgtaact gcgtttcttg ctgtcctctg tcaggggttt gatcaccggt  17760 

ttcttctgag gcttctcgac ctcgggttgc gcagcggggg cggcagcttc tgccgctgcc  17820 

tcggcctcag cgcgcttctc ctccgcccgt gtggcaaagg tgtcgccgcg aatggcatga  17880 

tcgttcatgt cctccaccgg ctgcattgcc gcggctgccg cgttggagtt ctcttccgcg  17940 

ccgctgccac tgttgttgcc gccgcctgcg ccatccccgc cctgttcggt gtcatctttt  18000 

aagcttgcct ggtaggcgtc cacatccaac agtgcgggaa tgttaccacc ctccaggtca  18060 

tcgtaggtga tcctaaagcc ctcctggaag ggttgccgct tgcggatgcc caacaagttg  18120 

ctcaggcggc tgtgggtgaa gtccaccccg catcctggca gcaaaatgat gtctggatgg  18180 

aaggcttcgt ttgtatatac cccaggcatg acaagaccag tgactgggtc aaaccccagt  18240 

ctgaagttgc gggtgtcaaa ctttaccccg atgtcgcttt ccagaacccc gttctgcctg  18300 

cccactttca agtagtgctc cacgatcgcg ttgttcataa ggtctatggt catggtctcg  18360 

gagtagttgc cctcgggcag cgtgaactcc acccactcat atttcagctc cacctgtttg  18420 

tccttagtaa gcgagcgcga caccatcacc cgcgccttaa acttattggt aaacatgaac  18480 

tcgttcacat ttggcatgtt ggtatgcagg atggttttca ggtcgccgcc ccagtgcgaa  18540 

cggtcgtcaa gattgatggt ctgtgtgctt gcctcccccg ggctgtagtc attgttttga  18600 

atgaccgtgg ttagaaagtt gctgtggtcg ttctggtagt tcagggatgc cacatccgtt  18660 

gacttgttgt ccacaaggta cacacgggtg gtgtcgaata ggggtgccaa ctcagagtaa  18720 

cggatgctgt ttctcccccc ggtaggccgc aggtaccgcg gaggcacaaa cggcgggtcc  18780 

aggggagcat cgaaggggga acccagcgcc gccgccactg gcgccgcgct caccacgctc  18840 

tcgtaggagg gaggaggacc ttcctcatac atcgccgcgc gctgcatact aaggggaata  18900 

caagaaaacc aacgctcggt gccatggcct tggtgagttt tttattttgc atcatgcttt  18960 

tttttttttt ttttaaaaca ttctccccag cctggggcga aggtgcgcaa acgggttgcc  19020 

actccctccc aaatccagga cgctgctgtc gtctgccgag tcatcgtcct cccacaccag  19080 

accccgctga cggtcgtgcc tttgacgacg ggtgggcggg cgcgggccgg gcacatccct  19140 

gtgctcctgc gcatacgtct tccatctact catcttgtcc actaggctct ctatcccgtt  19200 

gttgggaaat gccggaggca ggttcttttc gcgctgcggc tgcagcagcg agttgtttag  19260 

gtactcctcc tcgcccagca ggcgcgggcg ggtggtgcga gtgctggtaa aagaccctat  19320 

caagcttgga aatgggctac tcgcatctga ccgcggggcc gcagcgccta gatcggacaa  19380 

gctgcttggc ctgcggaagc tttcctttcg cagcgccgcc tctgcctgct cgcgctgttg  19440 

caactctagc agggtctgcg gttgcgggga aaacacgctg tcgtctatgt cgtcccagag  19500 

gaatccatcg ttaccctcgg gcacctcaaa tcccccggtg tagaaaccag ggggcggtag  19560 

ccagtgcggg ttcaagatgg cattggtgaa atactcgggg ttcacggcgg ccgcgcgatg  19620 

caagtagtcc attaggcgat tgataaacgg ccggtttgag gcatacatgc ccggttccat  19680 

gttgcgcgcg gtcatgtcca gcgccacgct gggcgttacc ccgtcgcgca tcaggttaag  19740 

gctcacgctc tgctgcacat agcgcaagat gcgctcctcc tcgctgttta aactgtgcaa  19800 

cgaggggatc ttctgccgcc ggttggtcag caggtagttc agggttgcct ccaggctgcc  19860 

cgtgtcctcc tgccccagcg cgcggctgac acttgtaatc tcctggaaag tatgctcgtc  19920 

cacatgcgcc tgacctatgg cctcgcggta cagtgtcagc aagtgaccta ggtatgtgtc  19980 

ccgggacacg ctgccactgt ccgtgaaggg cgctattagc agcagcaaca ggcgcgagtt  20040 

gggcgtcagc aagctagaca cggtcgcgcg gtcgcctgtg ggagcccgca ccccccacag  20100 

cccctgcaag ttcttgaaag cctggctcag gtttacggtc tgcaggcctt gtctactggt  20160 

ctggaaaaaa tagtctggcc cggactggta cacctcactt tgcggtgtct cagtcaccat  20220 

tagccgcagt gcgctcacaa agttggtgta gtcctcctgt ccccgcggca cgttggcggg  20280 

ctgtgtactc aggaaggcgt ttagtgcaac catggagccc aggttgccct gctgctgcgc  20340 

gcgctcacgc tgcgccacgg cctcgcgcac atcccccacc agccggtcca ggttggtctg  20400 

cacgttgccg ctgttgtaac gagccacgcg ctgaagcagc gcgtcgtaga ccaggccggc  20460 

ctcatcgggc cggatggccc tgttttcggc cagcgcgttt acgatcgcca gcaccttctc  20520 

gtgcgtgggg tttgcgcgcg ccgggaccac cgcttccaga attgcggaga gccggttggc  20580 

ctgcggctgc tgccggaacg cgtcagggtt acgcgcagtc agcgacatga tgcggtccat  20640 

gacctggcgc cagtcgtccg tggagttaag gccggacggc tggctctgca gcgccgcccg  20700 

caccgccggg tccgttgcgt cttgcatcat ctgatcagaa acatcaccgc ttagtactcg  20760 

ccgtcctctg gctcgtactc atcgtcctcg tcatattcct ccacgccgcc gacgttgcca  20820 

gcgcgcgcgg gtgccaccgc cagcccaggt ccggccccag ctgcctccag ggcgcgtcgg  20880 

cttggggccc agcgcaggtc agcgcccgcg tcaaagtagg actcggcctc tctatcgccg  20940 

ctgcccgtgc cagccagggc cctttgcagg ctgtgcatca gctcgcggtc gctgagctcg  21000 

cgccgccggc tcacgctcac ggccttgtgg atgcgctcgt tgcgataaac gcccaggtcg  21060 

tcgctcaagg taagcacctt caacgccatg cgcatgtaga acccctcgat ctttacctcc  21120 

ttgtctatgg gaacgtaagg ggtatggtat atcttgcggg cgtaaaactt gcccagactg  21180 

agcatggaat agttaatggc ggccaccttg tcagccaggc tcaagctgcg ctcctgcacc  21240 

actatgctct gcagaatgtt tatcaaatcg agcagccagc ggccctcggg ctctactatg  21300 

tttagcagcg catccctgaa tgcctcgttg tccctgctgt gctgcactat aaggaacagc  21360 

tgcgccatga gcggcttgct atttgggttt tgctccagcg cgcttacaaa gtcccacaga  21420 

tgcatcagtc ctatagccac ctcctcgcgc gccacaagcg tgcgcacgtg gttgttaaag  21480 

cttttttgaa agttaatctc ctggttcacc gtctgctcgt acgcggttac caggtcggcg  21540 

gccgccacgt gtgcgcgcgc gggactaatc ccggtccgcg cgtcgggctc aaagtcctcc  21600 

tcgcgcagca accgctcgcg gttcaggcca tgccgcaact cgcgccctgc gtggaacttt  21660 

cgatcccgca tctcctcggg ctcctctccc tcgcggtcgc gaaacaggtt ctgccgcggc  21720 

acgtacgcct cgcgcgtgtc acgcttcagc tgcacccttg ggtgtcgctc aggagagggc  21780 

gctcctagcc gcgccaggcc ctcgccctcc tccaagtcca ggtagtgccg ggcccggcgc  21840 

cgcgggggtt cgtaatcacc atctgccgcc gcgtcagccg cggatgttgc ccctcctgac  21900 

gcggtaggag aaggggaggg tgccctgcat gtctgccgct gctcttgctc ttgccgctgc  21960 

tgaggagggg ggcgcatctg ccgcagcacc ggatgcatct gggaaaagca aaaaaggggc  22020 

tcgtccctgt ttccggagga atttgcaagc ggggtcttgc atgacgggga ggcaaacccc  22080 

cgttcgccgc agtccggccg gcccgagact cgaaccgggg gtcctgcgac tcaacccttg  22140 

gaaaataacc ctccggctac agggagcgag ccacttaatg ctttcgcttt ccagcctaac  22200 

cgcttacgcc gcgcgcggcc agtggccaaa aaagctagcg cagcagccgc cgcgcctgga  22260 

aggaagccaa aaggagcgct cccccgttgt ctgacgtcgc acacctgggt tcgacacgcg  22320 

ggcggtaacc gcatggatca cggcggacgg ccggatccgg ggttcgaacc ccggtcgtcc  22380 

gccatgatac ccttgcgaat ttatccacca gaccacggaa gagtgcccgc ttacaggctc  22440 

tccttttgca cggtctagag cgtcaacgac tgcgcacgcc tcaccggcca gagcgtcccg  22500 

accatggagc actttttgcc gctgcgcaac atctggaacc gcgtccgcga ctttccgcgc  22560 

gcctccacca ccgccgccgg catcacctgg atgtccaggt acatctacgg atatcatcgc  22620 

cttatgttgg aagacctcgc ccccggagcc ccggccaccc tacgctggcc cctctaccgc  22680 

cagccgccgc cgcacttttt ggtgggatat cagtacctgg tgcggacttg caacgactac  22740 

gtctttgact caagggctta ctcgcgtctc aggtacaccg agctctcgca gccgggtcac  22800 

cagaccgtta actggtccgt tatggccaac tgcacttaca ccatcaacac gggcgcatac  22860 

caccgctttg tggacatgga tgacttccag tctaccctca cgcaggtgca gcaggccata  22920 

ttagccgagc gcgttgtcgc cgacctggcc ctgcttcagc cgatgagggg cttcggggtc  22980 

acacgcatgg gaggaagagg gcgccaccta cggccaaact ccgccgccgc cgtagcgata  23040 

gatgcaagag atgcaggaca agaggaagga gaagaagaag tgccggtaga aaggctcatg  23100 

caagactact acaaagacct gcgccgatgt caaaacgaag cctggggcat ggccgaccgc  23160 

ctgcgcattc agcaggccgg acccaaggac atggtgcttc tgtcgaccat ccgccgtctc  23220 

aagaccgcct actttaatta catcatcagc agcacctccg ccagaaacaa ccccgaccgc  23280 

cacccgctgc cgcccgccac ggtgctcagc ctaccttgcg actgtgactg gttagacgcc  23340 

tttctcgaga ggttttccga tccggtcgat gcggactcgc tcaggtccct cggtggcgga  23400 

gtacctacac aacaattgtt gagatgcatc gttagcgccg tatccctgcc gcacggcagc  23460 

cccccgccaa cccataaccg ggacatgacg ggcggcgtct tccaactgcg cccccgcgag  23520 

aacggccgcg ccgtcaccga gaccatgcgc cgtcgccgcg gggagatgat cgagcgcttt  23580 

gtcgaccgcc tcccggtgcg ccgtcgtcgc cgccgtgtcc cccctccccc accgccgcca  23640 

gaagaagaag aagaagggga ggcccttatg gaagaggaga ttgaagaaga agaggcccct  23700 

gtagcctttg agcgcgaggt gcgcgacact gtcgccgagc tcatccgtct tctggaggag  23760 

gagttaaccg tgtcggcgcg caactcccag tttttcaact tcgccgtgga cttctacgag  23820 

gccatggagc gccttgaggc cttgggggat atcaacgaat ccacgttgcg acgctgggtt  23880 

atgtacttct tcgtggcaga acacaccgcc accaccctca actacctctt tcagcgcctg  23940 

cgaaactacg ccgtcttcgc ccggcacgtg gagctcaatc tcgcgcaggt ggtcatgcgc  24000 

gcccgcgatg ccgaaggggg cgtggtctac agccgcgtct ggaacgaggg aggcctcaac  24060 

gccttctcgc agctcatggc ccgcatctcc aacgacctcg ccgccaccgt ggagcgagcc  24120 

ggacgcggag atctccagga ggaagagatc gagcagttca tggccgaaat cgcctatcaa  24180 

gacaactcag gagacgtgca ggagattttg cgccaggccg ccgtcaacga caccgaaatt  24240 

gattctgtcg aactctcttt caggttcaag ctcaccgggc ccgtcgtctt cacgcagagg  24300 

cgccagattc aggagatcaa ccgccgcgtc gtcgcgttcg ccagcaacct ccgcgcgcag  24360 

caccagctcc tgcccgcgcg cggcgccgac gtgcccctgc cccctctccc ggcgggtccc  24420 

gagccccccc tacctccggg ggcccgcccg cgtcaccgct tttagatgca tcatccaagg  24480 

acacccccgc ggcccaccgc ccgccgcgcg gtaccgtagt cgcgccgcgg ggatgcggcc  24540 

tcttgcaagt catcgacgcc gccaccaacc agcccctgga aatcaggtat cacctggacc  24600 

tagcccgcgc cctgacccgg ctatgcgagg taaacctgca ggagctcccg cctgacctgt  24660 

cgccgcggga gctccagacc atggacagct cccatctgcg cgatgttgtc atcaagctcc  24720 

gaccgccgcg cgcggacatc tggactttgg gctcgcgcgg cgtggtggtc cgatccacca  24780 

taactcccct cgagcagcca gacggtcaag gacaagcagc cgaagtagaa gaccaccagc  24840 

caaacccgcc aggcgagggg ctcaaattcc cactctgctt ccttgtgcgc ggtcgtcagg  24900 

tcaacctcgt gcaggatgta cagcccgtgc accgctgcca gtactgcgca cgtttttaca  24960 

aaagccagca cgagtgttcg gcccgtcgca gggacttcta ctttcaccac atcaacagcc  25020 

actcctccaa ctggtggcgg gagatccagt tcttcccgat cggctcgcat cctcgcaccg  25080 

agcgtctctt tgtcacctac gatgtagaga cctatacttg gatgggggcc tttgggaagc  25140 

agctcgtgcc cttcatgctg gttatgaagt tcggcggaga tgagcctctg gtgaccgccg  25200 

cgcgagacct agccgtggac cttggatggg accgctggga acaagacccg cttaccttct  25260 

actgcatcac cccagaaaaa atggccatag gtcgccagtt taggaccttt cgcgaccacc  25320 

tgcaaatgct aatggcccgt gacctgtgga gctcattcgt cgcttccaac cctcatcttg  25380 

cagactgggc cctgtcagaa cacgggctca gctcccctga ggagctcacc tacgaggaac  25440 

ttaaaaaatt gccctccatc aagggcaccc cgcgcttctt ggaactttac atcgtgggcc  25500 

acaacatcaa cggcttcgac gagatcgtgc tcgccgccca ggtaattaac aaccgttccg  25560 

aggtgccggg acccttccgc atcacacgca actttatgcc tcgcgcggga aagatacttt  25620 

tcaacgatgt caccttcgcc ctgccaaacc cgcgttccaa aaagcgcacg gactttttgc  25680 

tctgggagca gggcggatgc gacgacactg acttcaaata ccagtacctc aaagtcatgg  25740 

ttagggacac ctttgcgctc acccacacct cgctccggaa ggccgcgcag gcatacgcgc  25800 

tacccgtaga aaagggatgc tgcgcctacc aggccgtcaa ccagttctac atgctaggct  25860 

cttaccgttc ggaggccgac gggtttccga tccaagagta ctggaaagac cgcgaagagt  25920 

ttgtcctcaa ccgcgagctg tggaaaaaaa agggacagga taagtatgac atcatcaagg  25980 

aaaccctgga ctactgcgcc ctagacgtgc aggtcaccgc cgagctggtc aacaagctgc  26040 

gcgactccta cgcctccttc gtgcgtgacg cggtaggtct cacagacgcc agcttcaacg  26100 

tcttccagcg tccaaccata tcatccaact cacatgccat cttcaggcag atagtcttcc  26160 

gagcagagca gcccgcccgt agcaacctcg gtcccgacct cctcgctccc tcgcacgaac  26220 

tatacgatta cgtgcgcgcc agcatccgcg gtggaagatg ctaccctaca tatcttggaa  26280 

tactcagaga gcccctctac gtttacgaca tttgcggcat gtacgcctcc gcgctcaccc  26340 

accccatgcc atggggtccc ccactcaacc catacgagcg cgcgcttgcc gcccgcgcat  26400 

ggcagcaggc gctagacttg caaggatgca agatagacta cttcgacgcg cgcctgctgc  26460 

ccggggtctt taccgtggac gcagaccccc cggacgagac gcagctagac ccactaccgc  26520 

cattctgttc gcgcaagggc ggccgcctct gctggaccaa cgagcgccta cgcggagagg  26580 

tagccaccag cgttgacctt gtcaccctgc acaaccgcgg ttggcgcgtg cacctggtgc  26640 

ccgacgagcg caccaccgtc tttcccgaat ggcggtgcgt tgcgcgcgaa tacgtgcagc  26700 

taaacatcgc ggccaaggag cgcgccgatc gcgacaaaaa ccaaaccctg cgctccatcg  26760 

ccaagttgct gtccaacgcc ctctacgggt cgtttgccac caagcttgac aacaaaaaga  26820 

ttgtcttttc tgaccagatg gacgcggcca ccctcaaagg catcaccgcg ggccaggtga  26880 

atatcaaatc ctcctcgttt ttggaaactg acaatcttag cgcagaagtc atgcccgctt  26940 

ttgagaggga gtactcaccc caacagctgg ccctcgcaga cagcgatgcg gaagagagtg  27000 

aggacgaacg cgcccccacc cccttttata gccccccttc aggaacaccc ggtcacgtgg  27060 

cctacaccta taaaccaatc accttccttg atgccgaaga gggcgacatg tgtcttcaca  27120 

ccctggagcg agtggacccc ctagtggaca acgaccgcta cccctcccac ttagcctcct  27180 

tcgtgctggc ctggacgcga gccttcgtct cagagtggtc cgagtttcta tacgaggagg  27240 

accgcggaac accgctcgag gacaggcctc tcaagtctgt atacggggac acggacagcc  27300 

ttttcgtcac cgagcgtgga caccggctca tggaaaccag aggtaagaaa cgcatcaaaa  27360 

agcatggggg aaacctggtt tttgaccccg aacggccaga gctcacctgg ctcgtggaat  27420 

gcgagaccgt ctgcggggcc tgcggcgcgg atgcctactc cccggaatcg gtatttctcg  27480 

cgcccaagct ctacgccctt aaaagtctgc actgcccctc gtgcggcgcc tcctccaagg  27540 

gcaagctgcg cgccaagggc cacgccgcgg aggggctgga ctatgacacc atggtcaaat  27600 

gctacctggc cgacgcgcag ggcgaagacc ggcagcgctt cagcaccagc aggaccagcc  27660 

tcaagcgcac cctggccagc gcgcagcccg gagcgcaccc cttcaccgtg acccagacta  27720 

cgctgacgag gaccctgcgc ccgtggaaag acatgaccct ggcccgtctg gacgagcacc  27780 

gactactgcc gtacagcgaa agccgcccca acccgcgaaa cgaggagata tgctggatcg  27840 

agatgccgta gagcacgtga ccgagctgtg ggaccgcctg gaactgcttg gtcaaacgct  27900 

caaaagcatg cctacggcgg acggcctcaa accgttgaaa aactttgctt ccttgcaaga  27960 

actgctatcg ctgggcggcg agcgccttct ggcgcatttg gtcagggaaa acatgcaagt  28020 

cagggacatg cttaacgaag tggcccccct gctcagggat gacggcagct gcagctctct  28080 

taactaccag ttgcagccgg taataggtgt gatttacggg cccaccggct gcggtaagtc  28140 

gcagctgctc aggaacctgc tttcttccca gctgatctcc cctaccccgg aaacggtttt  28200 

cttcatcgcc ccgcaggtag acatgatccc cccatctgaa ctcaaagcgt gggaaatgca  28260 

aatctgtgag ggtaactacg cccctgggcc ggatggaacc attataccgc agtctggcac  28320 

cctccgcccg cgctttgtaa aaatggccta tgacgatctc atcctggaac acaactatga  28380 

cgttagtgat cccagaaata tcttcgccca ggccgccgcc cgtgggccca ttgccatcat  28440 

tatggacgaa tgcatggaaa atctcggagg tcacaagggc gtctccaagt tcttccacgc  28500 

atttccttct aagctacatg acaaatttcc caagtgcacc ggatacactg tgctggtggt  28560 

tctgcacaac atgaatcccc ggagggatat ggctgggaac atagccaacc taaaaataca  28620 

gtccaagatg catctcatat ccccacgtat gcacccatcc cagcttaacc gctttgtaaa  28680 

cacttacacc aagggcctgc ccctggcaat cagcttgcta ctgaaagaca tttttaggca  28740 

ccacgcccag cgctcctgct acgactggat catctacaac accaccccgc agcatgaagc  28800 

tctgcagtgg tgctacctcc accccagaga cgggcttatg cccatgtatc tgaacatcca  28860 

gagtcacctt taccacgtcc tggaaaaaat acacaggacc ctcaacgacc gagaccgctg  28920 

gtcccgggcc taccgcgcgc gcaaaacccc taaataaaga cagcaagaca cttgcttgat  28980 

caaaatccaa acagagtctg gtttttattt atgttttaaa ccgcattggg aggggaggaa  29040 

gccttcaggg cagaaacctg ctggcgcaga tccaacagct gctgagaaac gacattaagt  29100 

tcccgggtca aagaatccaa ttgtgccaaa agagccgtca acttgtcatc gcgggcggat  29160 

gaacgggaag ctgcactgct tgcaagcggg ctcaggaaag caaagtcagt cacaatcccg  29220 

cgggcggtgg ctgcagcggc tgaagcggcg gcggaggctg cagtctccaa cggcgttcca  29280 

gacacggtct cgtaggtcaa ggtagtagag tttgcgggca ggacggggcg accatcaatg  29340 

ctggagccca tcacattctg acgcaccccg gcccatgggg gcatgcgcgt tgtcaaatat  29400 

gagctcacaa tgcttccatc aaacgagttg gcgctcatgg cggcggctgc tgcaaaacag  29460 

atacaaaact acatgagacc cccaccttat atattctttc ccacccttaa gccccgccca  29520 

tcgatggcaa acagctatta tgggtattat gggtgctagc gacatgaggt tgccccgtat  29580 

tcagtgtcgc tgatttgtat tgtctgaagt tgtttttacg ttaagttgat gcagatcaat  29640 

taatacgata cctgcgtcat aattgattat ttgacgtggt ttgatggcct ccacgcacgt  29700 

tgtgatatgt agatgataat cattatcact ttacgggtcc tttccggtga tccgacaggt  29760 

tacggggcgg cgacctcgcg ggttttcgct atttatgaaa attttccggt ttaaggcgtt  29820 

tccgttcttc ttcgtcataa cttaatgttt ttatttaaaa taccctctga aaagaaagga  29880 

aacgacaggt gctgaaagcg aggctttttg gcctctgtcg tttcctttct ctgtttttgt  29940 

ccgtggaatg aacaatggaa gttaacggat ccaggccgcg agcaaaaggc cagcaaaagg  30000 

ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg  30060 

agcatcacaa aaatcaacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat  30120 

accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta  30180 

ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct  30240 

gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc  30300 

ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa  30360 

gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg  30420 

taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag  30480 

tatttggtat ctgcgctctg ccaaagccag ttaccttcgg aaaaagagtt ggtagctctt  30540 

gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta  30600 

cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc  30660 

agtggaacga aaactcacgt taagggattt tggtcatcag attatcaaaa aggatcttca  30720 

cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa  30780 

cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat  30840 

ttcgttcatc catagttgcc tgactccccg tagtgtagat aactacgata cgggagggct  30900 

taccatccgg ccccagtgct gcaatgatac cgcgtgaccc acgctcaccg gctcctgatt  30960 

tatcagcaat aaaccagcca gccggaagtg ccgagcgcag aagtggtcct gcaactttat  31020 

ccgcctccat ccagtctatt agttgttgcc gggaagctag agtaagtagt tcgccagtta  31080 

atagttttcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg  31140 

gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt  31200 

tgtgcaaaaa agcggttagc tccttcggtc ctccgatagt tgtcagaagt aagttggccg  31260 

cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg  31320 

taagatgctt ttctgtgact ggtgagtatt caaccaagaa tacgggataa taccgcgcca  31380 

catagcagaa ctttaaaagt gctcatcatt gggaaacgtt cttcggggcg aaaactctca  31440 

aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgcgcacc caagtgatct  31500 

tctgcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc  31560 

gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactttt cctttttcaa  31620 

tattattgaa gcatttatca gggttattgt ctcatcagcg gatacatatt tg          31672 

 
           
             4  
             30365  
             DNA  
             Artificial Sequence  
             
               derived from Adenovirus  
             
           
            4 

gaatcggcca gcgcgaatta actataacgg tcctaaggta gcgtcatcat cataatatac     60 

cttattttgg attgaagcca atatgataat gagggggtgg agtttgtgac gtggcgcggg    120 

gcgtgggaac ggggcgggtg acgtaggttt tagggcggag taacttgcat gtattgggaa    180 

ttgtagtttt tttaaaatgg gaagttacgt atcgtgggaa aacggaagtg aagatttgag    240 

gaagttgtgg gttttttggc tttcgtttct gggcgtaggt tcgcgtgcgg ttttctgggt    300 

gttttttgtg gactttaacc gttacgtcat tttttagtcc tatatatact cgctctgtac    360 

ttggcccttt ttacactgtg actgattgag ctggtgccgt gtcgagtggt gttttttaat    420 

aggttttttt actggtaagg ctgactgtta tggctgccgc tgtggaagcg ctgtatgttg    480 

ttctggagcg ggagggtgct attttgccta ggataacttc gtataatgta tgctatacga    540 

agttatggcg cgccagatct gtttgtcacg cccgcacctg gttttgcttc aggaaatatg    600 

actacgtccg gcgttccatt tggcatgaca ctacgaccaa cacgatctcg gttgtctcgg    660 

cgcactccgt acagtaggga tcgcctacct ccttttgaga cagagacccg cgctaccata    720 

ctggaggatc atccgctgct gcccgaatgt aacactttga caatgcacaa cgtgagttac    780 

gtgcgaggtc ttccctgcag tgtgggattt acgctgattc aggaatgggt tgttccctgg    840 

gatatggttc tgacgcggga ggagcttgta atcctgagga agtgtatgca cgtgtgcctg    900 

tgttgtgcca acattgatat catgacgagc atgatgatcc atggttacga gtcctgggct    960 

ctccactgtc attgttccag tcccggttcc ctgcagtgca tagccggcgg gcaggttttg   1020 

gccagctggt ttaggatggt ggtggatggc gccatgttta atcagaggtt tatatggtac   1080 

cgggaggtgg tgaattacaa catgccaaaa gaggtaatgt ttatgtccag cgtgtttatg   1140 

aggggtcgcc acttaatcta cctgcgcttg tggtatgatg gccacgtggg ttctgtggtc   1200 

cccgccatga gctttggata cagcgccttg cactgtggga ttttgaacaa tattgtggtg   1260 

ctgtgctgca gttactgtgc tgatttaagt gagatcaggg tgcgctgctg tgcccggagg   1320 

acaaggcgtc tcatgctgcg ggcggtgcga atcatcgctg aggagaccac tgccatgttg   1380 

tattcctgca ggacggagcg gcggcggcag cagtttattc gcgcgctgct gcagcaccac   1440 

cgccctatcc tgatgcacga ttatgactct acccccatgt aggcgtggac ttccccttcg   1500 

ccgcccgttg agcaaccgca agttggacag cagcctgtgg ctcagcagct ggacagcgac   1560 

atgaacttaa gcgagctgcc cggggagttt attaatatca ctgatgagcg tttggctcga   1620 

caggaaaccg tgtggaatat aacacctaag aatatgtctg ttacccatga tatgatgctt   1680 

tttaaggcca gccggggaga aaggactgtg tactctgtgt gttgggaggg aggtggcagg   1740 

ttgaatacta gggttctgtg agtttgatta aggtacggtg atcaatataa gctatgtggt   1800 

ggtggggcta tactactgaa tgaaaaatga cttgaaattt tctgcaattg aaaaataaac   1860 

acgttgaaac ataacatgca acaggttcac gattctttat tcctgggcaa tgtaggagaa   1920 

ggtgtaagag ttggtagcaa aagtttcagt ggtgtatttt ccactttccc aggaccatgt   1980 

aaaagacata gagtaagtgc ttacctcgct agtttctgtg gattcactag tgccattaag   2040 

tgtaatggta agtatcatag gtttagtttt atcaccatgc aagtaaactt gactgacaat   2100 

gttattttta gcagtttgac tttgggtttt tggataggct agaaggttag gcataaatcc   2160 

aactgcattt gtgtatggat ttgcattagt tgagttccca tttctaaagt tccagtaatg   2220 

ttttttaagt gaggagttct ccattagaac accgttttgg tcaaatctaa ggaatatact   2280 

aacacttgca acggtgcctg tcatggatga aagatctcca gatacagcca aagcagctac   2340 

agtagctagt acttgactcc cacattttgt aagaaccaaa gtaaatttgc agtcattatc   2400 

tgaatgaatt ctgcagttag gagatgggtc tggggttgtc cacagggtaa gtttgtcatc   2460 

atttttgttt cctattgtaa tggcccctga gttgtcaaag cttaaacccg ctccaagttt   2520 

agtaatcatg gcaccgtttt cattgtaatc aatgccagag ccaattttag tttttattgg   2580 

gttgatatct ggagactcag atgtgtttgt atcaaactcc agaccctttc ctgcatttat   2640 

agctatggca gtattatcaa agtttagtcc actggatttt tttatgctaa cttccagttt   2700 

tttagtattg tttgatgcat taaaaaggta taggcctctg ttatagttta tgtccaagtt   2760 

atgagatgca ttaatataca ggggtccctg ccccagttta agacgtagtt ttgtttgagc   2820 

atcaaatggg taatccacat ctagaattaa caagttgtta tttatacgca tgccaccgcc   2880 

cgttttaatt tccatgttgt ttgatgaatc ataaccaata gctcctgcaa ctttggttct   2940 

aagggagttt tgttcaacgg tgacacctgg tccagtaact actgttagtg tatcggagtt   3000 

ttgtgctact tgcaaaggac cgcttatttt aattcctatt tttccattat ttacataaat   3060 

aggatcttcc atgttaatgc ccaagctacc cgtggcagta gttagcgggg gtgatgcagt   3120 

tacagtaagg gtgtcgctgt cactgccaga gaggggggct gatgtttgca gggctagctt   3180 

tccatctgac actgtaatgg gccctttagt agcaatgctt agtttggagt cttgcacggt   3240 

cagtggggct tgtgactgta cgctaagagc gccgctagta actatcagag gagcggtggt   3300 

tgccactgtt agggcgcctg aggtaattgt aagtggtgcg gaggtgtcca aacttatgtt   3360 

tgactttgtt tttttaagtg gctgagtaac agtggttaca ttttgggagg tgaggtttcc   3420 

ggccttgtct agggtaagac cgctgcccat tttaagcgca agcatgccgt gggaggtgtc   3480 

caaaggttcg gagacgcgta gagagagaac tccaggggga ctttcttgga aaccattggg   3540 

tgaaacaaat ggaggggtaa gaaagggcac agttggaggc ccggtttctg tgtcatatgg   3600 

atacacgggg ttgaaggtgt cttcagacgg tctggcgcgt ttcatctgca acaatatgaa   3660 

gatagtgggt gcggagggac aagaacatga ggaatttgac atcccattta aactttggag   3720 

aaagtttgca gctaaaaggc ggctgagata ccagagttgg gaggaaggaa aggaggtgat   3780 

gctgaataag ctggacaaag atttgctgac tgattttaag taagtaattt attcagtcgt   3840 

agccgtccgc cgagtctttc accgcgtcaa agttgggaat aaactggtcc gggtagtggc   3900 

cgggaggtcc agaaaagggg ttgaagtaaa ccgaaggcac gaactcctca ataaattgta   3960 

gagttccaat gcctccggag cgcggctccg aggacgaggt ctgcagagtt aggatcgcct   4020 

gacggggcgt aaatgaagag cggccagcgc cgccgatctg aaatgtcccg tccggacgga   4080 

gaccaagaga ggagctcacc gactcgtcgt tgagctgaat acctcgccct ctgattttca   4140 

ggtgagttat accctgcccg ggcgaccgca ccctgtgacg aaagccgccc gcaagctgcg   4200 

cccctgagtt agtcatctga acttcggcct gggcgtctct gggaagtacc acagtggtgg   4260 

gagcgggact ttcctggtac accagggcag cgggccaact acggggatta aggttattac   4320 

gaggtgtggt ggtaatagcc gcctgttcga ggagaattcg gtttcggtgg gcgcggattc   4380 

cgttgacccg ggatatcatg tggggtcccg cgctcatgta gtttattcgg gttgagtagt   4440 

cttgggcagc tccagccgca agtcccattt gtggctggta actccacatg tagggcgtgg   4500 

gaatttcctt gctcataatg gcgctgacga caggtgctgg cgccgggtgt ggccgctgga   4560 

gatgacgtag ttttcgcgct taaatttgag aaagggcgcg aaactagtcc ttaagagtca   4620 

gcgcgcagta tttgctgaag agagcctccg cgtcttccag cgtgcgccga agctgatctt   4680 

cgcttttgtg atacaggcag ctgcgggtga gggagcgcag agacctgttt tttattttca   4740 

gctcttgttc ttggcccctg ctttgttgaa atatagcata cagagtggga aaaatcctat   4800 

ttctaagctc gcgggtcgat acgggttcgt tgggcgccag acgcagcgct cctcctcctg   4860 

ctgctgccgc cgctgtggat ttcttgggct ttgtcagagt cttgctatcc ggtcgccttt   4920 

gcttctgtgt gaccgctgct gttgctgccg ctgccgctgc cgccggtgca gtaggggctg   4980 

tagagatgac ggtagtaatg caggatgtta cgggggaagg ccacgccgtg atggtagaga   5040 

agaaagcggc gggcgaagga gatgttgccc ccacagtctt gcaagcaagc aactatggcg   5100 

ttcttgtgcc cgcgccacga gcggtagcct tggcgctgtt gttgctcttg ggctaacggc   5160 

ggcggctgct tagacttacc ggccctggtt ccagtggtgt cccatctacg gttgggtcgg   5220 

cgaacaggca gtgccggcgg cgcctgagga gcggaggttg tagcgatgct gggaacggtt   5280 

gccaatttct ggggcgccgg cgaggggaat gcgaccgagg gtgacggtgt ttcgtctgac   5340 

acctcttcgg cctcggaagc ttcgtctagg ctgtcccagt cttccatcat ctcctcctcc   5400 

tcgtccaaaa cctcctctgc ctgactgtcc cagtattcct cctcgtccgt gggtggcggc   5460 

ggcggcagct gcagcttctt tttgggtgcc atcctgggaa gcaagggccc gcggctgctg   5520 

atagggctgc ggcggcgggg ggattgggtt gagctcctcg ccggactggg ggtccaggta   5580 

aaccccccgt ccctttcgta gcagaaactc ttggcgggct ttgttgatgg cttgcaattg   5640 

gccaaggatg tggccctggg taatgacgca ggcggtaagc tccgcatttg gcgggcggga   5700 

ttggtcttcg tagaacctaa tctcgtgggc gtggtagtcc tcaggtacaa atttgcgaag   5760 

gtaagccgac gtccacagcc ccggagtgag tttcaacccc ggagccgcgg acttttcgtc   5820 

aggcgaggga ccctgcagct caaaggtacc gataatttga ctttcgctaa gcagttgcga   5880 

attgcagacc agggagcggt gcggggtgca taggttgcag cgacagtgac actccagtag   5940 

gccgtcaccg ctcacgtctt ccatgatgtc ggagtggtag gcaaggtagt tggctagctg   6000 

cagaaggtag cagtgacccc aaagcggcgg agggcattca cggtacttaa tgggcacaaa   6060 

gtcgctagga agcgcacagc aggtggcggg cagaattcct gaacgctcta ggataaagtt   6120 

cctaaagttt tgcaacatgc tttgactggt gaagtctggc agaccctgtt gcagggtttt   6180 

aagcaggcgt tcggggaaga taatgtccgc caggtgcgcg gccacggagc gctcgttgaa   6240 

ggccgtccat aggtccttca agttttgctt tagcagcttc tgcagctcct ttaggttgcg   6300 

ctcctccagg cattgctgcc acacgcccat ggccgtttgc caggtgtagc acagaaataa   6360 

gtaaacgcag tcgcggacgt agtcgcggcg cgcctcgccc ttgagcgtgg aatgaagcac   6420 

gttttgcccg aggcggtttt cgtgcaaaat tccaaggtag gagaccaggt tgcagagctc   6480 

cacgttggaa attttgcagg cctggcgcac gtagccctgg cgaaaggtgt agtgcaacgt   6540 

ttcctctagc ttgcgctgca tctccgggtc agcaaagaac cgctgcatgc actcaagctc   6600 

cacggtaaca agcactgcgg ccatcattag cttgcgtcgc tcctccaagt cggcaggctc   6660 

gcgcgtctca agccagcgcg ccagctgctc atcgccaact gcgggtaggc cctcctcggt   6720 

ttgttcttgc aagtttgcat ccctctccag gggtcgtgca cggcgcacga tcagctcgct   6780 

catgactgtg ctcataacct tggggggtag gttaagtgcc gggtaggcaa agtgggtgac   6840 

ctcgatgctg cgtttcagca cggctaggcg cgcgttgtca ccctcaagtt ccaccagcac   6900 

tccacagtga ctttcatttt cgctgttttc ttgttgcaga gcgtttgccg cgcgtttctc   6960 

gtcgcgtcca agaccctcaa agatttttgg cacttcgtcg agcgaggcga tatcaggtat   7020 

gacagcgccc tgccgcaagg ccagctgctt gtccgctcgg ctgcggttgg cacggcagga   7080 

taggggtatc ttgcagtttt ggaaaaagat gtgataggtg gcaagcacct ctggcacggc   7140 

aaatacgggg tagaagttga ggcgcgggtt gggctcgcat gtgccgtttt cttggcgttt   7200 

ggggggtacg cgcggtgaga acaggtggcg ttcgtaggca aggctgacat ccgctatggc   7260 

gaggggcaca tcgctgcgct cttgcaacgc gtcgcagata atggcgcact ggcgctgcag   7320 

atgcttcaac agcacgtcgt ctcccacatc taggtagtcg ccatgccttt ggtccccccg   7380 

cccgacttgt tcctcgtttg cctctgcgtc gtcctggtct tgctttttat cctctgttgg   7440 

tactgagcga tcctcgtcgt cttcgcttac aaaacctggg tcctgctcga taatcacttc   7500 

ctcctcctca agcgggggtg cctcgacggg gaaggtggta ggcgcgttgg cggcatcggt   7560 

ggaggcggtg gtggcgaact caaagggggc ggttaggctg tcctccttct cgactgactc   7620 

catgatcttt ttctgcctat aggagaagga aatggccagt cgggaagagg agcagcgcga   7680 

aaccaccccc gagcgcggac gcggtgcggc gcgacgtcca ccaaccatgg aggacgtgtc   7740 

gtccccgtcg ccgtcgccgc cgcctccccg cgcgccccca aaaaagcggc tgaggcggcg   7800 

tctcgagtcc gaggacgaag aagactcgtc acaagatgcg ctggtgccgc gcacacccag   7860 

cccgcggcca tcgacctcga cggcggattt ggccattgcg tccaaaaaga aaaagaagcg   7920 

cccctctccc aagcccgagc gcccgccatc cccagaggtg atcgtggaca gcgaggaaga   7980 

aagagaagat gtggcgctac aaatggtggg tttcagcaac ccaccggtgc taatcaagca   8040 

cggcaaggga ggtaagcgca cggtgcggcg gctgaatgaa gacgacccag tggcgcgggg   8100 

tatgcggacg caagaggaaa aggaagagtc cagtgaagcg gaaagtgaaa gcacggtgat   8160 

aaacccgctg agcctgccga tcgtgtctgc gtgggagaag ggcatggagg ctgcgcgcgc   8220 

gttgatggac aagtaccacg tggataacga tctaaaggca aacttcaagc tactgcctga   8280 

ccaagtggaa gctctggcgg ccgtatgcaa gacctggcta aacgaggagc accgcgggtt   8340 

gcagctgacc ttcaccagca acaagacctt tgtgacgatg atggggcgat tcctgcaggc   8400 

gtacctgcag tcgtttgcag aggtaaccta caagcaccac gagcccacgg gctgcgcgtt   8460 

gtggctgcac cgctgcgctg agatcgaagg cgagcttaag tgtctacacg ggagcattat   8520 

gataaataag gagcacgtga ttgaaatgga tgtgacgagc gaaaacgggc agcgcgcgct   8580 

gaaggagcag tctagcaagg ccaagatcgt gaagaaccgg tggggccgaa atgtggtgca   8640 

gatctccaac accgacgcaa ggtgctgcgt gcatgacgcg gcctgtccgg ccaatcagtt   8700 

ttccggcaag tcttgcggca tgttcttctc tgaaggcgca aaggctcagg tggcttttaa   8760 

gcagatcaag gctttcatgc aggcgctgta tcctaacgcc cagaccgggc acggtcacct   8820 

tctgatgcca ctacggtgcg agtgcaactc aaagcctggg catgcaccct ttttgggaag   8880 

gcagctacca aagttgactc cgttcgccct gagcaacgcg gaggacctgg acgcggatct   8940 

gatctccgac aagagcgtgc tggccagcgt gcaccacccg gcgctgatag tgttccagtg   9000 

ctgcaaccct gtgtatcgca actcgcgcgc gcagggcgga ggccccaact gcgacttcaa   9060 

gatatcggcg cccgacctgc taaacgcgtt ggtgatggtg cgcagcctgt ggagtgaaaa   9120 

cttcaccgag ctgccgcgga tggttgtgcc tgagtttaag tggagcacta aacaccagta   9180 

tcgcaacgtg tccctgccag tggcgcatag cgatgcgcgg cagaacccct ttgattttta   9240 

aacggcgcag acggcaaggg tggggggtaa ataatcaccc gagagtgtac aaataaaaac   9300 

atttgccttt attgaaagtg tctcctagta cattattttt acatgttttt caagtgacaa   9360 

aaagaagtgg cgctcctaat ctgcgcactg tggctgcgga agtagggcga gtggcgctcc   9420 

aggaagctgt agagctgttc ctggttgcga cgcagggtgg gctgtacctg gggactgtta   9480 

agcatggagt tgggtacccc ggtaataagg ttcatggtgg ggttgtgatc catgggagtt   9540 

tggggccagt tggcaaaggc gtggagaaac atgcagcaga atagtccaca ggcggccgag   9600 

ttgggcccct gcacgctttg ggtggacttt tccagcgtta tacagcggtc gggggaagaa   9660 

gcaatggcgc tacggcgcag gagtgactcg tactcaaact ggtaaacctg cttgagtcgt   9720 

tggtcagaaa agccaaaggg ctcaaagagg tagcatgttt ttgagcgcgg gttccaggca   9780 

aaggccatcc agtgtacgcc cccagtctcg cgaccggccg tattgactat ggcgcaggcg   9840 

agcttgtgtg gagaaacaaa gcctggaaag cgcttgtcat aggtgcccaa aaaatatggc   9900 

ccacaaccaa gatctttgac aatggctttc agttcctgct cactggagcc catggcggca   9960 

gctgttgttg atgttgcttg cttcttttat gttgtggcgt tgccggccga gaagggcgtg  10020 

cgcaggtaca cggtctcgat gacgccgcgg tgcggctggt gcacacggac cacgtcaaag  10080 

acttcaaaca aaacataaag aagggtgggc tcgtccatgg gatccacctc aaaagtcatg  10140 

tctagcgcgt gggcggagtt ggcgtagaga aggttttggc ccaggtctgt gagtgcgccc  10200 

atggacataa agttactgga gaatgggatg cgccaaaggg tgcgatcgca aagaaacttt  10260 

ttctgggtaa tactgtcaac cgcggttttg cctattagtg ggtagggcac gttggcgggg  10320 

taagcctgtc cctcgcgcat ggtgggagcg aggtagccta cgaatcctga gttgttatgc  10380 

tggtgaagaa ttccaacctg ctgatactcc ttgtatttag tatcgtcaac cacttgccgg  10440 

ctcatgggct ggaagtttct gaagaacgag tacatgcggt ccttgtagct ttctggaatg  10500 

tagaagccct ggtagccaat attgtagttg gccaacatct gcaccaggaa ccagtccttg  10560 

gtcatgttgc actgagctac gttgtagccc tccccgtcaa ctgagcgttt aatctcaaac  10620 

tcattgggag taagcaggcg gtcgttgccc ggccagctaa cagaagagtc aaaggtaatg  10680 

gccaccttct taaaggtgtg attaagatag aaggttccgt caaggtatgg tatggagcca  10740 

gagtaggtgt agtaagggtc gtagcctgat cccagggaag gggtttcctt tgtcttcaag  10800 

cgtgtgaagg cccaaccgcg aaatgctgcc cagttgcgcg atgggatgga gatgggcacg  10860 

ttggtggcgt tggcgggtat ggggtatagc atgttggcgg cggaaaggta gtcattaaag  10920 

gactggtcgt tggtgtcatt tctgagcatg gcttccagcg tggaggccgt gttgtgggcc  10980 

atggggaaga aggtggcgta aagacaaatg ctgtcaaact taatgctagc cccgtcaact  11040 

ctaagatcgt ttcccagaga gctctgcaga accatgttaa catccttcct gaagttccat  11100 

tcatatgtat atgagcctgg caggaggagg aggtttttaa tggcaaaaaa cttttggggc  11160 

acctgaatgt gaaagggcac gtagcggccg tttcccaaca acatggagcg ataacggagg  11220 

cccgcattgc ggtggtggtt aaagggatta acgttgtcca tgtagtccag agaccagcgc  11280 

gccccaaggt taatgtagca gtctacaagc ccgggagcca ccactcgctt gttcatgtag  11340 

tcgtaggtgt tggggttgtc agatatttcc acattggtgg ggttgtattt tagcttgtct  11400 

ggcaggtaca gcgcaatatt ggagtaaagg aaatttctcc ataggttggc atttaggtta  11460 

atttccatgg caaagttgtt acccactcct atttcattac gtgttgcaaa agtttcatct  11520 

tttgtccatg tagtatctcc attatcgcct gagccattgc cattagcctt aatagcttga  11580 

taggtgtcag ttaccccaat acccccaaga ggaaaacaat aatttggcaa ttcatcctca  11640 

gttccatggt tttcaatgat tctaacatct ggatcatagc tgtctacagc ctgattccac  11700 

atagaaaaat atctggttct atcacctatg gaatcaagca agagttgata ggacagctct  11760 

gtgtttctgt cttgcaaatc taccacggca tttagctgcg atgcctgacc agcaagaaca  11820 

cccatgttgc cagtgctgtt ataatacatt aggccaataa aattgtccct gaaagcaatg  11880 

taattgggtc tgtttggcat agattgttga cccaacatag ctttagaatt ttcatcacct  11940 

tttccaggtt tgtaagacag atgtgtgtct ggggtttcca tatttacatc ttcactgtac  12000 

aaaaccactt ttggtttagt agcattgcct tgccggtcgt tcaaagaggt agtatttgag  12060 

aagaattgca agtcaacctt tggaagaggc accccttttt catccggaac cagaacggat  12120 

tgaccaccaa aaggatttgt aggcctggca taagatccat agcatggttt catgggagtt  12180 

gtttttttaa gcactctccc tcctgccgca ttagcatcag cttcgttcca ctgagattcg  12240 

ccaatttgag gttctggttg ataggaagga tctgcgtata caggtttagc ttgtgtttct  12300 

gcattgtctg atcctatttg tagcccgctt tttgtaattg tttctccaga caaaggagcc  12360 

tgggcataga catgtgtttt cttagtagcc tgatctcgag cgttttgctc ttcttcttcc  12420 

tcttcttcat cttcatcttc ctcttcttca tcctcggcaa ctgcccggcc gctatcttcg  12480 

gtttgttccc actcacagga gttaggagcg cccttgggag ctagagcgtt gtaggcagtg  12540 

ccggagtagg gcttaaaagt aggccccctg tccagcacgc cgcggatgtc aaagtacgtg  12600 

gaagccatat caagcacacg gttgtcaccc acagccaggg tgaaccgcgc tttgtacgag  12660 

tacgcggtat cctcgcggtc cacagggatg aaccgcagcg tcaaacgctg ggaccggtct  12720 

gtggttacgt cgtgcgtagg tgccaccgtg gggtttctaa acttgttatt caggctgaag  12780 

tacgtctcgg tggcgcgggc aaactgcacc agcccggggc tcaggtactc cgaggcgtcc  12840 

tggcccgaga tgtgcatgta agaccactgc ggcatcatcg aaggggtagc catcttggaa  12900 

agcgggcgca cggcggctca gcagctcctc tggcggcgac atggacgcat acatgacaca  12960 

tacgacacgt tagctattta gaagcatcgt cggcgcttca gggattgcac ccccagaccc  13020 

acgatgctgt tcagtgtgct ttgccagttg ccactggcta cgggccgcat cgatcgcgga  13080 

ccgctggcgg cacggcgcag ggacgcgcgg ctagggcggg ttacaacaac ggcggacggc  13140 

cctggcagca caggtttctg ctgggtgtca gcggggggag gcaggtccag cgttacaggt  13200 

gtgtgctggc ccagcactcc ggtagccatg ggcgcgatgg gacgggtggt gggcaggcct  13260 

tgctttagtg cctcctcgta cgagggaggc tcatctattt gcgtcaccag agtttcttcc  13320 

ctgtcgggcc gcggacgctt ttcgccacgc ccctctggag acactgtctc cacggccggt  13380 

ggaggctcct ctacgggagg gcggggatca agcttactgt taatcttatt ttgcactgcc  13440 

tggttggcca ggtccaccac cccgctaatg ccagaggcca ggccatctac caccttttgt  13500 

tggaaatttt gctctttcaa cttgtccctc agcatctggc ctgtgctgct gttccaggcc  13560 

ttgctgccat agttcttaat ggtggaaccg aaatttttaa tgccgctcca cagcgagccc  13620 

cagctgaagg cgccaccgct catattgctg gtgccgatat cttgccagtt tcccatgaac  13680 

gggcgcgagc cgtgtcgcgg ggccagagac gcaaagttga tgtcttccat tctacaaaat  13740 

agttacagga ccaagcgagc gtgagactcc agacttttta ttttgatttt tccacatgca  13800 

acttgttttt aatcagtgtc tctgcgcctg caaggccacg gatgcaattc cgggcacggc  13860 

gccaatcgcc gcggcgatca gtggaataag gaggggcagg ataccgccgc gcatgcgacg  13920 

gtgcgacgcg cgccgccgcc ggtggtgcgc acgacgcatg ccgcccgtca ggccgtggcc  13980 

ggccatgccc ctcctacggt gcattcttcc tcggaatccc ggcaccggga aacggaggcg  14040 

gcaggtgagg gccatatctg caagaaccac aaagaccggc ttttaaacga tgctggggtg  14100 

gtagcgcgct gttggcagca ccagggtcct gcctccttcg cgagccaccc tgcgcacgga  14160 

aatcggggcc agcacgggct ggcgacggcg acggcggcgg cgggttccag tggtggttcg  14220 

gcgtcgggta gtcgctcgtc ttctggggcg gtaggtgtag ccacgatagc cgggggtagg  14280 

cgcgatggaa ggatgtaggg catattcggg cagtagtgcg ctggcggtgc cgtacttcct  14340 

ggaacggcgc gggcgccggg gggctgaaac gcgaaacatc cacgggtccg tttgcacctc  14400 

cgtagaggtt ttggacgcgg ccgcagcggc cgcctgcacc gcggcatctg ccaccgccga  14460 

ggcaaccggg gacgtttgtg tctccatgcc ctctgtggca gtggcaatac tagtgctact  14520 

ggtggtgggt atctgaacgt ccacggtctg cacgcccagt cccggtgcca cctgcttgat  14580 

tggccgcacg cggacctcgg gctccagccc aggctccacg gtcatttttt ccaagacatc  14640 

ttccagtcgc tggcgcttgg gtaccatcag ctgcacggtg ggtgccaagt caccagactc  14700 

gcgctttagg ccgcgctttt cttcggacgg tgcaagcgtg ggcagcacct gctgcagtgt  14760 

cacgggcttt aggctaggtg ttgggttgcc ctcgtccagc ggcaacgcca acatgtcctt  14820 

atgccgcttt ccgtaggcaa actccccgag gcgctcgttg gcctgctcaa gcaggtcctc  14880 

gtcgccgtac acctcatcat acacgcgctt gtaggtgcgg gtggagcgct caccgggcgt  14940 

aaaaactacg gtggtgccgg gtcgcaaaac acgtcttacg cgtcgacctt tccactgtac  15000 

ccgccgcctg ggcgcggttg cgtgcagcag ttccacctcg tcgtcaagtt catcatcatc  15060 

atcatctttc tttttctttt tgacccgctt tagctttcgg ggcttgtaat cctgctcttc  15120 

cttcttcggg gggccataga tctccggcgc gatgacctgg agcatctctt ctttgatttt  15180 

gcgcttggac atagcttcgt tgcgcgccgc cgccgctgga tacatacaac agtacgagtc  15240 

taagtagttt tttcttgcaa tctagttgcg cggggggcgg gtgcgcacgg gcacgcgcag  15300 

gccgctaacc gagtcgcgca cccagtacac gttgcccctg cgaccctgag tcatagcact  15360 

aatggccgcg gctgctgcgg cggccgctcg tcgcctggac ctggggggca cagtgacaat  15420 

acccgcggcc agccttcgag cggcccgcat ggccgcccgt cggccggtgc gacgtgcgcg  15480 

gttaagcagg gccgccgccg cgcgttgggc ggcagtgccg ggtcggcggc ggtggcgacg  15540 

tgctacgcgc ctccgccgtc tcttcatttt agcataacgc cgggctccgc gcaccacggt  15600 

ctgaatggcc gcgtccactg tggacactgg tggcggcgtg ggcgtgtagt tgcgcgcctc  15660 

ctccaccacc gcgtcaatgg cgtcatcgac ggtggtgcgc ccagtgcggc cgcgtttgtg  15720 

cgcgccccag ggcgcgcggt agtgcccgcg cacgcgcact gggtgttggt cggagcgctt  15780 

ctttgccccg ccaaacatct tgcttgggaa gcgcaggccc cagcctgtgt tattgctggg  15840 

cgatataagg atggacatgt ttgctcaaaa agtgcggctc gataggacgc gcggcgagac  15900 

tatgcccagg gccttgtaaa cgtaggggca ggtgcggcgt ctggcgtcag taatggtcac  15960 

tcgctggact cctccgatgc tgttgcgcag cggtagcgtc ccgtgatctg tgagagcagg  16020 

aacgttttca ctgacggtgg tgatggtggg ggctggcggg cgcgccaaaa tctggttctc  16080 

gggaaagcga ttgaacacgt gggtcagaga ggtaaactgg cggatgagct gggagtagac  16140 

ggcctggtcg ttgtagaagc tcttggagtg cacgggcaac agctcggcgc ccaccaccgg  16200 

aaagttgctg atctggctcg tggagcggaa ggtcacgggg tcttgcatca tgtctggcaa  16260 

cgaccagtag acctgctccg agccgcaggt tacgtcagga gtgcaaagga gggtccatga  16320 

gcggatcccg gtctgagggt cgccgtagtt gtatgcaagg taccagctgc ggtactgggt  16380 

gaaggtgctg tcattgctta ttaggttgta actgcgtttc ttgctgtcct ctgtcagggg  16440 

tttgatcacc ggtttcttct gaggcttctc gacctcgggt tgcgcagcgg gggcggcagc  16500 

ttctgccgct gcctcggcct cagcgcgctt ctcctccgcc cgtgtggcaa aggtgtcgcc  16560 

gcgaatggca tgatcgttca tgtcctccac cggctgcatt gccgcggctg ccgcgttgga  16620 

gttctcttcc gcgccgctgc cactgttgtt gccgccgcct gcgccatccc cgccctgttc  16680 

ggtgtcatct tttaagcttg cctggtaggc gtccacatcc aacagtgcgg gaatgttacc  16740 

accctccagg tcatcgtagg tgatcctaaa gccctcctgg aagggttgcc gcttgcggat  16800 

gcccaacaag ttgctcaggc ggctgtgggt gaagtccacc ccgcatcctg gcagcaaaat  16860 

gatgtctgga tggaaggctt cgtttgtata taccccaggc atgacaagac cagtgactgg  16920 

gtcaaacccc agtctgaagt tgcgggtgtc aaactttacc ccgatgtcgc tttccagaac  16980 

cccgttctgc ctgcccactt tcaagtagtg ctccacgatc gcgttgttca taaggtctat  17040 

ggtcatggtc tcggagtagt tgccctcggg cagcgtgaac tccacccact catatttcag  17100 

ctccacctgt ttgtccttag taagcgagcg cgacaccatc acccgcgcct taaacttatt  17160 

ggtaaacatg aactcgttca catttggcat gttggtatgc aggatggttt tcaggtcgcc  17220 

gccccagtgc gaacggtcgt caagattgat ggtctgtgtg cttgcctccc ccgggctgta  17280 

gtcattgttt tgaatgaccg tggttagaaa gttgctgtgg tcgttctggt agttcaggga  17340 

tgccacatcc gttgacttgt tgtccacaag gtacacacgg gtggtgtcga ataggggtgc  17400 

caactcagag taacggatgc tgtttctccc cccggtaggc cgcaggtacc gcggaggcac  17460 

aaacggcggg tccaggggag catcgaaggg ggaacccagc gccgccgcca ctggcgccgc  17520 

gctcaccacg ctctcgtagg agggaggagg accttcctca tacatcgccg cgcgctgcat  17580 

actaagggga atacaagaaa accaacgctc ggtgccatgg ccttggtgag ttttttattt  17640 

tgcatcatgc tttttttttt tttttttaaa acattctccc cagcctgggg cgaaggtgcg  17700 

caaacgggtt gccactccct cccaaatcca ggacgctgct gtcgtctgcc gagtcatcgt  17760 

cctcccacac cagaccccgc tgacggtcgt gcctttgacg acgggtgggc gggcgcgggc  17820 

cgggcacatc cctgtgctcc tgcgcatacg tcttccatct actcatcttg tccactaggc  17880 

tctctatccc gttgttggga aatgccggag gcaggttctt ttcgcgctgc ggctgcagca  17940 

gcgagttgtt taggtactcc tcctcgccca gcaggcgcgg gcgggtggtg cgagtgctgg  18000 

taaaagaccc tatcaagctt ggaaatgggc tactcgcatc tgaccgcggg gccgcagcgc  18060 

ctagatcgga caagctgctt ggcctgcgga agctttcctt tcgcagcgcc gcctctgcct  18120 

gctcgcgctg ttgcaactct agcagggtct gcggttgcgg ggaaaacacg ctgtcgtcta  18180 

tgtcgtccca gaggaatcca tcgttaccct cgggcacctc aaatcccccg gtgtagaaac  18240 

cagggggcgg tagccagtgc gggttcaaga tggcattggt gaaatactcg gggttcacgg  18300 

cggccgcgcg atgcaagtag tccattaggc gattgataaa cggccggttt gaggcataca  18360 

tgcccggttc catgttgcgc gcggtcatgt ccagcgccac gctgggcgtt accccgtcgc  18420 

gcatcaggtt aaggctcacg ctctgctgca catagcgcaa gatgcgctcc tcctcgctgt  18480 

ttaaactgtg caacgagggg atcttctgcc gccggttggt cagcaggtag ttcagggttg  18540 

cctccaggct gcccgtgtcc tcctgcccca gcgcgcggct gacacttgta atctcctgga  18600 

aagtatgctc gtccacatgc gcctgaccta tggcctcgcg gtacagtgtc agcaagtgac  18660 

ctaggtatgt gtcccgggac acgctgccac tgtccgtgaa gggcgctatt agcagcagca  18720 

acaggcgcga gttgggcgtc agcaagctag acacggtcgc gcggtcgcct gtgggagccc  18780 

gcacccccca cagcccctgc aagttcttga aagcctggct caggtttacg gtctgcaggc  18840 

cttgtctact ggtctggaaa aaatagtctg gcccggactg gtacacctca ctttgcggtg  18900 

tctcagtcac cattagccgc agtgcgctca caaagttggt gtagtcctcc tgtccccgcg  18960 

gcacgttggc gggctgtgta ctcaggaagg cgtttagtgc aaccatggag cccaggttgc  19020 

cctgctgctg cgcgcgctca cgctgcgcca cggcctcgcg cacatccccc accagccggt  19080 

ccaggttggt ctgcacgttg ccgctgttgt aacgagccac gcgctgaagc agcgcgtcgt  19140 

agaccaggcc ggcctcatcg ggccggatgg ccctgttttc ggccagcgcg tttacgatcg  19200 

ccagcacctt ctcgtgcgtg gggtttgcgc gcgccgggac caccgcttcc agaattgcgg  19260 

agagccggtt ggcctgcggc tgctgccgga acgcgtcagg gttacgcgca gtcagcgaca  19320 

tgatgcggtc catgacctgg cgccagtcgt ccgtggagtt aaggccggac ggctggctct  19380 

gcagcgccgc ccgcaccgcc gggtccgttg cgtcttgcat catctgatca gaaacatcac  19440 

cgcttagtac tcgccgtcct ctggctcgta ctcatcgtcc tcgtcatatt cctccacgcc  19500 

gccgacgttg ccagcgcgcg cgggtgccac cgccagccca ggtccggccc cagctgcctc  19560 

cagggcgcgt cggcttgggg cccagcgcag gtcagcgccc gcgtcaaagt aggactcggc  19620 

ctctctatcg ccgctgcccg tgccagccag ggccctttgc aggctgtgca tcagctcgcg  19680 

gtcgctgagc tcgcgccgcc ggctcacgct cacggccttg tggatgcgct cgttgcgata  19740 

aacgcccagg tcgtcgctca aggtaagcac cttcaacgcc atgcgcatgt agaacccctc  19800 

gatctttacc tccttgtcta tgggaacgta aggggtatgg tatatcttgc gggcgtaaaa  19860 

cttgcccaga ctgagcatgg aatagttaat ggcggccacc ttgtcagcca ggctcaagct  19920 

gcgctcctgc accactatgc tctgcagaat gtttatcaaa tcgagcagcc agcggccctc  19980 

gggctctact atgtttagca gcgcatccct gaatgcctcg ttgtccctgc tgtgctgcac  20040 

tataaggaac agctgcgcca tgagcggctt gctatttggg ttttgctcca gcgcgcttac  20100 

aaagtcccac agatgcatca gtcctatagc cacctcctcg cgcgccacaa gcgtgcgcac  20160 

gtggttgtta aagctttttt gaaagttaat ctcctggttc accgtctgct cgtacgcggt  20220 

taccaggtcg gcggccgcca cgtgtgcgcg cgcgggacta atcccggtcc gcgcgtcggg  20280 

ctcaaagtcc tcctcgcgca gcaaccgctc gcggttcagg ccatgccgca actcgcgccc  20340 

tgcgtggaac tttcgatccc gcatctcctc gggctcctct ccctcgcggt cgcgaaacag  20400 

gttctgccgc ggcacgtacg cctcgcgcgt gtcacgcttc agctgcaccc ttgggtgtcg  20460 

ctcaggagag ggcgctccta gccgcgccag gccctcgccc tcctccaagt ccaggtagtg  20520 

ccgggcccgg cgccgcgggg gttcgtaatc accatctgcc gccgcgtcag ccgcggatgt  20580 

tgcccctcct gacgcggtag gagaagggga gggtgccctg catgtctgcc gctgctcttg  20640 

ctcttgccgc tgctgaggag gggggcgcat ctgccgcagc accggatgca tctgggaaaa  20700 

gcaaaaaagg ggctcgtccc tgtttccgga ggaatttgca agcggggtct tgcatgacgg  20760 

ggaggcaaac ccccgttcgc cgcagtccgg ccggcccgag actcgaaccg ggggtcctgc  20820 

gactcaaccc ttggaaaata accctccggc tacagggagc gagccactta atgctttcgc  20880 

tttccagcct aaccgcttac gccgcgcgcg gccagtggcc aaaaaagcta gcgcagcagc  20940 

cgccgcgcct ggaaggaagc caaaaggagc gctcccccgt tgtctgacgt cgcacacctg  21000 

ggttcgacac gcgggcggta accgcatgga tcacggcgga cggccggatc cggggttcga  21060 

accccggtcg tccgccatga tacccttgcg aatttatcca ccagaccacg gaagagtgcc  21120 

cgcttacagg ctctcctttt gcacggtcta gagcgtcaac gactgcgcac gcctcaccgg  21180 

ccagagcgtc ccgaccatgg agcacttttt gccgctgcgc aacatctgga accgcgtccg  21240 

cgactttccg cgcgcctcca ccaccgccgc cggcatcacc tggatgtcca ggtacatcta  21300 

cggatatcat cgccttatgt tggaagacct cgcccccgga gccccggcca ccctacgctg  21360 

gcccctctac cgccagccgc cgccgcactt tttggtggga tatcagtacc tggtgcggac  21420 

ttgcaacgac tacgtctttg actcaagggc ttactcgcgt ctcaggtaca ccgagctctc  21480 

gcagccgggt caccagaccg ttaactggtc cgttatggcc aactgcactt acaccatcaa  21540 

cacgggcgca taccaccgct ttgtggacat ggatgacttc cagtctaccc tcacgcaggt  21600 

gcagcaggcc atattagccg agcgcgttgt cgccgacctg gccctgcttc agccgatgag  21660 

gggcttcggg gtcacacgca tgggaggaag agggcgccac ctacggccaa actccgccgc  21720 

cgccgtagcg atagatgcaa gagatgcagg acaagaggaa ggagaagaag aagtgccggt  21780 

agaaaggctc atgcaagact actacaaaga cctgcgccga tgtcaaaacg aagcctgggg  21840 

catggccgac cgcctgcgca ttcagcaggc cggacccaag gacatggtgc ttctgtcgac  21900 

catccgccgt ctcaagaccg cctactttaa ttacatcatc agcagcacct ccgccagaaa  21960 

caaccccgac cgccacccgc tgccgcccgc cacggtgctc agcctacctt gcgactgtga  22020 

ctggttagac gcctttctcg agaggttttc cgatccggtc gatgcggact cgctcaggtc  22080 

cctcggtggc ggagtaccta cacaacaatt gttgagatgc atcgttagcg ccgtatccct  22140 

gccgcacggc agccccccgc caacccataa ccgggacatg acgggcggcg tcttccaact  22200 

gcgcccccgc gagaacggcc gcgccgtcac cgagaccatg cgccgtcgcc gcggggagat  22260 

gatcgagcgc tttgtcgacc gcctcccggt gcgccgtcgt cgccgccgtg tcccccctcc  22320 

cccaccgccg ccagaagaag aagaagaagg ggaggccctt atggaagagg agattgaaga  22380 

agaagaggcc cctgtagcct ttgagcgcga ggtgcgcgac actgtcgccg agctcatccg  22440 

tcttctggag gaggagttaa ccgtgtcggc gcgcaactcc cagtttttca acttcgccgt  22500 

ggacttctac gaggccatgg agcgccttga ggccttgggg gatatcaacg aatccacgtt  22560 

gcgacgctgg gttatgtact tcttcgtggc agaacacacc gccaccaccc tcaactacct  22620 

ctttcagcgc ctgcgaaact acgccgtctt cgcccggcac gtggagctca atctcgcgca  22680 

ggtggtcatg cgcgcccgcg atgccgaagg gggcgtggtc tacagccgcg tctggaacga  22740 

gggaggcctc aacgccttct cgcagctcat ggcccgcatc tccaacgacc tcgccgccac  22800 

cgtggagcga gccggacgcg gagatctcca ggaggaagag atcgagcagt tcatggccga  22860 

aatcgcctat caagacaact caggagacgt gcaggagatt ttgcgccagg ccgccgtcaa  22920 

cgacaccgaa attgattctg tcgaactctc tttcaggttc aagctcaccg ggcccgtcgt  22980 

cttcacgcag aggcgccaga ttcaggagat caaccgccgc gtcgtcgcgt tcgccagcaa  23040 

cctccgcgcg cagcaccagc tcctgcccgc gcgcggcgcc gacgtgcccc tgccccctct  23100 

cccggcgggt cccgagcccc ccctacctcc gggggcccgc ccgcgtcacc gcttttagat  23160 

gcatcatcca aggacacccc cgcggcccac cgcccgccgc gcggtaccgt agtcgcgccg  23220 

cggggatgcg gcctcttgca agtcatcgac gccgccacca accagcccct ggaaatcagg  23280 

tatcacctgg acctagcccg cgccctgacc cggctatgcg aggtaaacct gcaggagctc  23340 

ccgcctgacc tgtcgccgcg ggagctccag accatggaca gctcccatct gcgcgatgtt  23400 

gtcatcaagc tccgaccgcc gcgcgcggac atctggactt tgggctcgcg cggcgtggtg  23460 

gtccgatcca ccataactcc cctcgagcag ccagacggtc aaggacaagc agccgaagta  23520 

gaagaccacc agccaaaccc gccaggcgag gggctcaaat tcccactctg cttccttgtg  23580 

cgcggtcgtc aggtcaacct cgtgcaggat gtacagcccg tgcaccgctg ccagtactgc  23640 

gcacgttttt acaaaagcca gcacgagtgt tcggcccgtc gcagggactt ctactttcac  23700 

cacatcaaca gccactcctc caactggtgg cgggagatcc agttcttccc gatcggctcg  23760 

catcctcgca ccgagcgtct ctttgtcacc tacgatgtag agacctatac ttggatgggg  23820 

gcctttggga agcagctcgt gcccttcatg ctggttatga agttcggcgg agatgagcct  23880 

ctggtgaccg ccgcgcgaga cctagccgtg gaccttggat gggaccgctg ggaacaagac  23940 

ccgcttacct tctactgcat caccccagaa aaaatggcca taggtcgcca gtttaggacc  24000 

tttcgcgacc acctgcaaat gctaatggcc cgtgacctgt ggagctcatt cgtcgcttcc  24060 

aaccctcatc ttgcagactg ggccctgtca gaacacgggc tcagctcccc tgaggagctc  24120 

acctacgagg aacttaaaaa attgccctcc atcaagggca ccccgcgctt cttggaactt  24180 

tacatcgtgg gccacaacat caacggcttc gacgagatcg tgctcgccgc ccaggtaatt  24240 

aacaaccgtt ccgaggtgcc gggacccttc cgcatcacac gcaactttat gcctcgcgcg  24300 

ggaaagatac ttttcaacga tgtcaccttc gccctgccaa acccgcgttc caaaaagcgc  24360 

acggactttt tgctctggga gcagggcgga tgcgacgaca ctgacttcaa ataccagtac  24420 

ctcaaagtca tggttaggga cacctttgcg ctcacccaca cctcgctccg gaaggccgcg  24480 

caggcatacg cgctacccgt agaaaaggga tgctgcgcct accaggccgt caaccagttc  24540 

tacatgctag gctcttaccg ttcggaggcc gacgggtttc cgatccaaga gtactggaaa  24600 

gaccgcgaag agtttgtcct caaccgcgag ctgtggaaaa aaaagggaca ggataagtat  24660 

gacatcatca aggaaaccct ggactactgc gccctagacg tgcaggtcac cgccgagctg  24720 

gtcaacaagc tgcgcgactc ctacgcctcc ttcgtgcgtg acgcggtagg tctcacagac  24780 

gccagcttca acgtcttcca gcgtccaacc atatcatcca actcacatgc catcttcagg  24840 

cagatagtct tccgagcaga gcagcccgcc cgtagcaacc tcggtcccga cctcctcgct  24900 

ccctcgcacg aactatacga ttacgtgcgc gccagcatcc gcggtggaag atgctaccct  24960 

acatatcttg gaatactcag agagcccctc tacgtttacg acatttgcgg catgtacgcc  25020 

tccgcgctca cccaccccat gccatggggt cccccactca acccatacga gcgcgcgctt  25080 

gccgcccgcg catggcagca ggcgctagac ttgcaaggat gcaagataga ctacttcgac  25140 

gcgcgcctgc tgcccggggt ctttaccgtg gacgcagacc ccccggacga gacgcagcta  25200 

gacccactac cgccattctg ttcgcgcaag ggcggccgcc tctgctggac caacgagcgc  25260 

ctacgcggag aggtagccac cagcgttgac cttgtcaccc tgcacaaccg cggttggcgc  25320 

gtgcacctgg tgcccgacga gcgcaccacc gtctttcccg aatggcggtg cgttgcgcgc  25380 

gaatacgtgc agctaaacat cgcggccaag gagcgcgccg atcgcgacaa aaaccaaacc  25440 

ctgcgctcca tcgccaagtt gctgtccaac gccctctacg ggtcgtttgc caccaagctt  25500 

gacaacaaaa agattgtctt ttctgaccag atggacgcgg ccaccctcaa aggcatcacc  25560 

gcgggccagg tgaatatcaa atcctcctcg tttttggaaa ctgacaatct tagcgcagaa  25620 

gtcatgcccg cttttgagag ggagtactca ccccaacagc tggccctcgc agacagcgat  25680 

gcggaagaga gtgaggacga acgcgccccc accccctttt atagcccccc ttcaggaaca  25740 

cccggtcacg tggcctacac ctataaacca atcaccttcc ttgatgccga agagggcgac  25800 

atgtgtcttc acaccctgga gcgagtggac cccctagtgg acaacgaccg ctacccctcc  25860 

cacttagcct ccttcgtgct ggcctggacg cgagccttcg tctcagagtg gtccgagttt  25920 

ctatacgagg aggaccgcgg aacaccgctc gaggacaggc ctctcaagtc tgtatacggg  25980 

gacacggaca gccttttcgt caccgagcgt ggacaccggc tcatggaaac cagaggtaag  26040 

aaacgcatca aaaagcatgg gggaaacctg gtttttgacc ccgaacggcc agagctcacc  26100 

tggctcgtgg aatgcgagac cgtctgcggg gcctgcggcg cggatgccta ctccccggaa  26160 

tcggtatttc tcgcgcccaa gctctacgcc cttaaaagtc tgcactgccc ctcgtgcggc  26220 

gcctcctcca agggcaagct gcgcgccaag ggccacgccg cggaggggct ggactatgac  26280 

accatggtca aatgctacct ggccgacgcg cagggcgaag accggcagcg cttcagcacc  26340 

agcaggacca gcctcaagcg caccctggcc agcgcgcagc ccggagcgca ccccttcacc  26400 

gtgacccaga ctacgctgac gaggaccctg cgcccgtgga aagacatgac cctggcccgt  26460 

ctggacgagc accgactact gccgtacagc gaaagccgcc ccaacccgcg aaacgaggag  26520 

atatgctgga tcgagatgcc gtagagcacg tgaccgagct gtgggaccgc ctggaactgc  26580 

ttggtcaaac gctcaaaagc atgcctacgg cggacggcct caaaccgttg aaaaactttg  26640 

cttccttgca agaactgcta tcgctgggcg gcgagcgcct tctggcgcat ttggtcaggg  26700 

aaaacatgca agtcagggac atgcttaacg aagtggcccc cctgctcagg gatgacggca  26760 

gctgcagctc tcttaactac cagttgcagc cggtaatagg tgtgatttac gggcccaccg  26820 

gctgcggtaa gtcgcagctg ctcaggaacc tgctttcttc ccagctgatc tcccctaccc  26880 

cggaaacggt tttcttcatc gccccgcagg tagacatgat ccccccatct gaactcaaag  26940 

cgtgggaaat gcaaatctgt gagggtaact acgcccctgg gccggatgga accattatac  27000 

cgcagtctgg caccctccgc ccgcgctttg taaaaatggc ctatgacgat ctcatcctgg  27060 

aacacaacta tgacgttagt gatcccagaa atatcttcgc ccaggccgcc gcccgtgggc  27120 

ccattgccat cattatggac gaatgcatgg aaaatctcgg aggtcacaag ggcgtctcca  27180 

agttcttcca cgcatttcct tctaagctac atgacaaatt tcccaagtgc accggataca  27240 

ctgtgctggt ggttctgcac aacatgaatc cccggaggga tatggctggg aacatagcca  27300 

acctaaaaat acagtccaag atgcatctca tatccccacg tatgcaccca tcccagctta  27360 

accgctttgt aaacacttac accaagggcc tgcccctggc aatcagcttg ctactgaaag  27420 

acatttttag gcaccacgcc cagcgctcct gctacgactg gatcatctac aacaccaccc  27480 

cgcagcatga agctctgcag tggtgctacc tccaccccag agacgggctt atgcccatgt  27540 

atctgaacat ccagagtcac ctttaccacg tcctggaaaa aatacacagg accctcaacg  27600 

accgagaccg ctggtcccgg gcctaccgcg cgcgcaaaac ccctaaataa agacagcaag  27660 

acacttgctt gatcaaaatc caaacagagt ctggttttta tttatgtttt aaaccgcatt  27720 

gggaggggag gaagccttca gggcagaaac ctgctggcgc agatccaaca gctgctgaga  27780 

aacgacatta agttcccggg tcaaagaatc caattgtgcc aaaagagccg tcaacttgtc  27840 

atcgcgggcg gatgaacggg aagctgcact gcttgcaagc gggctcagga aagcaaagtc  27900 

agtcacaatc ccgcgggcgg tggctgcagc ggctgaagcg gcggcggagg ctgcagtctc  27960 

caacggcgtt ccagacacgg tctcgtaggt caaggtagta gagtttgcgg gcaggacggg  28020 

gcgaccatca atgctggagc ccatcacatt ctgacgcacc ccggcccatg ggggcatgcg  28080 

cgttgtcaaa tatgagctca caatgcttcc atcaaacgag ttggcgctca tggcggcggc  28140 

tgctgcaaaa cagatacaaa actacatgag acccccacct tatatattct ttcccaccct  28200 

taagccccgc ccatcgatgg caaacagcta ttatgggtat tatgggtgct agcgacatga  28260 

ggttgccccg tattcagtgt cgctgatttg tattgtctga agttgttttt acgttaagtt  28320 

gatgcagatc aattaatacg atacctgcgt cataattgat tatttgacgt ggtttgatgg  28380 

cctccacgca cgttgtgata tgtagatgat aatcattatc actttacggg tcctttccgg  28440 

tgatccgaca ggttacgggg cggcgacctc gcgggttttc gctatttatg aaaattttcc  28500 

ggtttaaggc gtttccgttc ttcttcgtca taacttaatg tttttattta aaataccctc  28560 

tgaaaagaaa ggaaacgaca ggtgctgaaa gcgaggcttt ttggcctctg tcgtttcctt  28620 

tctctgtttt tgtccgtgga atgaacaatg gaagttaacg gatccaggcc gcgagcaaaa  28680 

ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc  28740 

cgcccccctg acgagcatca caaaaatcaa cgctcaagtc agaggtggcg aaacccgaca  28800 

ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg  28860 

accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct  28920 

catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt  28980 

gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag  29040 

tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc  29100 

agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac  29160 

actagaagaa cagtatttgg tatctgcgct ctgccaaagc cagttacctt cggaaaaaga  29220 

gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc  29280 

aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg  29340 

gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat cagattatca  29400 

aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt  29460 

atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca  29520 

gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtagtgta gataactacg  29580 

atacgggagg gcttaccatc cggccccagt gctgcaatga taccgcgtga cccacgctca  29640 

ccggctcctg atttatcagc aataaaccag ccagccggaa gtgccgagcg cagaagtggt  29700 

cctgcaactt tatccgcctc catccagtct attagttgtt gccgggaagc tagagtaagt  29760 

agttcgccag ttaatagttt tcgcaacgtt gttgccattg ctacaggcat cgtggtgtca  29820 

cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca  29880 

tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat agttgtcaga  29940 

agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact  30000 

gtcatgccat ccgtaagatg cttttctgtg actggtgagt attcaaccaa gaatacggga  30060 

taataccgcg ccacatagca gaactttaaa agtgctcatc attgggaaac gttcttcggg  30120 

gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgcgc  30180 

acccaagtga tcttctgcat cttttacttt caccagcgtt tctgggtgag caaaaacagg  30240 

aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact  30300 

tttccttttt caatattatt gaagcattta tcagggttat tgtctcatca gcggatacat  30360 

atttg                                                              30365 

 
           
             5  
             33  
             DNA  
             Homo sapiens  
           
            5 

gcggaattcg gcttggtgac ttagagaaca gag                                  33 

 
           
             6  
             33  
             DNA  
             Homo sapiens  
           
            6 

gcgggatcct tgaacccgga ccctctcaca cta                                  33 

 
           
             7  
             20  
             DNA  
             Artificial Sequence  
             
               derived from Adenovirus  
             
           
            7 

actctcttcc gcatcgctgt                                                 20 

 
           
             8  
             21  
             DNA  
             Artificial Sequence  
             
               derived from Adenovirus  
             
           
            8 

cttgcgactg tgactggtta g                                               21 

 
           
             9  
             20  
             DNA  
             Artificial Sequence  
             
               derived from Adenovirus  
             
           
            9 

ccgcacccac tatcttcata                                                 20 

 
           
             10  
             20  
             DNA  
             Artificial Sequence  
             
               derived from Adenovirus  
             
           
            10 

ggtgtccaaa ggttcggaga                                                 20