Gene therapy for retinal diseases

Abstract

This invention provides viral vectors and compositions for use in retinal gene therapy. [Solution] The viral vector includes a transgene construct. In some embodiments, the transgene construct is a DNA sequence including a promoter element, an open reading frame of a retinal gene longer than 10 kb, and a terminator sequence. In other embodiments, the transgene construct is a DNA segment of a retinal gene longer than 10 kb.

Description

TECHNICAL FIELD

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the priority benefit of Provisional Application No. 63 / 149,122, filed on February 12, 2021, which is incorporated herein by reference in its entirety, and is a non-provisional application.

[0002] SEQUENCE LISTING SUBMISSION A Sequence Listing contained in a file named sequence listing, which is 110 kilobytes in size (measured on MS-Windows) and created on February 11, 2022, is provided herewith, and is incorporated herein by reference in its entirety.

[0003] The present disclosure relates to gene therapy for treating Usher syndrome type IIA, Usher syndrome type 2C, retinitis pigmentosa 39, retinitis pigmentosa 25, Wagner syndrome, or age-related macular degeneration. BACKGROUND ART

[0004] Usher syndrome type 2 is a leading cause of co-occurrence of blindness and hearing loss. Retinitis pigmentosa (RP) is a leading cause of hereditary blindness. Usher syndrome 2A (Ush2A) mutations are estimated to cause approximately 20% of retinitis pigmentosa cases, and may be the single largest cause. Symptomatic Usher 2A syndrome is accompanied by hearing loss in youth, and vision loss occurs from early to middle adulthood. Non-syndromic blindness can be caused by Ush2A mutations, in which hearing is normal but vision impairment still occurs.

[0005] Hemicecntin mutations causing Usher syndrome type 2C (Ush2C), RP39, RP25, Wagner syndrome, and age-related macular degeneration are similarly accompanied by photoreceptor cell loss, but these diseases are much rarer than Usher syndrome type IIA. However, all of these diseases result in vision loss, and involve mutations in genes that are too large for conventional gene therapy using adeno-associated virus.

[0006] Viral gene therapy vectors achieved the first major clinical success in treating X-linked severe combined immunodeficiency. Retroviral vectors achieved long-term immune rearrangement but caused life-threatening acute lymphoblastic leukemia in 30% of subjects. Other vector systems, such as adeno-associated virus (AAV) systems, have advanced in recent years, although they have limited capabilities and are associated with the development of hepatocellular carcinoma. Lentiviral vectors, through chromosomal integration, enable sustained transgene expression but can induce insertional mutagenesis. Adenovirus vectors (Ad vectors) transduce both dividing and non-dividing cells, resulting in rapid and efficient transgene expression. In their initial iterations, Ad vectors were hindered by the induced pre-existing immune response. To minimize the immune and inflammatory responses, it was necessary to remove all endogenous Ad genes from the Ad vector to create a complete deletion (fd) Ad vector. Capsid modification and anti-inflammatory drugs also reduced Ad-related inflammation, making gene therapy approaches feasible.

[0007] Gene therapy approaches for Usher type 2 syndrome, caused by Ush2A mutations, have been proposed to include altering RNA splicing to skip affected exons (QR-421a in ProQR) or fixing mutations using CRISPR / Cas9 editing. These approaches would require treatment tailored to individual mutations or mutated regions. As of 2018, UMD-USHbase lists 3083 mutations in Ush2A associated with blindness. The most common mutations account for only 38% of pathogenic mutations. Supplying a normal copy of the entire gene would be a ready-made solution that could potentially resolve all Ush2A mutations. Furthermore, potential side effects of CRISPR editing in vivo include cancer and increased photoreceptor death. These side effects would be avoided by episomal copying of the gene. Similarly, Ush2C, RP39, RP25, and hemisent mutations causing Wagner syndrome and age-related macular degeneration could be resolved with a single gene therapy approach for each gene, rather than requiring thousands of individual solutions.

[0008] Furthermore, retinal gene therapy may benefit from employing genome editing techniques to address dominant-negative effects of mutations in the genome copy of genes, deletions of large non-coding regions of genes crucial to retinal function, or the addition of compensatory gene constructs to resolve other issues. In such cases, it may be important to address the issues using homologous recombination-independent specific gene insertion (HITI) or homologous recombination (HR) using longer donor / template sequences than those possible with 5kb adeno-associated virus (AAV). Adenoviruses can provide templates up to 30kb in length and can further adapt to the mechanisms required for CRISPR editing in the same vector, thereby increasing the efficiency of the therapy.

[0009] Furthermore, the gene length of the key transcript in the case of Ush2A is approximately 15,000 (15kb) base pairs. The maximum capacity of AAV is approximately 4,000 base pairs. Lentiviruses are another common gene therapy vector (despite concerns about the safety of genome integration), but they have a maximum of 10,000 base pairs. [Overview of the Initiative]

[0010] In one embodiment, the present disclosure provides a viral vector. According to an embodiment of the present disclosure, the viral vector comprises a transgene construct, the transgene construct being one of the following: (a) a DNA sequence comprising a promoter element, an open reading frame of a retinal gene longer than 10 kb, and a terminator sequence; and (b) a DNA segment of a retinal gene longer than 10 kb.

[0011] In one embodiment, the vector completely deletes endogenous genes other than the ITR sequence. In another embodiment, the vector deletes enough endogenous adenovirus genes to make up to 10 kb of space for the cargo sequence. In a further embodiment, the viral vector is a mimivirus that has been completely or partially removed.

[0012] In another embodiment, the viral vector is an adenovirus that has been completely or partially removed. In one embodiment, the viral vector is capsidated to a capsid based on adenovirus type 5. In a further embodiment, the viral vector is capsidated to a capsid based on adenovirus type 6. In yet another embodiment, the viral vector is capsidated to an adenovirus-based capsid and modified to remove the RGD sequence from the capsid protein. In yet another embodiment, the viral vector is capsidated to a capsid based on adenoviruses of families A-G. In yet another embodiment, the transgene construct is a DNA sequence comprising (a) a promoter element, an open reading frame of the retinal gene longer than 10 kb, and a terminator sequence.

[0013] In one embodiment, the promoter element is a human rhodopsin kinase promoter. In another embodiment, the promoter element is a human rhodopsin kinase promoter. According to a further embodiment, the promoter element is a mouse rhodopsin kinase promoter. In a further embodiment, the promoter element is a ubiquitous CMV promoter. In yet another embodiment, the promoter element is an upstream non-coding sequence of the Usherin gene containing the native promoter of the Usherin gene. In another embodiment, the promoter element is a synthetic transducin alpha-subunit promoter. In yet another embodiment, the promoter element is a minimal sequence of a photoreceptor retinoid-binding protein promoter and a human transducin alpha-subunit promoter. In yet another embodiment, the promoter element is a chemically controlled tet-inducible promoter. In another embodiment, the promoter element is a chemically inhibited promoter. In another embodiment, the promoter element is a human L-opsin promoter. In yet another embodiment, the promoter element is a human retinosulin proximal promoter and a human photoreceptor retinoid-binding protein enhancer. In further embodiments, the promoter element is specific to the photoreceptor.

[0014] In one embodiment, the transgene construct is a nucleotide sequence that translates to Ush2A. In another embodiment, the transgene construct is a nucleotide sequence that translates to ADGV1. In yet another embodiment, the transgene construct is a nucleotide sequence that translates to the eyes shut homolog. In yet another embodiment, the transgene construct is a nucleotide sequence that translates to hemisentin. In yet another embodiment, the transgene construct is a nucleotide sequence that translates to versican.

[0015] In one embodiment, the transgene construct is (b) a DNA segment of a retinal gene longer than 10kb. In another embodiment, the DNA segment is at least a segment of the Ush2A gene. In yet another embodiment, the DNA segment is at least a segment of the ADGVl gene. In yet another embodiment, the DNA segment is at least a segment of the eyes shut homolog gene. In yet another embodiment, the DNA segment is at least a segment of the hemisentin gene. In yet another embodiment, the DNA segment is at least a segment of the versican gene.

[0016] In one embodiment, the introduced gene construct is (b) a DNA segment of a retinal gene longer than 10kb.

[0017] In one embodiment, the Disclosure provides a composition. According to an embodiment of the Disclosure, the composition comprises a viral vector and a packing plasmid, the viral vector comprising (a) a DNA sequence comprising a promoter element, an open read frame of a retinal gene longer than 10 kb, and a terminator sequence, and (b) a transgene construct selected from a DNA segment of a retinal gene longer than 10 kb.

[0018] In one embodiment, the present disclosure provides a method for treating a retinal disease. According to an embodiment of the present disclosure, the method for treating a retinal disease comprises providing a viral vector comprising a transgene construct, the transgene construct being one of (a) a DNA sequence comprising a promoter element, an open read frame of a retinal gene longer than 10 kb, and a terminator sequence, and (b) a DNA segment of a retinal gene longer than 10 kb.

[0019] In one embodiment, the retinal disease is selected from the group consisting of Usher syndrome type IIA, Usher syndrome type 2C, retinitis pigmentosa 39, retinitis pigmentosa 25, Wagner syndrome, and age-related macular degeneration.

[0020] Brief Description of the Sequence Listing SEQ ID NO: 1 shows the DNA sequence of USH2A.

[0021] SEQ ID NO: 2 shows the DNA sequence of USH2A downstream of the CMV promoter.

[0022] SEQ ID NO: 3 shows the DNA sequence of USH2A downstream of the rhodopsin kinase promoter. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] [Figure 1] Shows the components of an adenoviral vector. In particular, Figure 1A shows the design of a vector genome model according to an embodiment of the present disclosure, Figure 1B shows the design of a pPaC packaging expression plasmid according to an embodiment of the present disclosure, Figure 1C shows vector encapsidation by cotransfection into a host cell according to an embodiment of the present disclosure, and Figure 1D shows the release of the packaged vector according to an embodiment of the present disclosure. [Figure 2] Shows transduction of test cells using a hemophilia factor 8 gene transfer vector according to an embodiment of the present disclosure. [Figure 3] Shows transfection of mouse retinal cells for testing specific gene transcription. [Figure 4] Shows construction of the USH2A vector by (1) forced cloning and (2) stuffing via recombineering. DESCRIPTION OF EMBODIMENTS

[0024] Before describing any embodiments of the present disclosure in detail, it is to be understood that the present disclosure is not limited in its application to the details of construction and arrangement of components set forth in the following description or illustrated in the drawings. The present disclosure is capable of other embodiments and of being practiced or carried out in various ways. It is also understood that the expressions and terminology used herein are for the purpose of description and should not be regarded as limiting. The use of "including", "comprising" or "having" and variations thereof herein is meant to encompass the items listed thereafter and equivalents thereof as well as additional items. The use of "consisting essentially of" and "consists essentially of" and variations thereof herein is meant to encompass the items listed thereafter, equivalents thereof and additional items, provided that such equivalents and additional items do not materially alter the overall properties, use or manufacture. The use of "consisting of" and variations thereof herein is meant to include only the items listed thereafter and those items only.

[0025] Referring to the drawings, like numerals refer to like elements throughout. The terms first, second, etc. may be used herein to describe various elements, components, regions, and / or sections, but it will be understood that these elements, components, regions, and / or sections should not be limited by these terms. These terms are only used to distinguish one element, component, region and / or section from another element, component, region and / or section. Thus, a first element, component, region or section could be termed a second element, component, region or section without departing from the present disclosure.

[0026] Numerical ranges in this disclosure are approximations and may include values ​​outside the range unless otherwise indicated. Numerical ranges include all values ​​from and including the lower and higher values ​​(unless otherwise specified), in increments of one unit, provided that there is at least two units of separation between any lower and any higher value. For example, if a physical or other property, such as the amount of an ingredient by weight, is 10 to 100, then all individual values ​​such as 10, 11, 12, etc., and subranges such as 10 to 44, 55 to 70, 97 to 100 are intended to be explicitly listed. In the case of a range that includes an explicit value (e.g., the range from 1, or 2, or 3 to 5, or 6, or 7), any subrange between any two explicit values ​​is included (e.g., the above range 1 to 7 includes subranges 1 to 2, 2 to 6, 5 to 7, 3 to 7, 5 to 6, etc.). For ranges containing values ​​less than 1 or fractions greater than 1 (e.g., 1.1, 1.5, etc.), one unit is appropriately considered to be 0.0001, 0.001, 0.01, or 0.1. For ranges containing single-digit numbers less than 10 (e.g., 1 to 5), one unit is usually considered to be 0.1. These are merely examples of what is specifically intended, and all possible combinations of numbers between the listed minimum and maximum values ​​should be considered expressly stated in this disclosure.

[0027] Spatial terms such as “beneath,” “below,” “lower,” “above,” and “upper” may be used herein to facilitate descriptions of the relationship between one element or feature and another, as shown in the figures. It will be understood that spatially relative terms are intended to encompass different orientations depending on the orientation in use or example. For example, if the apparatus in the figure is turned upside down, an element described as being “below” or “beneath” another element or feature will be oriented “above” the other element or feature. Thus, the illustrative term “below” may encompass both upward and downward orientations. The apparatus may be oriented in other directions (rotated by 90° or in other orientations), and the spatially relative descriptors used herein will be interpreted accordingly.

[0028] As used herein, the term "and / or" includes any combination of one or more of the related enumerated items. For example, when used in a phrase such as "A and / or B," the phrase "and / or" is intended to include both A and B; A or B; A (alone); and B (alone). Similarly, when used in a phrase such as "A, B and / or C," the term "and / or" is intended to include each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0029] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which this disclosure belongs. Generally, the nomenclature used herein, as well as the laboratory procedures in cell culture, molecular genetics, nucleic acid chemistry, and hybridization described below, are well known and commonly used in the art. Standard techniques are used for recombinant nucleic acid methods, polynucleotide synthesis, and microbial culture and transformation (e.g., electroporation, lipofection). Generally, enzymatic reactions and purification steps are carried out according to the manufacturer's specifications. Techniques and procedures are generally carried out in accordance with conventional methods in the art and the various general references provided throughout this document (generally, see Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd ed. (1989), Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, which are incorporated herein by reference). Units, prefixes, and symbols may be shown in their SI-approved forms. Unless otherwise specified, nucleic acids are written from left to right in the 5' to 3' direction, and amino acid sequences are written from left to right in the amino to carboxyl direction. Amino acids may be referred herein by either a commonly known three-letter symbol or a single-letter symbol recommended by the IUPAC-IUB Biochemical Nomenclature Committee. Similarly, nucleotides may be referred by a commonly accepted single-letter code. Unless otherwise specified, software, electrical, and electronic terms used herein are as defined in The New IEEE Standard Dictionary of Electrical and Electronics Terms (5th sup.th edition, 1993).

[0030] As used herein, the term “adenovirus vector” includes any gene construct or viral construct based on an adenovirus that is used to introduce genetic material. As used herein, the terms “deletion adenovirus” or “deletion adenovirus vector” include any adenovirus or adenovirus vector having one or more endogenous genes or gene fragments deleted therefrom. In contrast, as used herein, the terms “complete deletion adenovirus” and “complete deletion adenovirus vector” include any adenovirus and adenovirus vector from which all endogenous adenovirus genes and genetic material are deleted, except for the internal terminal repeat (ITR) and packaging signal (Ψ). As used herein, the term “adenovirus vector genome” includes the genetic material found in an adenovirus vector.

[0031] A “coding sequence” or a sequence that “codes” a selected polypeptide is a nucleic acid molecule that, when controlled by appropriate regulatory sequences (or “regulatory elements”), is transcribed into a polypeptide in vivo (in the case of DNA) and translated (in the case of mRNA). The boundaries of a coding sequence are determined by a 5' (amino) terminal start codon and a 3' (carboxy) terminal translation termination codon. A transcription termination sequence may be located 3' of the coding sequence. The transcription and translation of a coding sequence are typically regulated by “regulatory elements” including, but not limited to, transcription promoters, transcription enhancer elements, Shine and Delagamo sequences, transcription termination signals, polyadenylation sequences (located 3' relative to the translation termination codon), sequences for optimizing translation initiation (located 5' relative to the coding sequence), and translation termination sequences.

[0032] As used herein, the terms “delete” or “missing” mean to delete, erase, or remove.

[0033] As used herein, the terms “deletion Ad(virus) vector,” “removed-,” “mini-,” “deletion-,” “.DELTA.” or “pseudovector” refer to linear vector modules having an ITR. These vectors may also encode several structural gene sequences and / or non-structural gene sequences and / or one or more target genes or transgenes.

[0034] The term "expression" refers to the transcription and / or translation of an endogenous gene, transgene, or coding region within a cell.

[0035] A “gene delivery vector,” “GDV,” “gene transfer vector,” or “gene transfer vehicle” is a composition comprising the packaged vector module of this disclosure.

[0036] As used herein, the term “gene expression construct” refers to a promoter, at least a fragment of the gene of interest, and a polyadenylation signal sequence. The vector modules of this disclosure may include a gene expression construct.

[0037] As used herein, the terms “target gene,” “GOI,” and “transgene” refer to a gene that encodes a gene whose function is medically important and which may not be a naturally occurring flaviviral gene. A target gene may exert its effect at the RNA or protein level. Examples of target genes include, but are not limited to, therapeutic genes, immunomodulatory genes, viral genes, bacterial genes, protein-producing genes, inhibitory RNA or proteins, and regulatory proteins.

[0038] "Genetic sequence" refers to the order of nucleotides. Genetic sequences can be regulated. Regulation of gene expression can be achieved by one of the following: (1) changes in gene structure: site-directed recombinases (e.g., Cre based on the Cre-loxP system) can activate gene expression by removing an insertion sequence between the promoter and the gene; (2) changes in transcription: either by induction (covered) or reduction of inhibition; (3) changes in mRNA stability by specific sequences incorporated into mRNA or by siRNA; and (4) changes in translation by sequences in mRNA. Deletion adenoviruses are also called "high-capacity" adenoviruses. These deletion adenoviruses can accommodate gene sequences up to 33 kb.

[0039] The term "heterogeneous" is typically used for any combination of DNA sequences that are not closely related in nature.

[0040] The term "homology" refers to the existence of a common ancestor between a pair of structures or genes.

[0041] A “host cell” or “packaging cell” is a cell capable of packaging an adenovirus or adenovirus vector genome or modified genome to produce viral particles. It may be manipulated to provide a deficiency gene product or its equivalent. Thus, a packaging cell can package an adenovirus genome into adenovirus particles. The production of such particles requires that the genome be replicated and that the proteins necessary for assembling the infectious virus be produced. The particles may also require specific proteins necessary for the maturation of the viral particles. Such proteins may be provided by a vector, a packaging construct, or a packaging cell. Exemplary host cells (HCs) that may be used to construct ap packaging cell lines according to this disclosure include, but are not limited to, A549, HeLa, MRC5, W138, CHO cells, Vero cells, human embryonic retinal cells, or any eukaryotic cell, as long as the host cell allows for the proliferation of adenovirus. Some host cell lines include adipocytes, chondrocytes, epithelial cells, fibroblasts, glioblastomas, hepatocytes, keratinocytes, leukemia cells, lymphohoblastoids, monocytes, macrophages, myoblasts, and neurons. Other cell types include, but are not limited to, cells derived from primary cell cultures, such as human primary prostate cells, human embryonic retinal cells, and human stem cells. Eukaryotic diploid and aneuploid cell lines are included within the scope of this disclosure. Packaging cells must be capable of expressing the products of the different constructs described herein at appropriate levels for those products in order to generate high-titer stocks of recombinant viral vectors.

[0042] "Immune response" refers to acquired immune responses, such as cellular or humoral immune responses.

[0043] As used herein, the terms “introduce” or “transfection” refer to the delivery of an expression vector to a host cell. A vector may be introduced into a cell by transfection, which typically means the insertion of heterologous DNA or RNA into a cell by physical means (e.g., calcium phosphate transfection, electroporation, microinjection, or lipofection); infection, which typically means introduction by an infectious pathogen, i.e., a virus; or transduction, which typically means stable infection of a cell by a virus, or the introduction of genetic material from one microorganism to another by a viral pathogen (e.g., a bacteriophage). The vector may be a plasmid, a virus, or another vehicle.

[0044] The terms “packaging construct” or “packaging expression plasmid” refer to an engineered plasmid construct of a circular double-stranded DNA molecule, the DNA molecule comprising a subset of adenovirus structural or non-structural genes under the control of at least a promoter. A “packaging construct” does not contain two or more ITRs or genetic information that would enable independent viral replication to infect, produce viral particles, and / or efficiently package this genetic material into the viral particles.

[0045] As used herein, the term "plasmid" refers to an extrachromosomal DNA molecule that can replicate independently of chromosomal DNA. In most cases, it is circular and double-stranded.

[0046] The term "promoter" refers to a regulatory region of DNA that promotes the transcription of a particular gene. Promoters typically contain a TATA box that can instruct RNA polymerase II to initiate RNA synthesis at the appropriate transcription start site of a particular coding sequence. Promoters may further contain other recognition sequences, generally located upstream of the TATA box or 5' relative to the TATA box, called upstream promoter elements, which affect the transcription initiation rate.

[0047] The term "transfection" refers to the instruction of cytogenetic material as DNA or RNA (e.g., the introduction of an isolated nucleic acid molecule or a construct of this disclosure). As used herein, the term "transduction" refers to the introduction of cellular DNA, either as DNA or using the GDV of this disclosure. The GDV of this disclosure can be transduced into target cells.

[0048] The term "vector" refers to a nucleic acid used to infect host cells into which polynucleotides can be inserted. Vectors are often replicons. Expression vectors enable the transcription of the nucleic acid inserted within them. Some common vectors include, but are not limited to, plasmids, cosmids, viruses, phages, recombinant expression cassettes, and transposons. The term "vector" can also refer to an element that facilitates the introduction of a gene from one location to another.

[0049] The term "vector module" refers to an adenovirus gene composition packaged within an adenovirus virion.

[0050] Embodiments of the present disclosure provide an adenovirus vector. The adenovirus vector comprises a transgene construct which is a DNA segment or a DNA sequence. In one embodiment, the adenovirus vector comprises a capsid. In one embodiment, the capsid is provided as a gene therapy for the treatment of Usher syndrome type IIA, Usher syndrome type 2C, retinitis pigmentosa39, retinitis pigmentosa25, Wagner syndrome, or age-related macular degeneration, or for the prevention of retinal photoreceptor degeneration caused by loss of Ush2A, ADGRV1, eyes shut homolog, hemisentin, or versican (VCAN) protein due to mutation. Viral vector

[0051] The vectors of this disclosure are based on viruses selected from the group consisting of adenoviruses and mimiviruses. Preferably, the viral vectors are based on adenoviruses. The viral vectors used in this disclosure are designed as complete deletion (FD) viruses, more specifically, FD adenoviruses packaged in the absence of helper viruses. The viral vector is one of two independently modifiable components of a capsid, the other being a circular packaging expression plasmid.

[0052] In one embodiment, the viral vector is an ad vector.

[0053] In one embodiment, the viral vector is an Ad vector that is completely deleted and does not contain a helper virus.

[0054] In one embodiment, an adenovirus vector 70, preferably an fd adenovirus vector, can receive a gene construct and has inverted terminal repeat sequences (ITRs) 72, 72 and a packaging signal (Ψ) 73, as shown in Figure 1. The deleted endogenous gene is replaced by a size-compensating stuffer 75. In the embodiments shown, these stuffers 75 are prepared from a fragment of the human gene 5-aminoimidazole-4-carboxamidorivonucleotide formyltransferase gene (ATIC). In further embodiments, but not limited to, other stuffer sequences such as human hypoxanthine-guanine phosphoribosyltransferase may be used.

[0055] The fd-based vector genome module is operated for transgene constructs of different lengths. In some embodiments, the fd-based vector genome module is operated for transgene constructs of 1 kb, or 5 kb, or 10 kb to 20 kb, or 25 kb, or 30 kb, or 35 kb, preferably 10 kb, or more than 10 kb, or 12 kb, or 15 kb, or 18 kb, or 20 kb to 22 kb, or 25 kb, or 28 kb, or 30 kb, or 32 kb, or less than 35 kb.

[0056] In one embodiment, the introduced gene construct 76 is a DNA sequence comprising a promoter element, an open reading frame of a retinal gene longer than 10 kb, and a terminator sequence.

[0057] In one embodiment, the promoter is specific to the photoreceptor. Non-limiting examples of photoreceptor-specific promoters include, but are not limited to, the human rhodopsin kinase promoter, the mouse rhodopsin kinase promoter, the ubiquitous CMV promoter, the ubiquitous CAG promoter, the upstream non-coding sequence of the Usherin gene containing the Usherin gene's native promoter, the synthetic transducing alpha-subunit promoter, the photoreceptor retinoid-binding protein promoter, the minimal sequence of the human transducing alpha-subunit promoter, the chemically controlled tet-inducible promoter, the human L-opsin promoter, the retinosuxin proximal promoter, the human interphotorceptor retinoid-bindin protein enhancer, and combinations thereof.

[0058] In certain embodiments, the promoter is selected from the group consisting of a human rhodopsin kinase promoter, a mouse rhodopsin kinase promoter, a ubiquitous CMV promoter, a ubiquitous CAG promoter, an upstream non-coding sequence of the Usherin gene containing the Usherin gene's native promoter, a synthetic transducin alpha-subunit promoter, a photoreceptor-retinoid-binding protein promoter and the minimal sequence of the human transducin alpha-subunit promoter, a chemically inhibited promoter, a human L-opsin promoter, and a human retinosuxin proximal promoter and a human photoreceptor-retinoid-binding protein enhancer.

[0059] In one embodiment, the open reading frame of the retinal gene is a nucleotide sequence. In one embodiment, the nucleotide sequence is translated into a photoreceptor-specific protein. Non-limiting examples of nucleotide sequences include those that translate into proteins encoded by genes selected from Ush2A, ADGV1, eyes shut homologs, hemisentins, versicans, and combinations thereof. In one embodiment, the nucleotide sequence is translated into proteins encoded by genes selected from the group consisting of Ush2A, ADGV1, eyes shut homologs, hemisentins, and versicans.

[0060] In one embodiment, the transgene construct is a DNA segment of a retinal gene longer than 10 kb. Non-limiting examples of suitable DNA segments of a retinal gene longer than 10 kb include the genomic sequence of the Ush2A gene, the genomic sequence of the ADGV1 gene, the genomic sequence of the eyes shut homolog gene, the genomic sequence of the hemisentin gene, the genomic sequence of the versican gene, and combinations thereof.

[0061] Different transgene expression cassettes have been successfully transferred to the fd-based genome vector module. Due to the episomal position of the Ad vector genome, recombination into the cellular genome is not expected and has not been observed.

[0062] Packing plasmid In one embodiment, a packing plasmid is provided for a viral vector.

[0063] The circular packaging expression plasmid (pPac), as shown in Figure 1, is developed to provide late genes (L1, L2, L3, L4, L5) in trans along with early genes E2 and E4. LITR, the packaging signal (Ψ), early genes E1 and E3, and protein IX are not present in the circular pPac.

[0064] In one embodiment, different packaging plasmids capsid the vector module into capsids of different serotypes, such as Ad2(pPaC2), Ad5(pPaC5), Ad6(pPaC6), Ad4(pPaE4), and Ad35(pPaB35).

[0065] Transfection In one embodiment, a viral vector and a packing plasmid are transfected into packaging cells.

[0066] In one embodiment, the packaging cell may contain one or more viral vectors and one or more plasmids. In a preferred embodiment, the packaging cell contains at least one, preferably two or more, more preferably three or more viral vectors and one packing plasmid.

[0067] Referring further to Figure 1, the viral vector 70 and plasmid 82 are introduced into a eukaryotic host cell or packaging cell 85 via cotransfection. In the specific embodiment shown, the viral vector 70 and plasmid 82 are cotransfected into packaging cells 85 using an optimized and standardized one-week cotransfection protocol.

[0068] In one embodiment, the viral vector is an adenovirus vector, particularly an fd adenovirus vector, and the packaging cells are derived from cell lines such as, but not limited to, human embryonic kidney cells (HEK293) and PerC.6 cells. The packaging cells required to package the fd adenovirus vector must be modified to express a gene encoded within the E1 region of the adenovirus vector. In a particular embodiment, the packaging cells are HEK293-derived Q7 packaging cells modified to express a gene encoded within the E1 region of the adenovirus vector.

[0069] Capsid According to embodiments of this disclosure, as shown in Figure 1, packing cells 85 containing a viral vector 70 and plasmid 82 are capsidized.

[0070] Packaging cells 85 containing the viral vector 70 and plasmid 82 are delivered with a capsid of an adenovirus or mimivirus serotype. In one embodiment, the capsid is modified to remove the RGD sequence from the capsid protein, based on an adenovirus. In one embodiment, the capsid is based on adenovirus families A-G. In one embodiment, the viral vector is delivered with capsids of adenovirus serotypes Ad2, Ad5, Ad6, and Ad35, and combinations thereof. In one embodiment, the viral vector is delivered with an Ad6 serotype capsid.

[0071] In the provided embodiments, the FD vector genome module is capsidized by a standardized cotransfection protocol of the vector genome module and packaging expression plasmid. Current technology provides 2,000 packages of viral genome per cell, and technological advancements may lead to improved production efficiency in the future. The packaged vector is then purified by column chromatography.

[0072] Treatment of retinal diseases In one embodiment, a viral vector is provided for the treatment of a retinal disease caused by a gene mutation. The viral vector may conform to any embodiment or combination of embodiments described herein. In a further embodiment, the viral vector is provided as a composition comprising at least one viral vector and at least one packing plasmid. In another embodiment, packaging cells are provided, each containing at least one viral vector and at least one on packing plasmid. In yet another embodiment, a capsid is provided, each containing at least one viral vector, preferably at least one packing plasmid. The viral vector, packing plasmid, and packaging cells may conform to any embodiment or combination of embodiments described herein. In a further embodiment, the packaging cells containing the viral vector and packing plasmid are capsidized into a capsid.

[0073] In one embodiment, a viral vector, composition, packaging cells and / or capsid are injected into the eye of a target, preferably a mammalian target, more preferably a human target. The viral vector is injected into the eye either subretinally or intravitreously to infect a photoreceptor and cause the photoreceptor to express a protein encoded by the transgene construct. In a further embodiment, a packing plasmid is provided for the viral vector. The viral vector and packing plasma may be transfected with packaging cells. The packaging cells may be capsidized with a capsid. The viral vector, packing plasmid, packaging cells and capsid may be independent according to any embodiment or combination of embodiments provided herein.

[0074] Examples and Case Studies The methods and compositions discussed herein enable the modification of mutations that cause retinal diseases and prevent photoreceptor death and blindness. The methods and compositions provided herein prevent, for example, the progression of Usher syndrome type IIA, Usher syndrome type 2C, retinitis pigmentosa, retinitis pigmentosa, Wagner syndrome, or age-related macular degeneration due to hemisentin mutations.

[0075] The viral vectors for gene therapy described herein are specifically engineered when it is necessary to deliver large transgenes. For example, a viral vector may deliver full-length human coagulation factor VIII cDNA (7kb) driven from a CMV pre-initial promoter / enhancer along with an immunosuppressive molecule for a total expression cassette of approximately 12kb. Fibroblasts transduced with such a vector produced factor VIII as both focus and shadow.

[0076] In one embodiment, a broad-spectrum CMV pre-early promoter / enhancer was used as a standard to measure the activity of a photoreceptor-specific human rhodopsin kinase (RK) promoter, for which a very potent variant has been identified that drives gene expression in both cone and rod cells. Since the universal CMV pre-early promoter / enhancer promoter functions in microglial cells and therefore can induce harmful inflammatory responses (Figure 3), it is used only as an indicator of RK activity. Enhanced green fluorescent protein (GFP) is used to test the promoter activity of different vectors.

[0077] Enhanced green fluorescent protein (GFP) genes were used as test genes in in vivo electroporation studies. Approximately 0.5 pi of a solution of each purified plasmid DNA in PBS (5 mg / ml) was injected into the subretinal space of neonatal mice using a Hamilton syringe equipped with a 32-gauge blunt-end needle. Electroporation was performed by applying five square pulses using a pulse generator. On day 21 post-electroporation, the mice were sacrificed and retinal sections were analyzed for vector uptake. As illustrated in Figure 3, both promoters drove GFP expression to similar levels in photoreceptor cells. The broadly reactive CMV promoter also induced GFP transgene expression in microglia. Therefore, vectors using the broadly reactive CMV promoter may be more likely to elicit an inflammatory response.

[0078] In this disclosure, useful promoters are incorporated to study FGP expression. Once the activity of the RK promoter is confirmed, its ability to drive USH2A expression in the photoreceptor is investigated. The design of these vector genomic molecules follows the design of a previously constructed CMV-GFP vector. The expression cassette will contain a specific RK promoter, a GFP sequence, and a human growth hormone polyadenylation site (HT-pA). The whole is synthesized with the necessary cloning elements so that it can be easily transferred to a viral vector, as shown in Figure 4. The integrity of the assembled construct is confirmed by sequencing.

[0079] For further experimental / case study purposes, the vector genome module will be packaged into human Ad5 (or other human serotypes) using a standard cotransfection protocol.

[0080] To investigate expression cassettes in transfection studies, three different test cell groups—HEK293, patient iPSCs, and Y79 retinoblastoma cells—will be used in in vitro studies. All experiments will be performed at least three times. HEK293 cells and undifferentiated patient iPSC cells, which do not continuously produce asherin, are well-suited for studying asherin protein production. To assess RK promoter activity, these cells will be cotransfected with plasmids encoding Crx and Sp1 transcription factors. Transgene transcription will be measured by RT-qPCR using various sets of USH2A probes, and protein production will be measured by immunostaining of fixed cells with polyclonal anti-asherin antibody. GFP expression levels will be measured by FACS. Y79 retinoblastoma cells constitutively express USH2A in a splice variant that skips exon 62. Vector-delivered transgene expression can be distinguished from constitutively transcribed USH2A using an exon 62-specific RT-qPCR probe. To investigate RK promoter activity in Y79 cells, providing transcription factors is not necessary. In the second step, a capsidized vector is used to transduce test cells at different infection multiplicities. Previous studies have observed the highest levels of transgene expression after 48 hours. The percentage of transgene expression and the level of transgene expression per cell in tissue culture cells are quantitatively analyzed by immunofluorescence in FACS.

[0081] This study will be conducted using GHFP and USH2A to deliver vector genome module plasmids and packaged vectors. While USH2 and autosomal recessive RP are autosomal recessive, they are not X-linked genetic disorders, and sex-related phenotypic differences have not been reported in humans or mice. Nevertheless, C57BI / 6 wild-type mice will be used for both sexes in GFP testing and Usha2a knockout (for USH2A testing). Four mice will be included per group for molecular, biochemical, and imaging studies, but eight mice per group will be included for electroretinogram recordings. A vector genome module lacking the transgene will be used as a negative control.

[0082] One-month-old mice are injected with a capsidized vector using a subretinal injection procedure. 2 × 10 9 A vector suspension with a concentration of IU / ml in a volume of 1 μl or less is delivered.

[0083] Mice are sacrificed 21 days after electroporation, when the photoreceptors have just completed their differentiation, or 7 days after transduction, when transgene expression has reached its maximum level and injection-related wounds have completely healed. Whole-mount retinas and retinal sections are analyzed for vector uptake. If effective transgene expression is observed, additional mice are added to investigate therapeutic effects, as well as inflammatory and toxic responses, at 3 and 6 months.

[0084] In addition to in situ PCR using ITR-specific probes to localize cells that have taken up vector DNA after electroporation and transduction, the GFP test gene is directly visualized. Usherin expression in treated knockout mice is detected by immunofluorescence and confocal microscopy. The number of transgene-expressing cells and the transgene expression intensity per cell are scored using the ImageJ program. Furthermore, GFP and USH2A expression in the retina is evaluated by semi-quantitative immunoblotting analysis. In parallel, retinal sections are examined immunohistochemistry for evidence of inflammatory and immune responses. Since asherin is essential for the integrity of the Ush2 protein complex at the photoreceptor, the inventors investigate the rescue of the USH2 protein complex in treated Ush2a knockout retinas. Mean values ​​are obtained from four sections per animal. Data from four mice per experimental and control group (eyes treated with simulated injection or control vector) are compared using Student's t-test and / or one-way ANOVA. Since Ush2a knockout mice do not show retinal degermation until 1 year of age, the inventors will assess retinal health in mice using electroretinography (ERG) and optical coherence tomography (OCT) 3 and 6 months after viral vector transduction, but will not assess rescue of retinal degeneration. Eight mice will be tested per experimental and control group based on the standard deviation of previous ERG and OCT data. Student's t-test and two-way ANOVA will be performed to analyze the ERG and OCT data.

[0085] While several embodiments and related methods have been described in detail in this specification, it should be clear that modifications and variations are possible, all of which fall within the original spirit and scope of the invention. Furthermore, since numerous modifications and changes are readily conceivable to those skilled in the art, it is undesirable to limit the invention to the exact configurations and operations illustrated and described, and therefore all suitable modifications and equivalents may be made within the scope of this disclosure. Sequence List SEQUENCE LISTING <110> Greffex, Inc. <120> GENE THERAPY FOR RETINAL DIEASES <130> 543346.22 <150> US 63 / 149,122 <151> 2021-02-12 <160> 3 <170> PatentIn version 3.5 <210> 1 <211> 15612 <212> DNA <213> Homo sapiens <400> 1 atgaattgtc ctgtactgag cctcggtagc ggctttttgt tccaagtcat agaaatgctc 60 atctttgcgt acttcgcgtc aatatctctt acagagtctc ggggtctctt cccaaggctt 120 gaaaatgtag gagcgtttaa aaaggtttcc atcgtaccaa cccaggctgt atgtggcctt 180 cctgaccgca gtactttttg ccactcttcc gccgctgctg agtcaattca gttttgcact 240 cagcggttct gcattcaaga ttgcccctat cggtcttctc acccaacata tacggcactg 300 ttttctgcgg ggcttagttc ctgtattacc cctgacaaga atgatcttca ccctaacgcg 360 cactcaaata gtgcatcatt tatcttcgga aaccacaaat cctgtttttc ttctcctccg 420 tcccccaaac tgatggcctc ctttacgttg gcggtttggc ttaaaccgga acaacaagga 480 gtaatgtgcg tgattgagaa gacggttgat ggacaaatcg tctttaagct gacgatatca 540 gagaaagaga cgatgtttta ttaccggacg gtgaatgggc tgcaaccccc gattaaagta 600 atgactctcg gtcggatcct cgtgaagaaa tggattcacc tgagtgtaca agtccatcaa 660 actaagattt cattttttat caacggagtc gagaaggacc ataccccatt taacgctagg 720 acattgagtg ggagcattac agatttcgcc tccggaacgg tacagatcgg ccaatccttg 780 aatggattgg aacaattcgt cggaagaatg caggacttcc ggctgtacca ggtagcactg 840 acaaatcgag agatccttga agtctttagt ggggacctgt tgcggctgca tgcccaatca 900 cactgccggt gtccaggttc tcacccgagg gtacacccgc tggctcagcg gtactgtata 960 ccaaatgatg ccggggacac agcagataac agagtcagtc gacttaaccc ggaggcacac 1020 cctctctcat tcgtaaacga caacgacgtg gggacttctt gggtgtctaa cgtcttcaca 1080 aatataacac aactgaatca aggcgtcacc atctccgttg acctcgagaa cgggcagtat 1140 caggtgttct atattattat acaatttttc agtccacagc ccacggagat acgaatccag 1200 cggaagaaag agaattccct ggactgggaa gactggcagt acttcgctcg caattgcggg 1260 gcgtttggca tgaagaataa cggagacctg gagaagcctg attccgtgaa ctgccttcag 1320 ctctcaaact tcacccccta ctcaagaggg aatgtgacat tctcaattct tactccgggc 1380 cccaattatc ggcccgggta caacaacttt tataacacgc cttccctgca agaatttgta 1440 aaggcaaccc aaatcagatt tcacttccat ggacaatact ataccactga aacagccgtt 1500 aatctgaggc accgctacta cgccgtggat gaaatcacga tcagtggacg gtgtcagtgc 1560 catggccacg ctgataactg cgacacaact agccaacat acagatgcct ctgcagccaa 1620 gaaagtttca ctgagggcct tcactgtgat agatgcctgc ccctttataa cgataagcct 1680 ttcaggcaag gagaccaagt ttatgcgttt aactgcaaac catgccaatg caacagccat 1740 tccaagtcct gtcattacaa catttccgtc gacccttttc catttgagca ttttagggga 1800 gggggcggag tatgtgatga ctgcgaacat aacactactg gtagaaactg tgagttgtgc 1860 aaagattact tctttcgcca agtgggtgcg gatccatccg cgatcgacgt atgtaagccc 1920 tgtgactgtg acaccgtcgg gacgcggaat ggatctatcc tttgtgatca aataggtggg 1980 caatgcaatt gtaaacgaca cgtatcaggt aggcagtgca accagtgtca aaatggattc 2040 tacaatctcc aggaactcga ccctgacgga tgtagtccat gcaactgtaa cacctctggc 2100 accgtagacg gtgacataac atgtcatcag aatagtggac agtgtaaatg taaggctaat 2160 gttatcgggc tcaggtgcga tcattgcaat ttcggtttta agttcctccg ctcatttaat 2220 gatgtagggt gtgaaccgtg tcagtgtaac cttcacggta gtgttaacaa gttttgtaac 2280 ccccattctg gtcagtgcga gtgcaagaag gaggccaagg ggttgcaatg cgatacttgc 2340 cgcgaaaatt tttatgggct tgatgtaacc aactgtaagg cgtgcgactg cgatactgca 2400 ggatctctgc caggaactgt gtgcaacgcg aaaactggac agtgcatttg taaaccgaat 2460 gtggaaggac gacagtgcaa caagtgcctg gagggaaatt tttatctccg gcagaacaac 2520 tcatttctct gtctcccatg taattgcgat aaaactggca ccattaatgg cagcttgctt 2580 tgtaataaaa gcactggaca atgcccctgc aagctgggag ttacggggct gcggtgtaac 2640 caatgcgaac cacaccggta caatctcacc atcgacaatt tccaacattg tcagatgtgt 2700 gaatgcgatt ccctcgggac actccctggc acgatatgtg accccattag tggccaatgc 2760 ctgtgtgtac ccaaccgaca gggtcgacga tgtaatcagt gtcagccagg attttacata 2820 agcccgggaa acgccacagg ttgcctgccc tgctcatgtc acacgacggg agcagtcaat 2880 cacatatgca atagcctcac tgggcaatgc gtgtgccaag acgcaagcat tgctggtcag 2940 cgctgtgatc agtgcaaaga tcactacttc gggtttgacc cacagacagg tcggtgccag 3000 ccgtgcaact gccaccttag cggtgcattg aacgaaacat gtcatttggt aacgggtcag 3060 tgcttttgca aacaatttgt aacgggttcc aagtgtgacg catgtgttcc ttctgccagc 3120 catcttgacg ttaacaatct ccttggttgt tctaagactc catttcagca accaccgcca 3180 agaggccagg tccaaagctc atcagccatc aacctctcat ggtcacctcc cgattcaccg 3240 aacgcgcact ggttgaccta ctcattgctc cgcgatggat tcgagattta taccaccgag 3300 gaccaatatc cgtattccat tcagtacttt ctggacactg acttgctgcc ctatactaaa 3360 tatagttatt atatcgagac gaccaatgtg cacggttcta cgcgatctgt ggccgtcaca 3420 tataagacta agcctggagt gcctgaaggt aatctgactt tgagctacat aatcccaatt 3480 ggatccgaca gcgtgaccct tacgtggacc acgctttcca accaatcagg accaattgaa 3540 aaatacatct tgtcttgcgc tccgttggcg gggggtcaac cctgcgtctc atatgagggc 3600 catgagacaa gtgcgactat atggaatctc gttccctttg caaagtatga cttttctgta 3660 caagcctgca catctggcgg atgcctgcat agtcttccaa tcacagttac cacagcacaa 3720 gcaccccccc agaggttgag cccgccaaag atgcaaaaga tttcttctac ggaactccat 3780 gtggagtgga gtcctccggc agaactcaat ggtattatca ttaggtatga actgtacatg 3840 cgacggctta gatctaccaa agagactacg agtgaggagt cacgcgtgtt tcagtcatct 3900 ggctggttgt caccccattc attcgtggag tctgcaaatg aaaacgcgtt gaagcctcct 3960 caaacaatga ctacgataac tggacttgag ccatatacta aatatgagtt tagagtactg 4020 gctgtcaaca tggccggctc cgtctcaagt gcatgggttt ccgagcgcac aggcgaatcc 4080 gcgcctgtct tcatgatacc ccccagtgtt tttcctctgt ctagttactc cctcaacata 4140 tcttgggaaa agcctgcgga caacgtgacc agaggtaagg tcgtgggcta cgatatcaac 4200 atgctgtcag agcagtcccc acagcagagc atcccgatgg ccttttcaca actcttgcat 4260 acggccaaat cacaagaact gtcatacacg gtcgagggac ttaaacctta tcggatctac 4320 gagtttacga tcacgctgtg caactcagtc gggtgtgtca cttctgcgag cggtgcaggc 4380 cagacgcttg cagcagcgcc agcgcagctc aggccccctt tggtcaaggg aataaattct 4440 actacgatcc atctccggtg gtttcctccg gaggaactga atggccctag tcccatttat 4500 cagctcgaaa ggagagagtc ttcactgcct gcactcatga ccactatgat gaaaggaatc 4560 cgctttattg gaaacgggta ctgcaaattc cccagctcca cgcaccctgt gaatacggat 4620 tttacaggga tcaaggcctc ttttagaacg aaagtgccgg agggcctcat cgttttcgct 4680 gctagccccg gcaaccaaga ggagtatttc gcccttcagt tgaagaaggg aaggctgtac 4740 ttcctgttcg atcctcaggg aagtccagtt gaggttacga caacaaatga ccatggtaaa 4800 caatacagcg atggtaaatg gcacgagata atcgcgatta gacaccaggc gtttgggcag 4860 atcacactcg acggcattta cactggatcc agtgcaattc tcaacggtag cacagttatt 4920 ggcgacaata cgggggtctt cttgggcggt ctgccacgat catacactat acttcgcaag 4980 gatcctgaaa taatacagaa gggttttgtc ggatgcctta aggacgtgca ttttatgaaa 5040 aactacaatc cttccgctat atgggagcca ctggactggc agtcttcaga ggaacaaatc 5100 aacgtgtata attcttggga agggtgtccc gcgagcctta acgaaggggc acaatttttg 5160 ggggcaggtt tcctcgagtt gcacccatac atgtttcatg gggggatgaa tttcgaaatt 5220 tctttcaagt ttcggacgga ccaattgaac ggcttgcttc tgtttgtcta caacaaggac 5280 ggccctgatt tcctggccat ggagctcaaa agtgggatac tgaccttccg ccttaatact 5340 agtctggcgt ttacccaagt agatctgctg cttggccttt cttactgcaa cgggaaatgg 5400 aacaaagtga tcatcaaaaa agaaggctcc tttatttctg catctgtgaa tggcctgatg 5460 aagcatgcct ccgaaagtgg tgaccaaccc cttgtagtca acagcccggt atatgttggg 5520 gggatcccac aagaactctt gaacagttac cagcacttgt gcctcgaaca aggtttcggc 5580 ggatgcatga aggacgtgaa gtttacccgg ggggccgtag tcaaccttgc cagcgtatca 5640 tccggggccg tgcgggtcaa tttggacgga tgcctgtcta cagattccgc agttaattgt 5700 cgagggaatg attcaatact tgtatatcaa gggaaagaac aatcagtata cgaagggggc 5760 ctccaacctt ttacagagta tttgtataga gtcattgcaa gtcatgaagg gggatccgtt 5820 tactcagact ggtcacgggg ccgcacaacc ggcgccgcgc cacagtccgt ccccacacct 5880 tccagggtgc ggtcactgaa cggttacagc atcgaagtga cgtgggatga accagtagta 5940 aggggtgtga ttgaaaagta tatattgaaa gcttacagtg aggattccac acgcccccca 6000 aggatgccat ccgcgtctgc ggaattcgtt aatacttcca accttacagg cattttgaca 6060 ggactgctcc cttttaaaaa ttacgctgtc acacttactg cttgtaccct cgcggggtgc 6120 acagaaagta gtcatgcgct gaatatctcc accccgcagg aagcccctca agaggttcag 6180 ccgccggtcg caaagagtct tccgagtagt ctgctccttt cctggaaccc ccctaaaaaa 6240 gcaaacggta tcatcacaca atactgcctc tacatggacg ggcggttgat ttactctggt 6300 agtgaggaa actacatagt aaccgatctt gctgtcttta ctccacatca attcctcctt tctgcgtgta ctcatgtagg ctgcacgaac tcaagttggg tacttctcta tactgcccag ttgccacctg agcacgttga ctctcccgta ttgactgtat tggacagcag gaccattcac 6480. ttgccacctg attcaatgga agcagcctcg gaaataagc ggcatccttg aaaggtatgt tctctatatg tctaatcaca cccatgactt cacaatttgg agcgtcattt father agactgttt caggatcata tgttgcaata tgtactccct gggaataaat acttgataaa gctgggtgcg tgcaccggag gtggttgtac ggtgagcgag gcatctgagg cgcttacgga cgaggacatc 6720 ccagagggtg tacccgcccc gaaggcccac tcttacagcc ctgacagctt caatgtgagc tggactgaac ccgaataccc aaatggcgta ataaccagtt atggactgta cctcgatggt 6900. 6900. 6900. 6900. 6900. 6900. 6900. 6900. 6900 ttgcacagtt tccgggtcca ggcttgcacc gcgaaaggtt gcgccctggg tccgctcgtg 6960 gaaaatcgaa cactcgaggc cccacctgag gggaccgtga atgtgtttgt taagacacag 7020 ggaagtcgaa aagcgcatgt ccgatgggag gccccattta gacctaatgg cttgcttacg 7080 cactctgtgt tgttcacggg gatcttctac gtcgaccctg taggtaataa ttacacattg 7140 ctgaacgtca ccaaggtgat gtactccgga gaggaaacta acctttgggt actgattgac 7200 ggcctggtac cgtttacgaa ctatacagtt caagtgaaca tcagtaacag ccaagggtcc 7260 ctcattacgg atccgattac tatcgcgatg ccgccgggtg ctcctgatgg agttctcccc 7320 ccccgccttt catctgctac tcctacttca ttgcaggttg tgtggtcaac ccccgcgaga 7380 aacaacgcgc ctggtagccc cagatatcaa cttcaaatgc ggagcgggga ttccacccac 7440 ggtttcctcg agctcttttc caatccttca gcaagcttgt cctacgaagt ttctgatctc 7500 cagccatata ctgaatacat gttccgactg gtggcgagca atggatttgg gtcagcccac 7560 agttcttgga tcccctttat gaccgctgag gacaagccag ggcccgtggt tccgcccatc 7620 ctgctggatg tcaaaagtcg aatgatgctg gtcacgtggc aacatccgag aaagtccaac 7680 ggcgttataa ctcactacaa catttacctg catggtcggc tttacttgcg caccccagga 7740 aatgtaacga actgtaccgt tatgcacctc cacccttata ccgcctacaa gttccaggtc 7800 gaggcatgta cgtccaaagg atgctcactt agccccgaaa gccagaccgt ttggacgctt 7860 cctggggctc cggaagggat ccctagtccg gagctcttta gcgatacacc caccagtgtg 7920 ataatcagct ggcagccccc tactcatccg aatggtctcg tcgaaaattt cacgatagaa 7980 aggcgagtta aaggtaaaga ggaagtgacc accttggtta ctctgccccg gagccatagt 8040 atgcgcttta tagacaagac ctccgccctt agcccctgga ccaaatatga ataccgggtt 8100 cttatgtcca cgttgcatgg agggacaaat tccagcgctt gggtagaagt cactactcga 8160 ccaagccggc ccgcgggcgt tcaaccacct gttgttacgg tactggagcc tgatgctgtc 8220 caagttacgt ggaaaccccc actgatccag aatggtgaca tactgagtta cgagatacat 8280 atgccagacc ctcacataac attgacgaac gttacctctg cagtgctttc ccagaaagtt 8340 actcacctca ttccgtttac gaactatagt gtcacaatcg tggcctgtag cggtggtaac 8400 ggatatcttg gagggtgcac cgaaagcctc cctacttacg tgacgacgca cccgacagtt 8460 ccgcagaatg taggtcccct gtccgttatt ccgttgtccg aatcctatgt cgtgatatct 8520 tggcagcccc cctctaaacc taatggaccc aatctgaggt acgagctgct tcgacgcaag 8580 atccaacagc cattggcgtc caacccgccg gaagacctga atcggtggca caatatttat 8640 agtggcacgc aatggttgta cgaagacaag ggcttgtcca ggttcacgac ctacgagtat 8700 atgctttcg ttcataattc cgtcggattt accccgtctc gcgaagtgac cgtcaccacc 8760 ttggctggct tgccagaacg cggggcgaac ctcactgctt cagtcctgaa ccacaccgcg 8820 atagatgttc ggtgggctaa gccaactgtg caagatctcc agggcgaggt tgaatactat 8880 accttgtttt ggtcctctgc aacgtccaat gatagtctca agatattgcc ggatgttaac 8940 agtcacgtaa ttggacatct caaacctaac acagaatact ggatatttat ttccgtgttt 9000 aatggagtgc actccataaa cagtgctgga ctgcatgcga cgacttgtga tggtgagcca 9060 caggggatgc ttccccctga agtagttatc attaactcaa ccgcagtcag agtaatttgg 9120 acttctccaa gcaacccaaa tggagtagta accgagtact ctatttatgt gaacaataaa 9180 ctgtacaaga ccggcatgaa cgtacccggc tcatttattt tgagagacct gtccccgttt 9240 acgatttacg atatacaggt ggaggtctgt acaatatatg catgcgtcaa gtccaacgga 9300 acacaaatca ctaccgtcga agatacaccc tcagatattc ctacgccaac cattaggggt 9360 attacgtcta ggagtcttca aatagactgg gtgtcacccc ggaagcccaa cgggattatt 9420 ctcggatatg acctgttgtg gaagacttgg tatccgtgcg ccaaaacaca gaaattggtc 9480 caagatcagt cagatgaatt gtgtaaggcg gtgcggtgtc aaaaacccga gagtatctgc 9540 ggacacatct gttatagctc agaggcaaaa gtatgctgta acggtgttct gtacaatccc 9600 aaaccaggcc atcgctgttg tgaggagaaa tacatcccgt tcgtgctgaa ctcaacgggg 9660 gtctgttgcg gtggtcgcat tcaagaagcc cagcccaacc accagtgttg ctctggttac 9720 tacgcccgca tcctcccagg ggaggtatgt tgccctgacg aacaacacaa cagggtaagc 9780 gtaggcattg gcgactcctg ttgtggccgg atgccgtata gtactagcgg taaccaaatt 9840 tgctgcgcgg gaagacttca tgatggacac ggacagaaat gttgcggccg ccaaattgta 9900 tcaatgacc tcgaatgctg cggcggcgaa gaggcgtag tttacaatg gctgccgggg 9960 atgttctgtt gcggacaaga ctacgtaaat atgagcgata ctatttgctg ttctgcaagc 10020 tctggggagt caaggcccca cataaaaaag atgacccag taccagttaa gtgctgcgaa 10080 actgagctca ttcccaagtc acagaatgt tgcaatgggg taggatataa tccgttgaag 10140 tacgtctgct cagatagat ttcaccggt atgatgatga agagaccaa ggaatgtaga 10200 attctgtgcc ccgccagtat ggaagcaacg gaacactgtg gtcggtgcga ctttaatttc 10260 acgagtcaca tctgcactgt catccgcggc tcccatact caacggcaa ggcctctata 10320 gaggaaatgt gcagttccgc ggaggaaacc atacatacgg gaagcgtca tacgtattct 10380 tatactgacg tcaacctgaa gccctacatg acgtacgagt accgaatcag cgcgtggaat 10440 tcatacggcc gcggattgag taagcagtc agggctagaa cgaaggagga tgttccgcaa 10500 ggcgtaagcc cacccacttg gacgaaaata vakaaccttg aggatacaat tgtccttaat 10560 tggaggaagc cgatccagag taacggtcct attatttact acatctctct gcggaatggg 10620 height height height height height height height 10680 caagaataca gctaccagct gaagcatgc acagtggcgg gatgtgccac ctcaagcaaa 10740 gtagtagccg cgactaccca aggcttccg gagagtatac ttcccccag tatcaccgcg 10800 ctgagcgccg tagcattgca cctcagctgg tcagttccgg aaaaagtaa tggcgtcatt 10860 aaggaatatc aaatacggca ggtgggaaa ggccttattc acacgatac gaccgaccga 10920 cggcacata cagtgactgg tctcaccg tatacgaatt attcattcac actgaccgcc 10980 tgcacaagtg ctggatgtac cttagtgag cccttccttg ggcaactct gcaagcggcc 11040 ccagaaggcg ttgggtcac cccacgccat attattaca actctaccac ggttgagctc 11100 tactggtccc ttccggagaa acccaacggc ctggtatccc aatatcaact ctccagaaat 11160 ggtaacctgc tctttttggg cggtagtgag gaacaaaatt tcacggataa gaatctcgag 11220 cctaactcaa ggtatactta taaacttgag gtgaaaaccg gaggcggcag ttctgccagc 11280 gacgattata tagtacaaac acctatgtcc acaccggagg aaatctatcc accgtataac 11340 ataacagtga taggtcctta ctcaatattt gtagcgtgga tcccgccagg gatattgata 11400 cccgagattc ctgtagagta taacgtgttg ctcaatgatg gctccgtgac tccgcttgca 11460 ttctcagttg gacatcatca gagtacgctc cttgaaaacc ttacgccatt tactcaatat 11520 gaaatccgga tccaggcgtg ccagaacggt agctgtggtg tgtcctctcg gatgtttgtc 11580 aaaactcctg aagcggcacc aatggacctc aacgtcccg tgcttaaggc tttgggctca 11640 gcttgcatag aaatcaagtg gatgccacct gagaaaccaa atggtatcat aattaactat 11700 ttcatttacc ggcgccccgc agggatcgag gaggaaagcg tgctttttgt atggtctgaa 11760 ggggcactgg aattcatgga cgaaggagat accttgaggc catttaccct ttacgagtac 11820 cgcgtgagag cttgcaactc taaggggagc gtcgaaagct tgtggtctct cacgcagacc 11880 cttgaggctc ctccccaaga tttcccagcg ccgtgggccc aggcgacatc tgcacacagc 11940 gtgttgctta actggactaa acctgaaagc ccgaacggca tcatctcaca ttatagggtt 12000 gtttatcaag aacggccgga cgatccgact ttcaattctc ccacggtgca cgcctttaca 12060 gtaaagggta ctagccatca ggcccacctc tacggtctcg aaccattcac gacgtatcgc 12120 atcggcgttg tagcggccaa tcatgctggt gagatactta gtccgtggac gctgatacaa 12180 actctggagt ctagcccctc cggtttgcgc aacttcattg tggagcaaaa agagaatggt 12240 cgagctcttc tccttcaatg gtccgaaccc atgcgcacga atggcgtgat aaagacttat 12300 aatatattct cagatggctt ccttgaatac tcaggtttga accgacagtt cctttttagg 12360 aggcttgacc cattcactct ctacacgctg acgctcgaag cgtgtaccag ggccggttgt 12420 gcacattctg caccacagcc cctttggacg gatgaggctc cccctgattc tcaactcgcc 12480 ccaacggtgc atagcgtcaa gtcaacctca gtagagctta gttggtctga gccagtgaac 12540 cctaatggaa aaattataag atacgaagtg atacgccgat gtttcgaagg taaagcttgg 12600 ggcaatcaaa caatccaggc cgatgagaaa attgtattta cagaatataa caccgagagg 12660 aacacgttca tgtataacga taccgggctg caaccttgga ctcaatgtga atataaaatt 12720 tatacttgga attcagcggg ccatacctgc tcctcatgga acgtggtgcg gactctccaa 12780 gcaccgccgg agggcctttc accgcctgta atctcttacg tttccatgaa tcctcagaag 12840 ctgcttatct cctggatacc acccgaacaa agcaacggaa tcatacagtc ttatcggttg 12900 cagaggaacg agatgttgta tcctttttcc ttcgaccccg tcacttttaa ttatacagac 12960 gaggaactgt tgccatttag cacctattcc tacgcccttc aagcgtgtac gagcggtggc 13020 tgctccacat ccaaaccgac cagcattacc acacttgaag ccgctccaag cgaggtctct 13080 ccacctgacc tctgggccgt ttctgctacg caaatgaatg tgtgctggtc cccgcccacg 13140 gttcagaatg ggaaaatcac taaatacctc gtaaggtatg acaataaaga aagtctggct 13200 ggacaagggc tctgccttct ggtcagtcac ctgcagcctt atagtcaata taattttagt 13260 ctcgtagcgt gtaccaacgg cggctgtaca gcctctgttt caaaaagcgc ttggacaatg 13320 gaggccttgc ccgagaacat ggactctccg accctgcagg tgaccggatc agaatccata 13380 gaaatcacgt ggaagccacc acgcaatccg aacggacaga ttcgaagcta cgaattgcgc 13440 cgggatggga ctatcgtgta cacagggctg gagactcgat acagggactt caccctgact 13500 ccgggagtgg aatattcata tactgtcacg gccagtaact cacagggtgg aatcctgagt 13560 cctctggtga aagatcgcac tagtcccagt gcgccctctg gcatggagcc accgaaattg 13620 caagcaagag ggccccagga gatcctggtc aattgggatc ctcccgttag gactaacggt 13680 gacattatta actacacctt gtttatcagg gagttgttcg aaagggaaac caagattatc 13740 cacattaata ctacccataa cagcttcggc atgcagtcat atattgtaaa ccaattgaaa 13800 ccgttccacc gatacgagat taggatccag gcatgcacga ccttggggtg tgcatccagc 13860 gactggacgt ttattcaaac gccagaaatt gcgcctttga tgcagccgcc cccgcacctg 13920 gaggtacaga tggctccggg cgggttccaa cccacagtct ctcttctctg gaccggacct 13980 ctgcagccta atggaaaggt cctgtattat gaactttaca gacgacagat cgccactcaa 14040 ccccgcaaat ccaatcctgt gcttatctac aacgggtcta gcacgagctt cattgatagc 14100 gaattgctgc cttttaccga gtacgagtac caagtgtggg cagtgaactc agcgggggaaa 14160 gcgcttcct cttggacgtg gtgccggacg ggacctgcgc cacctgaggg gctgcgggca 14220 cccacgttcc atgttatcag ctcaacgcaa gctgtcgtaa atatctccgc tccaggtaaa 14280 ccgaacggta ttgtatccct ctataggttg ttctcttcct cagcacacgg cgctgaaaca 14340 gtactgagcg aaagcaatggc aacccagcag accctgcacg ggttgcaggc ctttacgaac 14400 tatagcattg gagtcgaagc gtgcacctgc ttcaactgtt gtagcaaagg cccccaccgcg 14460 gaactccgga cccatcctgc tccaccctct ggcctgtcct ccccacaaat agggaccttg 14520 gcctctcgca cagcgagttt ccgctggagt ccgccgatgt ttccaaatgg ggtcatacat 14580 tcatacgagc ttcagttcca tgtcgcgtgc ccccctgact ctgcgctgcc ttgcacccca 14640 agccaaatcg agactagga cacagggctg ggccagaaag ccagtcttgg gggtctgcaa 14700 ccctatacga cgtataagct tagggtagtc gcccacaacg aggtaggtag tacggccagt 14760 gaatggataa gtttcacgac acaaaaagag ttgccacagt accgggcccc gttctccgtg 14820 gacagcaacc tcagtgttgt atgtgtcaat tggtctgata cattccttct gaatgggcaa 14880 cttaaggaat atgttttgac agacgggggg agacgcgttt actctgggct tgacacgact 14940 ctgtatattc cacggaccgc ggataagact ttttttttc aggtgatttg cactactgac 15000 gaggggtctg ttaaaacccc gttgatacag tacgatacct caacgggatt gggtctcgtt 15060 cttacaaccc cggggaagaa aaagggcagc aggagcaaat ctacagagtt ctatagtgag 15120 ctgtggttta ttgtcctgat ggccatgttg ggtctcattc ttttggcgat atttctgtca 15180 ctcatattgc aaagaaagat acataaggag cttatatca gagagcgccc tcctttggtt 15240 cctttgcaaa agagaatgtc accgcttaac gtttatccgc ctggtgagaa tcacatggga 15300 ctggctgata ccaagatccc taggtctggt acacctgtgt ctatccgcag taaccgctcc 15360 gcttgtgtac tgcgaatacc gtcacagaat cagacaagtc tgacatacag ccaaggcagc 15420 ctccaccgga gtgtatctca actcatggac attcaggata agaaggtcct catggacaac 15480 agcttgtggg aagcgatcat gggtcacaac tctggcctct atgtagacga ggaagacctc 15540 atgaatgcaa taaaggattt ttcaagcgtg actaaagaac ggacgacttt cacggacacg 15600 cacttgtaat aa 15612 <210> 2 <211> 35289 <212> DNA <213> Homo sapiens <400> 2 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat 60 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc 120 ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg 180 cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc 240 cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct 300 gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct 360 tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga 420 tgacgaccat cagggacagc ttcaaggatc gctcgcggct cttaccagcc taacttcgat 480 cactggaccg ctgatcgtca cggcgattta tgccgcctcg gcgagcacat ggaacgggtt 540 ggcatggatt gtaggcgccg ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc 600 atggagccgg gccacctcga cctgaatgga agccggcggc acctcgctaa cggattcacc 660 actccaagaa ttggagccaa tcaattcttg cggagaactg tgaatgcgca aaccaaccct 720 tggcagaaca tatccatcgc gtccgccatc tccagcagcc gcacgcggcg catctcgggc 780 agcgttgggt cctggccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta 840 ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga 900 agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc 960 cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat gttccggatc 1020 tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc 1080 tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc cagttgttta 1140 ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat cgtgagcatc 1200 ctctctcgtt tcatcggtat cattaccccc atgaacagaa atccccctta cacggaggca 1260 tcagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag aagccagaca 1320 ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga catctgtgaa 1380 tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac 1440 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat 1500 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca 1560 gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag 1620 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 1680 gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 1740 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 1800 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 1860 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 1920 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 1980 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 2040 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 2100 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 2160 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 2220 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 2280 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct 2340 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 2400 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 2460 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 2520 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 2580 taaattaaaa atgaagtttt aaatcaatct aaagttata tgagtaaact tggtctgaca 2640 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 2700 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 2760 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 2820 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 2880 agtctattaa ttgttgccgg gaagctagag tagtagttc gccagttaat agtttgcgca 2940 acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 3000 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 3060 cggttagctc cttcggtcct ccgatcgttg tcaagtaa gttggccgca gtgttatcac 3120 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 3180 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 3240 gctcttgccc ggcgtcaca cgggataata ccgcgccaca tagcagact ttaaaagtgc 3300 tcatcattgg aaacgttct tcggggcga aactctcaag gatcttaccg ctgttgagat 3360 ccagttcgat gtaacccact cgtgcaccca actgatctc agcatcttt actttcacca 3420 gcgtttctgg gtgagcaaaa acaggaggc aaatgccgc aaaaaaggga ataagggcga 3480 cacggaaatg ttgaatactc atactctcc ttttcaata ttattgaagc atttatcagg 3540 gttattgtct catgagcgga tacatatttg aatgtatta gaaaaataa caatagggg 3600 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga 3660 cattaaccta taaaaatagg cgtatcacga ggcccttcg tctcaagaa ttacggcccc 3720 atatagttta aacatacatc atcaatata taccttattt tggattgaag ccaatatgat 3780 aatgaggggg tggagtttgt gacgtggcgc ggggcgtggg aacggggcgg gtgacgtagg 3840 ttttagggcg gagtaacttg tatgtgttgg gaattgtagt tttcttaaaa tgggaagtta 3900 cgtaacgtgg gaaaacggaa gtgacgattt gaggaagttg tgggtttttt ggctttcgtt 3960 tctgggcgta ggttcgcgtg cggttttctg ggtgtttttt gtggacttta accgttacgt 4020 cattttttag tcctatatat actcgctctg cacttggccc ttttttacac tgtgactgat 4080 tgagctggtg ccgtgtcgag tggtgttttt ttaataggtt ttctttttta ctggtaaggc 4140 tgactgttat ggctgccgct atgcccgggg ggaggcctaa tcggccttag cgatcgcgcg 4200 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 4260 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 4320 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 4380 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 4440 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 4500 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat 4560 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 4620 actttccaaa atgtcgtaac aactccgcccc cattgacgca aatgggcggt aggcgtgtac 4680 ggtgggaggt ctatataagc agagctggcg cgccagccgc caccatgaat tgtcctgtac 4740 tgagcctcgg tagcggcttt ttgttccaag tcatagaaat gctcatcttt gcgtacttcg 4800 cgtcaatatc tcttacagag tctcggggtc tcttcccaag gcttgaaaat gtaggagcgt 4860 ttaaaaaggt ttccatcgta ccaacccagg ctgtatgtgg ccttcctgac cgcagtactt 4920 tttgccactc ttccgccgct gctgagtcaa ttcagttttg cactcagcgg ttctgcattc 4980 aagattgccc ctatcggtct tctcacccaa catatacggc actgttttct gcggggctta 5040 gttcctgtat tacccctgac aagaatgatc ttcaccctaa cgcgcactca aatagtgcat 5100 catttatctt cggaaaccac aaatcctgtt tttcttctcc tccgtcccccc aaactgatgg 5160 cctcctttac gttggcggtt tggcttaaac cggaacaaca aggagtaatg tgcgtgattg 5220 agaagacggt tgatggacaa atcgtcttta agctgacgat atcagagaaa gagacgatgt 5280 tttattaccg gacggtgaat gggctgcaac ccccgattaa agtaatgact ctcggtcgga 5340 tcctcgtgaa gaaatggatt cacctgagtg tacaagtcca tcaaactaag atttcatttt 5400 ttatcaacgg agtcgagaag gaccataccc catttaacgc taggacattg agtgggagca 5460 ttacagattt cgcctccgga acggtacaga tcggccaatc cttgaatgga ttggaacaat 5520 tcgtcggaag aatgcaggac ttccggctgt accaggtagc actgacaaat cgagagatcc 5580 ttgaagtctt tagtggggac ctgttgcggc tgcatgccca atcacactgc cggtgtccag 5640 gttctcacc gagggtacac ccgctggctc agcggtactg tataccaaat gatgccgggg 5700 acacagcaga taacagagtc agtcgactta acccggaggc acaccctctc tcattcgtaa 5760 acgacaacga cgtggggact tcttgggtgt ctaacgtctt caacaatata acacaactga 5820 atcaaggcgt caccatctcc gttgacctcg agaacgggca gtatcaggtg ttctatatta 5880 ttatacaatt tttcagtcca cagcccacgg agatacgaat ccagcggaag aaagagaatt 5940 ccctggactg ggaagactgg footcttcg ctcgcaattg cggggcgttt ggcatgaaga 6000 ataacggaga cctggagaag cctgattccg tgaactgcct tcagctctca aacttcaccc 6060 cctactcaag agggaatgtg acattctcaa ttcttactcc gggccccaat tatcggcccg 6120 ggtacaacaa cttttataac acgcttccc tgcaagaatt tgtaaaggca acccaaatca 6180 gatttcactt ccatggacaa tactatacca ctgaaacagc cgttaatctg aggcaccgct 6240 actacgccgt ggatgaaatc acgatcagtg gacggtgtca gtgccatggc cacgctgata 6300 actgcgacac aactagccaa ccatacagat gcctctgcag ccaagaaagt ttcactgagg 6360 gccttcactg tgatagatgc ctgccccttt ataacgataa gcctttcagg caaggagacc 6420 aagtttatgc gtttaactgc aaaccatgcc aatgcaacag ccattccaag tcctgtcatt 6480 acaacatttc cgtcgaccct tttccatttg agcattttag gggagggggc ggagtatgtg 6540 atgactgcga acataacact actggtagaa actgtgagtt gtgcaaagat tacttctttc 6600 gccaagtggg tgcggatcca tccgcgatcg acgtatgtaa gccctgtgac tgtgacaccg 6660 tcgggacgcg gaatggatct atcctttgtg atcaaatagg tgggcaatgc aattgtaaac 6720 gacacgtatc aggtaggcag tgcaaccagt gtcaaaatgg attctacaat ctccaggaac 6780 tcgaccctga cggatgtagt ccatgcaact gtaacacctc tggcaccgta gacggtgaca 6840 taacatgtca tcagaatagt ggacagtgta aatgtaaggc taatgttatc gggctcaggt 6900 gcgatcattg caatttcggt tttaagttcc tccgctcatt taatgatgta gggtgtgaac 6960 cgtgtcagtg taaccttcac ggtagtgtta acaagttttg taacccccat tctggtcagt 7020 gcgagtgcaa gaaggaggcc aaggggttgc aatgcgatac ttgccgcgaa aatttttatg 7080 ggcttgatgt aaccaactgt aaggcgtgcg actgcgatac tgcaggatct ctgccaggaa 7140 ctgtgtgcaa cgcgaaaact ggacagtgca tttgtaaacc gaatgtggaa ggacgacagt 7200 gcaacaagtg cctggaggga aatttttatc tccggcagaa caactcattt ctctgtctcc 7260 catgtaattg cgataaaact ggcaccatta atggcagctt gctttgtaat aaaagcactg 7320 gacaatgccc ctgcaagctg ggagttacgg ggctgcggtg taaccaatgc gaaccacacc 7380 ggtacaatct caccatcgac aatttccaac attgtcagat gtgtgaatgc gattccctcg 7440 ggacactccc tggcacgata tgtgacccca ttagtggcca atgcctgtgt gtacccaacc 7500 gacagggtcg acgatgtaat cagtgtcagc caggatttta cataagcccg ggaaacgcca 7560 caggttgcct gccctgctca tgtcacacga cgggagcagt caatcacata tgcaatagcc 7620 tcactgggca atgcgtgtgc caagacgcaa gcattgctgg tcagcgctgt gatcagtgca 7680 aagatcacta cttcgggttt gacccacaga caggtcggtg ccagccgtgc aactgccacc 7740 ttagcggtgc attgaacgaa acatgtcatt tggtaacggg tcagtgcttt tgcaaacaat 7800 ttgtaacggg ttccaagtgt gacgcatgtg ttccttctgc cagccatctt gacgttaaca 7860 atctccttgg ttgttctaag actccatttc agcaaccacc gccaagaggc caggtccaaa 7920 gctcatcagc catcaacctc tcatggtcac ctcccgattc accgaacgcg cactggttga 7980 cctactcatt gctccgcgat ggattcgaga tttataccac cgaggaccaa tatccgtatt 8040 ccattcagta ctttctggac actgacttgc tgccctatac taaatatagt tattatatcg 8100 agacgaccaa tgtgcacggt tctacgcgat ctgtggccgt cacatataag actaagcctg 8160 gagtgcctga aggtaatctg actttgagct acataatccc aattggatcc gacagcgtga 8220 cccttacgtg gaccacgctt tccaaccaat caggaccaat tgaaaaatac atcttgtctt 8280 gcgctccgtt ggcggggggt caaccctgcg tctcatatga gggccatgag acaagtgcga 8340 ctatatggaa tctcgttccc tttgcaaagt atgacttttc tgtacaagcc tgcacatctg 8400 gcggatgcct gcatagtctt ccaatcacag ttaccacagc acaagcaccc ccccagaggt 8460 tgagcccgcc aaagatgcaa aagatttctt ctacggaact ccatgtggag tggagtcctc 8520 cggcagaact caatggtatt atcattagt atgaactgta catgcgacgg cttagatcta 8580 ccaaagagac tacgagtgag gagtcacgcg tgtttcagtc atctggctgg ttgtcacccc 8640 attcattcgt ggagtctgca aatgaaaacg cgttgaagcc tcctcaaaca atgactacga 8700 taactggact tgagccatat actaaatatg agtttagagt actggctgtc aacatggccg 8760 gctccgtctc aagtgcatgg gtttccgagc gcacaggcga atccgcgcct gtcttcatga 8820 taccccccag tgtttttcct ctgtctagtt actccctcaa catatcttgg gaaaagcctg 8880 cggacaacgt gaccagaggt aaggtcgtgg gctacgatat caacatgctg tcagagcagt 8940 ccccacagca gagcatcccg atggcctttt cacaactctt gcatacggcc aaatcacaag 9000 aactgtcata cacggtcgag ggacttaaac cttatcggat ctacgagttt acgatcacgc 9060 tgtgcaactc agtcgggtgt gtcacttctg cgagcggtgc aggccagacg cttgcagcag 9120 cgccagcgca gctcaggccc cctttggtca agggaataaa ttctactacg atccatctcc 9180 ggtggtttcc tccggagagaa ctgaatggcc ctagtcccat ttatcagctc gaaggagag 9240 agtcttcact gcctgcactc atgaccacta tgatgaaagg aatccgcttt attggaaacg 9300 ggtactgcaa attccccagc tccacgcacc ctgtgaatac ggattttaca gggatcaagg 9360 cctcttttag aacgaaagtg ccggagggcc tcatcgtttt cgctgctagc cccggcaacc 9420 aagaggagta tttcgccctt cagttgaaga agggaaggct gtacttcctg ttcgatcctc 9480 agggaagtcc agttgaggtt acgacaacaa atgaccatgg taaacaatac agcgatggta 9540 aatggcacga gataatcgcg attagacacc aggcgtttgg gcagatcaca ctcgacggca 9600 tttacactgg atccagtgca attctcaacg gtagcacagt tattggcgac aatacgggg 9660 tcttcttggg cggtctgcca cgatcataca ctatacttcg caaggatcct gaataatac 9720 agaagggttt tgtcggatgc cttaagggacg tgcattttat gaaaaactac aatccttccg 9780 ctatatggga gccactggac tggcagtctt cagaggaaca aatcaacgtg tataattctt 9840 gggaagggtg tcccgcgagc cttaacgaag gggcacaatt tttgggggca ggtttcctcg 9900 agttgcaccc atacatgttt catgggggga tgaatttcga aatttctttc aagtttcgga 9960 cggaccaatt gaacggcttg cttctgtttg tctacaacaa ggacggccct gatttcctgg 10020 ccatggagct caaaagtggg atactgacct tccgccttaa tactagtctg gcgtttaccc 10080 aagtagatct gctgcttggc ctttcttact gcaacgggaa atggaacaaa gtgatcatca 10140 aaaaagaagg ctcctttatt tctgcatctg tgaatggcct gatgaagcat gcctccgaaa 10200 gtggtgacca accccttgta gtcaacagcc cggtatatgt tggggggatc ccacaagaac 10260 tcttgaacag ttaccagcac ttgtgcctcg aacaaggttt cggcggatgc atgaaggacg 10320 tgaagtttac ccggggggcc gtagtcaacc ttgccagcgt atcatccggg gccgtgcggg 10380 tcaatttgga cggatgcctg tctacagatt ccgcagttaa ttgtcgaggg aatgattcaa tacttgtata tcaagggaa gacaatcag tatacgaagg gggcctccaa ccttttacag agtattgta taggtcatt gcaagtcatg aagggggatc cgtttactca gactggtcac ggggccgcac aaccggcgcc gcgccacagt ccgtccccac accttccagg gtgcggtcac 10620 tgaacggtta cagcatcgaa gtgacgtggg atgaaccagt agtaaggggt gtgattgaaa agtatatt gaaagcttac agtgaggatt ccacacgccc cccaaggatg ccatccgcgt ctgcggaatt cgttaatact tccaacctta caggcatttt gacaggactg ctccctttta aaaattacgc tgtcacactt actgcttgta ccctcgcggg gtgcacagaa agtagtcatg cgctgaatat ctccaccccg caggaagccc ctcaagaggt tcagccgccg gtcgcaaaga gtcttccgag tagtctgctc ctttcctgga acccccctaa aaaagcaaac ggtatcatca cacaatactg cctctacatg gacgggcggt tgatttactc tggtagtgag gaaaactaca 11040 tagtaaccga tcttgctgtc tttactccac atcaattcct cctttctgcg tgtactcatg 11100 taggctgcac gaactcaagt tgggtacttc tctatactgc ccagttgcca cctgagcacg 11160 ttgactctcc cgtattgact gtattggaca gcaggaccat tcacattcaa tggaagcagc 11220 ctcggaaaat aagcggcatc cttgaaaggt atgttctcta tatgtctaat cacacccatg 11280 acttcacaat ttggagcgtc atttataata gtacagaact gtttcaggat catatgttgc 11340 aatatgtact ccctgggaat aatacttga taaagctggg tgcgtgcacc ggaggtggtt 11400 gtacggtgag cgaggcatct gaggcgctta cggacgagga catcccagag ggtgtacccg 11460 ccccgaaggc ccactcttac agccctgaca gcttcaatgt gagctggact gaacccgaat 11520 acccaaatgg cgtaataacc agttatggac tgtacctcga tggtatattg atacataatt 11580 cttccgaact tagttatagg gcctacggat ttgctccgtg gagcttgcac agtttccggg 11640 tccaggcttg caccgcgaaa ggttgcgccc tgggtccgct cgtggaaaat cgaacactcg 11700 aggccccacc tgaggggacc gtgaatgtgt ttgttaagac acagggaagt cgaaaagcgc 11760 atgtccgatg ggaggcccca tttagaccta atggcttgct tacgcactct gtgttgttca 11820 cggggatctt ctacgtcgac cctgtaggta ataattacac attgctgaac gtcaccaagg 11880 tgatgtactc cggagaggaa actaaccttt gggtactgat tgacggcctg gtaccgttta 11940 cgaactatac agttcaagtg aacatcagta acagccaagg gtccctcatt acggatccga 12000 ttactatcgc gatgccgccg ggtgctcctg atggagttct ccccccccgc ctttcatctg 12060 ctactcctac ttcattgcag gttgtgtggt caacccccgc gagaaacaac gcgcctggta 12120 gccccagata tcaacttcaa atgcggagcg gggattccac ccacggtttc ctcgagctct 12180 tttccaatcc ttcagcaagc ttgtcctacg aagtttctga tctccagcca tatactgaat 12240 acatgttccg actggtggcg agcaatggat ttgggtcagc ccacagttct tggatcccct 12300 ttatgaccgc tgaggacaag ccagggcccg tggttccgcc catcctgctg gatgtcaaaa 12360 gtcgaatgat gctggtcacg tggcaacatc cgagaaagtc caacggcgtt ataactcact 12420 acaacattta cctgcatggt cggctttact tgcgcacccc aggaaatgta agaactgta 12480 ccgttatgca cctccaccct tataccgcct acaagttcca ggtcgaggca tgtacgtcca 12540 areatgctc acttagcccc gaaagccaga ccgtttggac gcttcctggg gctccggaag 12600 ggatccctag tccggagctc tttagcgata cacccaccag tgtgataatc agctggcagc 12660 cccctactca tccgaatggt ctcgtcgaaa atttcacgat agaaaggcga gttaaaggta 12720 aaggaagt gaccaccttg gttactctgc cccggagcca tagtatgcgc tttatagaca 12780 agacctccgc ccttagcccc tggaccaaat atgaataccg ggttcttatg tccacgttgc 12840 atggagggac aaattccagc gcttgggtag aagtcactac tcgaccaagc cggcccgcgg 12900 gcgttcaacc acctgttgtt acggtactgg agcctgatgc tgtccaagtt acgtggaaac 12960 ccccactgat ccagaatggt gacatactga gttacgagat acatatgcca gaccctcaca 13020 taacattgac gaacgttacc tctgcagtgc tttcccagaa agttactcac ctcattccgt 13080 ttacgaacta tagtgtcaca atcgtggcct gtagcggtgg taacggatat cttggagggt 13140 gcaccgaaag cctccctact tacgtgacga cgcacccgac agttccgcag aatgtaggtc 13200 ccctgtccgt tattccgttg tccgaatcct atgtcgtgat atcttggcag cccccctcta 13260 aacctaatgg acccaatctg aggtacgagc tgcttcgacg caagatccaa cagccattgg 13320 cgtccaaccc gccggaagac ctgaatcggt ggcacaatat ttatagtggc acgcaatggt 13380 tgtacgaaga caagggcttg tccaggttca cgacctacga gtatatgctt ttcgttcata 13440 attccgtcgg atttaccccg tctcgcgaag tgaccgtcac caccttggct ggcttgccag 13500 aacgcggggc gaacctcact gcttcagtcc tgaaccacac cgcgatagat gttcggtggg 13560 ctaagccaac tgtgcaagat ctccagggcg aggttgaata ctataccttg ttttggtcct 13620 ctgcaacgtc caatgatagt ctcaagatat tgccggatgt taacagtcac gtaattggac 13680 atctcaaacc taacacagaa tactggatat ttatttccgt gtttaatgga gtgcactcca 13740 taaacagtgc tggactgcat gcgacgactt gtgatggtga gccacagggg atgcttcccc 13800 ctgaagtagt tatcattaac tcaaccgcag tcagagtaat ttggacttct ccaagcaacc 13860 caaatggagt agtaaccgag tactctattt atgtgaacaa taaactgtac aagaccggca 13920 tgaacgtacc cggctcattt attttgagag acctgtcccc gtttacgatt tacgatatac 13980 aggtggaggt ctgtacaata tatgcatgcg tcaagtccaa cggaacacaa atcactaccg 14040 tcgaagatac accctcagat attcctacgc caaccattag gggtattacg tctaggagtc 14100 ttcaaataga ctgggtgtca ccccggaagc ccaacgggat tattctcgga tatgacctgt 14160 tgtggaagac ttggtatccg tgcgccaaaa cacagaaatt ggtccaagat cagtcagatg 14220 aattgtgtaa ggcggtgcgg tgtcaaaaac ccgagagtat ctgcggacac atctgttata 14280 gctcagaggc aaaagtatgc tgtaacggtg ttctgtacaa tcccaaacca ggccatcgct 14340 gttgtgagga gaaatacatc ccgttcgtgc tgaactcaac gggggtctgt tgcggtggtc 14400 gcattcaaga agcccagccc aaccaccagt gttgctctgg ttactacgcc cgcatcctcc 14460 caggggaggt atgttgccct gacgaacaac acaacagggt aagcgtaggc attggcgact 14520 cctgttgtgg ccggatgccg tatagtacta gcggtaacca aatttgctgc gcgggaagac 14580 ttcatgatgg acacggacag aaatgttgcg gccgccaaat tgtatcaaat gacctcgaat 14640 gctgcggcgg cgaagaaggc gtagtttaca ataggctgcc ggggatgttc tgttgcggac 14700 aagactacgt aaatatgagc gatactattt gctgttctgc aagctctggg gagtcaaagg 14760 cccacataaaa aaagaatgac ccagtaccag ttaagtgctg cgaaactgag ctcattccca 14820 agtcacagaa atgttgcaat ggggtaggat ataatccgtt gaagtacgtc tgctcagata 14880 agatttcaac cggtatgatg atgaaagaga ccaaggaatg tagaattctg tgccccgcca 14940 gtatggaagc aacggaacac tgtggtcggt gcgactttaa tttcacgagt cacatctgca 15000 ctgtcatccg cggctcccat aactcaactg gcaaggcctc tatagaggaa atgtgcagtt 15060 ccgcggagga aaccatacat acgggaagcg tcaatacgta ttcttatact gacgtcaacc 15120 tgaagcccta catgacgtac gagtaccgaa tcagcgcgtg gaattcatac ggccgcggat 15180 tgagtaaagc agtcagggct agaacgaagg aggatgttcc gcaaggcgta agcccaccca cttggacgaa aatagacaac cttgaggata caattgtcct taattggagg aagccgatcc aggtacgg tcctattatt tactacattc ttctgcgga tgggatag aggttcgag gcactagtct gagtttttcc gtattcagcc atttcaaga tacagctacc agctgaaagc atgcacagtg gcgggatgtg ccacctcaag caaagtagta gccgcgacta cccaaggcgt tccggagagt atacttccgc ccagtatcac cgcgctgagc gccgtagcat 15540 tgcacctcag ctggtcagtt ccggaaaaa gtaatggcgt cattagga tatcaatac ggcaggtggg aaaaggcctt attcacactg attacgaccga ccgacggcaa catacagtga ctggtctcca accgtatacg aattattcat tcacactgac cgcctgcaca agtgctggat gtacctctag tgagcccttc cttgggcaaa ctctgcaagc ggccccagaa ggcgtttggg 15780. tcaccccacg ccatattatt atcaactcta ccacggttga gctctactgg tcccttccgg agaacccaa cggcctggta tcccaatatc aactctccag aaatggtac ctgctctttt tgggcggtag tgaggaca aatttcacgg ataagaatct cgagcctaac tcaaggtata cttataaact tgaggtgaaa accggaggcg gcagttctgc cagcgacgat tatatagtac 16080. aaacacctat gtccacaccg gaggaaatct atccaccgta taacataaca gtgataggtc cttactcaat atttgtagcg tggatcccgc cagggatatt gatacccgag attcctgtagg agtataacgt gttgctcaat gatggctccg tgactccgct tgcattctca gttggacatc atcagagtac gctccttgaa aaccttacgc catttactca atatgaatc cggatccagg cgtgccagaa cggtagctgt ggtgtgtcct ctcggatgtt tgtcaaaact cctgaagcgg caccaatgga cctcaacagt cccgtgctta aggctttggg ctcagcttgc atagaaatca agtggatgcc acctgagaaa ccaaatggta tcataattaa ctatttcatt taccggcgcc 16440 ccgcagggat cgaggaggaa agcgtgcttt ttgtatggtc tgaaggggca ctggaattca 16500 tggacgaagg agataccttg aggccattta ccctttacga gtaccgcgtg agagcttgca 16560 actctaaggg gagcgtcgaa agcttgtggt ctctcacgca gacccttgag gctcctcccc 16620 aagatttccc agcgccgtgg gcccaggcga catctgcaca cagcgtgttg cttaactgga 16680 ctaaacctga aagcccgaac ggcatcatct cacattatag ggttgtttat caagaacggc 16740 cggacgatcc gactttcaat tctcccacgg tgcacgcctt tacagtaaag ggtactagcc 16800 atcaggccca cctctacggt ctcgaaccat tcacgacgta tcgcatcggc gttgtagcgg 16860 ccaatcatgc tggtgagata cttagtccgt ggacgctgat acaaactctg gagtctagcc 16920 cctccggttt gcgcaacttc attgtggagc aaaaagagaa tggtcgagct cttctccttc 16980 aatggtccga acccatgcgc acgaatggcg tgataaagac ttataatata ttctcagatg 17040 gcttccttga atactcaggt ttgaaccgac agttccttt taggaggctt gacccattca 17100 ctctctacac gctgacgctc gaagcgtgta ccagggccgg ttgtgcacat tctgcaccac 17160 agcccctttg gacggatgag gctccccctg attctcaact cgccccaacg gtgcatagcg 17220 tcaagtcaac ctcagtagag cttagttggt ctgagccagt gaaccctaat ggaaaaatta 17280 taagatacga agtgatacgc cgatgtttcg aaggtaaagc ttggggcaat caaacaatcc 17340 aggccgatga gaaaattgta tttacagaat ataacaccga gaggaacacg ttcatgtata 17400 acgataccgg gctgcaacct tggactcaat gtgaatataa aatttatact tggaattcag 17460 cgggccatac ctgctcctca tggaacgtgg tgcggactct ccaagcaccg ccggagggcc 17520 tttcaccgcc tgtaatctct tacgtttcca tgaatcctca gaagctgctt atctcctgga 17580 taccacccga acaaagcaac ggaatcatac agtcttatcg gttgcagagg aacgagatgt 17640 tgtatccttt ttccttcgac cccgtcactt ttaattatac agacgaggaa ctgttgccat 17700 ttagcaccta ttcctacgcc cttcaagcgt gtacgagcgg tggctgctcc acatccaaac 17760 cgaccagcat taccacactt gaagccgctc caagcgaggt ctctccacct gacctctggg 17820 ccgtttctgc tacgcaaatg aatgtgtgct ggtccccgcc cacggttcag aatgggaaaa 17880 tcactaaata cctcgtaagg tatgacaata aagaaagtct ggctggacaa gggctctgcc 17940 ttctggtcag tcacctgcag ccttatagtc aatataattt tagtctcgta gcgtgtacca 18000 acggcggctg tacagcctct gtttcaaaaa gcgcttggac aatggaggcc ttgcccgaga 18060 acatggactc tccgaccctg caggtgaccg gatcagaatc catagaaatc acgtggaagc 18120 caccacgcaa tccgaacgga cagattcgaa gctacgaatt gcgccgggat gggactatcg 18180 tgtacacagg gctggagact cgatacaggg acttcaccct gctccggga gtggaatatt catatactgt cacggccagt aactcacagg gtggaatcct gagtcctctg gtgaaagatc 18300 gcactagtcc cagtgcgccc tctggcatgg agccaccgaa attgcaagca agaggcccc aggagatcct ggtcaattgg gatcctcccg ttaggactaa cggtgacatt attaactaca ccttgtttat cagggagttg ttcgaaaggg aaaccaagat tatccacatt aatactaccc father cggcatgcag tcatatattg taaaccaatt gaaaccgttc caccgatacg attaggat ccaggcatgc acgaccttgg ggtgtgcatc cagcgactgg acgtttattc 18600 aaacgccaga aattgcgcct ttgatgcagc cgccccgca cctggaggta cagatggctc cgggcgggtt ccccc gtctctcttc tctggaccgg acctctgcag cctaatgga 18720. aggtcctgta ttatgaactt tacagacgac agatcgccac tcaaccccgc aaatccaatc ctgtgcttat ctacaacggg tctagcacga gcttcattga tagcgaattg ctgcctttta 18840 ccgagtacga gtaccaagtg tgggcagtga actcagcggg gaaagcgcct tcctcttgga 18900 cgtggtgccg gacgggacct gcgccacctg aggggctgcg ggcacccacg ttccatgtta 18960 tcagctcaac gcaagctgtc gtaaatatct ccgctccagg taaaccgaac ggtattgtat 19020 ccctctatag gttgttctct tcctcagcac acggcgctga aacagtactg agcgaaggaa 19080 tggcaaccca gcagaccctg cacgggttgc aggcctttac gaactatagc attggagtcg 19140 aagcgtgcac ctgcttcaac tgttgtagca aaggccccac cgcggaactc cggacccatc 19200 ctgctccacc ctctggcctg tcctccccac aaatagggac cttggcctct cgcacagcga 19260 gtttccgctg gagtccgccg atgtttccaa atggggtcat acattcatac gagcttcagt 19320 tccatgtcgc gtgcccccct gactctgcgc tgccttgcac cccaagccaa atcgagacta 19380 agtacacagg gctgggccag aaagccagtc ttgggggtct gcaaccctat acgacgtata 19440 agcttagggt agtcgcccac aacgaggtag gtagtacggc cagtgaatgg ataagtttca 19500 cgacacaaaa agagttgcca cagtaccggg ccccgttctc cgtggacagc aacctcagtg 19560 ttgtatgtgt caattggtct gatacattcc ttctgaatgg gcaacttaag gaatatgttt 19620 tgacagacgg ggggagacgc gtttactctg ggcttgacac gactctgtat attccacgga 19680 ccgcggataa gacttttttt tttcaggtga tttgcactac tgacgagggg tctgttaaaa 19740 ccccgttgat acagtacgat acctcaacgg gattgggtct cgttcttaca accccgggga 19800 agaaaaaggg cagcaggagc aaatctacag agttctatag tgagctgtgg tttattgtcc 19860 tgatggccat gttgggtctc attcttttgg cgatatttct gtcactcata ttgcaaagaa 19920 agatacataa ggagccttat atcagagagc gccctccttt ggttcctttg caaaagagaa 19980 tgtcaccgct taacgttat ccgcctggtg agaatcacat gggactggct gataccaaga 20040 tccctaggtc tggtacacct gtgtctatcc gcagtaaccg ctccgcttgt gtactgcgaa 20100 taccgtcaca gaatcagaca agtctgacat acagccaagg cagcctccac cggagtgtat 20160 ctcaactcat ggacattcag gataagaagg tcctcatgga caacagcttg tgggaagcga 20220 tcatgggtca caactctggc ctctatgtag acgaggaaga cctcatgaat gcaataaagg 20280 atttttcaag cgtgactaaa gaacggacga ctttcacgga cacgcacttg tataaggca 20340 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac 20400 cagccttgtc ctaataaaat tagttgcat cattttgtct gactaggtgt ccttctataa 20460 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag 20520 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc 20580 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt 20640 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac 20700 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc 20760 caaattgctg ggattacagg cgtgaaccac tggccggcca tcgatgtaag aaattatatt 20820 acaagctaaa gcaacagctg tgggtggaag aagagaagaa ataatgtgtt ttttctttct 20880 tctttgatgg caaactcaac agactctgcc taaagcagaa ctgtagttaa aaggagactg 20940 caacttcaga gttttccatt acctagttc tcaacactgg cagattttt taggagctag 21000 tatctcttgt gaataaacat aagaagttca gcacacagaa agaaatggtg ggccagcact 21060 aattaaaac aaaaatgagc cgggtacaat ggctcacccc catgacccca gcactttggg 21120 aggccgaggt gggtggatca caaggtcagg agttcgagat cagcctggtc aacatggtga 21180 aacccccgtc tctactaaaaacaaaaat tagctgggtg tggtggcgggg cgcatgtaat 21240 cccagctact agggaggctg aggcaggaga atcgcttga cctggaggt ggaggttgta 21300 gtgagccgaa accgcgccac tgcactccag cctggccac aagagcgaga ctccgtctca 21360 aaaaaaaaaaaaaaaaaaccc aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaoe / v-at-catg-tcct-tcatgggtta 21420 agatagtttt taagacctt gccaaatatt tactgagttg cctactacga tgttaagcac 21480 tgttctaggt attcaggata catcagtgaa aaaaccgtc cctgcaaccc tcttaaaaaa 21540 aagtcactgt ccttatggag cttactgggg aggcaataa aaataggca aataatagta 21600 tacataaaa gaaatatta ggtatttatc aaaatgagtt gaaaacttgc acacaaatga 21660 ttatagcaat ttgttcata aactaccaaa aactgtcctt gaagttttga tgcccttca 21720 tatgtccatg gataaactgt ggtacattca tacacagtga atattattc agcattaaga 21780 aataagctac caacccacaa aaagatgtgg aagaaactta atgcatgttg ttaactgaaa 21840 gtgcctaaaa gggctccata ctgtatgatc ccaactatct gacattctgg aaatgggtac 21900 agagtcagta aaaacagctg ggcatggtgg tgcatgcctg taatcccagc tacttgagag 21960 gccaaggcgg areactgct tgagtcccag gatatcaaaa ccagctcaga caacatagca 22020 agactccatt ccaaaaaaaa aaaaaagaaa agaaaaag ataacagtgg ttgcaaaggg 22080 ttggggagaa gggaaggag ggatgaggcg gaggagcaca agggagtttt agggcattga 22140 aactattctg catgatactg taatggggga gacatgccac tgtgcatttt tccaaataca 22200 22260 aatcggttca tcagttgtaa caaatacacc acactaatgc atgatgttaa caggagaaac 22320 tgcgggggaag cggggtatat ggaaattccc tataatcacc gtgtattttt tctgtaaacc 22380 tgtaactact ctaaaaaaat aaagcccgtt ttaaaaagaa cttgaaaaaa taattccccc 22440 aaccaaaaat caagtcaaat aaaatcactg tcctcatgaa atttatgcga gaggcacaga 22500 agaaaacaaa ctaggcaata aatagtaaac aatagaagta aaatatcccc taaggcatac 22560 aacaggcata ataaaacaaa acaccatatg tacaactccc ccatctccct cccactcctc 22620 atagatggtt taggctaagg ttgataagtg ggagaaaact tttggtttct gaaacatctc 22680 atagttaagc gtatctggat gggtgcggtg gctcatgcct gtaatcccca acactttggg 22740 aggccaaggt gggtggatca cctgaggttg ggagtttcag actggtctgg ccaatatggt 22800 gaaactctgt ctttactaaa aatacaaaaa ttagctgggc ctggtggtac atgcctgtaa 22860 tcccaactac tcaggaggct gaggcaggag atcacttgaa cctgggaggc ggaggttgtg 22920 gtgagccgag atcatgccac tgcactccag cctgggcgac agagcaagac tctgcctcaa 22980 aaaaaaaga gtatccggtt ccccttcaa tatcaattt tcaagacat taatactga 23040 atagaaagt gccagagaaa tgctgaaagt tgcttaatct agtaatagc tgaaataat 23100 aggaatttc ttctttccaa tcataagcaa accagtcatt aaaatctcag accatacac 23160 acaccacac aacccatct tgttcttgc ttcaatttg ttttagatac aatcgcaacc 23220 atacactctg taatacatc actgccaat actcaacca catgaaaagg caagtgcct 23280 tttctattgt ggaataagc ccatgttggg gagaaatcct gggaatctcg cacaagaaac 23340 gcttatttac gtgtccagggg ccgccctctgc catgtttacc acaataata ctttaaactt 23400 caaaagaaac gcacacacag tctaggaac atgaaggct tttatatagt ccaatgaaaa 23460 tgctttggaa atgaaaagt atagaactg acagtacatg gggattttt tgtctcagaa 23520 tcttaatgga gatataac tataagcagg aaataggtca gatatacag cagacaatc 23580 agtttcaaga agtcctacaa ggaaaaattt gtgcaaaatg tcaggttgct aattattcat 23640 tcagtaaaca tttagggaat gtgccaagca gtcttctaag ccctggggat tcagcgacaa 23700 acaaaaacag atgcagcttc tgctctcaaa agctgacatc ctggctgggc gctgtggctc 23760 atgcctgtaa tcccagcact ttgggaggcc gaggcaggca gatcacctga ggtcaggagt 23820 tcaaaaccag cctggccaac gtggtgaaac cctatctcta ctaaaaatac aaaattagcc 23880 aggtgtggtg gcaggcgcct gtagtcccag ctacttgaga ggccgaggca ggagaatcac 23940 ttcaacccgg gaggtggagt ttgcagcgag ccaagatcgc accactgcac tccagcctgg 24000 gcgaaaagag tgaaactcta tctcaaaaaa aaaaaaaaaa ggaagctgcc atcctaatgg 24060 gggaagacaa tttgaacaga ttgaatagaa acacatgcag caaaacaggg tgatgggatg 24120 aaagcatgat gggctgctac tttggtctaa gcggtcaagg aaggcttctc tgaggaggtg 24180 aggcctgaat aacaagaaac caggacggtg aagggcagag ggaagaattc tgggtgaagg ggacaaggag ccagaggcgc tggagtgga agagctgggc ccacttggag gcacagggcg 24300 gaggggctag cagcaggcgc aggagagct gctgaggggg cagataggag gcaggcaggc 24360 agcacagggt ttgtggggca gggcaaagtg cttaagaaaa gccaaggaat ggctttaagt agggtctgac atcatccgat ttccattttt aaaagattac tgtgctgtga agcccagaaa ggccgagaga aaacaaatta cttagattac ttcagcctac aggacttcac agggtgtctg tggggatttg ggagtgtta gtgtcaaagt gtatctgaga tctgaggcca ggtgtggtgg 24600. ctcacgcctg taatcccagc actctgggag gctgagatag gcagataact tgaggtcagg agtttgagac cagcctggcc aacatggtga aaccccatct ctactaaaaa cagaaaaact agccaggcat tgtggtgcgc acctgtaatc ccagctactc aggaggctga ggcaggagaa tcgcttgaac cagagattgc gccactgcac tccagcctgg gtgacagagc gagactccgt 24840 tgtctttatt taaaaaaaaa aaaaaaaaat ctatctgagg cagaacaata gaacgcggca 24900 aacgtgaaga aactttcaga ttctccccc acctggcacg ctgctcccct ccccatcgc 24960 tctctcagct gatgacaggg aaaggaagcc caggctgcac cccctcacca aatgctgacc 25020 gccccaggca ggaggaggc tgccttgct ctcagccaag gccagctgca ccggtagtgc 25080 ttgggattcc aaggcctcca tgcctgggac tctcacctct gtccctgccc ctactctccc 25140 ttgtaccaga gcaggacagg ctctcacagc ctaccttgg aacagaacag vakacacatt 25200 taaaaaccct ctctaaactc tacaaaccca gagaagtcc tactttctg ttccaagaaa 25260 cctctttatc gttgttagtt cttaacatca cattctctgct tggcctact tcattgggc 25320 ttctagatcc atattccaa ggcaacggct tcttacaata tgtgcaaac aacagtccc 25380 ttttctgttc tcaaatgcat tctaaaaggt tggaatttca caacttttct tctgctggtt 25440 cctccacagt acacctacca agcctttacc ccaaggcttt cttgaccctc ttctctataa 25500 aatccaggcc atcccatcct gtctcatgtc ttcaaaaaga tctacgtggc agagacattc 25560 caatctccat ctcccccatg acatcttttc agaactccag actactcaac ttctgtgctc 25620 ttcagtccaa atggcgtctc aaaccagatg gttatcagat ggctacagga ggcattacgg 25680 aatgtatcat cttctgaatg tgaaccacat cagatgcaat acagagaaac acacactctc 25740 caggcacagc aaaccatgcc acagaacgct cgggctttac ctgtgatggg aagcttgggc 25800 tgcagtgtgt acagctgggc aggggtctga tgtgggttca ttccatatct caagggcatc 25860 tgagatacgc ctttgctgta ctgtttcctg aaataatctg aaattgcttc atcatattgt 25920 gccgtatgag tgaatgccta caaaaagcgt aagttttgtg agacacgagc agcactttcc 25980 aaagtgggga caagatggtt ttctacgtcc acagcaaaca acatctcctc actattcagt 26040 gctatgtggt ggctgcacaa cacgatgaca ccatgaatga cagcaaagca ctgaatggcc 26100 caacaaatta cccctttcca attctttttc aatctttttt tttttttttg agacaaagtc 26160 tcactctgct gcccaggcta gagtgcagtg gtgtgatgtc agcttactgc aacttccacc 26220 tcctgggttc aagcgattct cctgcctcag tctcccgagt agctgggatt acaggcgtac 26280 cacaccacac cacactcggc taatttttgt attttcagta gagacagggt ttcaccacat 26340 tggccaggct ggtctcaaac tcctgacctc aggtgatccg cccgcctcgg cctcccaaag 26400 tgctaggatt acaggcgtga gccaccgcgc ccagctctct ttcaatcttt ttgacagaac 26460 acctctgtaa tagctggttt ctccattttt aacaaaagga tagcagctcc aagcaaagct 26520 ttggtaggta atgcaatcta ccaagaactg aatactgctt ggtctggcaa acactgcact 26580 ccatggatgt aacttacaat atcatgaaag tgcatcccac cttcaaggct aactggcgtc 26640 tagtctccaa ggaggtgtcc ttactctcgg agctctgcat ctccgtggac accaccacat 26700 agtcctctgg ttcacacacc actgtcactc gagcgtggtt tttggctgca gctctcagta 26760 aggttactcc acctttgaag aacaaaagac ttattgattt ttgtatgtcc ctatcagctt 26820 atgaaccgct tttaaataaa gtcattgtgc atgtacttcc attttaattc ctgacttaaa 26880 aatcaggaaa ataagctggg cgcagtggct catgcctgga atcccagcac tttgggaggc 26940 tgaggcaggt ggatcacctg aggtcaggag ttcgagacca ctctggccaa catggcgaaa 27000 ttccatctct ataaaaaata caaaaaaaat tagccaggcg tggtggtgtg cacctgtaat 27060 cccagctact caggaggctg aggcaagaga atcacttgag cccaggaggc agaggttgca 27120 gtcagccaaa actgtgccac tgtactccag cctgggtgac acagcaagac ttcatctcaa 27180 aaaaaaaaaaaaaataaaaatcaggaaaacccaga tgcatctctg 27240 catacaaaaaaagcttagaa attaataatg aaaaaaaaaataatg attattaagta 27300 atcccaaaac acatccaga agtagctatt ttctcttat ttcctcagta ataaaattca 27360 aaatgaaatc tcactaaaat ttaaataatc ctctcctctc agtctctaa aatggttttt 27420 ctgacttacc aatgtcaatt tgctccacag cctcctcac agttacacct ggagaagcca 27480 ctgtctttac aaagggatag agattgcagg spaactct aaaaaaaaaaaaaaaaaaactctg 27540 gctgtcatta aaagtccac tcatgtggtt tacagctatc taggaaaaaa acagtcagt 27600 actattaacg taacaaag ccatagagaa ctaatatata aactgtgagc ctgacatttt 27660 agaattctaa cgagtgtgtt aaaagtcag ggtgtcctgc tgggtcgag cagaggtaat 27720 ttagtcagca gtgctcccc ggatctgaat acacagtact ccctccacgt cacactgcaa 27780 cgctgctgct tgcataacct tagctaccca gaaccacca gccatgaaaa atttagaa 27840 cattttaata gaagtataaa gtacaagatt acagaataa ttctcatttt ggaatgtgat 27900 cactattatc cagaacatta gatttagat tctttttcca gtggattggc acatgatga 27960 aaaatatcag agcattctaa gggtgaaaca ttgttttgc cttcagagag gtcataaaaa 28020 tgatggtaac aggaatgcca aagaataat ttaaaataca gaagtaatct aaaattaca 28080 atcagtgaag tacagttcca ctcctaggcc tgtagtgcga ggagcttac gggcaggctc 28140 aaacaagcc taactgctga aaactatta aaaaaaaaaaaaaac agagaacag 28200 aattggaggg gaacatttt gcagctcatc tttagt ctactgccct cctactaaaa 28260 cacacaaaaaaactagagac cagtatctct taggtactt cacacaaaaa 28320 ctctcaacaa aacactaca aactgaatcc agaaaaatgc aaaaaccacc tgatcatctc 28380 aatagatgta caaaaaggat ctgacacaat tcaacaggta ttcatataga aacactcagc 28440 aaattaaaaa tagaagggaa cttactgaac atgacaaagt gtaagaaaaa cccacagcta 28500 acatcatact tgatggtgaa agactaaaaat acttgtctac tcaactcaga aacaagacaa 28560 gaggccaggc gcagtggctc atgcctataa tcccagcact ttgggaggct gagccaggca 28620 gatcacctga ggtcagaagt tcgagaccag cgtggccaat atggtaaaac cccatctcta 28680 ctaaaatac aaaaattagc cgggcaaggt ggcgcacgcc tgcaatccca gcactttggg 28740 aggctcagga ggatggatca actgaggttg gaagttcaag accagcctga ccaacatgga 28800 gaaaccccat ctctactaaa aatacaaaat tagccaggca aggtggcaca tgcctgtaac 28860 cccaggctga agcaagataa tcacttaaac ccgggaggcg gaggtcacag tgagccaaga 28920 tcacgcaact gcactccgcc ttgggcaaca agagcaaaac gccatcttaa aaaaaagaaa 28980 caaaacaata atgtccattc ttaccacttc tactgaacac tgtactagaa gttctagcca 29040 gagcaattag caaagaaaag aaaataaaag gcatcaggac tggaaagaaa gcagtgaagt 29100 tatatctatt tgcagatgat accacctagt atacagaaaa tcccaaattc acaaaaataa 29160 ccttttagca ctaataaatg agttcagcaa agccgcagga tatatgatca atatataaaa 29220 atcaattgta ttaaaatata cattagcagt gaatgagaaa aaaattaaga aaacaattct 29280 acttcacaag acaatgttaa agaaattaaa gacaccccat gttcacagac tggaagattt 29340 aatattctta ggatggtaac atttcccaaa ttgatctaca gattcaacac aatacctatg 29400 aaaattccaa ctgcgttgtt ttgcagaaat ggacaagctg attctaaaaa tcatgtggaa 29460 atgcaaggga ctctgaagag tcaaagtcat cttgaataaa aaaaacgaag ttggggccgg 29520 gcgcagtggc tcacgcctgt aatcctagca ctttgggagg ccgaggcggg cggatcacga 29580 gtcaggaga tcgagaccat cctggcgaac acggtgaaac cctgtctcta ctaaaaatac 29640 aaaaaaatta gctgcgcatg gtagcggcg cctatagtcc cagctactcg ggaggctgag 29700 gcaggagaag ggcgtgaacc cgggaggcag agcttgcagt gagccaagat cgcgccactg 29760 cactccagcc tgggcaacag agccagactc cgtctcaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaahaa 29820 aaaaaaaaaaaacaga aaatgaagtt ggaggactca cacttgctga ttcaaatt 29880 tgttaccagc ctatagtaat agtgtagtac tgcataca gcaggcacat ggaccaaggg 29940 tgccaagacc attcgattgg gcaagaaca gtctttcaa caatgatgc tgagacaact 30000 agacagccac atacaaata aaatagact tctatttcat gtctgtatat gaaaattac 30060 ttaaatgaa ccaagactt aagtataaga gctaacacta aaatactct ttaaaagaat 30120 gtacacacgc cgggtgcggt ggctcacact tgtaatccca gcactttggg agaccgaggc 30180 aggtggatca cctgaggtca ggagttcaag accagcctgg ccaacatgga gaaaccccgt 30240 ctctactaaa aacacaaaaa aattagctgg gcgtggtggc gggcacctgt aatcacagct 30300 acttgcgagg ctgaggcatg agaatcgctt gaacccagga ggcgaaggtt gcagtgagcc 30360 gagatcacgc cactgcactc cagcctgggc aacaagagcg aaactccatc tcaaaaaaaa 30420 aaaaaaaaag aaagaaaata tacacaaaaa tcttcatcac tgaggattag gcaatggttt 30480 cttagacatg acaccattaa cacaagcaac aaaagaaaaa ataaattcaa catcatcaaa 30540 attaaaaact tttgtgggcc gggcatggtg gctcaagcct gtaatcccag cactttggga 30600 ggccaaggcg ggcagataac ccgaggtcag gaattcaaga ccaacctggc caacatagtg 30660 aaaccctatc tctattaaaa atccaaaaaa attagccaag tgtggtggcg ggcacctgta 30720 accccagcta ctcatgagac tgaggcagaa tcgcttgaat ccgggaggtg gaggttgcag 30780 tgagccaaga tgacgccact gtgctccagc ctggggcgaga gtaagactcc gtctcaaaaa 30840 aaagaaaaaaaaaaaaaaa cttgtgatga tcaaggaaa ccatcaagaa aggctacac atccataga 30900 atgagaggaa ataacttac aactcataa taaacagata atacctcaa ctaaaaatgg 30960 acaagaggtc tcacagaca tttctccaa ggaggtaac aactggctag gaagcacacatc 31020 caacggtgtt ccacattatt tggaagcaa attataacca caatgagatg ccacttcaga 31080 acccctagaa tgactagaat caaaagaca gatactaacg tgtgaga tgtggagaaa 31140 ctggagcccc atccagcaca ctgctgctgg aaatgtaaaa tggtgcagca ctctgaaata 31200 cagggtcgca gttcctcaa agctaaacat agttaccata tgaccaagga attccactcc 31260 taggtatata cccaatgaa acgaatgtgt atgtccacac aaaaacttat acacaaatat 31320 ttatagctgc attattcata atattcaaa agtagaaca accaaatgt ccatcaaca 31380 atgaactgat aaacaacagg gtatgaccac acaatggatt atttttcagc actaaaaacg 31440 aaatgaacta ttgatactat gctataacat ggataaacct tgaaaacatg ctaagtgaaa 31500 gaagccagga acaaaaaaac noteattgt tagattctat ttatatgaaa atgtccacta 31560 caggcaaatc agacacgtaa caattggtgg ttgccgaggc tggaggacgt tggggaaatg 31620 gagaataaat gcgtatgggt atagggttta gggtgggggt gatgaatatg ttttaatatt 31680 ggatgtggca attaatatgg tttggatgtg tgtcctctcc aaatctcatt ttgcaatgtg 31740 atcccccaatg tggaggtgg ggcctggtgg gagatgattg ggtcacgggg gagatccctt 31800 atgaatggct tggtgccctc cccataatga gtcctcactc tgagttcatg tagagctggt 31860 tgtttaacag agccgggcag gccgggcgtg gtggctcaca cctaatccca gcactttggg 31920 aggccggggt gggtgcatca tttgaggtca ggagttccaa gagcagcctg accaacatgg 31980 tgaaacccca tctctactaa aaatgaaa aatgagttgg gcgtggtgac aaacgcctgt 32040 aatctcagct actcaggagg ctgaggcagg agaatcactt gaagacagga ggcagaggtt 32100 gcagtgagcc aaggtcatgc cactgaactc caacctgagc aacagagcga gaccctgtct 32160 caaaaataaa ttaaattaa ttaagataa aagttaaaaa gtaaaagagc tgggcaactc 32220 ctcctctccc tctctcttgc ttcttctctc atgggatatg ccagctcccc ctctgctttc 32280 cactgtgatt ataaacttcc tgaggccctc accagaagtt gatgctggca ccacacttct 32340 tgtactgtct atagaatcac gagccaaaat aaacctgttt tcataaatta cccagcttct 32400 agtattcctt tatagcaatg cagaacagac taacagta atggttgcac aacttctgtg 32460 aaccactgat ttgtacattt taaattggtg agttgtatgg tatgtggatt agatctcaat 32520 aaagctgtta cgtatattta agaaacag ccagtatctt tatgcaaata actcagttag 32580 ttgtgcaagt tgataattcc taatatgaaa acctgaaatc tgaaatgctc caaatctga 32640 aacttttga gcgtcacat cakacttgga ggaatgctc actgaagcat ttcggatttc 32700 tgagggacgc tcactggca tgttacctgc aaatattcca aaatctgaag aaatctgaaa 32760 tcagacac ttctgggtcc caagcattc agataggga tactcacct atacatacta 32820 agatttagat tgtaacttcc taaaaaactt cagtatgtat tttggtatat aaatctggcc 32880 aaaaccaaaat gcttacacac atacaaagtc acacagaagg tacgcagta agggtttatag 32940 tgagtaagaa aacacaggga caccaggttt ggcatagtct tgtcgttgg ttcgtaatgt 33000 gttaaaagtt taatttcagg ttttacctt atagattga aatcaagtct ggccatgtca 33060 gcattatctt ctggaatatt acgagctagg attcctatt taaaaaagtt aaacaaagtc 33120 attagcttca aaaaacaagt cagattagg atgtctttc aacatagtgt cttcaatcaa 33180 attaaatccc attgattaaa aaaaaaattt atttttttga gatggtctca ctctatcgcc 33240 caggctggat tgtagtggcg taatcacaac tcactatagc cctgacctcc agggctcaag 33300 tgatcctcct acctcagcct cccaagtagc tgggactaca ggaatgcatc accacaccca 33360 gctggtcttt tcaattttt aaagagaagg ggggtgttgc catcttgccc agggtgatct 33420 taaactcctg ggctcaagcg gttctcctgt ctcagctctc caagtgctgg gactacaggc 33480 aagagccacc atgcccagca tatattctt attaaagaa tatcaaacat gtagttgggg 33540 agttatcaaa caaaaaaatt aggattaaca gatatatctg gctggccaca gtggctcatg 33600 tctgtaatac cagcactttg ggaggctaag gtgggggatc gcttgagccc aggtgttcga 33660 gaccagcctg ggcaacaagg tgagaccccg tctctataaa aaatacaaaa attagctggg 33720 tgtggtggca catgcctata gtcccagcta cttgggaggc tgaggtggga gaactgcttg 33780 agcctgggaa gcagacagag gctccagcga gccatgatcg caccactgca ctccagcctg 33840 ggcaagagata taatcctgtc tcaaaaaaaaa aaaagagaga tatatttaaa aatctttttc 33900 ttcagttctt gatcccagtt tcttgagcaa taaatgtaa aaaaggtaaa aggaatttta 33960 caaaactgtt gctagttcca gaacagtcta tgcgtttctg taagtcaatg gcttgtctga 34020 tgatctgtag ccattcagct ttattcacca aattgtacca caactgacaa actaggaga 34080 taccagtttc ttgccgtttc ggtttatata caattcattc ttaggtgaac gaacaaatgt 34140 aaagcacttt gaaaaaaaaa attcctttgc aagtcagtta cgctgtgatg ctttcttcta 34200 agtttgaact attttaaaag tattttcata cgatgaaatc taaatatctt cagcaacttg 34260 agcaggaaac accagactct aaattaggta tcaggtcagg actaatagca agcagaggaa 34320 aacaatctga attcatcaag tagtaagaca agctggattt taaaccagtt ctaacgtatt 34380 atgaaatctg gcagcaactt cataacatta actttatatg taaatttctc ttgagccttt 34440 ttcttccttt attttaggac gctatcaact aaaatattcc tggaaaccac tgactgtttt 34500 tacattaaag ataccaacca cttaccagca tggactgcag gatgcaaagt tttcacacgt 34560 ccccccaaca tttcaggaaa tcccgtcaac tcagagacat ctctgaaaaa cagataaaca 34620 gaatggtact actggtgtgg ctgaattgta attttcacta ttcaaagcat ctactgtttt 34680 catttcaaga gattttgagc ctgaattcaa cgtgaaaatt ctgatttttt aaagtatttc 34740 actatatttt atttcttaag taaaattcta tttacttccg aaaaaaagtt tatttaattt 34800 atttcctaac tgttgtgtat gttccgtacg attaagggcg aattccagca cactggcggc 34860 cgttactagt ggatccaaca gtctccacgt aaacggtcaa agtccccgcg gccctagaca 34920 aatattacgc gctatgagta acacaaaatt attcagattt cacttcctct tattcagttt 34980 35040. tcccgcgaaa atggccaaat cttactcggt taccccaaa tttactacaa catccgccta aaaccgcgcg aaaattgtca cttcctgtgt acaccggcgc acaccaaaaa cgtcactttt 35100. 35160. gccacatccg tcgcttacat gtgttccgcc acacttgcaa catcacactt ccgccacact actacgtcac ccgccccgtt cccacgcccc gcgccacgtc acaaactcca ccccctcatt atcatattgg cttcaatcca aaataaggta tattattgat gatgattgtt taaacatgat gagctcagc 35289 <210> 3 <211> 35067 <212> DNA <213> Homo sapiens <400> 3 60. tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat 120. gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg 180 cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc 240 cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct 300 gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct 360 tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga 420 tgacgaccat cagggacagc ttcaaggatc gctcgcggct cttaccagcc taacttcgat 480 cactggaccg ctgatcgtca cggcgattta tgccgcctcg gcgagcacat ggaacgggtt 540 ggcatggatt gtaggcgccg ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc 600 atggagccgg gccacctcga cctgaatgga agccggcggc acctcgctaa cggattcacc 660 actccaagaa ttggagccaa tcaattcttg cggagaactg tgaatgcgca aaccaaccct 720 tggcagaaca tatccatcgc gtccgccatc tccagcagcc gcacgcggcg catctcgggc 780 agcgttgggt cctggccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta 840 ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga 900 agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc 960 cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat gttccggatc 1020 tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc 1080 tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc cagttgttta 1140 ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat cgtgagcatc 1200 ctctctcgtt tcatcggtat cattaccccc atgaacagaa atccccctta cacggaggca 1260 tcagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag aagccagaca 1320 ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga catctgtgaa 1380 tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac 1440 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat 1500 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca 1560 gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag 1620 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 1680 gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 1740 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 1800 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 1860 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 1920 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 1980 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 2040 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 2100 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 2160 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 2220 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 2280 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct 2340 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 2400 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 2460 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 2520 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 2580 taaattaaaa atgaagtttt aaatcaatct aaagttata tgagtaaact tggtctgaca 2640 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 2700 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 2760 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 2820 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 2880 agtctattaa ttgttgccgg gaagctagag tagtagttc gccagttaat agtttgcgca 2940 acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 3000 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 3060 cggttagctc cttcggtcct ccgatcgttg tcaagtaa gttggccgca gtgttatcac 3120 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 3180 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 3240 gctcttgccc ggcgtcaca cgggataata ccgcgccaca tagcagact ttaaaagtgc 3300 tcatcattgg aaacgttct tcggggcga aactctcaag gatcttaccg ctgttgagat 3360 ccagttcgat gtaacccact cgtgcaccca actgatctc agcatcttt actttcacca 3420 gcgtttctgg gtgagcaaaa acaggaggc aaatgccgc aaaaaaggga ataagggcga 3480 cacggaaatg ttgaatactc atactctcc ttttcaata ttattgaagc atttatcagg 3540 gttattgtct catgagcgga tacatatttg aatgtatta gaaaaataa caatagggg 3600 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga 3660 cattaaccta taaaaatagg cgtatcacga ggcccttcg tctcaagaa ttacggcccc 3720 atatagttta aacatacatc atcaatata taccttattt tggattgaag ccaatatgat 3780 aatgaggggg tggagtttgt gacgtggcgc ggggcgtggg aacggggcgg gtgacgtagg 3840 ttttagggcg gagtaacttg tatgtgttgg gaattgtagt tttcttaaaa tgggaagtta 3900 cgtaacgtgg gaaaacggaa gtgacgattt gaggaagttg tgggtttttt ggctttcgtt 3960 tctgggcgta ggttcgcgtg cggttttctg ggtgtttttt gtggacttta accgttacgt 4020 cattttttag tcctatatat actcgctctg cacttggccc ttttttacac tgtgactgat 4080 tgagctggtg ccgtgtcgag tggtgttttt ttaataggtt ttctttttta ctggtaaggc 4140 tgactgttat ggctgccgct atgcccgggg ggaggcctaa tcggccttag cgatcgctta 4200 gggccccaga agcctggtgg ttgtttgtcc ttctcagggg aaaagtgagg cggccccttg 4260 gaggaagggg ccgggcagaa tgatctaatc ggattccaag cagctcaggg gattgtcttt 4320 ttctagcacc ttcttgccac tcctaagcgt cctccgtgac cccggctggg atttagcctg 4380 gtgctgtgtc agccccggtc tcccaggggc ttcccagtgg tccccaggaa ccctcgacag 4440 ggcccggtct ctctcgtcca gcaagggcag ggacgggcca caggccaagg gcagccgcca 4500 ccatgaattg tcctgtactg agcctcggta gcggcttttt gttccaagtc atagaaatgc 4560 tcatctttgc gtacttcgcg tcaatatctc ttacagagtc tcggggtctc ttcccaaggc 4620 ttgaaaatgt aggagcgttt aaaaaggttt ccatcgtacc aacccaggct gtatgtggcc 4680 ttcctgaccg cagtactttt tgccactctt ccgccgctgc tgagtcaatt cagttttgca 4740 ctcagcggtt ctgcattcaa gattgcccct atcggtcttc tcacccaaca tatacggcac 4800 tgttttctgc ggggcttagt tcctgtatta cccctgacaa gaatgatctt caccctaacg 4860 cgcactcaaa tagtgcatca tttatcttcg gaaaccacaa atcctgtttt tcttctcctc 4920 cgtcccccaa actgatggcc tcctttacgt tggcggtttg gcttaaaccg gaacaacaag 4980 gagtaatgtg cgtgattgag aagacggttg atggacaaat cgtctttaag ctgacgatat 5040 cagagaaaga gacgatgttt tattaccgga cggtgaatgg gctgcaaccc ccgattaaag 5100 taatgactct cggtcggatc ctcgtgaaga aatggattca cctgagtgta caagtccatc 5160 aaactaagat ttcatttttt atcaacggag tcgagaagga ccatacccca tttaacgcta 5220 ggacattgag tgggagcatt acagatttcg cctccggaac ggtacagatc ggccaatcct 5280 tgaatggatt ggaacaattc gtcggaagaa tgcaggactt ccggctgtac caggtagcac 5340 tgacaaatcg agagatcctt gaagtcttta gtggggacct gttgcggctg catgcccaat 5400 cacactgccg gtgtccaggt tctcacccga gggtacaccc gctggctcag cggtactgta 5460 taccaaatga tgccggggac acagcagata acagagtcag tcgacttaac ccggaggcac 5520 accctctctc attcgtaaac gacaacgacg tggggacttc ttgggtgtct aacgtcttca 5580 caaatataac acaactgaat caaggcgtca ccatctccgt tgacctcgag aacgggcagt 5640 atcaggtgtt ctatattatt atacaatttt tcagtccaca gcccacggag atacgaatcc 5700 agcggaagaa agagaattcc ctggactggg aagactggca gtacttcgct cgcaattgcg 5760 gggcgtttgg catgaagaat aacggagacc tggagaagcc tgattccgtg aactgccttc 5820 agctctcaaa cttcaccccc tactcaagag ggaatgtgac attctcaatt cttactccgg 5880 gccccaatta tcggcccggg tacaacaact tttataacac gccttccctg caagaatttg 5940 taaaggcaac ccaaatcaga tttcacttcc atggacaata ctataccact gaaacagccg 6000 ttaatctgag gcaccgctac tacgccgtgg atgaaatcac gatcagtgga cggtgtcagt 6060 gccatggcca cgctgataac tgcgacacaa ctagccaacc atacagatgc ctctgcagcc 6120 aagaaagttt cactgagggc cttcactgtg atagatgcct gcccctttat aacgataagc 6180 ctttcaggca aggagaccaa gtttatgcgt ttaactgcaa accatgccaa tgcaacagcc 6240 attccaagtc ctgtcattac aacatttccg tcgacccttt tccatttgag cattttaggg 6300 gagggggcgg agtatgtgat gactgcgaac ataacactac tggtagaaac tgtgagttgt 6360 gcaaagatta cttctttcgc caagtgggtg cggatccatc cgcgatcgac gtatgtaagc 6420 cctgtgactg tgacaccgtc gggacgcgga atggatctat cctttgtgat caaataggtg 6480 ggcaatgcaa ttgtaaacga cacgtatcag gtaggcagtg caaccagtgt caaaatggat 6540 tctacaatct ccaggaactc gaccctgacg gatgtagtcc atgcaactgt aacacctctg 6600 gcaccgtaga cggtgacata acatgtcatc agaatagtgg acagtgtaaa tgtaaggcta 6660 atgttatcgg gctcaggtgc gatcattgca atttcggttt taagttcctc cgctcattta 6720 atgatgtagg gtgtgaaccg tgtcagtgta accttcacgg tagtgttaac aagttttgta 6780 acccccattc tggtcagtgc gagtgcaaga aggaggccaa ggggttgcaa tgcgatactt 6840 gccgcgaaaa tttttatggg cttgatgtaa ccaactgtaa ggcgtgcgac tgcgatactg caggatctct gccaggaact gtgtgcaacg cgaaaactgg acagtgcatt tgtaaaccga atgtggaagg acgacagtgc aacaagtgcc tggagggaa tttttatctc cggcagaaca actcatttct ctgtctccca tgtaattgcg ataaaactgg caccattaat ggcagcttgc tttgtaataa aagcactgga caatgcccct gcaagctggg agttacgggg ctgcggtgta accaatgcga accacaccgg tacaatctca ccatcgaca tttccaacat tgtcagatgt gtgaatgcga ttccctcggg acactccctg gcacgatatg tgaccccatt agtggccaat gcctgtgtgt acccaaccga cagggtcgac gatgtaatca gtgtcagcca ggattttaca tagcccggg aaacgccaca ggttgcctgc cctgctcatg tcacacgacg ggagcagtca atcacatatg caatagcctc actgggcaat gcgtgtgcca agacgcaagc attgctggtc 7440 agcgctgtga tcagtgcaaa gatcactact tcgggtttga cccacagaca ggtcggtgcc 7500 agccgtgcaa ctgccacctt agcggtgcat tgaacgaaac atgtcatttg gtaacgggtc 7560 agtgctttg caaacaattt gtaacgggtt ccaagtgtga cgcatgtgtt ccttctgcca 7620 gccatcttga cgttaacaat ctccttggtt gttctaagac tccatttcag caaccaccgc 7680 caagaggcca ggtccaaagc tcatcagcca tcaacctctc atggtcacct cccgattcac 7740 cgaacgcgca ctggttgacc tactcattgc tccgcgatgg attcgagatt tataccaccg 7800 aggaccaata tccgtattcc attcagtact ttctggacac tgacttgctg ccctatacta 7860 aatatagtta ttatatcgag acgaccaatg tgcacggttc tacgcgatct gtggccgtca 7920 catataagac taagcctgga gtgcctgaag gtaatctgac tttgagctac ataatcccaa 7980 ttggatccga cagcgtgacc cttacgtgga ccacgctttc caaccaatca ggaccaattg 8040 aaaaatacat cttgtcttgc gctccgttgg cggggggtca accctgcgtc tcatatgagg 8100 gccatgagac aagtgcgact atatggaatc tcgttccctt tgcaaagtat gacttttctg 8160 tacaagcctg cacatctggc ggatgcctgc atagtcttcc aatcacagtt accacagcac 8220 aagcaccccc ccagaggttg agcccgccaa agatgcaaaa gatttcttct acggaactcc 8280 atgtggagtg gagtcctccg gcagaactca atggtattat cattaggtat gaactgtaca 8340 tgcgacggct tagatctacc aaagagacta cgagtgagga gtcacgcgtg tttcagtcat 8400 ctggctggtt gtcaccccat tcattcgtgg agtctgcaaa tgaaaacgcg ttgaagcctc 8460 ctcaaacaat gactacgata actggacttg agccatatac taaatatgag tttagagtac 8520 tggctgtcaa catggccggc tccgtctcaa gtgcatgggt ttccgagcgc acaggcgaat 8580 ccgcgcctgt cttcatgata ccccccagtg tttttcctct gtctagttac tccctcaaca 8640 tatcttggga aaagcctgcg gaacgtga ccagaggtaa ggtcgtgggc tacgatatca 8700 acatgctgtc agagcagtcc ccacagcaga gcatcccgat ggccttttca caactcttgc 8760 atacggccaa atcacaagaa ctgtcataca cggtcgaggg acttaaaacct tatcggatct 8820 acgagtttac gatcacgctg tgcaactcag tcgggtgtgt cacttctgcg agcggtgcag 8880 gccagacgct tgcagcagcg ccagcgcagc tcaggccccc tttggtcaag ggaataatt 8940 ctactacgat ccatctccgg tggtttcctc cggaggaact gaatggccct agtcccattt 9000 atcagctcga aaggagag tcttcactgc ctgcactcat gaccactatg atgaaaggaa 9060 tccgctttat tggaaacggg tactgcaaat tccccagctc cacgcaccct gtgaatacgg 9120 attttacagg gatcaaggcc tcttttagaa cgaaagtgcc ggagggcctc atcgttttcg 9180 ctgctagccc cggcaaccaa gaggagtatt tcgccctca gttgagaag ggaggctgt 9240 acttcctgtt cgatcctcag ggaagtccag ttgaggttac gaccaacaat gaccatggta 9300 aacaatacag cgatggtaa tggcacgaga taatcgcgat tagacaccag gcgttttggggc 9360 agatcacact cgacggcatt tacactgat ccagtgcaat tctcaacggt agcacagtta 9420 ttggcgacaa tacggggggtc ttctggggcg gtctgccacg atcatacact atactcgca 9480 aggatcctga ataatacag aagggttttg tcggatgcct taggacgtg cattttatga 9540 aaaactacaa tccttccgct atatgggagc cactggactg gcagctctca gaggacaaaa 9600 tcaacgtgta taattctgg gaagggtgtc ccgcgagcct taacgaaggg gcacaatttt 9660 tggggcagg ttcctcgag ttgcaccat acatgtttca tggggtg aatttcgaaa 9720 tttctttcaa gtttcggacg gaccaattga acggcttgct tctgtttgtc tacaacagg 9780 acggccctga tttcctggcc atggagctca aaagtgggat actgaccttc cgccttaata 9840 ctagtctggc gtttacccaa gtagatctgc tgcttggcct ttcttactgc aacgggaaat 9900 ggaacaaagt gatcatcaaa aaagaaggct cctttatttc tgcatctgtg aatggcctga 9960 tgaagcatgc ctccgaaagt ggtgaccaac cccttgtagt caacagcccg gtatatgttg 10020 gggggatccc acaagaactc ttgaacagtt accagcactt gtgcctcgaa caaggtttcg 10080 gcggatgcat gaaggacgtg aagtttaccc ggggggccgt agtcaacctt gccagcgtat 10140 catccggggc cgtgcgggtc aatttggacg gatgcctgtc tacagattcc gcagttaatt 10200 gtcgagggaa tgattcaata cttgtatatc aagggaaaga acaatcagta tacgaagggg 10260 gcctccaacc ttttacagag tatttgtata gagtcattgc aagtcatgaa gggggatccg 10320 tttactcaga ctggtcacgg ggccgcacaa ccggcgccgc gccacagtcc gtccccacac 10380 cttccagggt gcggtcactg aacggttaca gcatcgaagt gacgtgggat gaaccagtag 10440 taagggtgt gattgaaag tatatattga aagcttacag tgaggattcc acacgccccc 10500 caggatgcc atccgcgtct gcggaattcg ttaatactc caacttaca ggcattttga 10560 caggactgct cccttttaa aattacgctg tcacacttac tgcttgtacc ctcgcggggt 10620 gcacagaaag tagtcatgcg ctgaatatct ccaccccgca ggaagcccct caagaggttc 10680 agccgccggt cgcaagagt cttccgagta gtctgctcct ttcctggaac ccccctaaaa 10740 aagcaaacgg tatcatcaca caatactgcc tctacatgga cggggcggttg attackctg 10800 gtagtgagga aaactacata gtaaccgatc ttgctgtctt tactccacat cavattcctcc 10860 tttctgcgtg tactcatgta ggctgcacga actcaagttg ggtactctc tatactccc 10920 agttgccacc tgagcacgtt gactctcccg tattgactgt attggacagc aggaccattc 10980 acattcaatg gaagcagcct cggaaaataa gcggcatcct tgaaaggtat gttctctata 11040 tgtctaatca cacccatgac ttcacaattt ggagcgtcat ttataatagt acagaactgt 11100 ttcaggatca tatgttgcaa tatgtactcc ctgggaataa atacttgata aagctgggtg 11160 cgtgcaccgg aggtggttgt acggtgagcg aggcatctga ggcgcttacg gacgaggaca 11220 tcccagaggg tgtacccgcc ccgaaggccc actcttacag ccctgacagc ttcaatgtga 11280 gctggactga acccgaatac ccaaatggcg taataaccag ttatggactg tacctcgatg 11340 gtatattgat acataattct tccgaactta gttatagggc ctacggattt gctccgtgga 11400 gcttgcacag tttccgggtc caggcttgca ccgcgaaagg ttgcgccctg ggtccgctcg 11460 tggaaaatcg aacactcgag gccccacctg aggggaccgt gaatgtgttt gttaagacac 11520 agggaagtcg aaaagcgcat gtccgatggg aggccccatt tagacctaat ggcttgctta 11580 cgcactctgt gttgttcacg gggatcttct acgtcgaccc tgtaggtaat aattacacat 11640 tgctgaacgt caccaaggtg atgtactccg gagaggaaac taacctttgg gtactgattg 11700 acggcctggt accgtttacg aactatacag ttcaagtgaa catcagtaac agccaagggt 11760 ccctcattac ggatccgatt actatcgcga tgccgccggg tgctcctgat ggagttctcc 11820 ccccccgcct ttcatctgct actcctactt cattgcaggt tgtgtggtca acccccgcga 11880 gaaacaacgc gcctggtagc cccagatatc aacttcaaat gcggagcggg gattccaccc 11940 acggtttcct cgagctcttt tccaatcctt cagcaagctt gtcctacgaa gtttctgatc 12000 tccagccata tactgaatac atgttccgac tggtggcgag caatggattt gggtcagccc 12060 acagttcttg gatccccttt atgaccgctg aggacaagcc agggcccgtg gttccgccca 12120 tcctgctgga tgtcaaaagt cgaatgatgc tggtcacgtg gcaacatccg agaaagtcca 12180 acggcgttat aactcactac aacatttacc tgcatggtcg gctttacttg cgcaccccag 12240 gaaatgtaac gaactgtacc gttatgcacc tccaccctta taccgcctac aagttccagg 12300 tcgaggcatg tacgtccaaa ggatgctcac ttagccccga aagccagacc gtttggacgc 12360 ttcctggggc tccggaaggg atccctagtc cggagctctt tagcgataca cccaccagtg 12420 tgataatcag ctggcagccc cctactcatc cgaatggtct cgtcgaaaat ttcacgatag 12480 aaaggcgagt taaaggtaaa gaggaagtga ccaccttggt tactctgccc cggagccata 12540 gtatgcgctt tatagacaag acctccgccc ttagcccctg gaccaaatat gaataccggg 12600 ttcttatgtc cacgttgcat ggagggacaa attccagcgc ttgggtagaa gtcactactc 12660 gaccaagccg gcccgcgggc gttcaaccac ctgttgttac ggtactggag cctgatgctg 12720 tccaagttac gtggaaaccc ccactgatcc agaatggtga catactgagt tacgagatac 12780 atatgccaga ccctcacata acattgacga acgttacctc tgcagtgctt tcccagaaag 12840 ttactcacct cattccgttt acgaactata gtgtcacaat cgtggcctgt agcggtggta 12900 acggatatct tggagggtgc accgaaagcc tccctactta cgtgacgacg cacccgacag 12960 ttccgcagaa tgtaggtccc ctgtccgtta ttccgttgtc cgaatcctat gtcgtgatat 13020 cttggcagcc cccctctaaa cctaatggac ccaatctgag gtacgagctg cttcgacgca 13080 agatccaaca gccattggcg tccaacccgc cggaagacct gaatcggtgg cacaatattt 13140 atagtggcac gcaatggttg tacgaagaca agggcttgtc caggttcacg acctacgagt 13200 atatgctttt cgttcataat tccgtcggat ttaccccgtc tcgcgaagtg accgtcacca 13260 ccttggctgg cttgccagaa cgcggggcga acctcactgc ttcagtcctg aaccacaccg 13320 cgatagatgt tcggtgggct aagccaactg tgcaagatct ccagggcgag gttgaatact 13380 ataccttgtt ttggtcctct gcaacgtcca atgatagtct caagatattg ccggatgtta 13440 acagtcacgt aattggacat ctcaaaccta acacagaata ctggatattt atttccgtgt 13500 ttaatggagt gcactccata aacagtgctg gactgcatgc gacgacttgt gatggtgagc 13560 cacaggggat gcttccccct gaagtagtta tcattaactc aaccgcagtc agagtaattt 13620 ggacttctcc aagcaaccca aatggagtag taaccgagta ctctatttat gtgaacaata 13680 aactgtacaa gaccggcatg aacgtacccg gctcatttat tttgagagac ctgtccccgt 13740 ttacgattta cgatatacag gtggaggtct gtacaatata tgcatgcgtc aagtccaacg 13800 gaacacaaat cactaccgtc gaagatacac cctcagatat tcctacgcca accattaggg 13860 gtattacgtc taggagtctt caaatagact gggtgtcacc ccggaagccc aacgggatta 13920 ttctcggata tgacctgttg tggaagactt ggtatccgtg cgccaaaaca cagaaattgg 13980 tccaagatca gtcagatgaa ttgtgtaagg cggtgcggtg tcaaaaaccc gagagtatct 14040 gcggacacat ctgttatagc tcagaggcaa aagtatgctg taacggtgtt ctgtacaatc 14100 ccaaaccagg ccatcgctgt tgtgaggaga aatacatccc gttcgtgctg aactcaacgg 14160 gggtctgttg cggtggtcgc attcaagaag cccagcccaa ccaccagtgt tgctctggtt 14220 actacgcccg catcctccca ggggaggtat gttgccctga cgaacaacac aacagggtaa 14280 gcgtaggcat tggcgactcc tgttgtggcc ggatgccgta tagtactagc ggtaaccaaa 14340 tttgctgcgc gggaagactt catgatggac acggacagaa atgttgcggc cgccaaattg 14400 tatcaaatga cctcgaatgc tgcggcggcg aagaaggcgt agtttacaat aggctgccgg 14460 ggatgttctg ttgcggacaa gactacgtaa atatgagcga tactatttgc tgttctgcaa 14520 gctctgggga gtcaaaggcc cacataaaaa agaatgaccc agtaccagtt aagtgctgcg 14580 aaactgagct cattcccaag tcacagaaat gttgcaatgg ggtaggatat aatccgttga 14640 agtacgtctg ctcagataag atttcaaccg gtatgatgat gaagagacc aaagtaatgta 14700 gaattctgtg ccccgccagt atggaagcaa cggaacactg tggtcggtgc gactttaatt 14760 tcacgagtca catctgcact gtcatccgcg gctcccataa ctcaactggc aaggcctcta 14820 taggagaat gtgcagttcc gcggagaaa ccatacatac gggaagcgtc aatacgtatt 14880 cttatactga cgtcaacctg aagccctaca tgacgtatga gtaccgaatc agcgcgtgga 14940 attcatacgg ccgcggattg agtaaagcag tcagggctag aacgaaggag gatgttccgc 15000 aaggcgtaag cccaccact tggacgaaaa tagacaacct tgaggataca attgtcctta 15060 attgggagaa gccgatccag agtaacggtc ctattattta ctacattctt ctgcggaatg 15120 ggatagagag gtttcgaggc actagtctga gtttttccga taggaaggt attcagccat 15180 ttcaagaata cagctaccag ctgaaagcat gcacagtggc gggatgtgcc acctcaagca 15240 aagtagtagc cgcgactacc caaggcgttc cggagagtat acttccgcccc agtatcaccg 15300 cgctgagcgc cgtagcattg cacctcagct ggtcagttcc ggaaaaaagt aatggcgtca 15360 ttaaggaata tcaaatacgg caggtgggaa aaggccttat tcacactgat acgaccgacc 15420 gacggcaaca tacagtgact ggtctccaac cgtatacgaa ttatcattc acactgaccg 15480 cctgcacaag tgctggatgt acctctagtg agcccttcct tgggcaaact ctgcaagcgg 15540 ccccagaagg cgtttgggtc accccacgcc atattattat caactctacc acggttgagc 15600 tctactggtc ccttccggag aaacccaacg gcctggtatc ccaatatcaa ctctccagaa 15660 atggtaacct gctctttttg ggcggtagtg aggaacaaaa tttcacggat aagaatctcg 15720 agcctaactc aaggtatact tataaacttg aggtgaaaac cggaggcggc agttctgcca 15780 gcgacgatta tatagtacaa acacctatgt ccacaccgga ggaaatctat ccaccgtata 15840 acataacagt gataggtcct tactcaatat ttgtagcgtg gatcccgcca gggatattga 15900 tacccgagat tcctgtagag tataacgtgt tgctcaatga tggctccgtg actccgcttg 15960 cattctcagt tggacatcat cagagtacgc tccttgaaaa ccttacgcca tttactcaat 16020 atgaaatccg gatccaggcg tgccagaacg gtagctgtgg tgtgtcctct cggatgtttg 16080 tcaaaactcc tgaagcggca ccaatggacc tcaacagtcc cgtgcttaag gctttgggct 16140 cagcttgcat agaaatcaag tggatgccac ctgagaaacc aaatggtatc ataattaact 16200 atttcattta ccggcgcccc gcagggatcg aggaggaaag cgtgcttttt gtatggtctg 16260 aaggggcact ggaattcatg gacgaaggag ataccttgag gccatttacc ctttacgagt 16320 accgcgtgag agcttgcaac tctaagggga gcgtcgaaag cttgtggtct ctcacgcaga 16380 cccttgaggc tcctccccaa gatttcccag cgccgtgggc ccaggcgaca tctgcacaca 16440 gcgtgttgct taactggact aaacctgaaa gcccgaacgg catcatctca cattataggg 16500 ttgtttatca agaacggccg gacgatccga ctttcaattc tcccacggtg cacgccttta 16560 footaaggg tactagccat caggcccacc tctacggtct cgaaccattc acgacgtatc 16620 gcatcggcgt tgtagcggcc aatcatgctg gtgagatact tagtccgtgg acgctgatac 16680 aaactctgga gtctagcccc tccggtttgc gcaacttcat tgtggagcaa aaagagaatg 16740 gtcgagctct tctccttcaa tggtccgaac ccatgcgcac gaatggcgtg ataaagactt 16800 samaatatt ctcagatggc ttccttgaat actcaggttt gaaccgacag ttccttttta 16860 ggaggcttga cccattcact ctctacacgc tgacgctcga agcgtgtacc agggccggtt 16920 gtgcacattc tgcaccacag cccctttgga cggatgaggc tccccctgat tctcaactcg 16980 ccccaacggt ccatagcgtc aagtcaacct catagagagct tagttggtct gagccagtga 17040 accctaatgg aaaaattata agatacgaag tgatacgccg atgtttcgaa ggtaaagctt 17100 ggggcaatca aacaatccag gccgatgaga aaattgtatt tacagaatat aacaccgaga 17160 ggaacacgtt catgtataac gataccgggc tgcaaccttg gactcaatgt gaatataaaa 17220 tttatacttg gaattcagcg ggccatacct gctcctcatg gaacgtggtg cggactctcc 17280 aagcaccgcc ggagggcctt tcaccgcctg taatctctta cgtttccatg aatcctcaga 17340 agctccttat ctcctggata ccacccgaac aaagcaacgg aatcatacag tcttatcggt 17400 tgcagaggaa cgagatgttg tatccttttt ccttcgaccc cgtcactttt aattatacag 17460 acgaggaact gttgccattt agcacctatt cctacgccct tcaagcgtgt akgagcggtg 17520 gctgctccac atccaaaccg accagcatta ccacacttga agccgctcca agcgaggtct 17580 ctccacctga cctctgggcc gtttctgcta cgcaaatgaa tgtgtgctgg tccccgccca 17640 cggttcagaa tgggaaaatc actaaatacc tcgtaaggta tgacaataaa gaaagtctgg 17700 ctggacaagg gctctgcctt ctggtcagtc acctgcagcc ttatagtcaa tataatttta 17760 gtctcgtagc gtgtaccaac ggcggctgta cagcctctgt ttcaaaaagc gcttggacaa 17820 tggaggcctt gcccgagaac atggactctc cgaccctgca ggtgaccgga tcagaatcca 17880 tagaaatcac gtggaagcca ccacgcaatc cgaacggaca gattcgaagc tacgaattgc 17940 gccgggatgg gactatcgtg tacacagggc tggagactcg atacagggac ttcaccctga 18000 ctccgggagt ggaatattca tatactgtca cggccagtaa ctcacagggt ggaatcctga 18060 gtcctctggt gaaagatcgc actagtccca gtgcgccctc tggcatggag ccaccgaaat 18120 tgcaagcaag agggccccag gagatcctgg tcaattggga tcctcccgtt aggactaacg 18180 gtgacattat taactacacc ttgtttatca gggagttgtt cgaaagggaa accaagatta tccacattaa tactacccat aacagcttcg gcatgcagtc atatattgta aaccaattga aaccgttcca ccgatacgag attaggatcc aggcatgcac gaccttgggg tgtgcatcca gcgactggac gtttattcaa acgccagaaa ttgcgccttt gatgcagccg cccccgcacc 18420. tggaggtaca gatggctccg ggcggggttcc aacccacagt ctctcttctc tggaccggac 18480 ctctgcagcc taatggaag gtcctgtatt atgaacttta cagacgacag atcgccactc aaccccgcaa atccaatcct gtgcttatct acaacgggtc tagcacgagc ttcattgata gcgaattgct gccttttacc gagtacgagt accaagtgtg ggcagtgaac tcagcgggga 18660. aagcgccttc ctcttggacg tggtgccgga cgggacctgc gccacctgag gggctgcggg 18720 cacccacgtt ccatgttatc agctcaacgc aagctgtcgt aaatatctcc gctccaggta 18780. aaccgaacgg tattgtatcc ctctataggt tgttctcttc ctcagcacac ggcgctgaaa 18840 cagtactgag cgaaggaatg gcaacccagc agaccctgca cgggttgcag gcctttacga 18900 actatagcat tggagtcgaa gcgtgcacct gcttcaactg ttgtagcaaa ggccccaccg 18960 cggaactccg gacccatcct gctccaccct ctggcctgtc ctccccacaa atagggacct 19020 tggcctctcg cacagcgagt ttccgctgga gtccgccgat gtttccaaat ggggtcatac 19080 attcatacga gcttcagttc catgtcgcgt gcccccctga ctctgcgctg ccttgcaccc 19140 caagccaaat cgagactaag tacacagggc tgggccagaa agccagtctt gggggtctgc 19200 aaccctatac gacgtataag cttagggtag tcgcccacaa cgaggtaggt agtacggcca 19260 gtgaatggat aagtttcacg acacaaaaag agttgccaca gtaccgggcc ccgttctccg 19320 tggacagcaa cctcagtgtt gtatgtgtca attggtctga tacattcctt ctgaatgggc 19380 aacttaagga atatgttttg acagacgggg ggagacgcgt ttactctggg cttgacacga 19440 ctctgtatat tccacggacc gcggataaga ctttttttt tcaggtgatt tgcactactg 19500 acgaggggtc tgttaaaacc ccgttgatac agtacgatac ctcaacgggga ttgggtctcg 19560 ttcttacaac cccggggaag aaaaagggca gcaggagcaa atctacagag ttctatagtg 19620 agctgtggtt tattgtcctg atggccatgt tgggtctcat tctttttggcg atatttctgt 19680 cactcatatt gcaaagaaag atacataagg agccttatat cagagagcgc cctcctttgg 19740 ttcctttgca aaagagaatg tcaccgctta acgtttatcc gcctggtgag aatcacatgg 19800 gactggctga taccaagatc cctaggtctg gtacacctgt gtctatccgc agtaaccgct 19860 ccgcttgtgt actgcgaata ccgtcacaga atcagacaag tctgacatac agccaaggca 19920 gcctccaccg gagtgtatct caactcatgg acattcagga taagaaggtc ctcatggaca 19980 acagcttgtg ggaagcgatc atgggtcaca actctggcct ctatgtagac gaggaagacc 20040 tcatgaatgc aataaaggat ttttcaagcg tgactaaaga acggacgact ttcacggaca 20100 cgcacttgta ataaggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt 20160 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga 20220 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt 20280 tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc 20340 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc 20400 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg 20460 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat 20520 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg gccggccatc 20580 gatgtagaa attackattac aagctaagc aacagctgtg ggtgaagaa gagagaaat 20640 aatgtgtttt ttctttctc tttgatgca aaccacag actctgccta aagcagaact 20700 gtagttaaaa ggagactgca acttcagagt ttccattac ctagttttctc aacactggca 20760 gattttttta ggagctagta tctcttgtga aaacata gagttcagc accagaaag 20820 aaatggtggg ccaccactaa ttaaaaacaa aaatgagccg ggtacaatgg ctcaccccca 20880 tgacccagc actttgggag gccgaggtgg gtggatcaca aggtcaggag tcgagatca 20940 gcctggtcaa catggtgaaa cccccgtctc tactaaaac aaaaatta gctgggtgtg 21000 gtggcgggcg catgtaatcc cagctactag ggaggctgag gcaggagaat cgcttgaacc 21060 tggaaggtgg aggttgtagt gagccgaaac cgcgccactg cactccagcc tggccaacaa 21120 gagcgagact ccgtctcaa aaaaaaaaa aaaaacccaaaaaaaaaaa tgtgtaattt 21180 catgtccttc atgggttaag atagttttta aagaccttgc caaatattta ctgagttgcc 21300. 21300. 21300. 21300. 21300. 21300. 21300. 21300 21360. tgcaaccctc ttaaaaaaaa gtcactgtcc ttatggagct tactggggag gcaaataaaa 21420. 21420. 21420. 21420. 21420. 21420. 21420. 21420. 21420 21480. aaacttgcac acaaatgatt atagcaattt tgttcataaa ctaccaaaaa ctgtccttga agttttgatg cccttcaata tgtccatgga taaactgtgg tacattcata cacagtgaaa tattattcag cattaagaa tagctacca accccacaaaa agatgtggaa gaaacttaat gcatgttgtt aactgaaagt gcctaaaagg gctccatact gtatgatccc aactatctga cattctggaa atgggtacag agtcagtaaa aacagctggg catggtggtg catgcctgta atcccagcta cttgagaggc caaggcgga ggactgcttg agtcccagga tatcaaaacc agctcagaca actagcaag actccattcc aaaaaaaaaaaaaaaag agaaaaagat 21840 aacagtggtt gcaagggtt gggagaagg gaaggggg atgaggcga ggagcacaagg 21900 ggagttttag ggcattgaaa ctattctgca tgatactgta atgggggaga catgccactg 21960 tgcatttttc caatacaca gatgtaca cacaagaat gatgttaaa ctatgaacct 22020 tggttaataa tgaatcaaa tcggttcatc agttgtaaca atacaccac actaatgcat 22080 gatgttaaca ggagaaactg cggggaagcg gggtatatgg aaattcccta taatcaccgt 22140 gtatttttc tgtaacctg taactactct aaaaaata agcccgtttt aaaagaact 22200 tgaaaaaata attccccaa ccaaaaatca agtcaataa atcactgtc ctcatgaat 22260 ttgcgaga ggcacagaag aaaaaaaact aggcaataa tagtaaaa taggtaa 22320 atatccccta aggcatacaa caggcataat aaaaaaaac accatagta caactccccc 22380 atctccctcc cactcctcat agatggttta ggctaaggtt gataagtggg agaaaacttt 22440 tggtttctga aacatctcat agttaagcgt atctggatgg gtgcggtggc tcatgcctgt 22500 aatccccaac actttgggag gccaaggtgg gtggatcacc tgaggttggg agttcagac 22560 tggtctggcc aatatggtga aactctgtct ttactaaaaa tacaaaaatt agctgggcct 22620 ggtggtacat gcctgtaatc ccaactactc aggaggctga ggcaggagat cacttgaacc 22680 tgggaggcgg aggttgtggt gagccgagat catgccactg cactccagcc tgggcgacag 22740 agcaagactc tgcctcaaaa aaaaaagagt atccggttcc ccttcaaata tcaaatttc 22800 aaagacatta ataactgaaa tagaaagtgc cagagaaatg ctgaaagttg cttaatctag 22860 taaatagctg aaataatag gaatttctct ctttccaatc ataagcaaac cagtcattaa 22920 aatctcagac acatacacac actcacacaa ccccatcttt gttcttgctt caatttttgtt 22980 ttagatacaa tcgcaaccat acactctgta ataacatcac tgccaaaatac tcaaaccaca 23040 tgaaaaggca aagtgccttt tctattgtgg aaataagccc atgttgggga gaaatcctgg 23100 gaatctcgca caagaaacgc ttattacgt gtccagggcc gcctctgcca tgtttaccac 23160 aataatact ttaaacttca aaagaaacgc acacacagtc taggaacaa tgaaggcttt 23220 tatatagtcc aatgaaaatg ctttggaaaa tgaaagtat aagaactgac agtacatggg 23280 gattttttg tctcagaatc ttaatggaga atataaacta taagcaggaa ataggtcaga 23340 tataacagca gacaaatcag tttcaagaag tcctacaagg aaaaatttgt gcaaaatgtc 23400 aggttgctaa ttatcattc agtaaacatt tagggaatgt gccaagcagt cttttaagcc 23460 ctggggattc agcgacaaac aaaaacagat gcagcttctg ctctcaaaag ctgacatcct 23520 ggctgggcgc tgtggctcat gcctgtaatc ccagcacttt gggaggccga ggcaggcaga 23580 tcacctgagg tcaggagttc aaaaccagcc tggcacgt ggtgaaaccc tatctctact 23640 aaaaatacaa attagccag gtgtggtggc aggcgctgt agtcccagct acttgagagg 23700 ccgaggcagg agaatcactt caacccggga ggtggagttt gcagcgagcc aagatcgcac 23760 cactgcactc cagcctgggc gaaagagtg aaacttac tcaaaaaaa aaaaaaaagg 23820 aagctgccat cctaatgggg gagacatt tgacagatt gagagaac acatgcagca 23880 aaacagggtg atgggatgaa agcatgatgg gctgctactt tggtctaagc ggtcaaggaa 23940 ggctctctg aggaggtgag gcctgataa caaaaacca ggacggtgaa gggcagagggg 24000 aagaatttctg gtgaagggg aaggagcc agaggcgctg gagtggaag agctgggccc 24060 acttggaggc acagggcgga ggggctagca gcaggcgcag ggagagctgc tgaggggca 24120 gataggaggc aggcaggcag cacaggtttt gtggggcagg gcaagtgct tagaaaagc 24180 caggaatgg ctttaagtag ggtctgacat catccgattt ccatttta agattactg 24240 tgctgtgaag cccagaaagg ccgagagaaaaaattact tagatta cagcctacag 24300 gactcacag ggtgtctgtg gggatttggg aagtgttagt gtcaaagtgt atctgagatc 24360 tgaggccagg tgtgtggct cacgcctgta atcccagcac tctgggaggc tgagataggc 24420 agatacttg aggtcaggag tttgagacca gcctggccaa catggtgaaa cccatctct 24480 actaaaaaca gaaaaactag ccaggcattg tggtgcgcac ctgtaatccc agctactcag 24540 gaggctgagg caggagaatc gcttgaacca gagattgcgc cactgcactc cagcctggggt 24600 gagagagcga gactccgttg tcttaattta aaaaaaaa aaaaaaatct atctgaggca 24660 gaacaataga acgcggcaaa cgtgaagaaa ctttcagatt ctcctcccac ctggcacgct 24720 gctcccctcc cccatcgctc tctcagctga tgacaggga aggaagccca ggctgcaccc 24780 cctcaccaaa tgctgaccgc cccaggcagg aaagggctg cccttgctct cagccaaggc 24840 cagctgcacc ggtagtgctt gggattccaa ggcctccatg cctgggactc tcacctctgt 24900 ccctgcccct actctccctt gtaccagagc aggacaggct ctcacagcct acctttggaa 24960 cagaacagga cacacattta aaaaccctct ctaaactcta caaacccaga gaaagtccta 25020 ctttcttgtt ccaagaaacc tctttatcgt tgttagttct taacatcaca tttctgcttt 25080 ggcctacttc aattgggctt ctagatccaa tattccaagg caacggcttc ttacaatatg 25140 tgcaaaacaa acagtccctt ttctgttctc aaatgcattc taaaaggttg gaatttcaca 25200 acttttcttc tgctggttcc tccacagtac acctaccaag cctttacccc aaggctttct 25260 tgaccctctt ctctataaaa tccaggccat cccatcctgt ctcatgtctt caaaaagatc 25320 tacgtggcag agacattcca atctccatct cccccatgac atcttttcag aactccagac 25380 tactcaactt ctgtgctctt cagtccaaat ggcgtctcaa accagatggt tatcagatgg 25440 ctacaggagg cattacggaa tgtatcatct tctgaatgtg aaccacatca gatgcaatac 25500 agaaacac acactctcca ggcacagcaa accatgccac agaacgctcg ggctttacct 25560 gtgatgggaa gcttgggctg cagtgtgtac agctggggcag gggtctgatg tgggttcatt 25620 ccatatctca agggcatctg agatacgcct ttgctgtact gtttcctgaa ataatctgaa 25680 attgcttcat catattgtgc cgtatgagtg aatgcctaca aaaagcgtaa gttttgtgag 25740 acacgagcag cactttccaa agtggggca agatggttt ctacgtccac agcaaac 25800 atctcctcac tattcagtgc tatgtggtgg ctgcacaaca cgatgacacc atgaatgaca 25860 gcaaagcact gaatggccca acaaattacc cctttccaat tctttttcaa tctttttttt 25920 tttttttgag acaaagtctc actctgctgc ccaggctaga gtgcagtggt gtgatgtcag 25980 cttactgcaa cttccacctc ctgggttcaa gcgattctcc tgcctcagtc tcccgagtag 26040 ctgggattac aggcgtacca caccacacca cactcggcta atttttgtat tttcagtaga 26100 gacagggttt caccacattg gccaggctgg tctcaaactc ctgacctcag gtgatccgcc 26160 cgcctcggcc tcccaaagtg ctaggattac aggcgtgagc caccgcgccc agctctcttt 26220 caatcttttt gacagaacac ctctgtaata gctggtttct ccatttttaa caaaaggata 26280 gcagctccaa gcaaagcttt ggtaggtaat gcaatctacc aagaactgaa tactgcttgg 26340 tctggcaaac actgcactcc atggatgtaa cttacaatat catgaaagtg catcccacct 26400 tcaaggctaa ctggcgtcta gtctccaagg aggtgtcctt actctcggag ctctgcatct 26460 ccgtggacac caccacatag tcctctggtt cacacaccac tgtcactcga gcgtggtttt 26520 tggctgcagc tctcagtaag gttactccac ctttgaagaa caaaagactt attgattttt 26580 gtatgtccct atcagcttat gaaccgcttt taaataagt cattgtgcat gtacttccat 26640 tttaattcct gacttaaaa tcaggaaat aagctggcg cagtggctca tgcctggaat 26700 cccagcactt tgggaggctg agcaggtgg atcacctgag gtcaggagtt cgagaccact 26760 ctggccaaca tggcgaatt ccatcttat aaaaaataca aaaaaaatta gccaggcgtg 26820 gtggtgtgca cctgtaatcc cagctactca ggaggctgag gcagagaat cacttgagcc 26880 caggaggcag aggttgcagt cagccaaaac tgtgccactg tactccagcc tggtgacac 26940 agcaagactt catctcaaa aaaaaaat aaaaaaat tcaggaaaat 27000 aacccagatg catctctgca tacaaaaaaa gcttagaaa tataaatgaa aaaaagaaaa 27060 tataaatgaa ttatagtaat cccaaaacac aatccagaag tagctatttt ctttctattt 27120 cctcagtaat aaaattcaa atgaatctc actaaaattt aaataatcct ctcctctcag 27180 tcttctaaaa tggttttct gacttaccaa tgtcaatttg ctccacagcc tcccacacag 27240 ttacacctgg agaagccact gtctttacaa agggatagag attgcaggca acactcact 27300 aaaacaaaga cgaatctggc tgtcattaaa aagtccactc atgtggttta cagctatcta 27360 ggaaaaaaac aagtcagtac tattaacgta aaaaaagcc atagagaact atatataa 27420 ctgtgagcct gatattag aattctaacg agtgtgtta aaagtcaggg tgtcctgctg 27480 gggtcgagca gaggtaattt agtcagcagg tgctccccgg atctgaatac acagtactcc 27540 ctccacgtca cactgcaacg ctgctgcttg catacctta gctaccaccaga accaccaagc 27600 catgaaaaat ttatagaaca ttttataga agtataaagt acagattac agatacatt 27660 ctcattttgg aatgtgatca ctattatcca gaacattaga atttagattc ttttccagt 27720 ggattggcac atgatgaaa atatcagag cattctaagg gtgaacatt tgttttgcct 27780 tcagagaggt cataaaaatg atggtacag gatgccaaa agataattt aaatacaga 27840 agtaatctaa aattaacaat cagtgaagta cagttccact cctaggcctg tagtgcgagg 27900 agcttacgg gcaggctcca aacaagccta actgctgaaa actattaaa aaaaaaaaaa 27960 aaaaaacag agaacagaa taggaggga aacatttgc agctcatctt tattaagtct 28020 actgccctcc tactaaaacc agacaatgt cacaagaaa ctagagacca gtatctctta 28080 ggtatcttca cacaaaact cacacaaaa ctgaatccag aaaaatgcaa 28140 aaaccacctg atcatctca tagagtaca aaaaggatct gatacattc aacaggtatt 28200 catatagaaa cactcagcaaaaaaaa gagggaact tactgaacat gaaagtgt 28260 aagaaaaacc cacagctaac atcatacttg atggtgaaag actaaaatac ttgtctactc 28320 aactcagaaa caacacaga ggccaggcgc agtggctcat gcctataatc ccaccacttt 28380 gggaggctga gccaggcaga tcacctgagg tcagaagttc gagaccagcg tggccaatat 28440 ggtaaaaccc catctctact aaaaatacaa aaattagccg ggcaaggtgg cgcacgcctg 28500 caatcccagc actttgggag gctcaggagg atggatcaac tgaggtttgga agttcaagac 28560 cagcctgacc aacatggaga aaccccatct ctactaaaa tacaaaatta gccaggcaag 28620 gtggcacatg cctgtaaccc caggctgaag cagataatc acttaaaccc gggaggcgga 28680 ggtcacagtg agccaagatc acgcaactgc actccgcctt gggcaacaag agcaaaacgc 28740 catcttaaaa aaaagaaaca aaacaataat gtccattctt accacttcta ctgaacactg 28800 tactagaagt tctagccaga caattagca aagaaaaga aataaaggc atcaggactg 28860 gaagaaagc agtgaagtta tatctatttg cagatgatac cacctagtat acagaaaatc 28920 ccaaattcac aaaaataacc ttttagcact aataaatgag ttcagcaaag ccgcaggata 28980 tatgatcaat atataaaaat caattgtatt aaataca ttagcagtga atgaaaaa 29040 aattaagaaa acaattctac ttcacaagac aatgttaaag aaattaaaga caccccatgt 29100 tcacagactg gaagatttaa tattcttagg atggtaacat ttcccaaatt gatctacaga 29160 ttcaacacaa tacctatgaa aattccaact gcgttgtttt gcagaaatgg acaagctgat 29220 tctaaaaatc atgtggaaat gcaagggact ctgaagagtc aaagtcatct tgaataaaaa 29280 aaacgaagtt ggggccgggc gcagtggctc acgcctgtaa tcctagcact ttgggaggcc 29340 gaggcgggcg gatcacgagg tcaggagatc gagaccatcc tggcgaacc ggtgaaaccc 29400 tgtctctact aaaaatacaa aaaatagc tgcgcatggt agcgggcgcc tatagtccca 29460 gctactcggg aggctgaggc aggagaaggg cgtgaacccg ggaggcagag cttgcagtga 29520 gccaagatcg cgccactgca ctccagcctg ggcaacagag ccagactccg tctcaaaaac 29580 aaaacaaaac aaaacaaaca aacaaacaaa aacagaaaa atgaagttgg aggactcaca 29640 cttgctgatt tcaaaatttg ttaccagcct atagtaatag tgtagtactg gcataacagc 29700 aggcacatgg accaagggtg ccaagaccat tcgattgggc aaagaacagt cttttcaaca 29760 aatgatgctg agaactag acagccacat aaaaaaa ataagacttc tatttcatgt 29820 ctgtatatga aaattaactt aaaatgaacc aaagacttaa gtataagagc taacactaaa 29880 ataactctta aaagaaatgt acacacgccg ggtgcggtgg ctcacacttg taatcccagc 29940 actttgggag accgaggcag gtggatcacc tgaggtcagg agttcaagac cagcctggcc 30000 aacatggaga aaccccgtct ctactaaaaaa cacaaaaaaa ttagctgggc gtggtggcgg 30060 gcacctgtaa tcacagctac ttgcgaggct gaggcatgag aatcgcttga acccaggagg 30120 cgaaggttgc agtgagccga gatcacgcca ctgcactcca gcctgggcaa caagagcgaa 30180 actccatctc aaaaaaaaaa aaaaaaagaa agaaaata cacaaaaatc ttcatcactg 30240 aggattaggc aatggttct tegcatgac accattaca caagcaaaaaaaaat 30300 aaattcaaca tcatcaaat taaaaacttt tgtggccgg gcatgtgc tcaagcctgt 30360 aatcccagca ctttgggagg ccaggcggg cagataaccc gaggtcagga attcagacc 30420 aacctggcca acatagtgaa accctatctc tattaaaaat ccaaaaaaat tagccaagtg 30480 tggtggcggg cacctgtaac cccagctact catgagactg agcagaatc gcttgaatcc 30540 gggaggtgga gggttgcagtg agccagatg acgccactgt gctccagcct gggcgagagt 30600 aagactccgt ctcaaaaaa agaaaaaact tgtgatgatc aaggaaacc atcaagaag 30660 gctaacacaa tccatagaat gagagaat aaacttacaa ctcaataata aacataaa 30720 tacctcaact aaaaatggac agaggtctc aacagacatt tctccaagg aggtaaaca 30780 ctggctagga agcacatcca acggtgttcc acattattg gaagcaaat tataaccaca 30840 atgagatgcc acttcagaac ccctagaatg actagaatca aaagacaga tagtaacgtt 30900 gatgaggatg tggagaact ggagccccat ccagcacact gctgctggaa atgtaaaatg 30960 gtgcagcact ctgaaataca gggtcgcagt tcccaaag ctaaacatag ttaccatatg 31020 accaaggaat tccactccta ggtatatacc ccaatgaaac gatgtgtat gtccacacaa 31080 aaacttatac acaatatttt atagctgcat tattcataat attcaaag tagaacaac 31140 ccaaatgtcc atcaaat gaactgata acaagggt atgaccac atgattat 31200 tttcagcac taaaaacgaa atgaactatt gatactatgc tataacatgg ataaccttg 31260 aaaacatgct aagtgaaaga agccaggaac aaaaaacca tatattgtta gattctattt 31320 attgaaaat gtccactaca ggcaatcag acacgtaaca attggtggtt gccgaggctg 31380 gaggacgttg gggaaatgga gaataaatgc gtagggtat agggttttagg gtggggttga 31440 tgaatatgtt ttaatattgg atgtggcaat taatatggtt tggatgtgtg tcctctccaa 31500 atctcatttt gcaatgtgat cccaatgtt ggaggtgggg cctggtggga gatgattggg 31560 tcacggggga gatcccttat gatggcttg gtgccctccc cataatgagt cctcactctg 31620 agttcatgta gagctggttg ttacagag ccggcaggc cgggcgtggt ggctcacacc 31680 taatcccagc actttggg gccggggtgg gtgcatcatt tgggtcagg agttccaga 31740 gcagcctgac siacatggtg aaacccatc tctaaaa atagtaaaa tgagttgggc 31800 gtggtgacaa acgcctgtaa tctcagctac tcaggaggct gaggcaggag atcacttga 31860 acacaggagg cagaggttgc agtgagccaa ggtcatgcca ctgaactcca acctgagcaa 31920 cagagcgaga ccctgtctca aaaaaaatt aaaaaaaa gttaaaaagt 31980 aaaagagctg ggcaactcct cctctccctc tctcttgctt cttctctcat gggatatgcc 32040 agctccccct ctgctttcca ctgtgattat aaacttcctg aggccctcac cagaagttga tgctggcacc acacttcttg tactgtctat agaatcacga gccaaaataa acctgttttc fathers cagcttctag fathers tagcaatgca gaacagacta acacagtaat ggttgcacaa cttctgtgaa ccactgattt gtacatttta aattggtgag ttgtatggta tgtggattag atctcaataa agctgttacg father agaaacagcc agtatcttta 32400. tgcaataac tcagttagtt gtgcaagttg fatherccta fatheraaac ctgaaatctg aaatgctcca aaatctgaaa ctttttgagc gtcaacatga cacttggagg aaatgctcac tgaagcattt cggatttctg agggacgctc aactggcatg ttacctgcaa atattccaaa atctgaaga atctgaaatc caagacactt ctgggtccca agcatttcag ataagggata ctcaacctat acatactaag atttagattg taacttccta aaaaacttca gtatgtattt 32640 tggtataataa atctggccaa accaaaatgc ttacacacat acaaagtcac acagaaggta 32700 cgcagtaaag gtttatagtg agtaagaaaa cacagggaca ccaggtttgg catagtcttt 32760 gtcgttggtt cgtaatgtgt taaaagttta atttcaggtt tttaccttat aagattgaaa 32820 tcaagtctgg ccatgtcagc attatcttct ggaatattac gagctaggat tcctaattta 32880 aaaaagttaa acaaagtcat tagcttcaaa aaacaagtca gattaaggaa tgtctttcaa 32940 catagtgtct tcaatcaaat taaatcccat tgattaaaaa aaaaatttat ttttttgaga 33000 tggtctcact ctatcgccca ggctggattg tagtggcgta atcacaactc actatagccc 33060 tgacctccag ggctcaagtg atcctcctac ctcagcctcc caagtagctg ggactacagg 33120 aatgcatcac cacacccagc tggtctttc aattttttaa agaaaggggg ggtgttgcca 33180 tcttgcccag ggtgatctta aactcctggg ctcaagcggt tctcctgtct cagctctcca 33240 agtgctggga ctacaggcaa gagccaccat gcccagcata atattcttaa ttaaagaata 33300 tcaaacatgt agttggggag tttcaaaca aaaaaattag gattaacaga tatatctggc 33360 tggccacagt ggctcatgtc tgtaatacca gcactttggg aggctaaggt gggggatcgc 33420 ttgagcccag gtgttcgaga ccagcctggg caacaaggtg agaccccgtc tctataaaaa 33480 atacaaaaat tagctgggtg tggtggcaca tgcctatagt cccagctact tgggaggctg 33540 aggtgggaga actgcttgag cctgggaagc agacagaggc tgcagcgagc catgatcgca 33600 ccactgcact ccagcctggg caagagtata atcctgtctc aaaaaaaaa aagagagata 33660 tatttaaaaa tctttttctt cagttcttga tcccagtttc ttgagcaata aaatgtaaaa 33720 aaggtaaaag gaatttaca aaactgttgc tagttccaga acagtctatg cgtttctgta 33780 agtcaatggc ttgtctgatg atctgtagcc attcagcttt attcaccaaa ttgtaccaca 33840 actgacaaac taggaggata ccagtttctt gccgtttcgg tttatataca attcattctt 33900 aggtgaacga acaaatgtaa agcactttga aaaaaaaaat tcctttgcaa gtcagttacg 33960 ctgtgatgct ttcttctaag tttgaactat tttaaaagta ttttcatacg atgaaatcta 34020 aatatcttca gcaacttgag caggaaacac cagactctaa attaggtatc aggtcaggac 34080 taatagcaag cagaggaaaa caatctgaat tcatcaagta gtaagacaag ctggatttta 34140 aaccagttct aacgtattat gaaatctggc agcaacttca taacattaac tttatatgta 34200 aatttctctt gagccttttt cttcctttat tttaggacgc tatcaactaa aatattcctg 34260 gaaaccactg actgttttta cattaaagat accaaccact taccagcatg gactgcagga 34320 tgcaaagttt tcacacgtcc ccccaacatt tcaggaaatc ccgtcaactc agagacatct 34380 ctgaaaaaca gataaaacaga atggtactac tggtgtggct gattgtaat ttcactatt 34440 caaagcatct actgttttca ttcagaga tttgagcct gattcaacg tgaaaatttct 34500 gatttttaa agtatttcac tatattttat ttcttaagta aaattctatt tacttccgaa 34560 aaaaagttta tttaatttat ttcctactg ttgtgtagt tccgtacgat taagggcgaa 34620 ttccagcaca ctggcggccg ttactagtgg atccacagt ctccacgtaa acggtcaaag 34680 tccccgcggc cctagacaaa tattacgcgc tatgagtac acaaattat tcagatttca 34740 cttcctctta ttcagttttc ccgcgaaaat ggccaaatct tactcggtta cgcccaatt 34800 tactacaaca tccgcctaaa accgcgcgaa aattgtcact tcctgtgtac accggcgcac 34860 accaaaaacg tcactttgc cacatccgtc gcttacatgt gttccgccac acttgcaaca 34920 tcacacttcc gccacactac tacgtcaccc gcccgttcc cacgccccgc gccacgtcac 34980 aaactccacc ccctcattat catattggct tcaatccaaa ataaggtata ttattgatga 35040 tgattgttta aacatgatga gctcagc 35067

Claims

1. A viral vector comprising a transgene construct, wherein the transgene construct is one of the following: (a) a DNA sequence comprising a promoter element, an open reading frame of a retinal gene longer than 10 kb, and a terminator sequence; and (b) a DNA segment of a retinal gene longer than 10 kb.

2. The viral vector according to claim 1, wherein the vector completely lacks endogenous genes other than the ITR sequence.

3. The viral vector according to claim 2, wherein the vector has a deletion of sufficient endogenous adenovirus genes to make space for up to 10 kb for a cargo sequence.

5. The viral vector according to claim 1, wherein the viral vector is a mimivirus that has been completely or partially removed.

6. The viral vector according to claim 1, wherein the viral vector is an adenovirus from which all or part of the virus has been removed.

7. The viral vector according to claim 6, wherein the viral vector is capsidized into a capsid based on adenovirus type 5.

8. The viral vector according to claim 6, wherein the viral vector is capsidized into a capsid based on adenovirus type 6.

9. The viral vector according to claim 6, wherein the viral vector is capsidized into an adenovirus-based capsid and modified to remove the RGD sequence from the capsid protein.

10. The viral vector according to claim 6, wherein the viral vector is capsidated to a capsid based on adenoviruses of families A to G.

11. The viral vector according to claim 6, wherein the introduced gene construct is a DNA sequence comprising (a) a promoter element, an open reading frame of a retinal gene longer than 10 kb, and a terminator sequence.

12. The viral vector according to claim 11, wherein the promoter element is a human rhodopsin kinase promoter.

13. The viral vector according to claim 11, wherein the promoter element is a human rhodopsin kinase promoter.

14. The viral vector according to claim 11, wherein the promoter element is a mouse rhodopsin kinase promoter.

15. The viral vector according to claim 11, wherein the promoter element is a ubiquitous CMV promoter.

16. The viral vector according to claim 11, wherein the promoter element is an upstream non-coding sequence of the Usherin genome from nucleotides XXX to YYY.

17. The viral vector according to claim 11, wherein the promoter element is a synthetic transducin alpha-subunit promoter.

18. The viral vector according to claim 11, wherein the promoter element is the minimum sequence of a photoreceptor-retinoid-binding protein promoter and a human transducin alpha-subunit promoter.

19. The viral vector according to claim 11, wherein the promoter element is a chemically controlled tet-inducible promoter.

20. The viral vector according to claim 11, wherein the promoter element is a chemically inhibited promoter.

21. The viral vector according to claim 11, wherein the promoter element is a human L-opsin promoter.

22. The viral vector according to claim 11, wherein the promoter element is a human retinosulin proximal promoter and a human photoreceptor-retinoid binding protein enhancer.

23. The viral vector according to claim 11, wherein the promoter element is specific to a photoreceptor.

24. The viral vector according to claim 11, wherein the introduced gene construct is a nucleotide sequence that translates to Ush2A.

25. The viral vector according to claim 11, wherein the introduced gene construct is a nucleotide sequence that translates to ADGV1.

26. The viral vector according to claim 11, wherein the introduced gene construct is a nucleotide sequence that translates to an eyes shut homolog.

27. The viral vector according to claim 11, wherein the introduced gene construct is a nucleotide sequence that translates to hemisentin.

28. The viral vector according to claim 11, wherein the introduced gene construct is a nucleotide sequence that translates to versican.

29. The viral vector according to claim 6, wherein the introduced gene construct is (b) a DNA segment of a retinal gene longer than 10 kb.

30. The viral vector according to claim 29, wherein the DNA segment is at least a segment of the Ush2A gene.

31. The viral vector according to claim 29, wherein the DNA segment is at least a segment of the ADGVl gene.

32. The viral vector according to claim 29, wherein the DNA segment is at least a segment of the eyes shut homolog gene.

33. The viral vector according to claim 29, wherein the DNA segment is at least a segment of the hemisentin gene.

34. The viral vector according to claim 29, wherein the DNA segment is at least a segment of the versican gene.

35. The viral vector according to claim 5, wherein the introduced gene construct is (b) a DNA segment of a retinal gene longer than 10 kb.

36. A composition comprising a viral vector and a packing plasmid, wherein the viral vector comprises (a) a DNA sequence comprising a promoter element, an open reading frame of a retinal gene longer than 10 kb, and a terminator sequence, and (b) a transgene construct selected from a DNA segment of a retinal gene longer than 10 kb.

37. A method for treating a retinal disease, comprising providing a viral vector comprising a transgene construct, wherein the transgene construct is one of the following: (a) a DNA sequence comprising a promoter element, an open reading frame of a retinal gene longer than 10 kb, and a terminator sequence; and (b) a DNA segment of a retinal gene longer than 10 kb.

38. The method according to claim 37, wherein the retinal disease is selected from the group consisting of Usher syndrome type IIA, Usher syndrome type 2C, retinitis pigmentosa 39, retinitis pigmentosa 25, Wagner syndrome, and age-related macular degeneration.

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