Rapid and deterministic generation of microglia from human pluripotent stem cells

Abstract

To provide a method for generating microglia from stem cells.SOLUTION: Disclosed is a method including the following steps of: a) targeted insertion of a nucleotide sequence encoding a transcriptional regulatory factor into a first safe harbor locus on a genome and b) targeted insertion of a coding sequence of a transcription factor PU.1 into a second safe harbor locus on the genome, in which the gene is functionally linked to an inducing promoter regulated by the transcriptional regulatory factor; and expression of PU.1 and culture of stem cells obtained from steps a) and b) while being exposed to a growth factor or a small molecule, the growth factor or the small molecule mimicking signal transduction during at least one stage from the embryonic development of microglia, or from the proliferation, differentiation, or polarization of adult-type microglia. Further, the present invention relates to microglia obtained by the method, and various applications thereof.SELECTED DRAWING: Figure 5A

Description

Technical Field

[0001] Field of the Invention The present invention relates to a method for generating microglia from stem cells, the method comprising the steps of: targeted insertion of a nucleotide sequence encoding a transcriptional regulatory factor protein into a first safe harbor site on the genome; and targeted insertion of a coding sequence of the transcription factor PU.1 into a second safe harbor site on the genome, wherein the gene is operably linked to an inducible promoter regulated by the transcriptional regulatory factor protein; expression of PU.1; and culturing of the stem cells obtained from steps a) and b) while exposing to at least one of a growth factor or a small molecule, wherein the at least one of the growth factor or the small molecule reproduces signal transduction between at least one of the stages of embryonic development of microglia or the stages of proliferation, differentiation, or polarization of adult microglia. Further, the present invention relates to microglia obtained by the method of the present invention and various uses thereof.

Background Art

[0002] Background of the Invention Microglia are resident immune cells in the central nervous system (CNS) [Schafer et al., 2015]. They have their origin in the early yolk sac macrophages that arise during the first wave of primitive hematopoiesis in early embryogenesis. Primitive yolk sac macrophages spread via the bloodstream as soon as the circulatory system is established and colonize the developing CNS. In contrast to tissue-resident macrophages in other organs, microglia are not replaced by fetal monocytes during later stages of embryogenesis [McGrath et al., 303; Ginhoux et al., 2010; Gomez Perdiguero et al., 2015]. After the microglial population is established during early embryogenesis, microglia maintain themselves throughout life by local proliferation and are not replaced by cells derived from the bone marrow [Reu et al., 2017]. Microglia are uniformly distributed throughout the brain and spinal cord and play important roles in the development, maintenance, plasticity, and defense of the CNS [Schafer et al., 2015]. In a healthy CNS, “resting” homeostatic microglia are highly ramified cells with small cell bodies and thin cellular processes. These processes of microglia are motile and continuously sample their environment to scan for internal or external danger signals (such as incoming pathogens or signals locally produced by damaged or dying cells). Detection of such signals triggers microglial activation, which includes complete changes in microglial morphology, gene expression, and function. Upon activation, microglia retract their processes and transform into an amoeboid-like appearance. They actively migrate towards CNS injury along chemotactic gradients and secrete inflammatory cytokines.

[0003] Through a wide repertoire of cell surface receptors, including those for neurotransmitters and cytokines, microglia communicate with neurons, other glial cells, and peripheral immune system cells [Kettenmann et al., 2011]. Given their diverse functions and their unique positioning as a representative immune system in the healthy CNS, it is not surprising that microglia are involved in the onset and progression of many neurological diseases [Ransohoff et al., 2016].

[0004] Very recently, single-cell transcriptome profiling of microglia in a mouse model of Alzheimer's disease (AD) and in mouse models of other neurodegenerative diseases, including aging, amyotrophic lateral sclerosis, or tauopathy-related frontotemporal lobar degeneration (FTLD-tau), has revealed a pro-inflammatory transcriptomic signature in a small subset of microglia called the microglial neurodegenerative phenotype (MGnD) [Krasemann et al., 2017] or disease-associated microglia (DAM) [Keren-Saul et al., 2017]. The conversion of microglia from a homeostatic phenotype to a disease-associated phenotype is thought to occur as a response to changes in brain homeostasis in neurodegeneration and to follow a unique transcriptional program that is temporally and spatially regulated [Krasemann et al., 2017; Keren-Shaul et al., 2017; Butovsky et al., 1998]. In most cases, it remains unclear whether these cells have a protective or a disease-inducing / worsening function. The availability of healthy and diseased human microglia in vitro and in vivo can facilitate the identification of factors involved in both their beneficial and harmful functions and can facilitate the development of strategies to restore the homeostatic microglial signature or to induce the DAM microglial signature. This point could lead the inventors to target microglia for the treatment of neurodegenerative diseases.

[0005] The isolation or in vitro derivation of many human cell types remains challenging and inefficient. In particular, cells of the human CNS, including microglia, are particularly difficult to obtain. Previously, low-efficiency isolation from neurosurgical samples or postmortem brain tissue was the only means of access. Human pluripotent stem cells (hPSCs) are, in theory, an unlimited and renewable source from which all cell types of the human organism can be generated [Thomson et al., 1998]. This revolutionary discovery that human dermal fibroblasts can be readily converted into human induced pluripotent stem cells (hiPSCs) with the same properties as embryonic stem cells enables the generation of autologous and customized cell types for applications in regenerative medicine. For several important applications, including disease modeling, drug discovery, and cell transplantation, large-scale production of mature human cell types from hPSCs is required. Recently, the first hPSC differentiation protocol for the generation of microglia was published. This was based on first forming embryoid bodies (EBs) that were cultured for several months in the same, “neuroglial differentiation medium, the component concentrations of which were adjusted to match those of human cerebrospinal fluid,” supplemented with interleukin (IL)-34 and colony-stimulating factor 1 (CSF-1) [Muffat et al., 2016]. This promising publication provided a sophisticated medium composition for the final maturation and maintenance of human microglia. However, the wide applicability of this protocol appears to be hampered by the long duration of the protocol, the ill-defined early stages of differentiation (i.e., the EB-based intermediate stage hardly adheres to the principles of an embryo), and the requirement for several mechanical manipulation steps for cell purification.Subsequently, several other groups demonstrated the generation of microglia-like cells from hPSCs by classical differentiation approaches that are similar but different [Abud et al., 2017; Takata et al., 2017; Haenseler et al., 2017; Pandya et al., 2017; Douvaras et al., 2017]. Nevertheless, the in vitro induction of specific human cell types, including microglia, in the amounts and purity required for downstream applications remains a challenge, and alternative methods are currently being explored [Cohen et al., 2011]. A more recent manufacturing strategy, as contrasted with classical differentiation, is direct cell reprogramming [Ladewig et al., 2013]. This refers to directly converting any cell type (typically skin fibroblasts) into another cell type without proceeding through pluripotent intermediates. Although it provides a rapid route for cell generation from readily available cell types, the yield and purity of the desired cell population remain low and insufficient [Zhang et al., 2013]. Recently, a third route, called "forward programming," has been proposed to produce mature human cell types at unprecedented speed and efficiency [Zhang et al., 2013].

[0006] Forward programming, as a method for directly converting pluripotent stem cells, including hPSCs, into mature cell types, is recognized as a powerful strategy for inducing human cells. This involves the forced expression of key lineage transcription factors (or non-coding RNAs, including IncRNAs and microRNAs) to convert stem cells into specific mature cell types. Currently available forward programming protocols are mainly based on lentiviral transduction of cells, which can cause irregular expression or complete silencing of randomly inserted inducible cassettes. This creates a situation where additional purification steps are required to isolate the subpopulation expressing the required transcription factors. Therefore, further improvement of these methods is clearly needed.

[0007] Any improvement of the described method must ensure that the stable transcription of the genetic material contained within the inducible cassette, such as the transgene, is resistant to silencing and other negative effects associated with the integration site. Silencing can be caused by multiple epigenetic mechanisms, including DNA methylation or histone modification. Using prior art methods based on lentiviral transduction, the resulting cells are a heterogeneous population with transgenes that are either fully expressed, partially expressed, or silenced. This is clearly not desirable for many applications. Viral vectors have been shown to have a tendency to integrate their genetic material into transcriptionally active areas of the genome, thus increasing the potential oncogenic events due to mutagenesis upon insertion. For many applications, it is desirable to control the transcription of the inserted genetic material in cells such that the inducible cassette can be initiated as needed and transcribed at a specific level, including high levels. This cannot be achieved if the insertion of the inducible cassette is random in the genome.

[0008] The problem that microglia are involved in several serious diseases and are also involved with brain tissue in a state where their isolation from living tissue is difficult has been addressed in several publications. To overcome this problem, human stem cells are used, for example, by [Muffat et al., 2016] under defined culture conditions or by co-culturing with neurons derived from stem cells [Haenseler et al., 2017; Takata et al., 2017] to generate microglia or microglia-like cells. These methods rely solely on exposure to growth factors and cytokines to differentiate the stem cells into microglia.

[0009] Furthermore, in virtually all central nervous system diseases, including neurodegenerative diseases, neuroinflammatory or autoimmune diseases, autoantibody-mediated encephalitis or infectious diseases, neurovascular diseases, stroke, traumatic brain injury, and cancer, microglia play an important role, but the exact mechanisms underlying their role in various diseases remain unknown, so the need for this particular cell type is huge. The prior art agrees that microglia derived from stem cells actually reproduce the disease phenotype of the original patient [Muffat et al., 2016; Abud et al., 2017; Takata et al., 2017]. Based on this finding, the huge scientific gap regarding the involvement of microglia in several diseases can be overcome by generating microglia from stem cells. However, classical protocols for differentiating stem cells are very time-consuming and the results are not satisfactory.

[0010] Accordingly, the inventors of the present invention have developed a rapid method for generating microglia from stem cells by using stable introduction of an inducible cassette into the genome of the stem cells and further enabling control of the transcription of the inducible cassette and the transcription factors inserted thereby. The potential for these transcription factors to function as reprogramming factors for microglia generation was not previously known and is the unique finding of the inventors. This enables people to generate a pure microglia population that expresses all of the surface markers and RNAs observed in the native microglia population. Furthermore, this method can be used to differentiate microglia from human iPS cells of patients with neurodegenerative diseases and thus enables analysis of cell populations that would otherwise remain completely unavailable for medical testing. Thus, there is a strong need for manufacturing mature human microglia from easily accessible sources. Accordingly, the technical problem underlying this application is to meet these needs. The technical problem is solved by providing embodiments that are reflected in the claims provided below, described in the description, and illustrated in the examples and drawings.

Summary of the Invention

[0011] The inventors of the present invention have developed a method for generating microglia from stem cells.

[0012] The present invention relates to a method for generating microglia from stem cells, the method comprising the following steps: a) targeted insertion of a nucleotide sequence encoding a transcriptional regulatory factor protein into a first safe harbor site on the genome; and b) targeted insertion of the coding sequence of the transcription factor PU.1 (SEQ ID NO: 1) into a second safe harbor site on the genome, the gene being operably linked to an inducible promoter regulated by the transcriptional regulatory factor protein; expression of PU.1 (SEQ ID NO: 2); and c) culturing the stem cells obtained from steps a) and b) while exposing them to at least one growth factor or small molecule, wherein the at least one growth factor or small molecule reproduces signal transduction between at least one of the stages of embryonic development of microglia or the stages of proliferation, differentiation, or polarization of adult microglia.

[0013] In one aspect of the method of the present invention, the at least one growth factor or small molecule is selected from the group consisting of: activin A (SEQ ID NO: 7), BMP4 (SEQ ID NO: 8), FGF (SEQ ID NO: 9), VEGF-A (SEQ ID NO: 10), LY294002, CHIR99021, SCF (SEQ ID NO: 11), IL-3 (SEQ ID NO: 12), IL-6 (SEQ ID NO: 13), CSF1 (SEQ ID NO: 14), IL-34 (SEQ ID NO: 15), CSF2 (SEQ ID NO: 16), CD200 (SEQ ID NO: 17), CX3CL1 (SEQ ID NO: 18), TGFβ1 (SEQ ID NO: 19), and IDE1.

[0014] In a further aspect of the method of the present invention, the at least one growth factor is CSF1 (SEQ ID NO: 14) or IL-34 (SEQ ID NO: 15).

[0015] In an additional aspect of the method of the present invention, the at least one small molecule is CHIR99021, LY294002, or IDE1.

[0016] In another aspect of the method of the present invention, the first and second safe harbor sites on the genome are different.

[0017] In a further aspect of the method of the present invention, the method further comprises the insertion and expression of the coding sequence (SEQ ID NO: 3) of the gene of the transcription factor CEBPB.

[0018] In another aspect of the method of the present invention, the method further comprises the insertion and expression of the coding sequence (SEQ ID NO: 4) of the gene of the transcription factor RUNX1.

[0019] In a further aspect of the method of the present invention, the method further comprises the insertion and expression of the coding sequence (SEQ ID NO: 5) of the gene of the transcription factor IRF8.

[0020] In another aspect of the method of the present invention, the method further comprises the insertion and expression of the coding sequence (SEQ ID NO: 6) of the gene of the transcription factor SALL1.

[0021] In an additional aspect of the method of the present invention, the transcriptional regulatory factor protein is a reverse tetracycline transactivator (rtTA) (SEQ ID NO: 20), and its activity is controlled by doxycycline or tetracycline.

[0022] In another aspect of the method of the present invention, the inducible promoter comprises a Tet-responsive element (TRE) (SEQ ID NO: 21).

[0023] In a further aspect of the method of the present invention, the first and second safe harbor sites on the genome are selected from the group consisting of: the hROSA26 locus (SEQ ID NO: 22), the AAVS1 locus (SEQ ID NO: 23), the CLYBL gene (SEQ ID NO: 24), the CCR5 gene (SEQ ID NO: 25), the HPRT gene (SEQ ID NO: 26), or a gene having site ID 325 (SEQ ID NO: 27) on chromosome 8, a gene having site ID 227 (SEQ ID NO: 28) on chromosome 1, a gene having site ID 229 (SEQ ID NO: 29) on chromosome 2, a gene having site ID 255 (SEQ ID NO: 30) on chromosome 5, a gene having site ID 259 (SEQ ID NO: 31) on chromosome 14, a gene having site ID 263 (SEQ ID NO: 32) on the X chromosome, a gene having site ID 303 (SEQ ID NO: 33) on chromosome 2, a gene having site ID 231 (SEQ ID NO: 34) on chromosome 4, a gene having site ID 315 (SEQ ID NO: 35) on chromosome 5, a gene having site ID 307 (SEQ ID NO: 36) on chromosome 16, a gene having site ID 285 (SEQ ID NO: 37) on chromosome 6, a gene having site ID 233 (SEQ ID NO: 38) on chromosome 6, a gene having site ID 311 (SEQ ID NO: 39) on chromosome 134, a gene having site ID 301 (SEQ ID NO: 40) on chromosome 7, a gene having site ID 293 (SEQ ID NO: 41) on chromosome 8, a gene having site ID 319 (SEQ ID NO: 42) on chromosome 11, a gene having site ID 329 (SEQ ID NO: 43) on chromosome 12, a gene having site ID 313 (SEQ ID NO: 44) on the X chromosome.

[0024] In another aspect of the method of the present invention, the stem cells are pluripotent stem cells, induced pluripotent stem cells (iPSCs), neural progenitor cells, hematopoietic stem cells, or embryonic stem cells (ESCs).

[0025] In a further aspect of the method of the present invention, the stem cells are human or mouse stem cells.

[0026] The present invention also relates to microglial cells obtained by any of the methods according to the present invention, wherein preferably the microglia express at least one of the microglial surface proteins selected from the group consisting of: ITGAM (CD11B) (SEQ ID NO: 45), ITGAX (CD11C) (SEQ ID NO: 46), CD14 (SEQ ID NO: 47), CD16 (SEQ ID NO: 48), ENTPD1 (CD39) (SEQ ID NO: 49), PTPRC (CD45) (SEQ ID NO: 50), CD68 (SEQ ID NO: 51), CSF1R (CD115) (SEQ ID NO: 52), CD163 (SEQ ID NO: 53), CX3CR1 (SEQ ID NO: 54), TREM2 (SEQ ID NO: 55), P2RY12 (SEQ ID NO: 56), TMEM119 (SEQ ID NO: 57), and HLA-DR (SEQ ID NO: 58).

[0027] In a further aspect of the present invention, the microglial cells are for use in therapy.

[0028] Furthermore, the present invention is directed to the use of such microglial cells according to the present invention for the in vitro diagnosis of diseases. Preferably the disease is selected from the group consisting of: diseases of the central nervous system, preferably neurodegenerative diseases; more preferably Alzheimer's disease, Parkinson's disease, frontotemporal dementia, or amyotrophic lateral sclerosis; neuroinflammatory or autoimmune diseases, preferably multiple sclerosis, autoimmune encephalitis, or infectious diseases, neurovascular diseases; preferably stroke, vasculitis; traumatic brain injury, and cancer.

[0029] Furthermore, the present invention is directed to the use of such microglial cells according to the present invention for in vitro co-culture with brain organoids. [Invention 1001] A method for generating microglia from stem cells, comprising the following steps: (a) Targeted insertion of a nucleotide sequence encoding a transcriptional regulatory factor protein into a first safe harbor site on the genome; and (b) Targeted insertion of the coding sequence of transcription factor PU.1 (SEQ ID NO: 1) into a second safe harbor site on the genome, wherein the gene is functionally linked to an inducible promoter regulated by the transcriptional regulatory factor protein; expression of PU.1 (SEQ ID NO: 2); and (c) Culturing the stem cells obtained from steps (a) and (b) while exposing them to at least one growth factor or small molecule, wherein the at least one growth factor or small molecule reproduces signal transduction between at least one of the stages of embryonic development of microglia or the stages of proliferation, differentiation, or polarization of adult microglia. [Invention 1002] The method of Invention 1001, wherein at least one growth factor or small molecule is selected from the group consisting of: activin A (SEQ ID NO: 7), BMP4 (SEQ ID NO: 8), FGF (SEQ ID NO: 9), VEGF-A (SEQ ID NO: 10), LY294002, CHIR99021, SCF (SEQ ID NO: 11), IL-3 (SEQ ID NO: 12), IL-6 (SEQ ID NO: 13), CSF1 (SEQ ID NO: 14), IL-34 (SEQ ID NO: 15), CSF2 (SEQ ID NO: 16), CD200 (SEQ ID NO: 17), CX3CL1 (SEQ ID NO: 18), TGFβ1 (SEQ ID NO: 19), and IDE1. [Invention 1003] The method of Invention 1001 or 1002, wherein at least one growth factor is CSF1 (SEQ ID NO: 14) or IL-34 (SEQ ID NO: 15). [Invention 1004] Any method of the present invention, wherein at least one of the small molecules is CHIR99021, LY294002, or IDE1. [Invention 1005] Any method of the present invention, wherein the first and second safe harbor sites on the genome are different. [Invention 1006] Any method of the present invention, further comprising insertion and expression of the coding sequence of the gene of transcription factor CEBPB (SEQ ID NO: 3). [Invention 1007] Any method of the present invention, further comprising insertion and expression of the coding sequence of the gene of transcription factor RUNX1 (SEQ ID NO: 4). [Invention 1008] Any method of the present invention, further comprising insertion and expression of the coding sequence of the gene of transcription factor IRF8 (SEQ ID NO: 5). [Invention 1009] Any method of the present invention, further comprising insertion and expression of the coding sequence of the gene of transcription factor SALL1 (SEQ ID NO: 6). [Invention 1010] Any method of the present invention, wherein the transcriptional regulatory factor protein is reverse tetracycline transactivator (rtTA) (SEQ ID NO: 20), and its activity is controlled by doxycycline or tetracycline. [Invention 1011] Any method of the present invention, wherein the inducible promoter contains a Tet-responsive element (TRE) (SEQ ID NO: 21). [Invention 1012] Any method of the present invention, wherein the first and second safe harbor sites on the genome are selected from the group consisting of: hROSA26 locus (SEQ ID NO: 22), AAVS1 locus (SEQ ID NO: 23), CLYBL gene (SEQ ID NO: 24), CCR5 gene (SEQ ID NO. 25), HPRT gene (SEQ ID NO. 26), or a gene having site ID 325 (SEQ ID NO: 27) on chromosome 8, a gene having site ID 227 (SEQ ID NO: 28) on chromosome 1, a gene having site ID 229 (SEQ ID NO: 29) on chromosome 2, a gene having site ID 255 (SEQ ID NO: 30) on chromosome 5, a gene having site ID 259 (SEQ ID NO: 31) on chromosome 14, a gene having site ID 263 (SEQ ID NO: 32) on the X chromosome, a gene having site ID 303 (SEQ ID NO: 33) on chromosome 2, a gene having site ID 231 (SEQ ID NO: 34) on chromosome 4, a gene having site ID 315 (SEQ ID NO: 35) on chromosome 5, a gene having site ID 307 (SEQ ID NO: 36) on chromosome 16, a gene having site ID 285 (SEQ ID NO: 37) on chromosome 6, a gene having site ID 233 (SEQ ID NO: 38) on chromosome 6, a gene having site ID 311 (SEQ ID NO: 39) on chromosome 134, a gene having site ID 301 (SEQ ID NO: 40) on chromosome 7, a gene having site ID 293 (SEQ ID NO: 41) on chromosome 8, a gene having site ID 319 (SEQ ID NO: 42) on chromosome 11, a gene having site ID 329 (SEQ ID NO: 43) on chromosome 12, and a gene having site ID 313 (SEQ ID NO: 44) on the X chromosome. [Inventive concept 1013] Any method of the present invention, wherein the stem cells are pluripotent stem cells, induced pluripotent stem cells (iPSCs), neural progenitor cells, hematopoietic stem cells, or embryonic stem cells (ESCs). [Inventive concept 1014] Any method of the present invention, wherein the stem cell is a human or mouse stem cell. [Invention 1015] Microglia obtained by any method of Inventions 1001 - 1014, preferably microglia expressing at least one of the microglia surface proteins selected from the group consisting of: ITGAM (CD11B) (SEQ ID NO: 45), ITGAX (CD11C) (SEQ ID NO: 46), CD14 (SEQ ID NO: 47), CD16 (SEQ ID NO: 48), ENTPD1 (CD39) (SEQ ID NO: 49), PTPRC (CD45) (SEQ ID NO: 50), CD68 (SEQ ID NO: 51), CSF1R (CD115) (SEQ ID NO: 52), CD163 (SEQ ID NO: 53), CX3CR1 (SEQ ID NO: 54), TREM2 (SEQ ID NO: 55), P2RY12 (SEQ ID NO: 56), TMEM119 (SEQ ID NO: 57), and HLA - DR (SEQ ID NO: 58). [Invention 1016] Microglia of Invention 1015 for use in therapy. [Invention 1017] Use of the microglia of Invention 1015 or 1016 for in vitro diagnosis of a disease. [Invention 1018] Use of the microglia of Invention 1017, wherein the disease is selected from the group consisting of: diseases of the central nervous system, preferably neurodegenerative diseases; more preferably Alzheimer's disease, Parkinson's disease, frontotemporal dementia, or amyotrophic lateral sclerosis; neuroinflammatory diseases or autoimmune diseases, preferably multiple sclerosis, autoimmune encephalitis, or infectious diseases, neurovascular diseases; preferably stroke, vasculitis; traumatic brain injury, and cancer. [Invention 1019] Use of the microglia of Invention 1015 or 1016 for in vitro culture with brain organoids.

Brief Description of the Drawings

[0030]

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[0031] The following abbreviations are used: T2A: T2A peptide (ribosome skipping signal), puroR: puromycin resistance gene, pA: polyadenylation signal, CAG: constitutive CAG promoter, TRE3GV: Tet-responsive element, HA-R, HA-L: homology arms (right, left), AmpR: ampicillin resistance gene, ori: origin of replication, NeoR: neomycin resistance gene, KanR: kanamycin resistance gene. **DETAILED DESCRIPTION OF THE INVENTION**

[0032] Detailed Description of the Invention The present invention relates to a method for generating microglia from stem cells, the method comprising the following steps: a) targeted insertion of a nucleotide sequence encoding a transcriptional regulatory factor protein into a first safe harbor site on the genome; and b) targeted insertion of the coding sequence of the transcription factor PU.1 (SEQ ID NO: 1) into a second safe harbor site on the genome, wherein the gene is operably linked to an inducible promoter regulated by the transcriptional regulatory factor protein; expression of PU.1 (SEQ ID NO: 2); and c) culturing the stem cells obtained from steps a) and b) while exposing them to at least one growth factor or small molecule, wherein the at least one growth factor or small molecule reproduces signal transduction between at least one of the stages of embryonic development of microglia or the stages of proliferation, differentiation, or polarization of adult microglia.

[0033] In one aspect, the present invention relates to a method for generating microglia from stem cells, the method comprising the following steps: a) targeted insertion of a nucleotide sequence encoding a transcriptional regulatory factor protein into a first safe harbor site on the genome; and b) targeted insertion of the coding sequence of the transcription factor PU.1 (SEQ ID NO: 1) into a second safe harbor site on the genome, wherein the gene is operably linked to an inducible promoter regulated by the transcriptional regulatory factor protein; expression of PU.1 (SEQ ID NO: 2); and c) culturing the stem cells obtained from steps a) and b) while exposing them to at least one growth factor or small molecule, wherein the at least one growth factor or small molecule reproduces signal transduction between at least one of the stages of embryonic development of microglia or the stages of proliferation, differentiation, or polarization of adult microglia.

[0034] In one aspect, the present invention relates to a method for generating microglia from stem cells, the method comprising the following steps: a) targeted insertion of a nucleotide sequence encoding a transcriptional regulatory factor protein into a first safe harbor site on the genome; and b) targeted insertion of the coding sequence of the transcription factor PU.1 (SEQ ID NO: 1) into a second safe harbor site on the genome, the gene being operably linked to an inducible promoter regulated by the transcriptional regulatory factor protein; expression of PU.1 (SEQ ID NO: 2); and c) culturing the stem cells obtained from steps a) and b) while exposing them to at least one growth factor or small molecule, wherein the at least one growth factor or small molecule reproduces signaling during at least one stage of embryonic development of microglia.

[0035] In one aspect, the present invention relates to a method for generating microglia from stem cells, the method comprising the following steps: a) targeted insertion of a nucleotide sequence encoding a transcriptional regulatory factor protein into a first safe harbor site on the genome; and b) targeted insertion of the coding sequence of the transcription factor PU.1 (SEQ ID NO: 1) into a second safe harbor site on the genome, the gene being operably linked to an inducible promoter regulated by the transcriptional regulatory factor protein; expression of PU.1 (SEQ ID NO: 2); and c) culturing the stem cells obtained from steps a) and b) while exposing them to at least one growth factor or small molecule, wherein the at least one growth factor or small molecule reproduces signaling during at least one stage of embryonic development of microglia.

[0036] In one aspect, the invention also relates to a method for generating microglia from stem cells, the method comprising the following steps: a) targeted insertion of a nucleotide sequence encoding a transcriptional regulatory factor protein into a first safe harbor site on the genome; and b) targeted insertion of the coding sequence of the transcription factor PU.1 (SEQ ID NO: 1) into a second safe harbor site on the genome, the gene being operably linked to an inducible promoter regulated by the transcriptional regulatory factor protein; expression of PU.1 (SEQ ID NO: 2); and c) culturing the stem cells obtained from steps a) and b) while exposing them to at least one growth factor or small molecule, the at least one growth factor or small molecule reproducing signaling during at least one stage of the differentiation of adult microglia.

[0037] In a further aspect, the invention relates to a method for generating microglia from stem cells, the method comprising the following steps: a) targeted insertion of a nucleotide sequence encoding a transcriptional regulatory factor protein into a first safe harbor site on the genome; and b) targeted insertion of the coding sequence of the transcription factor PU.1 (SEQ ID NO: 1) into a second safe harbor site on the genome, the gene being operably linked to an inducible promoter regulated by the transcriptional regulatory factor protein; expression of PU.1 (SEQ ID NO: 2); and c) culturing the stem cells obtained from steps a) and b) while exposing them to at least one growth factor or small molecule, the at least one growth factor or small molecule reproducing signaling during at least one stage of the polarization of adult microglia.

[0038] In another aspect, the present invention relates to a method for generating microglia from stem cells, the method comprising the following steps: a) targeted insertion of a nucleotide sequence encoding a transcriptional regulatory factor protein into a first safe harbor site on the genome; and b) targeted insertion of the coding sequence of the transcription factor PU.1 (SEQ ID NO: 1) into a second safe harbor site on the genome, the gene being operably linked to an inducible promoter regulated by the transcriptional regulatory factor protein; expression of PU.1 (SEQ ID NO: 2); and c) culturing the stem cells obtained from steps a) and b) while exposing them to at least one growth factor or small molecule, the at least one growth factor or small molecule mimicking the embryonic development of microglia.

[0039] In a further aspect, the present invention relates to a method for generating microglia from stem cells, the method comprising the following steps: a) targeted insertion of a nucleotide sequence encoding a transcriptional regulatory factor protein into a first safe harbor site on the genome; and b) targeted insertion of the coding sequence of the transcription factor PU.1 (SEQ ID NO: 1) into a second safe harbor site on the genome, the gene being operably linked to an inducible promoter regulated by the transcriptional regulatory factor protein; expression of PU.1 (SEQ ID NO: 2); and c) culturing the stem cells obtained from steps a) and b) while exposing them to at least one growth factor or small molecule, the at least one growth factor or small molecule mimicking the signaling between at least one of the stages of embryonic development of microglia or the stages of proliferation, differentiation, or polarization of adult microglia.

[0040] In a further aspect, the invention relates to a method for generating microglia from stem cells, the method comprising the steps of: a) targeted insertion of a nucleotide sequence encoding a transcriptional regulatory factor protein into a first safe harbor site on the genome; and b) targeted insertion of the coding sequence of the transcription factor PU.1 (SEQ ID NO: 1) into a second safe harbor site on the genome, the gene being operably linked to an inducible promoter regulated by the transcriptional regulatory factor protein; expression of PU.1 (SEQ ID NO: 2); and c) culturing the stem cells obtained from steps a) and b) while exposing them to at least one growth factor or small molecule, the at least one growth factor or small molecule mimicking signaling during at least one stage of embryonic development of microglia.

[0041] In one aspect, the invention also relates to a method for generating microglia from stem cells, the method comprising the steps of: a) targeted insertion of a nucleotide sequence encoding a transcriptional regulatory factor protein into a first safe harbor site on the genome; and b) targeted insertion of the coding sequence of the transcription factor PU.1 (SEQ ID NO: 1) into a second safe harbor site on the genome, the gene being operably linked to an inducible promoter regulated by the transcriptional regulatory factor protein; expression of PU.1 (SEQ ID NO: 2); and c) culturing the stem cells obtained from steps a) and b) while exposing them to at least one growth factor or small molecule, the at least one growth factor or small molecule recapitulating embryonic development of microglia in vitro.

[0042] When used within the scope of the present invention, the term "microglia" refers to a mature cell type, which is a unique cell population in the central nervous system. As defined in Comparative Anatomy and Histology, "Microglia are the resident histiocytic-type cells and the key innate immune effectors of the CNS. They are often described as either resting (i.e., ramified) or activated, but these terms fail to convey the dynamic remodeling of their fine processes and constitutive immunosurveillance activity. (…) Evidence suggests that early microglia are derived from yolk sac progenitors." (Hagan et al., 2012). The intended microglia develop during the early stages of the embryo and reside in the brain throughout adult life.

[0043] When used within the scope of the present invention, the term "generation of microglia" refers to the generation of mature cells (microglia) from stem cells, meaning the generation of cells obtained by any of the methods of the present invention described herein.

[0044] When used within the scope of the present invention, the term "stem cell" means a cell type that can divide to produce additional cells or can develop into cells with specific purposes. In the present invention, the stem cells used can be pluripotent stem cells. Pluripotent stem cells have the ability to differentiate into most cells in the body. There are several sources of pluripotent stem cells. Embryonic stem cells (ES cells) are pluripotent stem cells derived from the inner cell mass of the blastocyst, which is an early-stage pre-implantation embryo. Induced pluripotent stem cells (iPSCs) are adult cells that have been genetically reprogrammed into an embryonic stem cell-like state by forcibly expressing genes and factors important for maintaining the characteristics that determine embryonic stem cells. In 2006, it was shown that adult cells could be converted into pluripotent stem cells by introducing four specific genes encoding transcription factors (Takahashi et al., 2006), but in subsequent studies, the number of genes required decreased / changed. Oct-3 / 4 and several members of the Sox gene family have been identified as potentially important transcriptional regulators involved in the induction process. Additional genes, including several members of the Klf family, several members of the Myc family, Nanog, and LIN28, can increase the induction efficiency. Examples of genes that can be included in the reprogramming factors are Oct3 / 4, Sox2, Soxl, Sox3, Soxl5, Soxl7, Klf4, Klf2, c-Myc, N-Myc, L-Myc, Nanog, Lin28, Fbxl5, ERas, ECAT15-2, Tell, β-catenin, Lin28b, Sall1, Sall4, Esrrb, Nr5a2, Tbx3, and Glisl, and these reprogramming factors can be used alone or in combination of two or more of them.

[0045] When cells modified by the insertion of an inducible cassette are used in a human patient, it may be preferable for the cells to be iPSCs derived from that individual. The use of such autologous cells can obviate the need to match the cells to the recipient. Alternatively, commercially available iPSCs can be used, which are known to those skilled in the art. Alternatively, the cells can be tissue-specific stem cells, which may also be autologous or provided. Suitable cells include epiblast stem cells, induced neural stem cells, and other tissue-specific stem cells.

[0046] In some embodiments of the methods of the invention, the stem cells used may preferably be embryonic stem cells or stem cell lines. A number of embryonic stem cell lines are currently available, such as, for example, WA01 (HI), WA09 (H9), KhES-1, KhES-2, and KhES-3. Stem cell lines derived without destroying the embryo are available. The present invention is not extended to any method involving the destruction of a human embryo.

[0047] As used within the scope of the present invention, the term "targeted insertion" means insertion into a safe harbor (GSH) site on the genome, which is preferably a specific insertion into the sequence of the GSH as described elsewhere. Any suitable technique for inserting a polynucleotide into a specific sequence can be used, and several are described in the art. Suitable techniques include any method known to those skilled in the art that allows introduction of a cleavage at the desired position and then recombination of the vector into the gap. Thus, the first important step in a genome modification specific to a target site is the creation of a double-strand DNA break (DSB) at the genomic locus to be modified. Different mechanisms of cellular repair can be exploited to repair the DSB and introduce the desired sequence, and these are the more error-prone non-homologous end joining repair (NHEJ); and homologous recombination repair (HR) which can be used to insert an inducible cassette and which involves a donor DNA template.

[0048] Several techniques exist that enable the generation of customized site-specific DSBs in the genome. Many of these involve the use of customized endonucleases, such as zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), or the clustered regularly interspaced short palindromic repeat (CRISPR) / CRISPR-associated protein (CRISPR / Cas9) system (Gaj et al., 2013). Zinc finger nucleases are artificial enzymes created by fusing a zinc finger DNA-binding domain to the nuclease domain of the restriction enzyme FokI. The latter has a non-specific cleavage domain that must dimerize to cut DNA. This means that two ZFN monomers are required to enable dimerization of the FokI domain and cut DNA. The DNA-binding domain can be designed to target any genomic sequence of interest, which can be a tandem array of Cys2His2 zinc fingers, each of which recognizes a consecutive 3-nucleotide sequence in the target sequence. To allow optimal dimerization of the FokI domain, the two binding sites are separated by 5-7 bp. Thus, the enzyme can cut DNA at a specific site, and target specificity is increased by ensuring that two adjacent DNA-binding events must occur to achieve a double-strand break. Transcription activator-like effector nucleases, or TALENs, are dimeric transcription factor / nucleases. They are created by fusing a TAL effector DNA-binding domain to a DNA cleavage domain (nuclease). Transcription activator-like effectors (TALENs) can be designed to bind to virtually any desired DNA sequence and, when combined with a nuclease, can cut DNA at specific positions.TAL effectors are proteins secreted by Xanthomonas bacteria, and their DNA-binding domains contain repeats of a highly conserved 33-34 amino acid sequence, where the 12th and 13th amino acids are different. These two positions are highly variable and show a strong correlation with the recognition of specific nucleotides. This clear relationship between the amino acid sequence and DNA recognition enables the design of specific DNA-binding domains by selecting combinations of repeat segments containing appropriate residues at these two variable positions. Thus, TALENs are composed of an array of 33-35 amino acid modules, each of which targets a single nucleotide. By selecting an array of modules, almost any sequence can be targeted. The nuclease used can be FokI or its derivatives.

[0049] Three types of CRISPR mechanisms have been identified, and type II is the most studied among them. The CRISPR / Cas9 system (type II system) utilizes Cas9 nuclease to create double-stranded breaks in DNA at sites determined by short guide RNAs. The CRISPR / Cas system is a prokaryotic immune system that confers resistance to foreign genetic elements. CRISPR is a segment of prokaryotic DNA containing short repeats of nucleotide sequences. Each repeat is followed by a short segment of "protospacer DNA" derived from previous exposure to foreign genetic elements. CRISPR spacers recognize and cleave foreign genetic elements using RNA interference. The CRISPR immune response occurs in the following two stages: biosynthesis of CRISPR-RNA (crRNA), and interference guided by crRNA. CrRNA molecules are composed of variable sequences transcribed from protospacer DNA and CRISP repeats. Each crRNA molecule then hybridizes with a second RNA known as trans-activating CRISPR RNA (tracrRNA), and ultimately, both of these together form a complex with nuclease Cas9. The section of crRNA encoded by protospacer DNA causes Cas9 to cleave the complementary target DNA sequence when they are adjacent to a short sequence known as the protospacer adjacent motif (PAM). This natural system has been designed and utilized to introduce DSB cleavage at specific sites in genomic DNA in many other applications. In particular, the CRISPR type II system derived from Streptococcus pyogenes can be used. Briefly, the CRISPR / Cas9 system contains two components that are delivered into cells to effect genome editing: Cas9 nuclease itself, and a small guide RNA (gRNA). The gRNA is a fusion of a customized site-specific crRNA (directed at the target sequence) and a standardized tracrRNA.

[0050] When a DSB is generated, a donor template having homology to the target locus is supplied. The DSB can be repaired by a homology-directed repair (HDR) pathway that allows for accurate insertion. Derivatives from this system are also possible. Mutant forms of Cas9 are available, such as Cas9D10A which has only nickase activity. This means that the mutant form cleaves only one DNA strand and does not activate NHEJ. Alternatively, when a homology repair template is provided, DNA repair is carried out only via the high-precision HDR pathway. Cas9D10A can be used as a pair of Cas9 complexes designed to nick adjacent DNA, together with two types of sgRNAs complementary to the adjacent region in the opposite strand of the target site, which can be particularly beneficial. Elements for creating double-stranded DNA breaks can be introduced in one or more vectors, such as plasmids, for expression in cells. Thus, any method for creating a specific target-type double-stranded break in the genome to enable the insertion of a nucleotide sequence / gene / inducible cassette can be used in the method of the present invention. The method of the present invention may preferably utilize any one or more of ZFN, TALEN, and / or the CRISPR / Cas9 system, or any derivatives thereof, for inserting a gene / inducible cassette.

[0051] Once a DSB has been created by any suitable means, as described below, the gene / inducible cassette for insertion can be supplied in any suitable manner. The gene / inducible cassette, and related genetic materials, form donor DNA for repairing the DNA at the DSB, and this is inserted using the standard cellular repair machinery / pathways. As described above, which pathway is used to repair the damage depends on how the cleavage is initiated. However, this is also within the knowledge of those skilled in the art.

[0052] As used within the scope of the present invention, the term "gene" means a linear sequence of nucleotides along a segment of DNA that provides coded instructions for synthesizing RNA which, when translated into protein, results in the expression of hereditary characteristics.

[0053] As used within the scope of the present invention, the term "nucleotide sequence" refers to a sequence of bases in a DNA segment that forms a gene as defined above.

[0054] When used within the scope of the present invention, the term "transcription regulatory factor protein" refers to a protein that binds to DNA, preferably a protein that binds sequence-specifically to a DNA site located within or near a promoter, and that either promotes the binding of the transcription apparatus to the promoter and as a result promotes the transcription of a DNA sequence (a transcriptional activator) or interferes with this process (a transcriptional repressor). Such entities are also known as transcription factors. The DNA sequences to which transcription regulatory factor proteins bind are called transcription factor binding sites or response elements, and these are found within or near the promoter of the DNA sequence being regulated. Response elements are part of the present invention. Transcriptional activator proteins bind to response elements and promote gene expression. Such proteins are preferred for controlling the expression of inducible cassettes in the methods of the present invention. Transcriptional repressor proteins bind to response elements and prevent gene expression. Transcription regulatory factor proteins can be activated or inactivated by several mechanisms, which include the binding of substances, interactions with other transcription factors (e.g., homodimerization or heterodimerization) or co-regulatory proteins, phosphorylation, and / or methylation. Transcription regulatory factors can be controlled by activation or inactivation. When the transcription regulatory factor protein is a transcriptional activator protein, it is preferred that the transcriptional activator protein requires activation. This activation can occur via any suitable means, but preferably the transcription regulatory factor protein is activated by the addition of a foreign substance to the stem cell. The supply of the foreign substance to the stem cell is controllable, and thus the activation of the transcription regulatory factor protein is controllable. Such transcription regulatory factor proteins are also called inducible transcription regulatory factor proteins.

[0055] When used within the scope of the present invention, the term "transcription factor" refers to a protein that binds to DNA, preferably a protein that binds sequence-specifically to a DNA site located within or near a promoter, and that promotes the binding of the transcription apparatus to the promoter and, as a result, promotes the transcription of a DNA sequence (transcription activator), or that interferes with this process (transcription repressor). In the context of the present invention, a transcription factor is a desired genetic sequence, preferably a DNA sequence that translocates into a cell together with an inducible cassette. The introduction of an inducible cassette into the genome has the potential to change the phenotype of a cell by adding a genetic sequence that enables gene expression. The method of the present invention provides for the controllable transcription of the genetic sequences of a set of transcription factors within an inducible cassette in a cell.

[0056] Master regulators are one or more of the following: transcription factors, transcriptional regulators, cytokine receptors, or signaling molecules, etc. A master regulator is a gene that affects the lineage of the cells that express it. A network of master regulators may be required to determine the lineage of a cell. As used herein, a master regulator gene that is expressed at the onset of a developing lineage or cell type is involved in the specification of that lineage by regulating a plurality of downstream genes either directly or via a cascade of changes in gene expression. When a master regulator is expressed, it has the ability to re-specify the fate of cells that are fated to form other lineages. Transcription factors that may be used in the method of the present invention include PU.1 (SEQ ID NO: 2) (gene is SPI1, SEQ ID NO: 1), CEBPB (SEQ ID NO: 3), RUNX1 (SEQ ID NO: 4), IRF8 (SEQ ID NO: 5), and SALL1 (SEQ ID NO: 6).

[0057] When used within the scope of the present invention, the term "PU.1" (SEQ ID NO: 2) refers to the hematopoietic transcription factor PU.1, Spi-1 proto-oncogene, 31 kDa Transforming Protein, transcription factor PU.1, Spleen Focus Forming Virus (SFFV) proviral integrated oncogene Spi1, Spleen Focus Forming Virus (SFFV) proviral integrated oncogene, or the transcription factor also known as 31 kDa Transforming Protein, SFPI1, SPI-1, SPI-A, PU.1, or OF, where "SPI1" refers to its gene (SEQ ID NO: 1) (Spi-1 proto-oncogene), which encodes an ETS domain transcription factor that activates gene expression during the development of myeloid and B lymphocytes.

[0058] When used within the scope of the present invention, the term "safe harbor site on the genome" means a genetic site into which a genetic material can be inserted without causing harmful effects to the cell and into which transcription of the inserted genetic material is possible. Those skilled in the art may use these simplified criteria and / or more formal criteria to identify appropriate GSHs. Specific insertion into a safe harbor site (GSH) on the genome is preferred over random integration into the genome because such insertion is expected to be a safer genomic modification and is less likely to have unwanted secondary effects, such as silencing natural gene expression or causing mutations that result in cancerous cell types. Thus, a safe harbor site on the genome is a locus within the genome into which a gene or other genetic material can be inserted without harmful effects to the cell or to the inserted genetic material. Most beneficial are GSH sites where the expression of the inserted gene sequence is not disrupted by either read-through expression from adjacent genes or expression of inducible cassettes and where interference with the endogenous transcription program is minimal. More formal criteria have been proposed to assist in determining whether a particular locus is a GSH site (Pellenz et al., 2019). These criteria include the following sites: (i) > 300 kb from any cancer-related gene on the pan-cancer gene list, (ii) > 300 kb from any miRNA / other functional small RNA, (iii) > 50 kb from the 5' end of any gene, (iv) > 50 kb away from any origin of replication, (v) > 50 kb away from any ultra-conserved element, (vi) low transcriptional activity (no mRNA in ± 25 kb), (vii) not within a copy number polymorphism region, (viii) within open chromatin (DHS signal in ± 1 kb), and (ix) unique (one copy on the human genome). Since the GSHs that have already been identified do not meet all of these criteria, not all of these proposed criteria necessarily have to be met.A suitable GSH preferably can meet at least 3, 4, 5, 6, 7, or 8 of these criteria, and most preferably can meet 9 of these criteria.

[0059] In the method of the present invention, the insertion occurs in different GSHs. At least two GSHs are required. The first GSH is modified by the insertion of a transcriptional regulatory factor protein. The second GSH is modified by the insertion of an inducible cassette, where the cassette contains a coding sequence operably linked to an inducible promoter. Other genetic materials can also be inserted into either or both of these elements. The genetic sequence operably linked to the inducible promoter within the inducible cassette is preferably a DNA sequence. The genetic sequence of the inducible cassette preferably encodes an RNA molecule and is thus capable of being transcribed. Transcription is controlled using the inducible promoter. The RNA molecule can be of any sequence, but is preferably mRNA encoding a protein, shRNA, or gRNA.

[0060] The first GSH can be any suitable GSH site. Optionally, this is a GSH having a constitutively expressed endogenous promoter that constitutively expresses the inserted transcriptional regulatory factor. A suitable GSH is the hROSA26 site with respect to human cells. In a further aspect of the present invention, the inserted transcriptional regulatory factor protein operably linked to the promoter is operably linked to a constitutive promoter. The constitutive promoter can be used, for example, upon insertion at the hROSA26 site.

[0061] As used within the scope of the present invention, the term "inducible promoter" means a nucleotide sequence that initiates and regulates the transcription of a polynucleotide. An "inducible promoter" is a nucleotide sequence in which the expression of a genetic sequence operably linked to the promoter is controlled by an analyte, cofactor, regulatory protein, etc. In one embodiment of the method of the present invention, the control is affected by a transcriptional regulatory factor protein. The terms "promoter" or "control element" are intended to include the full-length promoter region and functional (e.g., controlling transcription or translation) segments of these regions. A gene encoding a transcriptional regulatory factor protein is preferably operably linked to a constitutive promoter. Alternatively, the first GSH can be selected such that it already has a constitutive promoter capable of driving the expression of the gene for the transcriptional regulatory factor protein and any associated genetic material. Constitutive promoters ensure persistent and high-level gene expression. Commonly used constitutive promoters include the human β-actin promoter (ACTB), cytomegalovirus (CMV) promoter, elongation factor-la (EFla) promoter, phosphoglycerate kinase (PGK) promoter, and ubiquitin C (UbC) promoter. The CAG promoter is a strong synthetic promoter often used to drive high-level gene expression.

[0062] As used within the scope of the present invention, the term "culturing" means the growth of microorganisms such as bacteria and yeast, or human cells, plant cells, or animal cells under appropriate conditions that ensure growth, as known to those skilled in the art.

[0063] When used within the scope of the present invention, the term "growth factor" means a signaling molecule that controls the activity of cells in an autocrine, paracrine, or endocrine manner. As used herein, the term "growth factor" in the context of the present invention may be used interchangeably with "cytokine". Growth factors or cytokines are produced by various cell types of an organism and exert their biological functions by binding to specific receptors and activating the associated downstream signaling pathways. They also regulate gene transcription in the nucleus and ultimately stimulate biological responses including regulatory cell processes such as cell division, cell survival, cell differentiation, adhesion, and migration.

[0064] When used within the scope of the present invention, the term "small molecule" means a bioactive molecule that is produced naturally or artificially, can diffuse through the cell membrane, and can regulate a signaling pathway. Small molecules preferably used within the scope of the present invention can inhibit phosphatidylinositol 3-kinase (PI3K) and glycogen synthase kinase 3, such as LY294002 and CHIR99021, respectively.

[0065] When used within the scope of the present invention, the term "replicate signaling" means to mimic, imitate, or resemble the function of secreted molecules, such as growth factors and / or chemokines, that affect cells in a natural environment, and thereby enable the generation of microglia by these actions.

[0066] When used within the scope of the present invention, the term "mimic signaling" means to mimic, imitate, resemble, or replicate the function of secreted molecules, such as growth factors and / or chemokines, that affect cells in a natural environment, and thereby enable the generation of microglia by these actions.

[0067] When used within the scope of the present invention, the term "microglial embryonic development" refers to the stepwise transition of pluripotent stem cells into mature microglial cells, following the outcome of differentiation during microglial development, from the embryo at the pre-implantation blastocyst stage through human embryonic, fetal, and postnatal development until a fully established and self-maintaining microglial population is reached.

[0068] When used within the scope of the present invention, the term "proliferation of adult microglia" means any cell division process that results in mature microglial cells.

[0069] When used within the scope of the present invention, the term "differentiation of adult microglia" means the differentiation of cells in the state of microglial progenitor cells into adult microglial cell types that encompass the typical characteristics of quiescent / resting microglial cells.

[0070] When used within the scope of the present invention, the term "polarization of adult microglia" means the response of mature microglial cells to extracellular stimuli brought about by the extracellular environment, signals from damaged neurons and glial cells respectively, or exposure to plasma proteins due to blood-brain barrier dysfunction. This response of microglia includes the movement of microglial cells to the site of injury, and the response can have either a neuroprotective or neurotoxic effect.

[0071] Furthermore, in one aspect of the method of the present invention, at least one of the growth factors or small molecules is selected from the group consisting of: activin A (SEQ ID NO: 7), BMP4 (SEQ ID NO: 8), FGF (SEQ ID NO: 9), VEGF-A (SEQ ID NO: 10), LY294002, CHIR99021, SCF (SEQ ID NO: 11), IL-3 (SEQ ID NO: 12), IL-6 (SEQ ID NO: 13), CSF1 (SEQ ID NO: 14), IL-34 (SEQ ID NO: 15), CSF2 (SEQ ID NO: 16), CD200 (SEQ ID NO: 17), CX3CL1 (SEQ ID NO: 18), TGFβ1 (SEQ ID NO: 19), and IDE1.

[0072] Activin A (SEQ ID NO: 7), when used in the present invention, means activin β-A chain, EDF, erythroid differentiation protein, FRP, FSH releasing protein, INHBA, inhibin β-A chain, inhibin β-1. The protein encoded by this gene is a member of the transforming growth factor β (TGF-β) family of proteins produced by pluripotent stem cells, endoderm, and mesoderm.

[0073] BMP4 (SEQ ID NO: 8), when used in the present invention, means bone morphogenetic protein 4, which is also known as ZYME, BMP2B, or BMP2B1. The protein encoded by this gene is a member of the bone morphogenetic protein family, which is part of the transforming growth factor β superfamily.

[0074] FGF (SEQ ID NO: 9), when used in the present invention, means fibroblast growth factor. The protein encoded by this gene is a member of a family of cell signaling proteins, for example, as described in Hui et al., 2018.

[0075] VEGF-A (SEQ ID NO: 10), when used in the present invention, means vascular endothelial growth factor A, which is also known as VPF, VEGF, or MVCD1. The protein encoded by this gene is a member of the PDGF / VEGF growth factor family and is a heparin-binding protein. This growth factor induces the proliferation and migration of vascular endothelial cells and is essential for both physiological and pathological angiogenesis.

[0076] LY294002, when used in the present invention, means a potent cell-permeable inhibitor of phosphatidylinositol 3-kinase (PI3K), which acts on the ATP-binding site of the enzyme (Vlahos et al., 1994). Its chemical structure is shown below. TIFF2025087820000002.tif32128

[0077] CHIR99021, when used in the present invention, means an aminopyrimidine derivative that is a very potent inhibitor of glycogen synthase kinase 3, which inhibits GSK3β (IC 50 = 6.7 nM) and inhibits GSK3α (IC 50 = 10 nM) and functions as a WNT activator. Its chemical structure is shown below. TIFF2025087820000003.tif37128

[0078] SCF (SEQ ID NO: 11), when used in the present invention, means stem cell factor, which is also known as Kit ligand, mast cell growth factor, or Steel factor. The protein encoded by this gene is an early-acting cytokine that plays an important role in the regulation of embryonic and adult hematopoiesis.

[0079] IL-3 (SEQ ID NO: 12), as used in the present invention, means interleukin-3, MCGF (mast cell growth factor), multi-CSF, HCGF, P cell stimulating factor, MGC79398, or MGC79399. The protein encoded by this gene is a cytokine that promotes proliferation.

[0080] IL-6 (SEQ ID NO: 13), as used in the present invention, means interleukin 6, which is also known as B cell stimulating factor 2, CTL differentiation factor, hybridoma growth factor, interferon-β-2, interleukin-6, IFN-β-2, IFNB2, BSF-2, CDF, interferon β2, B cell differentiation factor, interferon β2, interleukin BSF-2, BSF2, HGF, or HSF. The protein encoded by this gene is a cytokine that functions in inflammation and B cell maturation.

[0081] CSF1 (SEQ ID NO: 14), as used in the present invention, means colony stimulating factor 1, which is also known as colony stimulating factor 1 (macrophage), macrophage colony-stimulating factor 1, macrophage colony stimulating factor 1, ranimostim, CSF-1, MCSF, M-CSF, and the protein encoded by this gene is a cytokine that controls macrophage production, differentiation, and function.

[0082] IL-34 (SEQ ID NO: 15), as used in the present invention, means interleukin 34, which is also known as C16 or f77. The protein encoded by this gene is a cytokine that promotes monocyte and macrophage differentiation and survival through the colony stimulating factor 1 receptor.

[0083] CSF2 (SEQ ID NO: 16), as used in the present invention, means colony stimulating factor 2, which is also known as sargramostim, colony stimulating factor 2 (granulocyte-macrophage), granulocyte macrophage colony-stimulating factor, molgramostin, molgramostim, GMCSF, CSF, granulocyte macrophage colony-stimulating factor, granulocyte-macrophage colony stimulating factor, colony stimulating factor, GM-CSF. The protein encoded by this gene is a cytokine that controls the production, differentiation, and function of granulocytes and macrophages.

[0084] CD200 (SEQ ID NO: 17), as used in the present invention, means the CD200 gene, which is also known as the CD200 molecule, CD200 antigen, antigen identified by the monoclonal antibody MRC OX-2, OX-2 membrane glycoprotein, MOX1, MOX2, OX-2, or MRC. The protein encoded by this gene is a type I membrane glycoprotein containing two extracellular immunoglobulin domains, one transmembrane domain, and one cytoplasmic domain.

[0085] CX3CL1 (SEQ ID NO: 18), as used in the present invention, means the CX3CL1 gene, which is also known as C-X3-C motif chemokine ligand 1, Small Inducible Cytokine Subfamily D (Cys-X3-Cys), Member 1 (Fractalkine, Neurotactin), Chemokine (C-X3-C motif) ligand 1, CX3C membrane-anchored chemokine, Small Inducible Cytokine D1, C-X3-C motif chemokine 1, Neurotactin, Fractalkine, or SCYD1, NTT, Small Inducible Cytokine Subfamily D (Cys-X3-Cys), Member 1, C3Xkine, ABCD-3, CXC3C, CXC3, NTN, or FKN. The protein encoded by this gene belongs to the CX3C subgroup of chemokines, which are characterized by the number of amino acids located between conserved cysteine residues.

[0086] TGFβ1 (SEQ ID NO: 19), as used in the present invention, means transforming growth factor β1, which is also known as transforming growth factor β-1 proprotein, preprotransforming growth factor β-1, TGFB, transforming growth factor β1, transforming growth factor β-1, Latency-Associated Peptide, TGF-β-1 of Camurati-Engelmann disease, IBDIMDE, TGFβ, DPD1, CED, or LAP. The protein encoded by this gene is a secreted ligand of the TGF-β (transforming growth factor β) protein superfamily.

[0087] In a further aspect of the method of the present invention, at least one of the growth factors is CSF1 (SEQ ID NO: 14) or IL-34 (SEQ ID NO: 15). In a further aspect of the method of the present invention, at least one of the growth factors is CSF1 (SEQ ID NO: 14). In a further aspect of the method of the present invention, at least one of the growth factors is IL-34 (SEQ ID NO: 15).

[0088] In an additional aspect of the method of the present invention, at least one of the small molecules is CHIR99021, LY294002, or IDE1.

[0089] As used in the present invention, LY294002 means a potent cell-permeable inhibitor of phosphatidylinositol 3-kinase (PI3K), which acts on the ATP-binding site of the enzyme (Vlahos et al., 1994). Its chemical structure is shown below. TIFF2025087820000004.tif32128

[0090] As used in the present invention, CHIR99021 means an aminopyrimidine derivative that is a potent inhibitor of glycogen synthase kinase 3, which inhibits GSK3β (IC 50 = 6.7 nM), and inhibits GSK3α (IC 50 = 10 nM), and functions as a WNT activator. Its chemical structure is shown below. TIFF2025087820000005.tif37128

[0091] As used in the present invention, IDE1 means an inducer of embryonic endoderm; a small molecule that activates the TGF-β pathway and can be used as an alternative to the growth factor TGF-β. Its chemical structure is shown below. TIFF2025087820000006.tif19128

[0092] In another aspect of the method of the present invention, the first and second safe harbor sites on the genome are different.

[0093] In a further aspect of the method of the present invention, the method further comprises the insertion and expression of the coding sequence (SEQ ID NO: 3) of the gene of the transcription factor CEBPB.

[0094] CEBPB (SEQ ID NO: 3), when used in the present invention, means CCAAT enhancer-binding protein β, which is also known as CCAAT enhancer-binding protein β, CCAAT / enhancer-binding protein (C / EBP) β, interleukin 6-dependent DNA-binding protein, CCAAT / enhancer-binding protein β, nuclear factor of interleukin 6, transcription factor 5, nuclear factor NF-IL6, TCF5, liver-enriched transcriptional activator protein, CCAAT / enhancer-binding protein β, liver-enriched inhibitory protein, transcription factor C / EBP β, liver activator protein, C / EBP-β, C / EBP β, IL6DBP, NF-IL6, TCF-5, LAP, or LIP. This intronless gene encodes a transcription factor containing a basic leucine zipper (bZIP) domain.

[0095] In another aspect of the method of the present invention, the method further comprises the insertion and expression of the coding sequence (SEQ ID NO: 4) of the gene of the transcription factor RUNX1.

[0096] RUNX1 (SEQ ID NO: 4), when used in the present invention, refers to Runt Related Transcription Factor 1, Runt-Related Transcription Factor 1, Polyomavirus Enhancer Binding Protein 2 alpha B subunit, SL3 / AKV Core Binding Factor alpha B subunit, SL3-3 Enhancer Factor 1 alpha B subunit, Acute Myeloid Leukemia 1 protein, Cancer Gene AML-1, PEBP2-alpha B, PEA2-alpha B, CBFA2, AML1, Core Binding Factor Runt Domain alpha Subunit 2 Core Binding Factor Subunit alpha-2, AML1-EVI-1 fusion protein, Aml1 cancer gene of acute myeloid leukemia, CBF-alpha-2, AML1-EVI-1, PEBP2 alpha, CBF2 alpha, PEBP2aB, AMLCR1, or EVI-1. The protein encoded by this gene is the alpha subunit of CBF and is thought to be involved in the progression of normal hematopoiesis.

[0097] In a further aspect of the method of the present invention, the method further comprises the insertion and expression of the coding sequence (SEQ ID NO: 5) of the transcription factor IRF8 gene.

[0098] IRF8 (SEQ ID NO: 5), when used in the present invention, refers to Interferon Regulatory Factor 8, which is also known as Interferon Consensus Sequence-Binding Protein 1, H-ICSBP, ICSBP1, ICSBP, IRF-8, Interferon Consensus Sequence-Binding Protein, IMD32A, IMD32B, or Interferon consensus sequence-binding protein (ICSBP). It is a transcription factor of the Interferon (IFN) Regulatory Factor (IRF) family.

[0099] In another aspect of the method of the present invention, the method further comprises the insertion and expression of the coding sequence (SEQ ID NO: 6) of the gene for the transcription factor SALL1.

[0100] When used in the present invention, SALL1 (SEQ ID NO: 6) means Spalt Like Transcription Factor 1, which is also known as zinc finger protein Spalt-1, zinc finger protein SALL1, zinc finger protein 794, Sal-like protein 1, ZNF794, Sal-1, testicular appendage secretory protein Li 89, Spalt-Like Transcription Factor 1, Sal (Drosophila) -like 1, Sal-like 1 (Drosophila), HEL-S-89, HSAL1, HSal1, SAL1, or TBS. The protein encoded by this gene is a zinc finger transcriptional repressor and may be part of the NuRD histone deacetylase complex (HDAC).

[0101] In an additional aspect of the method of the present invention, the transcriptional regulatory factor protein is a reverse tetracycline transactivator (rtTA) (SEQ ID NO: 20), and its activity is controlled by doxycycline or tetracycline.

[0102] When used within the scope of the present invention, the term "reverse tetracycline trans-activator (rtTA)" means a transcriptional activator protein that is induced by tetracycline or a derivative thereof. Tetracycline-controlled transcriptional activation is a method of inducibly expressing a gene, where transcription is reversibly initiated or stopped in the presence of the antibiotic tetracycline or one of its derivatives (e.g., the more stable doxycycline). In this system, the transcriptional activator protein can be the tetracycline-responsive transcriptional activator protein (rtTa), or a derivative thereof. The transcriptional regulatory protein of the present invention can be rtTA. The rtTA protein can bind to DNA at specific TetO operator sequences. Several repeats of such TetO sequences are placed upstream of a minimal promoter (e.g., the CMV promoter, etc.) and together form the tetracycline response element (TRE) (SEQ ID NO: 21). This system has two formats, depending on whether the addition of tetracycline or a derivative activates (Tet-On) or inactivates (Tet-Off) the rtTA protein. In the Tet-ON system, doxycycline activates the rtTA protein, which can also be used in one aspect of the method of the present invention.

[0103] The Tet-On system consists of two elements: (1) a constitutively expressed tetracycline-responsive transcriptional activator protein (rtTA), and an rtTA-responsive inducible promoter (tetracycline response element, TRE). Since tetracycline or a more stable derivative thereof containing doxycycline (dox) can bind to this, it causes activation of rtTA, enables rtTA to bind to the TRE sequence, and induces the expression of TRE-regulated genes. In the method of the present invention, it is preferable to use this. Therefore, the transcriptional regulatory factor protein of the method of the present invention can be a tetracycline-responsive transcriptional activator protein (rtTA), which can be activated or inactivated by an antibiotic tetracycline or one of its derivatives supplied externally. When the transcriptional regulatory factor protein is rtTA, the inducible promoter inserted into the second GSH site contains a tetracycline response element (TRE). The substance supplied externally can be an antibiotic tetracycline or one of its derivatives such as doxycycline, preferably tetracycline or doxycycline.

[0104] Variants of the rtTA protein, and modified rtTA proteins can be used in the method of the present invention. These can include the Tet-On Advanced transactivator (also known as rtTA2S-M2), and Tet-On 3G (also known as rtTA-V16, derived from rtTA2S-S2).

[0105] In another aspect of the method of the present invention, the inducible promoter contains a tetracycline response element (TRE) (SEQ ID NO: 21).

[0106] As used within the scope of the present invention, the term "Tet-responsive element (TRE)" means the bacterial TetO sequence, consisting of 19 bp repeated 7 times and separated by a spacer sequence, together with a minimal promoter. Since the minimal promoter can be any suitable promoter, variants and modifications of the TRE sequence are available. Preferably, the minimal promoter shows no expression or a minimal level of expression in the absence of rtTA binding. Thus, the inducible promoter inserted into the second GSH may contain a TRE. The basic genetic principles underlying the present invention are also illustrated in FIG. 2, which shows the different GSH sites (hROSA26 and AAVS1), as well as the rtTA (SEQ ID NO: 20) and TRE (SEQ ID NO: 21) to be integrated.

[0107] In a further aspect of the method of the present invention, the first and second safe harbor sites on the genome are selected from the group consisting of: the hROSA26 locus (SEQ ID NO: 22), the AAVS1 locus (SEQ ID NO: 23), the CLYBL gene (SEQ ID NO: 24), the CCR5 gene (SEQ ID NO. 25), the HPRT gene (SEQ ID NO. 26), or a gene having site ID 325 (SEQ ID NO: 27) on chromosome 8, a gene having site ID 227 (SEQ ID NO: 28) on chromosome 1, a gene having site ID 229 (SEQ ID NO: 29) on chromosome 2, a gene having site ID 255 (SEQ ID NO: 30) on chromosome 5, a gene having site ID 259 (SEQ ID NO: 31) on chromosome 14, a gene having site ID 263 (SEQ ID NO: 32) on the X chromosome, a gene having site ID 303 (SEQ ID NO: 33) on chromosome 2, a gene having site ID 231 (SEQ ID NO: 34) on chromosome 4, a gene having site ID 315 (SEQ ID NO: 35) on chromosome 5, a gene having site ID 307 (SEQ ID NO: 36) on chromosome 16, a gene having site ID 285 (SEQ ID NO: 37) on chromosome 6, a gene having site ID 233 (SEQ ID NO: 38) on chromosome 6, a gene having site ID 311 (SEQ ID NO: 39) on chromosome 134, a gene having site ID 301 (SEQ ID NO: 40) on chromosome 7, a gene having site ID 293 (SEQ ID NO: 41) on chromosome 8, a gene having site ID 319 (SEQ ID NO: 42) on chromosome 11, a gene having site ID 329 (SEQ ID NO: 43) on chromosome 12, a gene having site ID 313 (SEQ ID NO: 44) on the X chromosome.Preferably, in a further aspect of the method of the present invention, the first and second safe harbor sites on the genome are selected from the group consisting of: hROSA26 locus (SEQ ID NO: 22), AAVS1 locus (SEQ ID NO: 23), CLYBL gene (SEQ ID NO: 24), CCR5 gene (SEQ ID NO. 25), HPRT gene (SEQ ID NO. 26). More preferably, the first and second safe harbor sites on the genome are selected from the group consisting of the hROSA26 locus (SEQ ID NO: 22) and the AAVS1 locus (SEQ ID NO: 23).

[0108] By searching for sites where the virus is naturally integrated without interfering with natural gene expression, additional sites can be identified. With respect to the method of the present invention, several GSH sites can be used and are described in more detail below.

[0109] The adeno-associated virus integration site 1 locus (AAVS1) (SEQ ID NO: 23) is located within the regulatory subunit 12C (PPP1R12C) gene of protein phosphatase 1 on human chromosome 19, and the gene is expressed uniformly and ubiquitously in human tissues. This site acts as an integration locus specific for AAV serotype 2 and has thus been identified as a promising GSH. AAVS1 has been shown to be a transcriptionally favorable environment because it contains an open chromatin structure and a natural chromosomal insulator that can render an inducible cassette resistant to silencing. No harmful effects on cells due to disruption of the PPP1R12C gene are known. Furthermore, the inducible cassette inserted at this site remains transcriptionally active in many diverse cell types. Thus, AAVS1 is considered a GSH and has been widely used for targeted gene recombination in the human genome.

[0110] The hROSA26 locus (SEQ ID NO: 22) was identified based on sequence similarity to the mouse GSH (ROSA26 - reverse oriented splice acceptor site #26). Although orthologous loci have been identified in humans, this locus is not generally used for the insertion of inducible cassettes. The inventors of the present invention use a targeting system specific to the hROSA26 locus and were thus able to insert genetic material into this locus. The hROSA26 locus (SEQ ID NO: 22) is located on chromosome 3 (3p25.3) and can be found in the Ensembl database (GenBank: CR624523). The exact genomic coordinates of the integration site are 3:9396280 - 9396303:Ensembl. The integration site is located within the open reading frame (ORF) of the long non - coding RNA (reverse strand) of THUMPD3. Since the hROSA26 locus has an endogenous promoter, the inserted genetic material may well utilize this endogenous promoter or, alternatively, may be inserted operably linked to a promoter.

[0111] Intron 2 of the citrate lyase beta - like (CLYBL) gene (SEQ ID NO: 24) on the long arm of chromosome 13 has been identified as a suitable GSH because it is one of the identified integration hotspots for phage - derived phiC31 integrase. Studies have demonstrated that inducible cassettes randomly inserted into this locus are stable and express. Insertion of inducible cassettes at this GSH has been shown not to disrupt neighboring gene expression (Cerbibi et al., 2015). Thus, CLYBL provides a GSH that can be used in the methods of the present invention.

[0112] CCR5 (SEQ ID NO: 25) is a gene encoding a major co-receptor for HIV-1, located on chromosome 3 (position 3p21.31). Interest in using this site as a GSH appears to have no harmful effects, but it arises from null mutations in this gene that confer resistance to HIV-1 infection. Zinc finger nucleases targeting exon 3 have been developed, enabling the insertion of genetic material at this locus. Considering that the natural function of CCR5 has not yet been elucidated, the site remains a putative GSH but can be used in the methods of the present invention. The hypoxanthine-guanine phosphoribosyltransferase (HPRT) gene encodes a transferase enzyme that plays a central role in the production of purine nucleotides through the purine salvage pathway. This has been discussed as a GSH site. Insertion at this site may be more appropriate for mature cell types, such as for modifications for gene therapy. GSHs have been identified in other organisms and include the ROSA26, HRPT, and Hippll (Hll) loci in mice.

[0113] The mammalian genome may contain GSH sites based on pseudo attP sites. For such sites, the hiC31 integrase, a recombinase derived from Streptomyces phage, has been developed as a non-viral insertion tool because of its ability to integrate a plasmid carrying an attB site, which contains an inducible cassette, into the pseudo attP site. GSHs also exist in the genomes of plants, and the modification of plant cells can be used in the methods of the present invention. GSHs have been identified in the genome of rice (Cantos et al., 2014).

[0114] The following SHS sites can be used in any of the methods of the present invention. They were published by Pellenz et al., 2019 and meet 5 out of the 9 criteria listed above: Site ID 325 (SEQ ID NO: 27) at chromosome 8: 68,720,172 - 68,720,191; Site ID 227 (SEQ ID NO: 28) at chromosome 1: 231,999,396 - 231,999,415; Site ID 229 (SEQ ID NO: 29) at chromosome 2: 45,708,354 - 45,708,373; Site ID 255 (SEQ ID NO: 30) at chromosome 5: 19,069,307 - 19,069,326; Site ID 259 (SEQ ID NO: 31) at chromosome 14: 92,099,558 - 92,099,577; Site ID 263 (SEQ ID NO: 32) at chromosome X: 12,590,812 - 12,590,831; Site ID 303 (SEQ ID NO: 33) at chromosome 2: 77,263,930 - 77,263,949; Site ID 317 (SEQ ID NO: 60) at chromosome 2: 77,263,930 - 77,263,949; Site ID 231 (SEQ ID NO: 34) at chromosome 4: 58,976,613 - 58,976,632; Site ID 315 (SEQ ID NO: 35) at chromosome 5: 7,577,728 - 7,577,747; Site ID 307 (SEQ ID NO: 36) at chromosome 16: 19,323,777 - 19,323,796; Site ID 285 (SEQ ID NO: 37) at chromosome 6: 89,574,320 - 89,574,339; Site ID 233 (SEQ ID NO: 38) at chromosome 6: 114,713,905 - 114,713,924; Site ID 311 (SEQ ID NO: 39) at chromosome 6: 134,385,946 - 134,385,965; Site ID 301 (SEQ ID NO: 40) at chromosome 7: 113,327,685 - 113,327,704; Site ID 293 (SEQ ID NO: 41) at chromosome 8: 40,727,927 - 40,727,946;Locus ID 319 (SEQ ID NO: 42) at 32,680,546 - 32,680,565 on chromosome 11; Locus ID 329 (SEQ ID NO: 43) at 126,152,581 - 126,152,600 on chromosome 12; Locus ID 313 (SEQ ID NO: 44) at 16,059,732 - 16,059,751 on the X chromosome.;

[0115] In another aspect of the method of the present invention, the stem cells are pluripotent stem cells, induced pluripotent stem cells (iPSCs), neural progenitor cells, hematopoietic stem cells, or embryonic stem cells (ESCs).

[0116] Within the scope of the present invention, the term "pluripotent stem cells" is used as defined above.

[0117] When used within the scope of the present invention, the term "neural progenitor cells" refers to the state of bipotent cells between pluripotent stem cells and mature somatic cells. Usually, this cell state is determined to become specialized cell types such as neurons, oligodendrocytes, and astrocytes.

[0118] Within the scope of the present invention, the term "induced pluripotent stem cells (iPSCs)" is used as defined above.

[0119] When used within the scope of the present invention, the term "hematopoietic stem cells" refers to stem cells that produce blood cells. This particular type of bipotent stem cell can produce any type of blood cell, but has lost the ability to produce other cell types.

[0120] Within the scope of the present invention, the term "embryonic stem cells (ESCs)" is used as defined above.

[0121] In a further aspect of the method of the present invention, the stem cells are human or mouse stem cells.

[0122] When used within the scope of the present invention, the term "human or mouse stem cells" means cells derived from humans or mice. However, the stem cells used in the methods of the present invention can be any cells of humans or animals. These are preferably mammalian cells, for example, cells derived from rodents such as mice and rats; cells derived from marsupials such as kangaroos and koalas; cells derived from non-human primates such as bonobos, chimpanzees, squirrel monkeys, rhesus monkeys, and apes; cells derived from camelids such as camels and llamas; cells derived from domestic animals such as horses, pigs, cows, buffalo, bison, goats, sheep, deer, reindeer, donkeys, otters, yaks, chickens, ducks, and turkeys; cells derived from domesticated animals such as cats, dogs, rabbits, and guinea pigs. The cells are preferably human cells. In one aspect, the cells are preferably derived from domestic animals. The type of cells used in the methods of the present invention varies depending on the use of the cells after the insertion of the genetic material into the GSH site.

[0123] The present invention also relates to microglial cells obtained by any of the methods according to the present invention, wherein preferably the microglia express at least one of the microglial surface proteins selected from the group consisting of: ITGAM (CD11B) (SEQ ID NO: 45), ITGAX (CD11C) (SEQ ID NO: 46), CD14 (SEQ ID NO: 47), CD16 (SEQ ID NO: 48), ENTPD1 (CD39) (SEQ ID NO: 49), PTPRC (CD45) (SEQ ID NO: 50), CD68 (SEQ ID NO: 51), CSF1R (CD115) (SEQ ID NO: 52), CD163 (SEQ ID NO: 53), CX3CR1 (SEQ ID NO: 54), TREM2 (SEQ ID NO: 55), P2RY12 (SEQ ID NO: 56), TMEM119 (SEQ ID NO: 57), and HLA-DR (SEQ ID NO: 58).

[0124] Thus, microglia are further defined by expressing at least one of the following surface proteins: ITGAM (CD11B) (SEQ ID NO: 45), ITGAX (CD11C) (SEQ ID NO: 46), CD14 (SEQ ID NO: 47), CD16 (SEQ ID NO: 48), ENTPD1 (CD39) (SEQ ID NO: 49), PTPRC (CD45) (SEQ ID NO: 50), CD68 (SEQ ID NO: 51), CSF1R (CD115) (SEQ ID NO: 52), CD163 (SEQ ID NO: 53), CX3CR1 (SEQ ID NO: 54), TREM2 (SEQ ID NO: 55), P2RY12 (SEQ ID NO: 56), TMEM119 (SEQ ID NO: 57), and HLA-DR (SEQ ID NO: 58). These proteins are defined as follows.

[0125] As used in the present invention, ITGAM (CD11B) means integrin subunit αM, which is the gene encoding the integrin αM chain. Integrin is a heterodimeric integral membrane protein composed of an α chain and a β chain. Its protein sequence is shown in SEQ ID NO: 45.

[0126] As used within the scope of the present invention, ITGAX (CD11C) means integrin subunit αX, and the gene encodes the protein of the integrin αX chain. Its protein sequence is shown in SEQ ID NO: 46.

[0127] As used in the present invention, CD14 means monocyte differentiation antigen CD14, and the protein encoded by this gene is a surface antigen that is selectively expressed in monocytes / macrophages. Its protein sequence is shown in SEQ ID NO: 47.

[0128] CD16, as used in the present invention, means FCGR3A, the Fc fragment of IgG receptor IIIa, and this gene encodes a receptor for the Fc portion of immunoglobulin G and is involved in the removal of antigen-antibody complexes from the circulation and other antibody-dependent responses. Its protein sequence is shown in SEQ ID NO: 48.

[0129] ENTPD1 (CD39), as used in the present invention, means ectonucleoside triphosphate diphosphohydrolase 1, and the protein encoded by this gene is a plasma membrane protein that hydrolyzes extracellular ATP and ADP to AMP. Its protein sequence is shown in SEQ ID NO: 49.

[0130] PTPRC (CD45), as used in the present invention, means receptor-type protein tyrosine phosphatase type C, and the protein encoded by this gene is a member of the protein tyrosine phosphatase (PTP) family. Its protein sequence is shown in SEQ ID NO: 50.

[0131] CD68, as used in the present invention, means the CD68 antigen, and this gene encodes an 110-kD transmembrane glycoprotein that is highly expressed in human monocytes and tissue macrophages. Its protein sequence is shown in SEQ ID NO: 51.

[0132] CSF1R (CD115), as used in the present invention, means colony-stimulating factor 1 receptor, and the protein encoded by this gene is the receptor for colony-stimulating factor 1, a cytokine that controls the production, differentiation, and function of macrophages. Its protein sequence is shown in SEQ ID NO: 52.

[0133] When used in the present invention, CD163 means the CD163 antigen, and the protein encoded by this gene is a member of the scavenger receptor cysteine-rich (SRCR) superfamily and is expressed exclusively in monocytes and macrophages. Its protein sequence is shown in SEQ ID NO: 53.

[0134] When used in the present invention, CX3CR1 means C-X3-C motif chemokine receptor 1, and the protein encoded by this gene is the receptor for fractalkine. Its protein sequence is shown in SEQ ID NO: 54. Fractalkine is a transmembrane protein and is a chemokine involved in leukocyte adhesion and migration.

[0135] When used in the present invention, TREM2 means Triggering Receptor Expressed On Myeloid Cells 2, which is expressed in myeloid cells, and this gene encodes a membrane protein that forms a receptor signaling complex together with a TYRO protein tyrosine kinase-binding protein. Its protein sequence is shown in SEQ ID NO: 55.

[0136] When used in the present invention, P2RY12 means purinergic receptor P2Y12, and the product of this gene belongs to the family of G protein-coupled receptors. Its protein sequence is shown in SEQ ID NO: 56.

[0137] When used in the present invention, TMEM119 means transmembrane protein 119, which is a gene encoding a protein. Its related pathways include microglial activation during neuroinflammation. Its protein sequence is shown in SEQ ID NO: 57.

[0138] As used in the present invention, HLA-DR means DRα and DRβ of class II of the major histocompatibility complex gene complex, and both HLA-DRA and HLA-DRB1 are paralogs of the HLA class II α chain. The protein sequence is shown in SEQ ID NO: 58.

[0139] In a further aspect, the invention also includes microglial cells according to the invention for use in therapy.

[0140] As used in the present invention, the term "treatment" means any form of treating a disease or an undesirable health condition in a biological, animal, or human. This may also include gene therapy. This can be defined as the intentional insertion of foreign DNA into the nucleus of a cell for therapeutic purposes. Such a definition includes: the supply of one or more genes to a cell to provide a wild-type version of a defective gene, the addition of a gene regarding an RNA molecule that interferes with the expression of a target gene (which may have a defect), the supply of suicide genes (such as thymidine kinase (HSV-tk), an enzyme of herpes simplex virus that converts the harmless prodrug ganciclovir (GCV) into a cytotoxic agent, and cytosine deaminase (CD), etc.), DNA vaccines (including adoptive cell immunotherapy) for immunization or cancer treatment, and any other supply of genes to a cell for therapeutic purposes. In addition, mature microglia can be used directly for transplantation into the human or animal body. Alternatively, microglia may also be used as a test material for research, which includes the effects of drugs on gene expression and the interaction of drugs with specific genes. Microglia for research may be relevant to the use of an inducible cassette having a genetic sequence of unknown function for the purpose of studying the controllable expression of the genetic sequence. In addition, this may enable the use of microglia to produce large amounts of desired substances, such as growth factors or cytokines.

[0141] Furthermore, the present invention also, in one aspect, encompasses the use of such microglial cells according to the invention for the in vitro diagnosis of diseases. Preferably, the disease is selected from the group consisting of: diseases of the central nervous system, preferably neurodegenerative diseases; more preferably Alzheimer's disease, Parkinson's disease, frontotemporal dementia, or amyotrophic lateral sclerosis; neuroinflammatory diseases or autoimmune diseases, preferably multiple sclerosis, autoimmune encephalitis, or infectious diseases, neurovascular diseases; preferably stroke, vasculitis; traumatic brain injury, and cancer.

[0142] Furthermore, the present invention encompasses the use of such microglial cells according to the invention for in vitro culture with brain organoids.

[0143] As used within the scope of the present invention, the term "organoid" means an in vitro 3D organ model derived from cells (mainly stem cells), and this, when combined with the microglia produced by the present invention, becomes a powerful tool for medical diagnosis to test the involvement of microglia with other cells of the brain and their interaction with such other cells.

[0144] As used herein, the singular forms "a", "an", and "the" are understood to include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a reagent" includes one or more of such various reagents, and reference to "a method" includes reference to equivalent steps and methods known to those skilled in the art that may be modified or substituted with respect to the methods described herein.

[0145] Unless otherwise specified, the term "at least" preceding a series of elements is understood to refer to all elements in that series. The term "at least one", unless specifically defined otherwise, refers to one or more, which may be, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more. One of ordinary skill in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the present invention.

[0146] As used anywhere in this specification, the term "and / or" includes the meanings "and", "or", and "all, or any other combination of the elements connected by that term".

[0147] The terms "less than" or, conversely, "more than" do not include units of measure.

[0148] For example, less than 20 means less than the specified numerical value. Similarly, "more than" or "greater than" means more or larger than the specified number, for example, more than 80% means more or larger than 80% of the specified number.

[0149] Throughout this specification and the claims appended hereto, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", are to be interpreted as including the stated integer or step or group of integers or steps but not as excluding any other integer or step or group of integers or steps. As used herein, the term "comprising" may be replaced with the term "containing" or "including", or, as sometimes used herein, the term "having". As used herein, "consisting of" excludes any element, step, or ingredient not specified.

[0150] The term "comprising" means "including but not limited to". The terms "comprising" and "including but not limited to" are used interchangeably.

[0151] The term "about" means plus or minus 10%, preferably plus or minus 5%, more preferably plus or minus 2%, and most preferably plus or minus 1%.

[0152] Throughout the description and claims of this specification, unless the context requires otherwise, the singular form includes the plural. In particular, when the indefinite article is used, the specification is to be understood as contemplating both the plural and the singular, unless the context requires otherwise.

[0153] The present invention is not limited to the specific methodologies, protocols, materials, reagents, substances, etc. described herein and is thus understood to be modifiable. The terms used herein are for the purpose of describing particular embodiments only and are not intended to limit the invention of the present invention, which is defined only by the claims.

[0154] All publications cited in the text of this specification (including all patents, patent applications, scientific publications, instructions for use, etc.), whether before or after, are hereby incorporated by reference in their entirety. It should not be construed that this specification admits that the present invention is not eligible to precede such disclosure for the sake of prior inventions. In the event of any conflict or contradiction between this specification and the matter incorporated by reference, this specification shall prevail over any such matter.

[0155] The entire contents of all documents and patent documents cited in this specification are hereby incorporated by reference.

[0156] A better understanding of the present invention and its advantages can be obtained from the following examples, which are provided for illustrative purposes only and are not intended to limit the scope of the present invention in any way.

Examples

[0157] Examples of the Invention The following examples illustrate the present invention but should not be construed as limiting the scope of the present invention.

[0158] Example 1 Materials and Methods For the initial screening experiment, the first hROSA-CAG-rtTA hiPSC line was generated (nucleofection of three plasmids expressing a donor plasmid with CAS9 nickase, two hROSA26-specific guide RNAs (SEQ ID NO: 66 and SEQ ID NO: 67), and the CAG-rtTA expression cassette shown in FIGS. 2A and 4A; selection with antibiotics, clonal expansion, and characterization of clonal individual hiPSC colonies), followed by transient transfection of four vectors targeting AAVS1 (SEQ ID NO: 61 - SEQ ID NO: 64) (see also FIGS. 4B - 4E), enabling either rapid overexpression of PU.1 (SEQ ID NO: 2) alone or rapid overexpression in combination with any of three other transcription factors, RUNX1 (SEQ ID NO: 4), CEBPB (SEQ ID NO: 3), or IRF8 (SEQ ID NO: 5), in the form of a bicistronic expression cassette (FIGS. 4B - 4E) (SEQ ID NO: 61 - SEQ ID NO. 64). For screening purposes, the targeted cells were not clonally expanded, and overexpression was only induced in a subset of the cells.

[0159] Surprisingly, the initial screening experiment demonstrated that in all three cell lines expressing any of the three other candidate reprogramming factors in addition to PU.1 (SEQ ID NO: 2), myeloid and microglial lineage markers were rapidly induced, but not in wild-type control hiPSCs or cells expressing PU.1 (SEQ ID NO: 2) alone.

[0160] Explanation To develop a prototype protocol and establish appropriate readout parameters, the inventors decided to focus on the combinatorial overexpression of PU.1 (SEQ ID NO: 2) and CEBPB (SEQ ID NO: 3). Accordingly, fully validated, dual GSH-targeted inducible PU.1 + CEBPB hiPSCs were generated and clonally expanded.

[0161] Observation The addition of doxycycline resulted in a rapid loss of the expression of the pluripotency markers OCT4 (SEQ ID NO: 78) and NANOG (SEQ ID NO: 79), as well as the induction of both transgenes, in all cells (see Figure 6).

[0162] Example 2 Materials and Methods Briefly, hiPSCs were plated as single cells on Matrigel in pluripotency maintenance medium. After 2 days, the medium was changed to Dulbecco's Modified Eagle Medium (DMEM) / F12, which was supplemented with small molecules and growth factors that mimic the series of embryonic events outlined above, in addition to dox for the induction of the transgenes. Three days after induction, the adherent cells began to detach from the tissue culture plate and were found as single floating cells in the supernatant.

[0163] Explanation Subsequently, the inventors conducted longer screening experiments for up to 20 days to optimize the medium composition, during which cells were induced. Multicolor flow cytometry demonstrated a remarkably robust and rapid induction of myeloid cell surface markers (CD11b (SEQ ID NO: 45), CD14 (SEQ ID NO: 47), CD45 (SEQ ID NO: 50), CD163 (SEQ ID NO: 53), CX3CR1 (SEQ ID NO: 54)), which were selected as a screening panel for the induction of primitive macrophages and / or microglia. The inventors also found important differences depending on the culture conditions: induction occurred most rapidly and efficiently when overexpression of transcription factors was carried out together with a timed exposure to a series of extracellular cues (small molecules, growth factors) that mimic the following series of embryonic development: (1) patterning of pluripotent epiblasts (hiPSCs) towards extraembryonic mesoderm and hemangioblasts (of the posterior primitive streak), (2) induction of primary hematopoiesis and early macrophage progenitor cells, (3) differentiation into primitive yolk sac macrophages, (4) differentiation into microglia (see Figure 5).

[0164] Observation As demonstrated by flow cytometry, cells rapidly initiated the expression of typical myeloid surface proteins, including CD45 (SEQ ID NO: 50) (also known as PTPRC), CD11b (SEQ ID NO: 45) (also known as ITGAM), CD14 (SEQ ID NO: 47), and CX3CR1 (SEQ ID NO: 54) (see Figures 5B - 5C). All cells migrated into the supernatant by day 10 and were plated on poly-L-lysine (PLL)-coated tissue culture dishes in the final medium for the differentiation and maintenance of microglia, chemically defined by Muffat et al., 2016. Interestingly, differentiation into microglia occurred even more efficiently after doxycycline was removed on day 10 of the induction protocol, thus clearly demonstrating that the cell phenotype is independent of the persistent expression of the transgene.

[0165] As quantified by flow cytometry (see Fig. 5c) or as demonstrated by immunohistochemical analysis (see Fig. 5D), 6 - 10 days after differentiation and maturation without the transgene in adherent culture, virtually all cells expressed a broad range of proteins common to myeloid and specific to microglia, including CD39 (SEQ ID NO: 49), P2RY12 (SEQ ID NO: 56), TREM2 (SEQ ID NO: 55), and TMEM119 (SEQ ID NO: 57). Next, co - culture experiments were performed, in which we plated microglial progenitor cells onto a pure population of cortical neurons derived from isogenic hiPSCs generated according to protocols previously published by Zhang et al., 2013 and Pawlowski et al., 2017. Microglial cells had a more branched (i.e., more inactive) morphology compared to cells in monoculture (see Fig. 5E). Real - time qPCR analysis of hiPSCs and microglia in monoculture demonstrated down - regulation of pluripotency factors, MYB - independence (consistent with microglia of primitive yolk sac macrophage origin), and high expression of microglial core transcription factors, classical surface markers, and recently suggested unique signature genes of microglia (see Fig. 5F).

[0166] References: TIFF2025087820000007.tif217165TIFF2025087820000008.tif237165TIFF2025087820000009.tif237165TIFF2025087820000010.tif17165

[0167] Sequence Information SEQUENCE LISTING <110> Westfaelische Wilhelms - Universitaet Muenster <120> Rapid and deterministic generation of microglia from human pluripotent stem cells <150> EP 19176722.7 <151> 2019-05-27 <160> 87 <170> PatentIn version 3.5 <210> 1 <211> 810 <212> DNA <213> Homo sapiens <400> 1 atgttacagg cgtgcaaaat ggaagggttt cccctcgtcc cccctccatc agaagacctg 60 gtgccctatg acacggatct ataccaacgc caaacgcacg agtattaccc ctatctcagc 120 agtgatgggg agagccatag cgaccattac tgggacttcc acccccacca cgtgcacagc 180 gagttcgaga gcttcgccga gaacaacttc acggagctcc agagcgtgca gcccccgcag 240 ctgcagcagc tctaccgcca catggagctg gagcagatgc acgtcctcga tacccccatg 300 gtgccacccc atcccagtct tggccaccag gtctcctacc tgccccggat gtgcctccag 360 tacccatccc tgtccccagc ccagcccagc tcagatgagg aggagggcga gcggcagagc 420 cccccactgg aggtgtctga cggcgaggcg gatggcctgg agcccgggcc tgggctcctg 480 cctggggaga caggcagcaa gaagaagatc cgcctgtacc agttcctgtt ggacctgctc 540 cgcagcggcg acatgaagga cagcatctgg tgggtggaca aggacaaggg caccttccag 600 ttctcgtcca agcacaagga ggcgctggcg caccgctggg gcatccagaa gggcaaccgc 660 aagaagatga cctaccagaa gatggcgcgc gcgctgcgca actacggcaa gacgggcgag 720 gtcaagaagg tgaagaagaa gctcacctac cagttcagcg gcgaagtgct gggccgcggg 780 ggcctggccg agcggcgcca cccgccccac 810 <210> 2 <211> 270 <212> PRT <213> Homo sapiens <400> 2 Met Leu Gln Ala Cys Lys Met Glu Gly Phe Pro Leu Val Pro Pro Pro 1 5 10 15 Ser Glu Asp Leu Val Pro Tyr Asp Thr Asp Leu Tyr Gln Arg Gln Thr 20 25 30 His Glu Tyr Tyr Pro Tyr Leu Ser Ser Asp Gly Glu Ser His Ser Asp 35 40 45 His Tyr Trp Asp Phe His Pro His His Val His Ser Glu Phe Glu Ser 50 55 60 Phe Ala Glu Asn Asn Phe Thr Glu Leu Gln Ser Val Gln Pro Pro Gln 65 70 75 80 Leu Gln Gln Leu Tyr Arg His Met Glu Leu Glu Gln Met His Val Leu 85 90 95 Asp Thr Pro Met Val Pro Pro His Pro Ser Leu Gly His Gln Val Ser 100 105 110 Tyr Leu Pro Arg Met Cys Leu Gln Tyr Pro Ser Leu Ser Pro Ala Gln 115 120 125 Pro Ser Ser Asp Glu Glu Glu Gly Glu Arg Gln Ser Pro Pro Leu Glu 130 135 140 Val Ser Asp Gly Glu Ala Asp Gly Leu Glu Pro Gly Pro Gly Leu Leu 145 150 155 160 Pro Gly Glu Thr Gly Ser Lys Lys Lys Ile Arg Leu Tyr Gln Phe Leu 165 170 175 Leu Asp Leu Leu Arg Ser Gly Asp Met Lys Asp Ser Ile Trp Trp Val 180 185 190 Asp Lys Asp Lys Gly Thr Phe Gln Phe Ser Ser Lys His Lys Glu Ala 195 200 205 Leu Ala His Arg Trp Gly Ile Gln Lys Gly Asn Arg Lys Lys Met Thr 210 215 220 Tyr Gln Lys Met Ala Arg Ala Leu Arg Asn Tyr Gly Lys Thr Gly Glu 225 230 235 240 Val Lys Lys Val Lys Lys Lys Leu Thr Tyr Gln Phe Ser Gly Glu Val 245 250 255 Leu Gly Arg Gly Gly Leu Ala Glu Arg Arg His Pro Pro His 260 265 270 <210> 3 <211> 1038 <212> DNA <213> Homo sapiens <400> 3 atgcaacgcc tggtggcctg ggacccagca tgtctccccc tgccgccgcc gccgcctgcc 60 tttaaatcca tggaagtggc caacttctac tacgaggcgg actgcttggc tgctgcgtac 120 ggcggcaagg cggcccccgc ggcgcccccc gcggccagac ccgggccgcg cccccccgcc 180 ggcgagctgg gcagcatcgg cgaccacgag cgcgccatcg acttcagccc gtacctggag 240 ccgctgggcg cgccgcaggc cccggcgccc gccacggcca cggacacctt cgaggcggct 300 ccgcccgcgc ccgcccccgc gcccgcctcc tccgggcagc accacgactt cctctccgac 360 ctcttctccg acgactacgg gggcaagaac tgcaagaagc cggccgagta cggctacgtg 420 agcctggggc gcctgggggc cgccaagggc gcgctgcacc ccggctgctt cgcgcccctg 480 cacccaccgc ccccgccgcc gccgccgccc gccgagctca aggcggagcc gggcttcgag 540 cccgcggact gcaagcggaa ggaggaggcc ggggcgccgg gcggcggcgc aggcatggcg 600 gcgggcttcc cgtacgcgct gcgcgcttac ctcggctacc aggcggtgcc gagcggcagc 660 agcgggagcc tctccacgtc ctcctcgtcc agcccgcccg gcacgccgag ccccgctgac 720 gccaaggcgc ccccgaccgc ctgctacgcg ggggccgcgc cggcgccctc gcaggtcaag 780 agcaaggcca agaagaccgt ggacaagcac agcgacgagt acaagatccg gcgcgagcgc 840 aacaacatcg ccgtgcgcaa gagccgcgac aaggccaaga tgcgcaacct ggagacgcag 900 cacaaggtcc tggagctcac ggccgagaac gagcggctgc agaagaaggt ggagcagctg 960 tcgcgcgagc tcagcaccct gcggaacttg ttcaagcagc tgcccgagcc cctgctcgcc 1020 tcctccggcc actgctag 1038 <210> 4 <211> 1440 <212> DNA <213> Homo sapiens <400> 4 atggcttcag acagcatatt tgagtcattt ccttcgtacc cacagtgctt catgagagaa 60 tgcatacttg gaatgaatcc ttctagagac gtccacgatg ccagcacgag ccgccgcttc 120 acgccgcctt ccaccgcgct gagcccaggc aagatgagcg aggcgttgcc gctgggcgcc 180 ccggacgccg gcgctgccct ggccggcaag ctgaggagcg gcgaccgcag catggtggag 240 gtgctggccg accacccggg cgagctggtg cgcaccgaca gccccaactt cctctgctcc 300 gtgctgccta cgcactggcg ctgcaacaag accctgccca tcgctttcaa ggtggtggcc 360 ctaggggatg ttccagatgg cactctggtc actgtgatgg ctggcaatga tgaaaactac 420 tcggctgagc tgagaaatgc taccgcagcc atgaagaacc aggttgcaag atttaatgac 480 ctcaggtttg tcggtcgaag tggaagaggg aaaagcttca ctctgaccat cactgtcttc 540 acaaacccac cgcaagtcgc cacctaccac agagccatca aaatcacagt ggatgggccc 600 cgagaacctc gaagacatcg gcagaaacta gatgatcaga ccaagcccgg gagcttgtcc 660 ttttccgagc ggctcagtga actggagcag ctgcggcgca cagccatgag ggtcagccca 720 caccacccag cccccacgcc caaccctcgt gcctccctga accactccac tgcctttaac 780 cctcagcctc agagtcagat gcaggataca aggcagatcc aaccatcccc accgtggtcc 840 tacgatcagt cctaccaata cctgggatcc attgcctctc cttctgtgca cccagcaacg 900 cccatttcac ctggacgtgc cagcggcatg acaaccctct ctgcagaact ttccagtcga 960 ctctcaacgg cacccgacct gacagcgttc agcgacccgc gccagttccc cgcgctgccc 1020 tccatctccg acccccgcat gcactatcca ggcgccttca cctactcccc gacgccggtc 1080 acctcgggca tcggcatcgg catgtcggcc atgggctcgg ccacgcgcta ccacacctac 1140 ctgccgccgc cctaccccgg ctcgtcgcaa gcgcagggag gcccgttcca agccagctcg 1200 ccctcctacc acctgtacta cggcgcctcg gccggctcct accagttctc catggtgggc 1260 ggcgagcgct cgccgccgcg catcctgccg ccctgcacca acgcctccac cggctccgcg 1320 ctgctcaacc ccagcctccc gaaccagagc gacgtggtgg aggccgaggg cagccacagc 1380 aactccccga ccaacatggc gccctccgcg cgcctggagg aggccgtgtg gaggccctac 1440 <210> 5 <211> 1281 <212> DNA <213> Homo sapiens <400> 5 atgtgtgacc ggaatggtgg tcggcggctt cgacagtggc tgatcgagca gattgacagt 60 agcatgtatc caggactgat ttgggagaat gaggagaaga gcatgttccg gatcccttgg 120 aaacacgctg gcaagcaaga ttataatcag gaagtggatg cctccatttt taaggcctgg 180 gcagttttta aagggaagtt taaagaaggg gacaaagctg aaccagccac ttggaagacg 240 aggttacgct gtgctttgaa taagagccca gattttgagg aagtgacgga ccggtcccaa 300 ctggacattt ccgagccata caaagtttac cgaattgttc ctgaggaaga gcaaaaatgc 360 aaactaggcg tggcaactgc tggctgcgtg aatgaagtta cagagatgga gtgcggtcgc 420 tctgaaatcg acgagctgat caaggagcct tctgtggacg attacatggg gatgatcaaa 480 aggagccctt ccccgccgga ggcctgtcgg agtcagctcc ttccagactg gtgggcgcag 540 cagcccagca caggcgtgcc gctggtgacg gggtacacca cctacgacgc gcaccattca 600 gcattctccc agatggtgat cagcttctac tatgggggca agctggtggg ccaggccacc 660 accacctgcc ccgagggctg ccgcctgtcc ctgagccagc ctgggctgcc cggcaccaag 720 ctgtatgggc ccgagggcct ggagctggtg cgcttcccgc cggccgacgc catccccagc 780 gagcgacaga ggcaggtgac gcggaagctg ttcgggcacc tggagcgcgg ggtgctgctg 840 cacagcagcc ggcagggcgt gttcgtcaag cggctgtgcc agggccgcgt gttctgcagc 900 ggcaacgccg tggtgtgcaa aggcaggccc aacaagctgg agcgtgatga ggtggtccag 960 gtcttcgaca ccagccagtt cttccgagag ctgcagcagt tctataacag ccagggccgg 1020 cttcctgacg gcagggtggt gctgtgcttt ggggaagagt ttccggatat ggcccccttg 1080 cgctccaaac tcattctcgt gcagattgag cagctgtatg tccggcaact ggcagaagag 1140 gctgggaaga gctgtggagc cggctctgtg atgcaggccc ccgaggagcc gccgccagac 1200 caggtcttcc ggatgtttcc agatatttgt gcctcacacc agagatcatt tttcagagaa 1260 aaccaacaga tcaccgtcta a 1281 <210> 6 <211> 3975 <212> DNA <213> Homo sapiens <400> 6 atgtcgcgga ggaagcaagc gaagcctcaa catttccaat ccgaccccga agtggcctcg 60 ctcccccggc gagatggaga cacagaaaag ggtcaaccga gtcgccctac taagagcaag 120 gatgcccacg tctgtggccg gtgctgtgcc gagttctttg aattatcaga tcttctgctc 180 cacaagaaga actgtactaa aaatcaatta gttttaatcg taaatgaaaa tccagcctcc 240 ccacccgaaa ccttctcccc cagcccccct cctgataatc ctgatgaaca aatgaatgac 300 acagttaaca aaacagatca agtggactgc agcgaccttt cagaacacaa cggacttgac 360 agggaagagt ccatggaggt ggaggccccg gttgctaaca aaagcggcag cggcacttcc 420 agcggcagcc acagcagtac cgccccaagc agcagcagca gcagcagcag cagcagcggc 480 ggcggcggca gctcctccac aggtacctca gcgatcacaa cctctctacc tcaactcggg 540 gacctgacaa cactgggcaa cttctccgta atcaacagca acgtcatcat cgagaacctc 600 cagagcacca aggtggcggt ggcccagttc tcccaggaag cgaggtgcgg cggggcctct 660 gggggcaagc tggccgtccc agccctcatg gaacaactcc tagctctgca gcagcagcag 720 atccaccagc tgcaattgat cgaacagatt cgtcaccaaa tattgctgtt ggcttctcag 780 aatgcagact tgccaacatc ttctagtcct tctcaaggta ctttacgaac atctgccaac 840 cccttgtcca cgctaagttc ccatttatct cagcagctgg cagcagcagc tggattggca 900 cagagcctcg ccagccaatc tgccagcatt agtggtgtga aacagctacc cccaatccag 960 ctacctcaga gcagttctgg caacaccatc attccatcca acagcggctc ttctcccaat 1020 atgaacatat tggcagcggc agttaccacc ccgtcctctg aaaaagtggc ttcaagtgct 1080 ggggcctccc atgtcagcaa cccagcggtc tcatcatcgt cctcaccagc ttttgcaata 1140 agcagtttat taagtcctgc gtctaatcca cttctacctc agcaagcctc cgctaactcg 1200 gttttcccca gccctttgcc caacatcgga acaactgcag aggatttaaa ctccttgtct 1260 gccttggccc agcaaagaaa aagcaagcca ccaaatgtca ctgcctttga agcgaaaagt 1320 acttccgatg aggcattctt caaacacaag tgcaggttct gcgcgaaggt ctttgggagt 1380 gacagtgcct tgcagatcca cttgcgttcc cataccggag agaggccatt caagtgcaac 1440 atctgcggga acaggttctc caccaagggg aatctgaaag tccactttca gcgccacaaa 1500 gagaaatacc ctcatatcca gatgaacccc tatcctgtgc ctgagcattt ggacaatatc 1560 cccacgagta ctggcatccc atatggcatg tccatccctc cagagaagcc agtcaccagc 1620 tggctagaca ccaaaccagt cctgcctact ctgaccactt cagtcggcct gccgttgccc 1680 ccaaccctcc caagcctcat acccttcatc aagacggaag agccagcccc catccccatc 1740 agccattctg ccaccagccc cccaggctca gtcaaaagtg actccggggg ccctgagtca 1800 gccacaagaa acctaggtgg gctcccagag gaagccgaag ggtccactct gccaccctct 1860 ggtggcaaaa gcgaagagag tggcatggtc accaactcag tcccgacggc gagcagtagc 1920 gtcctgagct ccccagcggc agactgcggc cccgcgggca gtgccaccac cttcaccaac 1980 cctttgttgc cgctcatgtc cgagcagttc aaggccaagt ttccttttgg gggactcctg 2040 gactcagctc aggcatcaga gacgtccaag cttcagcaac tggtagaaaa cattgacaag 2100 aaggccactg accccaatga gtgcatcatc tgccaccggg ttctcagctg ccagagcgcc 2160 ttgaaaatgc actacaggac acacactggg gagaggccct ttaagtgtaa gatctgtggc 2220 cgggctttca ccacgaaagg gaatcttaaa acccactaca gtgtccatcg tgctatgccc 2280 ccgctcagag tccagcattc ctgccccatc tgccagaaga agttcacgaa cgctgtggtc 2340 ctgcagcagc acatccgaat gcatatggga ggccagatcc ccaacacccc agtccccgac 2400 agctactctg agtccatgga gtctgacaca ggttcctttg atgagaaaaa ttttgatgac 2460 ctagacaact tctctgatga aaacatggaa gactgtcctg agggcagcat ccctgataca 2520 cctaagtctg cagacgcctc ccaagacagc ttatcctctt cgcctttgcc cctcgagatg 2580 tcgagcatcg ctgctttgga aaatcagatg aagatgatca atgctggcct ggcagagcag 2640 ctacaggcca gcctgaagtc agtggagaat gggtccatcg agggggatgt cctgaccaat 2700 gattcatcct cagtgggtgg tgacatggaa agccaaagtg ctggcagccc agccatctca 2760 gagtctacct cttccatgca ggctctgtcc ccgtccaaca gcacgcagga gttccacaag 2820 tcacccagca ttgaggagaa accacagaga gcggtcccaa gcgagtttgc caatggtttg 2880 tctcccaccc cagtgaatgg tggggctttg gatttgacat ctagtcacgc agagaaaatc 2940 atcaaagaag attctttggg gatcctcttc ccttttagag accggggtaa atttaaaaac 3000 actgcttgtg acatttgtgg caaaacattt gcttgtcaga gtgccttgga cattcactat 3060 agaagtcata ccaaagagag accatttatt tgcacagttt gcaatcgtgg cttttccaca 3120 aagggtaatt tgaagcagca catgttgaca catcagatgc gagatctgcc atcccagctc 3180 tttgagccca gttccaacct tggccccaat cagaactcag cggtgattcc cgccaactcg 3240 ttgtcatctc tcatcaagac agaggtcaac ggcttcgtgc atgtttctcc tcaggacagt 3300 aaggacaccc ccaccagtca cgtcccgtct gggcctctgt cttcctctgc cacatcccca 3360 gttctgctcc ctgctctgcc caggagaact cccaagcagc actactgcaa cacatgtggc 3420 aaaaccttct cctcatcgag tgccctgcag attcacgaga gaactcacac tggagagaaa 3480 ccctttgctt gcactatttg tggaagagct ttcacgacta aaggcaatct taaggtacac 3540 atgggcactc acatgtggaa tagcacccct gcacgacggg gtcggcggct ctctgtggat 3600 ggccccatga catttctagg aggcaatccc gtcaagttcc cagaaatgtt ccagaaggat 3660 ttggcggcaa gatcaggaag tggggatcct tccagcttct ggaatcagta tgcagcagcg 3720 ctctccaacg ggctggcgat gaaggccaac gagatctccg tcattcagaa cggtggcatc 3780 cctccaattc ctggaagcct cggcagtggg aacagctcac ctgttagtgg gctgacggga 3840 aacctggaga ggctccagaa ctcagagccc aatgctcccc tggccggcct ggagaaaatg 3900 gcaagcagtg agaacggaac caacttccgc ttcacccgct tcgtggagga cagcaaggag 3960 atcgtcacga gttaa 3975 <210> 7 <211> 116 <212> PRT <213> Homo sapiens <400> 7 Gly Leu Glu Cys Asp Gly Lys Val Asn Ile Cys Cys Lys Lys Gln Phe 1 5 10 15 Phe Val Ser Phe Lys Asp Ile Gly Trp Asn Asp Trp Ile Ile Ala Pro 20 25 30 Ser Gly Tyr His Ala Asn Tyr Cys Glu Gly Glu Cys Pro Ser His Ile 35 40 45 Ala Gly Thr Ser Gly Ser Ser Leu Ser Phe His Ser Thr Val Ile Asn 50 55 60 His Tyr Arg Met Arg Gly His Ser Pro Phe Ala Asn Leu Lys Ser Cys 65 70 75 80 Cys Val Pro Thr Lys Leu Arg Pro Met Ser Met Leu Tyr Tyr Asp Asp 85 90 95 Gly Gln Asn Ile Ile Lys Lys Asp Ile Gln Asn Met Ile Val Glu Glu 100 105 110 Cys Gly Cys Ser 115 <210> 8 <211> 106 <212> PRT <213> Homo sapiens <400> 8 Lys Lys Asn Lys Asn Cys Arg Arg His Ser Leu Tyr Val Asp Phe Ser 1 5 10 15 Asp Val Gly Trp Asn Asp Trp Ile Val Ala Pro Pro Gly Tyr Gln Ala 20 25 30 Phe Tyr Cys His Gly Asp Cys Pro Phe Pro Leu Ala Asp His Leu Asn 35 40 45 Ser Thr Asn His Ala Ile Val Gln Thr Leu Val Asn Ser Val Asn Ser 50 55 60 Ser Ile Pro Lys Ala Cys Cys Val Pro Thr Glu Leu Ser Ala Ile Ser 65 70 75 80 Met Leu Tyr Leu Asp Glu Tyr Asp Lys Val Val Leu Lys Asn Tyr Gln 85 90 95 Glu Met Val Val Glu Gly Cys Gly Cys Arg 100 105 <210> 9 <211> 146 <212> PRT <213> Homo sapiens <400> 9 Pro Ala Leu Pro Glu Asp Gly Gly Ser Gly Ala Phe Pro Pro Gly His 1 5 10 15 Phe Lys Asp Pro Lys Arg Leu Tyr Cys Lys Asn Gly Gly Phe Phe Leu 20 25 30 Arg Ile His Pro Asp Gly Arg Val Asp Gly Val Arg Glu Lys Ser Asp 35 40 45 Pro His Ile Lys Leu Gln Leu Gln Ala Glu Glu Arg Gly Val Val Ser 50 55 60 Ile Lys Gly Val Cys Ala Asn Arg Tyr Leu Ala Met Lys Glu Asp Gly 65 70 75 80 Arg Leu Leu Ala Ser Lys Cys Val Thr Asp Glu Cys Phe Phe Phe Glu 85 90 95 Arg Leu Glu Ser Asn Asn Tyr Asn Thr Tyr Arg Ser Arg Lys Tyr Thr 100 105 110 Ser Trp Tyr Val Ala Leu Lys Arg Thr Gly Gln Tyr Lys Leu Gly Ser 115 120 125 Lys Thr Gly Pro Gly Gln Lys Ala Ile Leu Phe Leu Pro Met Ser Ala 130 135 140 Lys Ser 145 <210> 10 <211> 121 <212> PRT <213> Homo sapiens <400> 10 Ala Pro Met Ala Glu Gly Gly Gly Gln Asn His His Glu Val Val Lys 1 5 10 15 Phe Met Asp Val Tyr Gln Arg Ser Tyr Cys His Pro Ile Glu Thr Leu 20 25 30 Val Asp Ile Phe Gln Glu Tyr Pro Asp Glu Ile Glu Tyr Ile Phe Lys 35 40 45 Pro Ser Cys Val Pro Leu Met Arg Cys Gly Gly Cys Cys Asn Asp Glu 50 55 60 Gly Leu Glu Cys Val Pro Thr Glu Glu Ser Asn Ile Thr Met Gln Ile 65 70 75 80 Met Arg Ile Lys Pro His Gln Gly Gln His Ile Gly Glu Met Ser Phe 85 90 95 Leu Gln His Asn Lys Cys Glu Cys Arg Pro Lys Lys Asp Arg Ala Arg 100 105 110 Gln Glu Asn Cys Asp Lys Pro Arg Arg 115 120 <210> 11 <211> 165 <212> PRT <213> Homo sapiens <400> 11 Met Glu Gly Ile Cys Arg Asn Arg Val Thr Asn Asn Val Lys Asp Val 1 5 10 15 Thr Lys Leu Val Ala Asn Leu Pro Lys Asp Tyr Met Ile Thr Leu Lys 20 25 30 Tyr Val Pro Gly Met Asp Val Leu Pro Ser His Cys Trp Ile Ser Glu 35 40 45 Met Val Val Gln Leu Ser Asp Ser Leu Thr Asp Leu Leu Asp Lys Phe 50 55 60 Ser Asn Ile Ser Glu Gly Leu Ser Asn Tyr Ser Ile Ile Asp Lys Leu 65 70 75 80 Val Asn Ile Val Asp Asp Leu Val Glu Cys Val Lys Glu Asn Ser Ser 85 90 95 Lys Asp Leu Lys Lys Ser Phe Lys Ser Pro Glu Pro Arg Leu Phe Thr 100 105 110 Pro Glu Glu Phe Phe Arg Ile Phe Asn Arg Ser Ile Asp Ala Phe Lys 115 120 125 Asp Phe Val Val Ala Ser Glu Thr Ser Asp Cys Val Val Ser Ser Thr 130 135 140 Leu Ser Pro Glu Lys Asp Ser Arg Val Ser Val Thr Lys Pro Phe Met 145 150 155 160 Leu Pro Pro Val Ala 165 <210> 12 <211> 133 <212> PRT <213> Homo sapiens <400> 12 Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr Ser Trp Val Asn Cys 1 5 10 15 Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys Gln Pro Pro Leu 20 25 30 Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu 35 40 45 Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn Arg Ala 50 55 60 Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile Leu Lys Asn 65 70 75 80 Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro 85 90 95 Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys Leu Thr 100 105 110 Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln Thr Thr Leu 115 120 125 Ser Leu Ala Ile Phe 130 <210> 13 <211> 184 <212> PRT <213> Homo sapiens <400> 13 Pro Val Pro Pro Gly Glu Asp Ser Lys Asp Val Ala Ala Pro His Arg 1 5 10 15 Gln Pro Leu Thr Ser Ser Glu Arg Ile Asp Lys Gln Ile Arg Tyr Ile 20 25 30 Leu Asp Gly Ile Ser Ala Leu Arg Lys Glu Thr Cys Asn Lys Ser Asn 35 40 45 Met Cys Glu Ser Ser Lys Glu Ala Leu Ala Glu Asn Asn Leu Asn Leu 50 55 60 Pro Lys Met Ala Glu Lys Asp Gly Cys Phe Gln Ser Gly Phe Asn Glu 65 70 75 80 Glu Thr Cys Leu Val Lys Ile Ile Thr Gly Leu Leu Glu Phe Glu Val 85 90 95 Tyr Leu Glu Tyr Leu Gln Asn Arg Phe Glu Ser Ser Glu Glu Gln Ala 100 105 110 Arg Ala Val Gln Met Ser Thr Lys Val Leu Ile Gln Phe Leu Gln Lys 115 120 125 Lys Ala Lys Asn Leu Asp Ala Ile Thr Thr Pro Asp Pro Thr Thr Asn 130 135 140 Ala Ser Leu Leu Thr Lys Leu Gln Ala Gln Asn Gln Trp Leu Gln Asp 145 150 155 160 Met Thr Thr His Leu Ile Leu Arg Ser Phe Lys Glu Phe Leu Gln Ser 165 170 175 Ser Leu Arg Ala Leu Arg Gln Met 180 <210> 14 <211> 159 <212> PRT <213> Homo sapiens <400> 14 Met Glu Glu Val Ser Glu Tyr Cys Ser His Met Ile Gly Ser Gly His 1 5 10 15 Leu Gln Ser Leu Gln Arg Leu Ile Asp Ser Gln Met Glu Thr Ser Cys 20 25 30 Gln Ile Thr Phe Glu Phe Val Asp Gln Glu Gln Leu Lys Asp Pro Val 35 40 45 Cys Tyr Leu Lys Lys Ala Phe Leu Leu Val Gln Asp Ile Met Glu Asp 50 55 60 Thr Met Arg Phe Arg Asp Asn Thr Pro Asn Ala Ile Ala Ile Val Gln 65 70 75 80 Leu Gln Glu Leu Ser Leu Arg Leu Lys Ser Cys Phe Thr Lys Asp Tyr 85 90 95 Glu Glu His Asp Lys Ala Cys Val Arg Thr Phe Tyr Glu Thr Pro Leu 100 105 110 Gln Leu Leu Glu Lys Val Lys Asn Val Phe Asn Glu Thr Lys Asn Leu 115 120 125 Leu Asp Lys Asp Trp Asn Ile Phe Ser Lys Asn Cys Asn Asn Ser Phe 130 135 140 Ala Glu Cys Ser Ser Gln Gly His Glu Arg Gln Ser Glu Gly Ser 145 150 155 <210> 15 <211> 230 <212> PRT <213> Homo sapiens <400> 15 Asn Glu Pro Leu Glu Met Trp Pro Leu Thr Gln Asn Glu Glu Cys Thr 1 5 10 15 Val Thr Gly Phe Leu Arg Asp Lys Leu Gln Tyr Arg Ser Arg Leu Gln 20 25 30 Tyr Met Lys His Tyr Phe Pro Ile Asn Tyr Lys Ile Ser Val Pro Tyr 35 40 45 Glu Gly Val Phe Arg Ile Ala Asn Val Thr Arg Leu Gln Arg Ala Gln 50 55 60 Val Ser Glu Arg Glu Leu Arg Tyr Leu Trp Val Leu Val Ser Leu Ser 65 70 75 80 Ala Thr Glu Ser Val Gln Asp Val Leu Leu Glu Gly His Pro Ser Trp 85 90 95 Lys Tyr Leu Gln Glu Val Glu Thr Leu Leu Leu Asn Val Gln Gln Gly 100 105 110 Leu Thr Asp Val Glu Val Ser Pro Lys Val Glu Ser Val Leu Ser Leu 115 120 125 Leu Asn Ala Pro Gly Pro Asn Leu Lys Leu Val Arg Pro Lys Ala Leu 130 135 140 Leu Asp Asn Cys Phe Arg Val Met Glu Leu Leu Tyr Cys Ser Cys Cys 145 150 155 160 Lys Gln Ser Ser Val Leu Asn Trp Gln Asp Cys Glu Val Pro Ser Pro 165 170 175 Gln Ser Cys Ser Pro Glu Pro Ser Leu Gln Tyr Ala Ala Thr Gln Leu 180 185 190 Tyr Pro Pro Pro Pro Trp Ser Pro Ser Ser Pro Pro His Ser Thr Gly 195 200 205 Ser Val Arg Pro Val Arg Ala Gln Gly Glu Gly Leu Leu Pro His His 210 215 220 His His His His His His 225 230 <210> 16 <211> 128 <212> PRT <213> Homo sapiens <400> 16 Met Ala Pro Ala Arg Ser Pro Ser Pro Ser Thr Gln Pro Trp Glu His 1 5 10 15 Val Asn Ala Ile Gln Glu Ala Arg Arg Leu Leu Asn Leu Ser Arg Asp 20 25 30 Thr Ala Ala Glu Met Asn Glu Thr Val Glu Val Ile Ser Glu Met Phe 35 40 45 Asp Leu Gln Glu Pro Thr Cys Leu Gln Thr Arg Leu Glu Leu Tyr Lys 50 55 60 Gln Gly Leu Arg Gly Ser Leu Thr Lys Leu Lys Gly Pro Leu Thr Met 65 70 75 80 Met Ala Ser His Tyr Lys Gln His Cys Pro Pro Thr Pro Glu Thr Ser 85 90 95 Cys Ala Thr Gln Ile Ile Thr Phe Glu Ser Phe Lys Glu Asn Leu Lys 100 105 110 Asp Phe Leu Leu Val Ile Pro Phe Asp Cys Trp Glu Pro Val Gln Glu 115 120 125 <210> 17 <211> 210 <212> PRT <213> Homo sapiens <400> 17 Gln Val Gln Val Val Thr Gln Asp Glu Arg Glu Gln Leu Tyr Thr Pro 1 5 10 15 Ala Ser Leu Lys Cys Ser Leu Gln Asn Ala Gln Glu Ala Leu Ile Val 20 25 30 Thr Trp Gln Lys Lys Lys Ala Val Ser Pro Glu Asn Met Val Thr Phe 35 40 45 Ser Glu Asn His Gly Val Val Ile Gln Pro Ala Tyr Lys Asp Lys Ile 50 55 60 Asn Ile Thr Gln Leu Gly Leu Gln Asn Ser Thr Ile Thr Phe Trp Asn 65 70 75 80 Ile Thr Leu Glu Asp Glu Gly Cys Tyr Met Cys Leu Phe Asn Thr Phe 85 90 95 Gly Phe Gly Lys Ile Ser Gly Thr Ala Cys Leu Thr Val Tyr Val Gln 100 105 110 Pro Ile Val Ser Leu His Tyr Lys Phe Ser Glu Asp His Leu Asn Ile 115 120 125 Thr Cys Ser Ala Thr Ala Arg Pro Ala Pro Met Val Phe Trp Lys Val 130 135 140 Pro Arg Ser Gly Ile Glu Asn Ser Thr Val Thr Leu Ser His Pro Asn 145 150 155 160 Gly Thr Thr Ser Val Thr Ser Ile Leu His Ile Lys Asp Pro Lys Asn 165 170 175 Gln Val Gly Lys Glu Val Ile Cys Gln Val Leu His Leu Gly Thr Val 180 185 190 Thr Asp Phe Lys Gln Thr Val Asn Lys Gly Val Asp His His His His 195 200 205 His His 210 <210> 18 <211> 76 <212> PRT <213> Homo sapiens <400> 18 Gln His His Gly Val Thr Lys Cys Asn Ile Thr Cys Ser Lys Met Thr 1 5 10 15 Ser Lys Ile Pro Val Ala Leu Leu Ile His Tyr Gln Gln Asn Gln Ala 20 25 30 Ser Cys Gly Lys Arg Ala Ile Ile Leu Glu Thr Arg Gln His Arg Leu 35 40 45 Phe Cys Ala Asp Pro Lys Glu Gln Trp Val Lys Asp Ala Met Gln His 50 55 60 Leu Asp Arg Gln Ala Ala Ala Leu Thr Arg Asn Gly 65 70 75 <210> 19 <211> 112 <212> PRT <213> Homo sapiens <400> 19 Ala Leu Asp Thr Asn Tyr Cys Phe Ser Ser Thr Glu Lys Asn Cys Cys 1 5 10 15 Val Arg Gln Leu Tyr Ile Asp Phe Arg Lys Asp Leu Gly Trp Lys Trp 20 25 30 Ile His Glu Pro Lys Gly Tyr His Ala Asn Phe Cys Leu Gly Pro Cys 35 40 45 Pro Tyr Ile Trp Ser Leu Asp Thr Gln Tyr Ser Lys Val Leu Ala Leu 50 55 60 Tyr Asn Gln His Asn Pro Gly Ala Ser Ala Ala Pro Cys Cys Val Pro 65 70 75 80 Gln Ala Leu Glu Pro Leu Pro Ile Val Tyr Tyr Val Gly Arg Lys Pro 85 90 95 Lys Val Glu Gln Leu Ser Asn Met Ile Val Arg Ser Cys Lys Cys Ser 100 105 110 <210> 20 <211> 747 <212> DNA <213> Homo sapiens <400> 20 atgtctagac tggacaagag caaagtcata aactctgctc tggaattact caatggagtc 60 ggtatcgaag gcctgacgac aaggaaactc gctcaaaagc tgggagttga gcagcctacc 120 ctgtactggc acgtgaagaa caagcgggcc ctgctcgatg ccctgccaat cgagatgctg 180 gacaggcatc atacccactc ctgccccctg gaaggcgagt catggcaaga ctttctgcgg 240 aacaacgcca agtcataccg ctgtgctctc ctctcacatc gcgacggggc taaagtgcat 300 ctcggcaccc gcccaacaga gaaacagtac gaaaccctgg aaaatcagct cgcgttcctg 360 tgtcagcaag gcttctccct ggagaacgca ctgtacgctc tgtccgccgt gggccacttt 420 acactgggct gcgtattgga ggaacaggag catcaagtag caaaagagga aagagagaca 480 cctaccaccg attctatgcc cccacttctg aaacaagcaa ttgagctgtt cgaccggcag 540 ggagccgaac ctgccttcct tttcggcctg gaactaatca tatgtggcct ggagaaacag 600 ctaaagtgcg aaagcggcgg gccgaccgac gcccttgacg attttgactt agacatgctc 660 ccagccgatg cccttgacga ctttgacctt gatatgctgc ctgctgacgc tcttgacgat 720 tttgaccttg acatgctccc cgggtaa 747 <210> 21 <211> 350 <212> DNA <213> Homo sapiens <400> 21 gagtttactc cctatcagtg atagagaacg tatgaagagt ttactcccta tcagtgatag 60 agaacgtatg cagactttac tccctatcag tgatagagaa cgtataagga gtttactccc 120 tatcagtgat agagaacgta tgaccagttt actccctatc agtgatagag aacgtatcta 180 cagtttactc cctatcagtg atagagaacg tatatccagt ttactcccta tcagtgatag 240 agaacgtatg tcgaggtagg cgtgtacggt gggcgcctat aaaagcagag ctcgtttagt 300 gaaccgtcag atcgcctgga gcaattccac aacacttttg tcttatactt 350 <210> 22 <211> 1797 <212> DNA <213> Homo sapiens <400> 22 gctcgaaacc ggacggagcc attgctctcg cagagggagg agcgcttccg gctagcctct 60 tgtcgccgat tggccgtttc tcctcccgcc gtgtgtgaaa acacaaatgg cgtattctgg 120 ttggagtaaa gctcctgtca gttacgccgt cgggagtacg cagccgctta gcgactctcg 180 cgttgccccc tgggtggggc gggtaggtag gtggggtgta gagatgctgg gtgtgcgggc 240 gcggccggcc tcctgcggcg ggaggggagg gtcagtgaaa tcggctctgg cgcgggcgtc 300 ctcccaccct ccccttcctt cgggggagtc ggtttacccg ccgcctgctt gtcttcgaca 360 cctgattggc tgtcgaagct gtgggaccgg gcccttgcta ctggctcgag tctcacatga 420 gcgaaaccac tgcgcggggc gcgggggtgg cggggaggcg ggcgttggta cggtcctccc 480 cgaggccgag cgccgcagtg tctggccccg cgcccctgcg caacgtggca ggaagcgcgc 540 gctggaggcg ggggcgggct gccggccgag acttctggat ggcggcggcc gcggctccgc 600 cccgggttcc caccgcctga agggcgagac aagcccgacc tgctacaggc actcgtgggg 660 gtgggggagg agcgggggtc ggtccggctg gtttgtgggt gggaggcgct tgttctccaa 720 aaaccggcgc gagctgcaat cctgagggag ctgcggtgga ggaggtggag agaaggccgc 780 acccttctgg gcagggggag gggagtgccg caataccttt atgggagttc tctgctgcct 840 cccgtcttgt aaggaccgcc ctgggcctgg aagaagccct ccctcctttc ctcctcgcgt 900 gatctcgtca tcgcctccat gtcgagtcgc ttctcgatta tgggcgggat tcttttgcct 960 aggcttaagg ggctaacttg gtccctgggc gttgccctgc aggggagtga gcagctgtaa 1020 gatttgaggg gcgactccga ttagtttatc ttcccacgga ctagagttgg tgtcgaggtt 1080 attgtaataa gggtggggta gggaaatgga gcttagtcat tcacctgggg ctgattttat 1140 gcaacgagac tgcggattat cactacttat catttttgga gcatttttct agagacagac 1200 ataaagcatg atcacctgag ttttatacca tttgagaccc ttgctgcacc accaaagtgt 1260 agcatcaggt taaatcttaa tagaaaaatt ttagcttttg cttgagaaac cagtgcttcc 1320 ctccctcacc ctctctcccc aggctctcta cccctttgca tccctaccag gcatcttagc 1380 aactctcact catacttgat cccattttcc atttgttgta cttgctcctc tagtattcag 1440 acatagcact agctttctcc ctctcttgat cttgggtagc ctggtgtctc gcgaaaccag 1500 acagattggt tccaccacaa attaaggctt gagctggggc ttgactctta cccagcagtg 1560 cttttattcc tccctagttc acgttcttaa atgtttatct tgattttcat tttatccttt 1620 ttccttagct gggattctgt ccctgaccgt cttcacagtc caggtgatct tgactactgc 1680 tttacagaga attggatctg aggttaggca acatctccct ttttcttcct ctaaatacct 1740 ctcatttctg ttcttaccag ttagtaactg atctcagatg cctgtgtgat agcttcc 1797 <210> 23 <211> 1642 <212> DNA <213> Homo sapiens <400> 23 tgctttctct gacctgcatt ctctcccctg ggcctgtgcc gctttctgtc tgcagcttgt 60 ggcctgggtc acctctacgg ctggcccaga tccttccctg ccgcctcctt caggttccgt 120 cttcctccac tccctcttcc ccttgctctc tgctgtgttg ctgcccaagg atgctctttc 180 cggagcactt ccttctcggc gctgcaccac gtgatgtcct ctgagcggat cctccccgtg 240 tctgggtcct ctccgggcat ctctcctccc tcacccaacc ccatgccgtc ttcactcgct 300 gggttccctt ttccttctcc ttctggggcc tgtgccatct ctcgtttctt aggatggcct 360 tctccgacgg atgtctccct tgcgtcccgc ctccccttct tgtaggcctg catcatcacc 420 gtttttctgg acaaccccaa agtaccccgt ctccctggct ttagccacct ctccatcctc 480 ttgctttctt tgcctggaca ccccgttctc ctgtggattc gggtcacctc tcactccttt 540 catttgggca gctcccctac cccccttacc tctctagtct gtgctagctc ttccagcccc 600 ctgtcatggc atcttccagg ggtccgagag ctcagctagt cttcttcctc caacccgggc 660 ccctatgtcc acttcaggac agcatgtttg ctgcctccag ggatcctgtg tccccgagct 720 gggaccacct tatattccca gggccggtta atgtggctct ggttctgggt acttttatct 780 gtcccctcca ccccacagtg gggccactag ggacaggatt ggtgacagaa aagccccatc 840 cttaggcctc ctccttccta gtctcctgat attgggtcta acccccacct cctgttaggc 900 agattcctta tctggtgaca cacccccatt tcctggagcc atctctctcc ttgccagaac 960 ctctaaggtt tgcttacgat ggagccagag aggatcctgg gagggagagc ttggcagggg 1020 gtgggaggga agggggggat gcgtgacctg cccggttctc agtggccacc ctgcgctacc 1080 ctctcccaga acctgagctg ctctgacgcg gccgtctggt gcgtttcact gatcctggtg 1140 ctgcagcttc cttacacttc ccaagaggag aagcagtttg gaaaaacaaa atcagaataa 1200 gttggtcctg agttctaact ttggctcttc acctttctag tccccaattt atattgttcc 1260 tccgtgcgtc agttttacct gtgagataag gccagtagcc agccccgtcc tggcagggct 1320 gtggtgagga ggggggtgtc cgtgtggaaa actccctttg tgagaatggt gcgtcctagg 1380 tgttcaccag gtcgtggccg cctctactcc ctttctcttt ctccatcctt ctttccttaa 1440 agagtcccca gtgctatctg ggacatattc ctccgcccag agcagggtcc cgcttcccta 1500 aggccctgct ctgggcttct gggtttgagt ccttggcaag cccaggagag gcgctcaggc 1560 ttccctgtcc cccttcctcg tccaccatct catgcccctg gctctcctgc cccttcccta 1620 caggggttcc tggctctgct ct 1642 <210> 24 <211> 291666 <212> DNA <213> Homo sapiens <400> 24 ctcggcgttc tcaccgctgc agaacggctg cagggggcag agtaaagagc tgctgtcagt 60 ttccctgccc agagagtgcc aacagctgct ctgacctatt gcaggcatgt gtgttccagg 120 atgcagcctt cctgcacttt gaccactcta gattaattgc taaccaagat atgcacatga 180 tggggctcac aaagtattag gcggtatttt aacaacaaaa gctgcgaaga aatccctaga 240 aatgttggct cttctggctg ggaaaacgca gtctgtcaac agttgcctca aaaactcaga 300 agctcagaga gttggattcg gtcaccagag aggcccctca aagaggcccc aaccattcag 360 atcctctgct ctgtccccca tgacagcggg gacgaaggtc ctaagccctg gccgggaggg 420 cagttgacct cgggactcgc cgcggggcgg agcgggtgct gggaggaagg agggcgtggc 480 tggcggggtg tggctggcgg gacggggcgg ggcgaagaca gagcgaggcg gggcctggga 540 cggggggggg gtcggtgtcg ccgtgggggc gtggaatgtg gggtgggcgt ggctaaaggc 600 aaagggcggg gcgcgcgcgt cgggaagatg gcgctacgtc tgctgcggag ggcggcgcgc 660 ggagctgcgg cggcggcgct gctgaggctg tgagtgcagg tccccgttcc ccgccttccc 720 ggcccggctc gccgcgggtc ggctcctgtt gcagccccgc gggccgggcg cggcctcccc 780 aagccctcac gggaacccag ccggacggaa gcaccatgcg cctcccacgc gctggctcga 840 cctcgtccaa ggtcgctgcc gcccgcgcct ccctttggga gggccctcgt tgcacccgga 900 aaatgggtac acaccttagg tcgaattacc tggactcagc cccaggaaac aaaactgcct 960 gcctccccca aatggcaaac gtgaacgcgc cagtctttga gaggcggctt gcctggtgaa 1020 gaacacctgc ggttcgttta acacagcggt tcccaaacgg tttggtctca ggacccgttt 1080 gaacacctgc ggttcgttta acacagcggt tcccaaacgg tttggtctca ggacccgttt 1080 acacgtctaa aaataatttg aggacccccc aaagcttttg cttatatcta ttgatttttg 1140 acacgtctaa aaataatttg aggacccccc aaagcttttg cttatatcta ttgatttttg 1140 tgcatgagga attaaaccag aaatttaaaa aatattcact taataacaaa cccattaact 1200 tgcatgagga attaaaccag aaatttaaaa aatattcact taataacaaa cccattaact 1200 tttaatacaa ataacacttt aatgcataac ttctcaaaaa aaaagtgaga agattggctc 1260 tttaatacaa ataacacttt aatgcataac ttctcaaaaa aaaagtgaga agattggctc 1260 atggttttta tatttttgaa aatttctttg gtttaataaa ggacagctgg agtctcatat 1320 atggttttta tatttttgaa aatttctttg gtttaataaa ggacagctgg agtctcatat 1320 cagctgtttt gaaggaaata tttgaaagaa attctgggct catgcagatg cagatattag 1380 cagctgtttt gaaggaaata tttgaaagaa attctgggct catgcagatg cagatattag 1380 ttgggaaagg gaggaccaag cagaccctct caacgggtct cagggacccc ctcaggtgtc 1440 ttgggaaagg gaggaccaag cagaccctct caacgggtct cagggacccc ctcaggtgtc 1440 tttggaccac actttgagaa ccgctggtct agcacttgcc catcgcctag gtacgtgcac 1500 tttggaccac actttgagaa ccgctggtct agcacttgcc catcgcctag gtacgtgcac 1500 atacatggtc tcatttggtt cccttgacca ccatgtgaag aagttgggtg tcatgtttat 1560 atacatggtc tcatttggtt cccttgacca ccatgtgaag aagttgggtg tcatgtttat 1560 aaagatgaaa aaacagggtc agaaaggtta aggattctca aagccctatg accagtaatg 1620 aaagatgaaa aaacagggtc agaaaggtta aggattctca aagccctatg accagtaatg 1620 agagagtttg gttcaaacac taagtgtatg gctcctggtc tggaacttct ttatttattt 1680 agagagtttg gttcaaacac taagtgtatg gctcctggtc tggaacttct ttatttattt 1680 atttttattt tttgagacag agtctcggtc tgtcacctag gctgtagtac agtggtgtaa 1740 atttttattt tttgagacag agtctcggtc tgtcacctag gctgtagtac agtggtgtaa 1740 tctcggctta ctgcaacctc cacctcctgg gttcaagcga ttctcatgcc tcagcctcct 1800 gagtggctgg cattacaggt gtgcaccacc acccctggct aatttttgta tttttagtag 1860 agacagagtt tcgccatgtt ggccaggctg ttctcgaact cctgacctca agggggtcca 1920 cccggatcag cctcccaaag tgctgggatt acaggcgtga gccaccatgg ccggcctggt 1980 ggtctggaac ttcttttttt tttttttttt ttttttttcc gagatggagt ttcgcttttg 2040 tcgcccaggc tggagtgcaa tggcgccatc tcagctgact gcaacctccg cctcccggat 2100 tcaagtgatt ctcctgccgc agcctccgga gtagttggga ttacagggcg tccgccacca 2160 tgcccggcta atttttgtat ttttaataga gacgaggttt caccatgttg gccaggctgg 2220 tctggagctc ctgacctcgg gtgatccacc tgcctcggcc tcccaaagtg ttgggattaa 2280 aggcgtgagc caccgtgccc ggcctgggtc tgaacttctt acgtcttctt gcactgtgac 2340 tcttcagtca cccatgtcct atgggaggac ccacctagtc ttatgcatcc cactttccca 2400 tttcctccgc tgcccctggc acaggggagt cacccctcgc cctagaggag cagctcaggc 2460 agtgtatcca ggctggtcca gaagggtcct tttctggcgc cttttgaggg ggccagcgtc 2520 agtgtatcca ggctggtcca gaagggtcct tttctggcgc cttttgaggg ggccagcgtc 2520 tgtgacctgt tcagtagcgg gtccagtgag cgcacacagt ccccacgcca gtacgctggg 2580 tgtgacctgt tcagtagcgg gtccagtgag cgcacacagt ccccacgcca gtacgctggg 2580 ggttaactca gtgctcctca cccgccttag cacactggtc tgttctcgac atgtctaccc 2640 ggttaactca gtgctcctca cccgccttag cacactggtc tgttctcgac atgtctaccc 2640 gctgaatata ggattcttga ttggcatttt ttttcctttg agcattttaa agcgtattat 2700 gctgaatata ggattcttga ttggcatttt ttttcctttg agcattttaa agcgtattat 2700 cgcactgtct ttttgcctcc attattctaa tgagaattca gacgttaatg ttattagggt 2760 cgcactgtct ttttgcctcc attattctaa tgagaattca gacgttaatg ttattagggt 2760 tctatgtaag tgatgagtct tttttttttt tttttttttt ttttttttgc ttgattttct 2820 tctatgtaag tgatgagtct tttttttttt tttttttttt ttttttttgc ttgattttct 2820 ctttgtcttg gtctttcggt ggttttatta tgatgtgtgt agatctgttt gagtttatcc 2880 ctttgtcttg gtctttcggt ggttttatta tgatgtgtgt agatctgttt gagtttatcc 2880 tcttggaagt ttattgagtt tcttggatgt gtatacatca aatttgggaa gtttttggct 2940 tcttggaagt ttattgagtt tcttggatgt gtatacatca aatttgggaa gtttttggct 2940 attgttctct caaaaaagtc tactctctta tctctctctt ctctttcgga ggctcccatt 3000 attgttctct caaaaaagtc tactctctta tctctctctt ctctttcgga ggctcccatt 3000 gcgtatatgt tggtgtagtt gatggaaccc cacaggtctt tgagactgtt ccattttctt 3060 gcgtatatgt tggtgtagtt gatggaaccc cacaggtctt tgagactgtt ccattttctt 3060 tctgttcctc aggctggaca atgtcaattg acctgtcttt gttttttttt tttttttttt 3120 tctgttcctc aggctggaca atgtcaattg acctgtcttt gttttttttt tttttttttt 3120 tttttttgag acagagtctt gctctgtcac caggctggag ttcagtggcg caatttctgc 3180 tttttttgag acagagtctt gctctgtcac caggctggag ttcagtggcg caatttctgc 3180 ccactgcaac ctctgactcc cgggttcaag cgattctcct gctccagctt cccgagtagt 3240 tgggattaca ggcacgcgcc accacgtcca gctaattttt gtatttttag tagagacggg 3300 gtttcaccat gttggccagg atggtctcga tcttctgacc tcgtgatcca cctgccttgg 3360 cctcccaaag tgctgggatt acaggtgtga gccaccgtgc ccagccgacc tgtttttagg 3420 tttgctgatt gctccgtctt ccaactcaaa tcattttcat tttaatccag aacttctttc 3480 tttttggaga atgagtctca ctctgtcacc caggctggag tgcagtggtg caatctgggc 3540 tcactacaac ctctgcctcc tgggctcaag ctgtcctccc acctcagcct cccaaatagc 3600 tgggactaca ggtgtgcacc actgcacctg gctaattgtt tctattcttt tgtagaaatc 3660 aggtttcgcc atgttgctca ggctggtctc gaacttggct caagcaatct gcctgcctct 3720 gtcgctcata gtattgggat tacaggcatg agccatcatg cccggccctt ttctatactt 3780 tctatctctt cattgatatt ctctatgtac tgaaacatcg ttctcatact ttcctttagt 3840 tctttttttc tttctttctc ttagagacgg agtctccagg ctggagtgca gtggtgagat 3900 cgtagctcac tgcagcctgc agctcctggg ctcaagtgat cctcctgcct cagcctcccc 3960 agtagctggg actacaggca cacaccacta tgcccagcta atttttaaat tttttgaaga 4020 gacagagtct tgcagtgttg ccaagggtgg tcttgaactg ctgggttcaa atgattttct 4080 tgccttggcc tccaaagcag tggaattcta ggcatgaacg actgcacctg gactccttca 4140 gttctttatg catagtttcc tttcgttttt tgaacatact ttaaatagat gatttaatta 4200 gtaagtccaa catctggact tcctcaggga cagtttgtac tgactacttt tttcctgtgt 4260 gtataggtca tactttcttt tcgttgcatg tcttgtatat ttttttgaaa actggacatt 4320 ttaagtaata taatggggca actctggaaa tcagatttct cccttttgcc agggtttatt 4380 gttgttgttg tttgcttagt gactttaata aactaattct gcattgtatt ctttgtcatg 4440 tgcaaccagc gaggtctctg cttggttagc ttagtagtct aacaattaga caaaggtttc 4500 cttaaattcc tggaaccagt aagtctccta gtctttgctg agggactctg tgtgtgtcag 4560 gacatgtctt caacattcag ccacagtttg tcctgcttcc acagagactg aaggttagct 4620 agacatgagc acttaaggtc tttccttagc cggtatacag ccctgcacat gtgcgtggcc 4680 tcctagagtc tcaggaacat gtcacaactt ttcagagctc tctatgggta tcttgttctc 4740 catttttgcc ttttaagtgt tttggttagc ctgttgttta ccctgttatc ttccatctca 4800 agtagctgtg atggtaaata agtacttctg attgtttgga caaatgccct ccgggaaaat 4860 actgttccca ctgtgcaagc tctgagtcag gtcaattaaa gacagtttta caagtagggt 4920 cttccaagga accacaagac aggtcaaata gtgacatttt ccagaaatgg ggctttcccc 4980 ctctagtggc tgccatgctg ctggttttca ctgtgatggg ggctattgat tttcaagtct 5040 accacagagc tggggaagag ggatagggta aactaaaccc ccacgacaat tgctgttctt 5100 actgagattc atacattttt cttgaataaa tgatccctat attgctgcaa gtctttaatt 5160 tctagaattc tgaaaaagtt tattctgaca atttttgcca atattcttgt tgattttatg 5220 gaggatagga ttttcaaagg tcctttttct gtagtttttg ctgaagtcag cctgcataga 5280 gttttgatcg agtggatttg aatttataaa tttttttttg catctgttgt gttgatcata 5340 aggcttttct tcttcagtgt gttaatacgg taaattacag tgattgactt tttaaaagtt 5400 aagctgacca ttgcattcct ggaataaacc ccactgattg tgatgtattg tttggattgc 5460 atgtgctaaa attttgttaa aattatttgt atctgtgttc ataaggaata atcgcctata 5520 gttttctgat attgccttgg tctgggtttg gtattaggat ggtgatggtc acataggatg 5580 tgttggagag ttcccgcttc tatttcatga aacaatttgt gtaaatttgg taagattttg 5640 cctgtaaatg tttgcacctg cagttttctt tgtggaaaaa gtgttaaact acaaatttag 5700 tttctttaat agacatattg ccattcagat tatctagttc ttcttgagta agctttggta 5760 gtttgtatct ttgaaggact atgtccattt catctaagtt gtcacattca cgtgcctgga 5820 gttgttcatg ctgactcctt atcctttttc ttacagctct acaaaggtat ataccacaca 5880 attcaccctt tgtaagtgtg tgattaaata atttctagta agtttatagg cccatcacta 5940 cagtccaatg ttaggccccc ctccaaattt ctatcatccc gcaaattctc ttgagcccat 6000 ttgcagttaa tccttacccc tgcctctagt ctgtcatctc cttactgttt gtgtcatcca 6060 tagtgatgtg tccttcttca tccttgatat ttgtaattct tttttctctg ttttttattt 6120 tatcacctgg cttgagttat tttgctcttt ttaaagaacc atctttttgt tttatcgatt 6180 tttctctttt ttttgttttc tatttcattg atttctgtct catctttatt atttcctacc 6240 ttcaccttac tttgggttta atttaccctt taccttttta aggtggaaat ttagataatt 6300 aattttagac ctttctactt tttttttttt tttttttgag acaaggtctt gctctgtcac 6360 ccaggctgga gtgcactgat gcaatcttgg ctcactgcaa cctccacatt ctaagctcaa 6420 acagtcctcc tgcctcagcc tcccaagtag ctgagactac aggcattagc caccatgtct 6480 ggctaatttt tgtatttttt ggagagacgg ggtttcgcca tgttgcccag gcttgtctca 6540 aactcctgga ttcaagcagt ctacccacct tggcctccga aagtgctggg attatggatg 6600 tgagccgcca cacccatccg gggtctactt tctaaatata agcactcagt aattacttta 6660 cagccatgct tggtggctca tgtctataat cccagcactt tgaaaggctg aggtgggagg 6720 atcacttgtg gccaggagtt caagtccaga ttgggtaaca tagtgagatg ctgtcactac 6780 aaaaaaaatt ttaaaagtta gctggatgtg gtggctcatg cctgtgatcc tagccacttg 6840 ggatgctgag gtgggaggat tccttgagcc tggggatttg gggtttcagt gagctgtgat 6900 gataccactg cactccagcc tgggtgacag agcaaagctc tattaaaaat agtgatgata 6960 ataataataa taataataat aataataata ataataacaa aattccctct aaggattgtt 7020 ttaattccct ctagcaaatt ttgcaagatt tctgatacct gtaatctctt ttcatttctt 7080 atgttgatac tctgtttctg ttttctaaag tattcatagt aagactccaa aagaaaaagg 7140 gctccctggc tgtatgtgtt cagtggtagc agccaaggaa tgtgccagga aggtacttca 7200 cattttgcag tgccgtggtg gagaggaatc tggtaaagaa aaaaccagat gaagacagtt 7260 tagagaggcg acaactacaa gctaagtacc tgagattctg ggtcaacagg aaggggacag 7320 tgtgtgagga agcagcaggg ccatggaagg tggctgaggg gcctggatgg ggttacctgt 7380 cagggtccca ataggccttt tgtagggttg gcttcatctt tgaagcctcc tgaggttacc 7440 gtgacaacca ctactgtccc tctgtactgc tgtgaggcca ctaagggaaa aagggaggga 7500 agggacccaa agggaggaca gaggtgggag aagagggagc gatggggaga gaaaggcaga 7560 gaacaatgga agcagtgagc gagaaagggg agaaacctca ctgaaaagga ggcagggtaa 7620 acatgactga gatggaaaaa tgatcacatt aaaaagaact cactcagagt aaggatgaat 7680 gtgtgttctt tgctacttga actggtatta aaaggccaat gtgaagtgtt agatatttta 7740 aaatgtttat tggtatcatt agaagttgac ttcagttgcc caaatgcatg gtgagaaagg 7800 ggatgtgtaa acaaaaatga tattttggat tgtgaggcag aaagtcattt taaatggccc 7860 tgggaggcag agggcccaat cctctcagtc gcctgggact gtcctgggtt tagcatggaa 7920 tgccttatgt tcccagaaac cccttaggtc ccagcaaacc aggatggcca ttcaccatag 7980 aggtggagaa gtaaagggct gtggatgaag acgggtactt attaggaaca gggaggagga 8040 tggagttgga ggggatgagt gcagggcagg atacagagcc cagaaggaaa ggaattgaca 8100 gaagctggga gaaagaataa aaagaccagt ggtccaagat ggtagaccac aggttcaaga 8160 actggagcaa gagtcttctg gaatggcctc cattctagaa ccgagagaga gtattcagaa 8220 tgctttaatg gagtaagaag ggccactgct tagagtgggg ctgaagagaa aagaaagaag 8280 gagaccctgc ggaagcggca ataccaggaa ggcagttgta tgcacatcat gcagaccagg 8340 agtagcctag gggtagggga catgcctggg ctggacagtg gaatttgtgg gaaatcagat 8400 ttccctgagc aaaaaccagt gggcagcagc caaactggtg agcaagaact ggctgtggtg 8460 gctgggcccg aattgggagc tacacctgcc cgggcaggtg tagctgcgag ggaggtctct 8520 gccaacaact cagctgtgcc gtgcctgctg aacggcctca ccttggcctc tggctctgtt 8580 gcagctgtgt ggggctcgtg ctcctacata ccgttatcct gtggtagagt tttaccgtgg 8640 accagagcca tctagaggac attgagttac ccacatactt tctgtgggag aggccaagga 8700 gtcagggggg gaggccggga cctgaagatc aactctttac tgcagagatc cctccctccc 8760 tctctctgtc ccatggatcc agagggaagt tgaggacagt ttcatgccac atgcttgacc 8820 cgactcaact gagtaactgt ggccactgag aagactctgt ccttgcaaat aactctcctt 8880 agtgcccata ttctgcagaa tgtttgagtg gcacgagtac ttcattcgtc tagaaatatt 8940 tcatggtagt cagttgctaa taacaggata tctgcaggta gtgtgttctc aggaggcaca 9000 cctgaagcct caaaggtgag gacatagaca tggctgtggg tgagcctgtg ccaagacatg 9060 acccatatcc atggggagtg ctctcagggc agctgacact gctcagcctc agcagtctcc 9120 agtgatagct tcggaacagc cacggtctga ctgatatact ttctgaaaac agcctttggt 9180 cacacagttt cctaactaaa tgtggtggtg gccactagaa gttgccattt tatgcgggac 9240 tccctgtgag gaagcccata gtgctggtag ctgaaattta ggaatgcagg tttctaagaa 9300 ggctcggaaa agcaagctgg gctccttggg gaggagctat tagtacgttt aattaaattt 9360 tttaacatat aaaaagttgt taattataaa aacaacattt ttttctaccc tgagtgcagc 9420 tggagctgtg ttctgccgtt acacaattcc tcattaattc cacacattgc ttacgatcct 9480 gtgaagttta gaattttatt aggcaataaa taagaaatca aggaaataat atttgtggtt 9540 tcatatcggt cacttgggta tgaatttaaa tccactgtaa ttcaaacagg tttagtttat 9600 ttcatgttgc tacttgaatg tgttatagcg tatcaattaa ataactcact tgaatgatgc 9660 agacttgaga ttcattccct ccttctttcc ttccttcctt tctcacccca tcctccctac 9720 cataaaccag gtttagaata gctgttaatt tagctgcttt gattttttgt tgttgtttat 9780 tttgaaacgg agtctcactc tgtcgctcag gctggagtgc agtggcacaa tctcagctca 9840 ctgcaacctc tgcctcctgg gttcaagcga tcctcctgcc tcagcctcct gagtagctgg 9900 aattacaggt gcacaccacc acgcacagct aattttttta tttttatttt ttatttttag 9960 tagagatggg gtttcaccat gttggtcagg ctggtcttga actcctggcc tcgtgatctg 10020 cctgccttgg tctcctgaag tgctgggatt ataggtgtga gccaccacgc ctggcttttt 10080 tttttttttt ttttttttta agactggtga agctatgttg cccagactgg actgcagtgg 10140 ctattcgcag gtgttatcat agcttactgc agccttgaca tcctgggcac cagtagcgct 10200 tcttcctcag tctctagtag ctgggactgc aggcatgtgc taccacaccc agctgatttt 10260 atgttttgtt tacatttaac gatgttaatc taggagcagt gatgatgttc actgttgttc 10320 cacatgactc tagcacatat atatttgtat gtgtataacc acagttgttc atttagtctg 10380 tttccaggtt ttcatttagt ctgttccagg ttcatttagt ctgtttccag acctaaaaag 10440 gcctgttggc taccgcgaca gccttggctg tcacactggg cccgcccatg ggtgtttctg 10500 taagtggcca tgaacctctt ctgaactaca ccacatgtta acatccttct cttagttagg 10560 tctttgagat tggcagaggt gctctattta ataataaagg aatgggctgg gcattggtgg 10620 ctcacacctg taattccagc actttgagag gctgaagtgg accgatcact tgaggtcaga 10680 agttcaagac cagcctggcc agcatggtga aaccccgtct ctactaaaaa tacaaaaatt 10740 agcctggtgt ggttgcgcat gcctgtagtc ccagctactc aggagactga ggcaggagaa 10800 ttgcttgaac ccaggaggtg gaggttgcag tgagccaaga tcatgccact gcactccagc 10860 ctgggtgaca gagcgcgact ccgtctcaat aataataata atagtaataa taataatata 10920 ataaaggaat gaaagtttcc ttagaaagta aggtttctat tttaatagaa tacttgagtg 10980 agcaagtctg tcttaactgt ttctcctcac tgtgtctaaa tgagaaggaa gaatgaccct 11040 ggtgtctgta tttgcaggac acttttcagt attaatattc agaaccccaa atattgaaaa 11100 gtgtaaaagg cagtacttgc tacttcattg tcccgttgct agccccagta aaatgagggt 11160 gtgtaaaagg cagtacttgc tacttcattg tcccgttgct agccccagta aaatgagggt 11160 cttagtttag cggaagactg tgtttactgt ttaactcaca gttcacgcta aaaattggca 11220 cttagtttag cggaagactg tgtttactgt ttaactcaca gttcacgcta aaaattggca 11220 cttttttttt tcctaataag ggcctctcta tggacctttt taaagtttat atggtcatta 11280 cttttttttt tcctaataag ggcctctcta tggacctttt taaagtttat atggtcatta 11280 aaaacatcat tgcacacata tactttgtac cactttggaa gtagtgcagg cttgtttgaa 11340 aaaacatcat tgcacacata tactttgtac cactttggaa gtagtgcagg cttgtttgaa 11340 agcatacact tgacttgtct tcttacttag tctcacagta ttctttgtga gaaggttgat 11400 agcatacact tgacttgtct tcttacttag tctcacagta ttctttgtga gaaggttgat 11400 atgcccacat tattctgtgg ccaaaaacaa aagtttagga aatgaaaaca gcaaacagaa 11460 atgcccacat tattctgtgg ccaaaaacaa aagtttagga aatgaaaaca gcaaacagaa 11460 aaccccatgc caagtgctgt tttcgtagtg aggcactgga gggcgctccg ctctagcaat 11520 aaccccatgc caagtgctgt tttcgtagtg aggcactgga gggcgctccg ctctagcaat 11520 gggagatgca cgcagtctct gcagtctgca ggctgccctc acgaacgcct ttcagaagat 11580 gggagatgca cgcagtctct gcagtctgca ggctgccctc acgaacgcct ttcagaagat 11580 gtaagagttt tttctgttca ttttaaggca aatgaaagac acatggatgg tagaggaaat 11640 gtaagagttt tttctgttca ttttaaggca aatgaaagac acatggatgg tagaggaaat 11640 atttctatac tgacctataa acggtccctt taaaagttta tgagaaactg aagaagtagc 11700 atttctatac tgacctataa acggtccctt taaaagttta tgagaaactg aagaagtagc 11700 tctgtgctcc agccctcctt atgacctcct ctctccctag cctgttgcag cctcaccggc 11760 tctgtgctcc agccctcctt atgacctcct ctctccctag cctgttgcag cctcaccggc 11760 ctcctcttct gggctgtgtc gtcagcctga agccctctgt ccctttgtgc ctcagctcaa 11820 atgtcacaaa tcagagaggt cttccttgac caccctgtac aacatagcct ccaccccgct 11880 aagcagcctc tgcactaccc tgtcccctta ctgctgtttt cattttttcc tattgcccca 11940 tttccaccag acatatcaca tgcttatttg tttattgtcc gcctgtatcc taccccagcc 12000 cccagatatt catgagagca gggacttggt ttattccatt gctactatat atccagaacc 12060 taaaacaatc cttggcagtg cctgatataa atttgcagaa ggaaaaaaga acagatttag 12120 gaatgaaata ttttgagtgt ttctaaagat ttagtggatt ttttttcttt attttggtaa 12180 attatacata acaaaattat cattttaact tttttttttt tgaggtggag tctagctctg 12240 cccccaggct ggagtgcagt ggtgcgatct cactcactgc aacctctgcc tcctgggttc 12300 aagcaactct cctgcctctg cctccgagta gctgggatta caggtgcccg ccaccacgcc 12360 tggctaattt tttgtatttt tagtagagac ggggtttcac catgttggcc aggctggtct 12420 cgaactcctg acctcaggtg atctacccgc ctcggcctcc caaagtgctg ggattacagg 12480 cgtgagccac catggccggc ccattttaac catctttaag tgtacagttt agtggcatta 12540 CGTGAGCCAC CATGGCCGGC CCATTTTAAC CATCTTTAAC TGTACAGTTT AGTGGCATTA 12540 agtacattca cattgttgta caactgtcac caccatccat cttcagaact tttttatctt 12600 AGTACATTCA CATTGTTGTA CAACTGTCAC CACCATCCAT CTTCA GAACT TTTTTATCTT 12600 cccacactga aaccctgtgc ccattaaaca ctaactccct gttcccttcc ccccacccag 12660 CCCACACTGA AACCCTGTGC CCATTA AACA CTAACTCCCT GTTCCCTTCC CCCACCCAG 12660 cccctggtaa ccaccattct cattattttc actgtgaatt tgactactct aggtacttca 12720 CCCCTGGTAA CCACCATTCT CATTATTTTC ACTGTGAATT TGACTACTCT AGGTACTTCA 12720 catacataga ctcacatatt tgtccttttg tgtttggctt atttcactca gcataatatc 12780 CATACATAGA CT CACATATT TGTCC TTTTG TGTTTGGCTT ATTT CACTCA GCATAATATC 12780 ttccaaggtg catccatgtt acagcaggta tctgattttc attcattttt aaggctgaat 12840 TTCCAAGGTG CATCCATGTT ACAGCAGGTAT CTGATTTTC ATTCATTTTT AAGGCTGAAT 12840 aatctattgt gtgcctattc catattttat ttatccatcc atctgttgat ggacatttga 12900 AATCTATTGT GTGCCTATTC CATATTTTAT TTATCCATCC ATCTGTTGAT GGACATTTGA 12900 gtatcactgc caccttttgg ctattgttag tgcatgttct atgtggagca cagaaatgtg 12960 GTATCACTGC CACCTTTTGG CTATTGTTAG TG CATGTTCT ATGTGGAGCA CAGAAATGTG 12960 atgtttaagt ttaagcctaa agtggatttc tttgaattat actagccagt cacagatcac 13020 ATGTTTAAGT TTAAGCCTAA AGTGGATTTCTTTGAATTAT ACTAGCCAGT CACAGATCAC 13020 tcctgggtag tactatggtt tgaatgtggt atcccctcca aaattcatgt tgaaacttaa 13080 TCCTGGG TAGT ACTATGGTT TGAATGTGGT ATCCCCTCCA AAATTCATGT TGA AACTTAA 13080 ttcccattgt ggtggtatta agaagtgggg ccttgtagga agtggttaag tcatgagggc 13140 TTCCCAT TGT GGTGGTATTA AGA AGTGGG GCCTTGTAGG AGTGGTTAAG TCATGAGGG C 13140 tgtgcctcat gaatgggtta gtatcattgg aaagaggctt cagagagagt tttccagtct 13200 tgcccttcca ccctgcgcca tgtgaggaca cagcaataag gcgccatatt ggaagcagag 13260 agcagccctc tgtagacacc aaggctggca ccttgatctt agattcccca gcctccagaa 13320 ctgtgggaag taagtttctg ttgtttataa attagcccag tgtgtaatac tttttaaaat 13380 agcagcataa acagactaag acaggtggtt tctactttta ggaaaatatt tttagttcat 13440 attaagatag aaacttattc tagctcctaa aaatcagaat tgttttaata tcctcaggtg 13500 ctctttaatt ttttcttaca attgtcaaaa acaagtttaa aatcaagtta ctaactctgc 13560 tataggcagg aagctcctgg atagatactc tggggttcaa gtccaatgct gacccgcagt 13620 tggcccttga ccttgttcaa ggcccttatc tagattgtgc cttatttttc tcacccatca 13680 aaaggaggat ggacaagttt cttcctgcca gttgacattc cagagtctag aaaggaatta 13740 ttgccctcaa gtaagacgac cttggggaat tcagaaataa aaagtatata gctttttgag 13800 ctctaagtta ttttggtcca tttgagctca atttgaccag agattgaaga taaaagctct 13860 gcctccccca aacttattgg tacagcctgc actcaattgg ttttaccgcc ttctcctgac 13920 attaagggag atgtttttgg ttgtgtggtg ttgacccttt aggtcctcag aaggggcggc 13980 accctcatgg tgtctctgcc tgcagcaggg agtgctacca ggcactttgt tccagtgact 14040 tatgatggac acagcacagc gcctaggggt taggattcta aaataattca ggtgtatgga 14100 gacgcagaac ctaaaagaac tcaggagaaa gagaaggtag atatcctaaa ttttagtctg 14160 attttgactc agaagtggat acctgatttt ggcctacatt tcacctcttt cagttttttt 14220 ctgctccatt tgggttccct gctcacttgg aaccctcaag ctggcttgca aatgcccaga 14280 gccacagtgc tctcataggg tctgcctgtt ctagggccca tcagccagca gtccagcagg 14340 gatgaccagt gagcccaaga aaacaagcag tgctgatgtc agatgtgtag cccttctcaa 14400 gtgctataca ttttaagaaa tgccttcact gggcatggtg gctcatgcct gtaatcccag 14460 cactttggga ggccaagatg ggcggatcac gaggtcagga gttggagacc aacctggcac 14520 aacatgataa aaccctgtct ctactaaaaa tacaaaaatt agctgaacgt ggtggtcaca 14580 cacctctaat ctcagctact caggaggctg aggcaggaga atcgcttaaa cccggaaggt 14640 ggaggttgca gtgagctgag atcctgccat tgcactccag cctgggtgac agtgagactc 14700 tgtctcaaaa aaagaaagaa agaaaaagag agagagagaa agagagagag agagagaaag 14760 aaagaaagaa aaagaaagaa agaaaaaaga aagaaaatgt cttcttcaga cacctcaact 14820 acaaaggagt cagccagtca agagaatttc tcctctacca ggaagagaat atgggtatag 14880 gagaagcttg gagtctctct gccacccctc atgttctctg gacaaagtcc caagctgtgc 14940 gtttgaatca tcagtgtcta cttgtcctgc agaacatcag gaatttacca ggtaatctgg 15000 tgacatgcct gcaccagagg actgggagaa tccttaccgt taaacctctt ctccccaccc 15060 ctgaacacat aaatacactt tttaagcttc ctaacgctgt actctgatca tggtgcccac 15120 ccccttggac agcataaatt tcccagtgac ctccaattac ctgccgctaa agtccaggcc 15180 ccttagtgtg gcatgtacgc acctcctcac ctatcctcca tctttccatt tccccccagg 15240 ccgcttacct gtggcccaca tgcaccactc accatgcccc acactctcga gtcatcagtg 15300 ccgcttacct gtggcccaca tgcaccactc accatgcccc acactctcga gtcatcagtg 15300 ccacttcctc ctggaagcct tctttccctg gacacctcag ccagccatct gtctcgtccc 15360 ccacttcctc ctggaagcct tctttccctg gacacctcag ccagccatct gtctcgtccc 15360 tgagagccct aacctctccg ctgacttttg tcacccttcc ctgtgttaag gtgatcgaga 15420 tgagagccct aacctctccg ctgacttttg tcacccttcc ctgtgttaag gtgatcgaga 15420 tgcagaactc ccagtggtct ttgatgcctc ctctcctctc ccacccacac cccatctgtt 15480 tgcagaactc ccagtggtct ttgatgcctc ctctcctctc ccacccacac cccatctgtt 15480 gccaggcctg ttgattttcc ccttcaggtg ccaactcttc tttcattgta ttagtttcct 15540 gccaggcctg ttgattttcc ccttcaggtg ccaactcttc tttcattgta ttagtttcct 15540 attgctgttg taacagatta tcgcaaacat ggtggtttaa attaagcttg tccctgcagc 15600 attgctgttg taacagatta tcgcaaacat ggtggtttaa attaagcttg tccctgcagc 15600 ctgtgggcca catgtgacct gggatgactt tgaatacgac ccaacacaaa tttgtgagct 15660 ctgtgggcca catgtgacct gggatgactt tgaatacgac ccaacacaaa tttgtgagct 15660 ttcttaaaac atgagatttt ttggcgattt tttttttttt ccctttagct tatcagctgt 15720 ttcttaaaac atgagatttt ttggcgattt tttttttttt ccctttagct tatcagctgt 15720 tgttagtgga ttttatgtgt ggtccaagac aattcttctt ccagtgtggg gccagggaag 15780 tgttagtgga ttttatgtgt ggtccaagac aattcttctt ccagtgtggg gccagggaag 15780 ccaaaaaaac tggacaccct tggttaagtt aacacagatt tgttaactta tgcttctgca 15840 ccaaaaaaac tggacaccct tggttaagtt aacacagatt tgttaactta tgcttctgca 15840 gtttggaagt ctgacctagg ttttgtgggc tgaaaccaag gtgtttacag ggatggagag 15900 gtttggaagt ctgacctagg ttttgtgggc tgaaaccaag gtgtttacag ggatggagag 15900 tgttctttct ggcagttcta ggggtgatct gtttgcttgc ctgtttcagc ttctagaggc 15960 tgttctttct ggcagttcta ggggtgatct gtttgcttgc ctgtttcagc ttctagaggc 15960 tgcccacatt ctttggctca tggctcctta tccatcttca aagccagacc cagtgggctg 16020 tgcccacatt ctttggctca tggctcctta tccatcttca aagccagacc cagtgggctg 16020 aattcttctc cccattgctg ccatctctcg ggttctgttg cttctgccac ccttttccac 16080 aattcttctc cccattgctg ccatctctcg ggttctgttg cttctgccac ccttttccac 16080 ttagaagcac ccttgtgatt acgttggccc cacctggata accagggcca gtcgccccaa 16140 ttagaagcac ccttgtgatt acgttggccc cacctggata accagggcca gtcgccccaa 16140 ctcagggtca gctgattagc aaccttcatt ccactttgcc atgtaacctg acatatcaat 16200 ctcagggtca gctgattagc aaccttcatt ccactttgcc atgtaacctg acatatcaat 16200 aaatactggg tataaggacg tggacatctt tgggggccat tattctgcat acccatccac 16260 aaatactggg tataaggacg tggacatctt tgggggccat tattctgcat acccatccac 16260 tgttatagat tcaggtttga actcttggaa tgaccatcta atgggctgag ttccctcatt 16320 tgttatagat tcaggtttga actcttggaa tgaccatcta atgggctgag ttccctcatt 16320 gcctctcatg ttatgtgctg ccagcataga tattaagatg tacattctat catctagacc 16380 gcctctcatg ttatgtgctg ccagcataga tattaagatg tacattctat catctagacc 16380 tgttggttcc aaaaacatct cttggatctg tccttttctt tttgtctcca ttgcccctac 16440 tgttggttcc aaaaacatct cttggatctg tccttttctt tttgtctcca ttgcccctac 16440 cctagtggca aacaatcctg tgaaaaagga ttgtttacat gtatgggtgc ccctccaggc 16500 cctagtggca aacaatcctg tgaaaaagga ttgtttacat gtatgggtgc ccctccaggc 16500 aatattcaag tgtactcttc gaggctgttt tagtacattc accatattgt gcaaccatca 16560 aatattcaag tgtactcttc gaggctgttt tagtacattc accatattgt gcaaccatca 16560 cctccatcta gttccaaaac attttcatcg ccctaaagga gaactctatc cattgagcgg 16620 cctccatcta gttccaaaac attttcatcg ccctaaagga gaactctatc cattgagcgg 16620 ccattcccca ttccctcctc tcctcaaccc ctggcaacca ccagtctgtc ttctgtctct 16680 ccattcccca ttccctcctc tcctcaaccc ctggcaacca ccagtctgtc ttctgtctct 16680 gtgcatttac cattcctttg tatggatata gcactacatt ttgtttattt tttaaattaa 16740 gtgcatttac cattcctttg tatggatata gcactacatt ttgtttattt tttaaattaa 16740 atttaatttt ttagagacag gatgtagctc tgtcatccag gttggagtat agtggcaccg 16800 atttaatttt ttagagacag gatgtagctc tgtcatccag gttggagtat agtggcaccg 16800 tcacccaggc tggagtgtag tggcatgatc atagctcact gtagccttgg actcctgggc 16860 tcacccaggc tggagtgtag tggcatgatc atagctcact gtagccttgg actcctgggc 16860 ttgagtgatt ccccctgcct caaccttcca agtagctagg actgcaggca cccaccacca 16920 ttgagtgatt ccccctgcct caaccttcca agtagctagg actgcaggca cccaccacca 16920 cacctggctc attttttaaa tttatttgtc agttgatgga catttgggct gtttccactt 16980 cacctggctc attttttaaa tttatttgtc agttgatgga catttgggct gtttccactt 16980 tttggtgatt gtgactagtg ctgctatgaa cattcatata caggtatttg ttttcagttc 17040 tttggtgatt gtgactagtg ctgctatgaa cattcatata caggtatttg ttttcagttc 17040 ctttaggttt atacctagga actgctgtgc catatagtaa ttctatgttt aactttttga 17100 ctttaggttt atacctagga actgctgtgc catatagtaa ttctatgttt aactttttga 17100 ggaaccacca aactgttttc cacagtggct gcaccatttt acattccacc agcaatgtac 17160 ggaaccacca aactgttttc cacagtggct gcaccatttt acattccacc agcaatgtac 17160 aagcgttctg atttctccac atcctcgcca acacttgcta ttttccattg tttttattgc 17220 aagcgttctg atttctccac atcctcgcca acacttgcta ttttccattg tttttattgc 17220 agccatccta gtagatgtga agttgtattc tcgcggtttc ttccaaatca tttttaaatg 17280 agccatccta gtagatgtga agttgtattc tcgcggtttc ttccaaatca tttttaaatg 17280 tacaaattgt ctaatttcac ttctctgctt cacatccttc ctattgcatt tataataaac 17340 cctggccccc atcttggact ccaaggagta tatgatcttc tccctcctac ctctttctct 17400 cactgccttc catgctcctc actcagtcca cttaccgagc atcattggcc ttctcctgtt 17460 ccctcaagca gggaaggcaa acacccatgg gcttttattt attatttctt tatttatttt 17520 ttaacacttc cctccttagg tccagagacc atgggctttg aggctcaggg gattcagatc 17580 caaggtccat ttagaagtaa atgatgagat gagttgggca ctgtggctca cgcctgtaat 17640 tagcactttg ggaggccgag gtgggcagat catgaggtca ggagatcgag actatcctgg 17700 ctaatacagt gaaaccccat ctctactaaa aattcaaaaa attagccggg catggtggtg 17760 catacctgta atcccagcta ctcgggaggc tgaggcagga gaatcgcttt aacctgggag 17820 gtgaaggttg cagtgagccg agatcgtgcc actgcactcc agcctgggca acagagtgag 17880 actccatctc aaaaaaaaaa aaaaaaaaag atgagatgat gcattgaatg ctaggtatgg 17940 cgtgtatgta gttcataaga acaccaataa acagtggatg taaaaagcaa catcagatag 18000 atacctaaag gccatgtctt tggaacaggg gtaaccacct gaagcatgtc ctcccttccc 18060 tgcctttttc ctccctcaga gcaccccgtt cttcattgca cttatgattt gtcaccagcc 18120 acgaatgtgt gcatctccct gttatgtttg gtctcctgca ctgggctgga aactccacgt 18180 tcacccttta tgcctgtgcc cagcacagtg cctgacacag taggcactga ttaaatgatt 18240 cttgaatgac tgcatcatca tatggtcaca gtaaacagaa tccatcacta attatctatt 18300 tgacatgagc gggtggaaag gcaaagtttt tatgcatgag tgtgggtgta tgtatgtttt 18360 cctagattat agatttgtct aaacattcct gctaatttta agatatcaaa tctgaagttt 18420 ccttggcttg aatttttgaa cataggccaa ggttgttttt agattcctca gtattctctc 18480 ttttgtgaaa gttttcaaaa gtataaacgg cgcatcttcc gtacaactgg tttacattgt 18540 tggtttatgc agttgagttg tgggcagagg ggaaacctca atgtctgaat gtggctttac 18600 cccagcgctg agatatcccc gggagagctt ggccagtctg agtccatgtg gcctaaagat 18660 cctccctagc cagttccctg tcccacctac attctgttta tttcagaact tctcacctgt 18720 tgaacacaca gcccttcctc aacacaggac cagctcagca tcgggaagct gtattttaag 18780 aaaatcatgg tcttccagct cagtggaggc atcttagcgg gatccaaaat aacattcgct 18840 tttatcttga aattctttct gtttttggtt tttccccaac tctcatcctt cccccacctt 18900 cacctcagac actccaggtt attactgaaa ttaagcatga tagaaagtgt ggactcatag 18960 gctcagttct gttccctcct tatggccagc gtgtggaaga gctcctctgc tctcatgcct 19020 tctccatcgg agctggttca tcaccttggc ctgtcttggg cggagtcccc gacttgggag 19080 ttgtgctgtt cgtttgctgt ccatcgggct gtaagctcct tgaggaccag gcaccatgtt 19140 tactcacccc tgtcccttct gggaacctgg gggaaactgc taactggcgt ggctctcaga 19200 ccttgcatgg ccccacacat tttttttctg tgggaaccac taagcattag caactcgcca 19260 gtgatcagca gggattagca gcgtttaggc tccgcagaca tttaatgtgc taattgagga 19320 gcccagggtt cggccaagga agtgatttac aaaggtggaa acgggactgt tgacacaggt 19380 gcatcttatt aattactggt ttggggaatt agtaaaaatg ttaataattt gagggattat 19440 tgttaatgaa ggataagaaa cttcacaacc atatggttta ctacttgatg cgattttccc 19500 ttttttaaaa aaatactttt aaatctgggt tttaggaagg acaagccatt tcttgcagaa 19560 cttattgatt gtattacata ggtaaatgga aaaatatttc ctagaacaat tttttgtaac 19620 agaaaattct aaaacatacc agtagtatga agtaagaggc acctatggca catctttgta 19680 ttattcagtc cctcctttat acattcttct gacaagaacg ggagtcatgc ctcccaggct 19740 ctgagtccca ggctccgagc attactgagg cagctcctaa gaagagagaa gggaagaaca 19800 tttcctgagg atctttgctg tgccaggttc cagaaggtgc tttggcgcat ctcacgtgag 19860 atacgggctc cttttgcctg ctttctaaga agggtgcggt gtgaggtggc tggctgcgtg 19920 gaagaggcct gggaggagta attccaccag tattgtgtct tagtcttcaa gtggccagtg 19980 cagtaccttg gcctgtggta gatcctgagt aaatgttgac tgaaacgaag atgccatggc 20040 cggtggtgaa gcagggatca gtgcaacaat gctgtgagaa tacagaggca ggggtggctt 20100 ttggagggca cctggatcat tcccagggaa gctgccctct gggcagagtt taaagtatgg 20160 cagagggaag agtgttcttg gcagagggca caggcccggt aggggagtta tgagcaggta 20220 attgtggagt atgggattcc caggggctgg gcatgacgtt gagagcgggg gaggttgagg 20280 ccagactgtg aagggattcc caggccagcc caggactgac ttcagattta catcctattg 20340 gcaacaagga gccagtggag atgtttaatc tctttcggcc tccatcaagt tagcctttca 20400 gtctgtaacc tggcctaagt gttagtccca tgtcatttga accccttgcc tttctctttt 20460 acttaaatta cattcacagt gcgctgtgaa acctaattgg aatagacaaa acgtggtgga 20520 actctgcagc ttatggggcc agttcatgct caccttaatg tgcttataat gtacatttaa 20580 atactgccag ttatttggag ctttcaggtg acttcacttc tctgtgtatt atgtacttcg 20640 cttcctacat ctgcagttaa tttgttaaga gaccttaatg agaaacttca aacagtttaa 20700 atgtcaatta ccaaaaaagt aaggttttca agttccagta atctacccga ttggtcatca 20760 gaatgtagtc ccaggtggat gtaatttaag acatgacatt ccagagcttc agttacaaag 20820 gaatgtagtc ccaggtggat gtaatttaag acatgacatt ccagagcttc agttacaaag 20820 aatttatata tttcgggact aataaactgg ctccagtggt ggctacactt gcaggattgt 20880 aatttatata tttcgggact aataaactgg ctccagtggt ggctacactt gcaggattgt 20880 ctgcagttat gactcatcta gtaccggcgc tgctcagaac tttctgtcca cggagttcac 20940 ctgcagttat gactcatcta gtaccggcgc tgctcagaac tttctgtcca cggagttcac 20940 actggttttg tgtttgggct ggtgcttttg aatctgaaaa aaagtgcata aaaatatgcc 21000 actggttttg tgtttgggct ggtgcttttg aatctgaaaa aaagtgcata aaaatatgcc 21000 agaaatgtag taaatattgt gatcatttca ttttttaatc cgacactttt gagaggaaaa 21060 agaaatgtag taaatattgt gatcatttca ttttttaatc cgacactttt gagaggaaaa 21060 gagtattcat cagtacttta tcatagattt aattaatggg ataatcttta atttctgact 21120 gagtattcat cagtacttta tcatagattt aattaatggg ataatcttta atttctgact 21120 cagacatctg gatttgtttc ttgccacccc ctcctttttt tctgtgttcc tttctcgtct 21180 cagacatctg gatttgtttc ttgccacccc ctcctttttt tctgtgttcc tttctcgtct 21180 ttatttttga tgaagataaa atacttaaag aaagcgttga atgtgatgat gagtattgtt 21240 ttatttttga tgaagataaa atacttaaag aaagcgttga atgtgatgat gagtattgtt 21240 ttaacagaaa gtaaaatcaa aagcatgcag aataaattaa ccttgcattt tctcggcagg 21300 ttaacagaaa gtaaaatcaa aagcatgcag aataaattaa ccttgcattt tctcggcagg 21300 tcagttagtg ggtgttatat ccttagttgt tggtgctaca aagtaaataa tttttggcag 21360 tcagttagtg ggtgttatat ccttagttgt tggtgctaca aagtaaataa tttttggcag 21360 ctggtttagt tttcaccact tctaagccta tgacaaatgg tcttggaaga tctttggtct 21420 ctggtttagt tttcaccact tctaagccta tgacaaatgg tcttggaaga tctttggtct 21420 taaaaaggaa aattttctgt gtgcgagttt taagttcagt tttgttgggt tttgaagtct 21480 tttttatgta agccacttat gtggcatttt gttttatttg aatttgaaat tttttttaag 21540 ttttaaagct agagtctgcc acattgaact gtttttcaca cttggcattt aaaagttaat 21600 ttgctttttc tgtttttaaa agagactaaa aaactaaggt ttcccctact aagcatttat 21660 tagtcaacca cttttgaaaa ttgaaaaatt agtttgtatg gttttagatt gtttttctcc 21720 caggagaaat aatgttgcta cataatctca gacctgcatt cacgtattta gcctgccata 21780 agtcactgtc aaaatgaaga aaatgctgaa agttagatat acctgcacat gtgaatttca 21840 tgtattttct ggtaatcaac tctgttttaa aggcttttgg gagttgtttt ctttcagaag 21900 cagcccactg gtgctgtatt tggcatgttg tgtgttatta aaacttcata taaatgcaga 21960 ggtactaaaa cgtgaagttt acaaaaccca tcggaacatt tttacttgtg tagttagcac 22020 gatcacttct ccaagtacaa taatctttgt gttgtgagcc tgtagacaaa actcgtgtct 22080 ggcatatctg attggaatgt gaaatttaat attttccact agttatttcc caacatgcaa 22140 agtgaactga tacccattaa atatatatat ttaagtaaaa atagagagga tcagtggcct 22200 aatcagcttt acagctgtga atttaggaag tgaattcaca acatccctct ctccagagag 22260 acgacttgat gacttactct cggtttcttt aagtgatata gtagaccttg tttggattaa 22320 tcagcagcct tgggagttgg agcagtcatt tgcaatattg agtggaaaag ttccttgtgc 22380 ttggaaaagt cagaaggcca ggagtgagag acatggcctc atcctttccc tggtggctcc 22440 tcacctgtga agttgcacca gggtggtaat aaaattgtag gagctcattt tatgtgaaac 22500 cattagcagc ctgggaattc aggactagct gcctttcata ctttaagaca ctggaaggtg 22560 tcatttataa gcatagagag cctgcaggaa ggcagaacag gcttgtaaac tggagcctga 22620 ggctctgggc agcaatcagt gtgtaataga gtttaaagtc ttaggttatc agaatcacaa 22680 atgggccttt aagtgttgtg ggcaaatccc aagaaacatc tgtctataaa aggcaggtac 22740 aatggccagt tatttcctcc tggttattgt gatattgtta ttttctgttt ctgcctttgt 22800 ggatgggagt ggggagagaa agggaggggc ccttgctaca agtgtaagaa taaaattgta 22860 ggatgggagt ggggagagaa agggaggggc ccttgctaca agtgtaagaa taaaattgta 22860 aatgtttgtg ttagttgaaa aaaaattgac tatggtaagt gtcgcagcta atgtagggtg 22920 aatgtttgtg ttagttgaaa aaaaattgac tatggtaagt gtcgcagcta atgtagggtg 22920 ttttcacctt gaaacagcga ctgccgccca gttcctgctg cttcgttcgc ggactctgaa 22980 ttttcacctt gaaacagcga ctgccgccca gttcctgctg cttcgttcgc ggactctgaa 22980 ttccactgct ggggcaggcg tgtgtgacag gcagagagag ccctggggtt gggggaacaa 23040 ttccactgct ggggcaggcg tgtgtgacag gcagagagag ccctggggtt gggggaacaa 23040 ggaagtgttt ccagttccca tttggcttat gcattggagg gttccccaat gtggggacag 23100 ggaagtgttt ccagttccca tttggcttat gcattggagg gttccccaat gtggggacag 23100 cgtccaggtt cttggcctgt ggaaatgaca aatggccctg acagggggct atcccagctt 23160 cgtccaggtt cttggcctgt ggaaatgaca aatggccctg acagggggct atcccagctt 23160 cagtgtttct gttcaccatc ctaagagtgg ttctttatct gaaaccatga agaaaaccag 23220 cagtgtttct gttcaccatc ctaagagtgg ttctttatct gaaaccatga agaaaaccag 23220 atgattagag aagaattcaa gaacccattc tgtagacatt gcctcctgac aagtacaaat 23280 atgattagag aagaattcaa gaacccattc tgtagacatt gcctcctgac aagtacaaat 23280 gcgtctgttg aaatctcagt gctccaagtc cccatctatc cgccttcccc ctcctccagg 23340 gcgtctgttg aaatctcagt gctccaagtc cccatctatc cgccttcccc ctcctccagg 23340 ttttagacag gttacctgat tgcccctggt tggccctcat cccctcccct atctgggcag 23400 ttttagacag gttacctgat tgcccctggt tggccctcat cccctcccct atctgggcag 23400 cgctctccct gctgtggggt tgggtggtcc tcctgctcct gggtcatatg gtatcgagag 23460 cgctctccct gctgtggggt tgggtggtcc tcctgctcct gggtcatatg gtatcgagag 23460 atgctgattc catctgcagc tccttctctg tactgtggac ttcactgctt gtctctttgg 23520 atgctgattc catctgcagc tccttctctg tactgtggac ttcactgctt gtctctttgg 23520 ccactaggaa gcatttattc tgattctcat gggcagcttg agaggcttcc cacagaggga 23580 agggtaggcc tttaatattt ctgtccttca tctcaaagac tgtttcttta aagtggacac 23640 agatgggatt gggctggaga gaggtggccc cagcccagga gatcatgttg tctgggtgaa 23700 accactgcac gctctggtcc ctgggctcct attgctcagg gttccttttt aaagcccact 23760 tgtcgagtct tctttcttta cagaggagat gttgtgctta ccctcttgtg aggatctcac 23820 agtgttctag cgcctctcct tggagatggt atttactggt ttgagggtga cctctgctaa 23880 tggacctgtt cagtcaccag gtacagggcc ttcagttgtg ttgtcgaaga ggctctgctc 23940 acccctgccc caccatatct gagaagtgcc cgaagttagt tggatgtggc ttctctgcct 24000 aggtagcctt gcacccctgg gaggatgggg gcctaagctg gaagtggatt tttctcatct 24060 agcagaagca cattcgatac tggatacaaa gccccatctt ctgtttcaga agcagcttgg 24120 agcatttctt gtttaactgt catgtaagcc ccttaagcaa tgcttttttc agagaggtat 24180 catcctgata aattgtcaaa tgcctctttc aaatccattc cttttaaaaa aaatcctctt 24240 ttctgtttat tctggagtca ctcatcatgc aaataatatc tttgtgagaa gtggtgttcc 24300 tggagttgga gaaggctctt tagtagggct gctggtcagt acagtccctg aaaaacatga 24360 ctatttataa cacaggcacc agagggttca gagacgtgct aaaagtggag aagttggggg 24420 caggggagtg ctggcaagga tgaactttgc ctatttcaca agttttgact aaagtagttc 24480 tgaataagaa aagcgtttga tgaaatctta cctagtataa agacatatta tttttggcct 24540 ctcttttttc tttcttcttt ttgttctttc acctttcccc tgtgacctcc cattccagct 24600 gagccctctt gttcccattt gccctgtatt gttgatctta accccagctc tgccctctct 24660 taaatcctgt tcttattctt ggagaagact acagtctccc gagctctttc tatatgccaa 24720 gcactgtact tttatacatt catattatca tgaggaaata ttcttttgcc tattttacag 24780 atgaagcaag tgagattcag ggaagttaag taatttatcc aaatttacat gcctactaag 24840 ggatggagca ggagccagac ccctgtttct gaagcatgtt tgctgaacca ctacacttta 24900 gccacctact gtggatacct ttggaaagga ggattctttt accgttacca aaaagatatt 24960 aataaaaata agtccaggtg tggtggctca tgcctgtaat cccagcactt tgggaggcag 25020 aggcaggtgg gttacttgag ctcaggagtt cgagaccagc ctgggcaaca tggtgaaatc 25080 ccgtctccac aaaaaataga aaaattagcc acgcatggtg gcgcatgcct gtagtgccag 25140 ctactccaga ggttgaggtg ggaggattgc ttgagcctgg gaggcagaag ttgcagtgac 25200 ctgagatagt gccactgcac tccagcctgg gtggcagagc cagaccctgt ctccaaatat 25260 ttatgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtgtatgt atacacatat ataatttata 25320 tgcaaatata taatatatac atatattaca tacatatatg tgtgtgtata tataatcaaa 25380 tgcccataaa ttgaagattc attaaataca gggatagtca taccatagta atgacgtagc 25440 tgttaaaaag aataaggtag ttcccataaa tggtagaata gcttgattgt actaatcccc 25500 ctgtatttta tgtagatttg taagattcac accaaatttt ttttttttga gttggagtct 25560 cgctcagtcg cccaggctgg agtgcagtgg cgtgatctcg gctcactgca acctccacct 25620 cccgggttca agcgattctt ctgcctcagc ctcctgagta gctgagacta caggcacgca 25680 ccaccacgcc cagctaattt ttgtattttt agtagagacg gggtttcacc atgttgacca 25740 gccaggatga tctcgatctc ttgacctcgt gatccacccg cctcagcctc ccaaagtgct 25800 gggattacag gcgtgagcca ccgcacctgg ccacacacaa agtattttta aaaactattg 25860 taagtcacca gaaagcaagc aaaatcaggc agaaactgga ggagaatcta cctcaaaaga 25920 caaactgcgg tggataaggt ttgcaggttt gtagctttaa cctgaaggta ctcccaaagc 25980 taattaagtg gccaaaagcc gcagccttac tgtattgcga tgttagaggg cagaggtaga 26040 aactggcaga gctggcagag cagtcagaaa attaagaggg aaaccccaga gaggagggag 26100 ctacagaggg cacaaacacc caaatctgca tatgaaattg cacaaatcct tggttggcta 26160 ctaaactatg tgtctgcaaa cctggtgaaa aagtagcagc tttaagcctg aaaaaaacta 26220 agcaaagatc ttggccactg cctaccatag ggaagacaga acttggtatt tgtgttcagc 26280 caagttagct gtctgctaga acaaaaatta gtcttcagag aaatgtgcag aatctggaat 26340 caagttagct gtctgctaga acaaaaatta gtcttcagag aaatgtgcag aatctggaat 26340 ctctataaca tatcgatcat catgtccagt gtcctatcag aaaccactag aggccgggca 26400 ctctataaca tatcgatcat catgtccagt gtcctatcag aaaccactag aggccgggca 26400 ttgtggctca tgcctgtaat cccagcactt tgggagacca aggcagacgg attgcctgag 26460 ttgtggctca tgcctgtaat cccagcactt tgggagacca aggcagacgg attgcctgag 26460 gtcagaagtt tgagaccagc ctggccaaca tagtgaaacc ccatctctac taaatataca 26520 gtcagaagtt tgagaccagc ctggccaaca tagtgaaacc ccatctctac taaatataca 26520 aaaaaaatta gccaggtgtg ctggtgggca cctgtagtcc cagctattca agaggctgag 26580 aaaaaaatta gccaggtgtg ctggtgggca cctgtagtcc cagctattca agaggctgag 26580 gcaagagaat cccttgaacc tgggaggcgg aggttgcagt gagccgagat tgtgccactg 26640 gcaagagaat cccttgaacc tgggaggcgg aggttgcagt gagccgagat tgtgccactg 26640 cactccagcc tggtgacaga gtgagactcc gtctcaaaaa aaacaaaaca aaaacaaaaa 26700 cactccagcc tggtgacaga gtgagactcc gtctcaaaaa aaacaaaaca aaaacaaaaa 26700 aaccccacta gatagaggta gaaatgggaa aaaagtagat aatagaagcc aaactcaaga 26760 aaccccacta gatagaggta gaaatgggaa aaaagtagat aatagaagcc aaactcaaga 26760 tgatagagat gttacaatta atagacaagg actttaaagc atctaccata aatacattca 26820 tgatagagat gttacaatta atagacaagg actttaaagc atctaccata aatacattca 26820 aagacttaat aaaaatgtat acacaatgaa tgaacagata gggagtatca gcttagaaat 26880 aagacttaat aaaaatgtat acacaatgaa tgaacagata gggagtatca gcttagaaat 26880 agtagctatg ggccaggtgc agtgtctcac acctgtaatc ccaacacttt gggggcccag 26940 agtagctatg ggccaggtgc agtgtctcac acctgtaatc ccaacacttt gggggcccag 26940 gtgggtggat cacttgaggc caggaattca tgaccagact ggccaacatg gcaaaaccct 27000 gtctctaact aaaaatacaa aaattagcca ggtgtgggag actgagacag gagaatctct 27060 tgaacgtggg aggcagaggt tgcagtgagc cgagatcata ccagtgcact ccagcctggg 27120 caacagagca agatcctgtc tcaaaaaaaa aaaaaaaaaa aaaaaaagag aagagaagaa 27180 aagaaataag gccaggtgtg gtagctaatg cctgtaatcc cagcactttg agaggcagaa 27240 gtgggcggat cacttgaggt caggagtttg ataccagcct gggcaacatg gcaaaacccc 27300 gtctctacta aaaatacaaa actgggcatg gtggcacgca tctgtaatcc cagctacttg 27360 ggaggctgag gcacaaaaat cgcttgagcc tgggaggtgg aggttgcagt gagccaagat 27420 ggcaccactg tactccagcc tgggtgacag tgcaggactc tgtctcaaaa aaaaaaaaaa 27480 aaatagctat gaaaaagagc tgaaaggtta gtggttgcca gaggttgagg ggaggaggga 27540 atgggaagtg attgtttaat gcatatgggg tttccttttg tgttgatgaa agtgttcttg 27600 agctagtggt gaggcttgta cagcactgtg aattttaaac tttaaaatgg tgaaaattgt 27660 gaattttatg aaaagagagg tgaaaataca tgaactaaat agaaattttt accagtaagc 27720 gaattttatg aaaagagagg tgaaaataca tgaactaaat agaaattttt accagtaagc 27720 tcaatagcag acaaagttaa ctgaaaaata gatcaatcgt atgtactcag tttgaggaaa 27780 tcaatagcag acaaagttaa ctgaaaaata gatcaatcgt atgtactcag tttgaggaaa 27780 agaaaaagat tgaagaaaag caaatagagc ctcagagacc tgtgggacag taccaagcag 27840 agaaaaagat tgaagaaaag caaatagagc ctcagagacc tgtgggacag taccaagcag 27840 tctaacatgt atagttggag ttccagaagc aaaaaagaga ttagggcaga aaaaatattt 27900 tctaacatgt atagttggag ttccagaagc aaaaaagaga ttagggcaga aaaaatattt 27900 gaggggataa tgaccagaaa tgttccaaat tcggtgaaat ccattaactc actgatccct 27960 gaggggataa tgaccagaaa tgttccaaat tcggtgaaat ccattaactc actgatccct 27960 taacaaacac tcagcaagat aaattaaaac acacacatgc acatgagagc cacattgctg 28020 taacaaacac tcagcaagat aaattaaaac acacacatgc acatgagagc cacattgctg 28020 aaatccaaaa ataagaaagt cctgaaagct gcctgagaaa aaaagaaggt gcacagaagg 28080 aaatccaaaa ataagaaagt cctgaaagct gcctgagaaa aaaagaaggt gcacagaagg 28080 aacaataata taactaacag ccgactgctt atcagaaaca gtggagagta gaaggtaaac 28140 aacaataata taactaacag ccgactgctt atcagaaaca gtggagagta gaaggtaaac 28140 gtattgaaag acaaaatatc aacccagggt tctacgtcca gcaaaactat ccttcaaaat 28200 gtattgaaag acaaaatatc aacccagggt tctacgtcca gcaaaactat ccttcaaaat 28200 tgaagcccaa ataaagacct ttataattaa acaaaaactg agagaatttg ttgcaaagaa 28260 tgaagcccaa ataaagacct ttataattaa acaaaaactg agagaatttg ttgcaaagaa 28260 tctgactatg agaaatgcta aaggaatttc tttaggctta agggaaataa ccccagatga 28320 tctgactatg agaaatgcta aaggaatttc tttaggctta agggaaataa ccccagatga 28320 taacttggat ctataaaaag aaatgaacca gaaatgatga agaaaataat cagatagatc 28380 catctgtgtt gatgtaaaca tgtaatagta ataataataa atatttatat agcaattatt 28440 tcaggaagga cacgcaagaa aactttaaca gtgataacct tccccgtgga aaaggggaag 28500 agattgagag cttggagtat gaaaaaggag actaaattat tattttgtgc ttcttttgat 28560 catttgaata ttttttaatg tggatgtatt acttattttt ttaaataaat ttttaaatgt 28620 catgaaatag ctaggaaaaa tagagatact tgatacaaga tgagaaaata atgttctgtt 28680 tctcccttgt aaggaatatg acagaaatcg agacattgta gactgggagt caggattggg 28740 tttctttcct actctgttag caggacactg tgtcacgtga caaaggctgt gaaggaggta 28800 tcagatcgcg ctctctggcc ttgggcacct tctttccatt gctaggtctt cttccccatc 28860 cccagacgtc cctgctcatt cccggagcgg cagctccagg aaggcacatc ttgctctaaa 28920 atggcctcct gaacactcct gttcccacac actgtctggc ttaatcttac tccaaggcag 28980 gcagctgcag agggtgccag ggctggttct gctcctgggt tcccatctcg ggtgatcagg 29040 acaccatcgt gtgctctgtc acccaagcca gcagcctgag agtccgcctt ttaactcacc 29100 ttgttaatga ccccaggtac gtttcattag gtcttcgtaa tgaaatgtac ctgggggtca 29160 ttaacaaggt caattgaccc aaatccttct caaatctgcc gcctttctct gtcatgattt 29220 ttataatctg caccacgccc cccacacccc caaccttttc caggatcaaa ttgagacttc 29280 tccagtgttt gcagagtgag gtcctgtctt tagcccaggc cttcctagcc tggcccctac 29340 ccaggtccaa ccttgttttc cacatgtgag ggtccaccct catctcccag tgaggtcagg 29400 tctggccatt cagcctcccc ttccacatgt tcagccctcc tggctttgcg cagcttattg 29460 ctttttcctg tgttccttcc ttgtcttcac atagcaaatc attctcacct ccagagatgc 29520 tgtgtcctat gtcctagaag ccttacctat tccctagcat gctgaactga gtggtcctgt 29580 cccggcctcc cataactctt gttaatgctc tgtcctactc ttggagagac catgaagact 29640 gggggaagag gacagggttt ggagttggac agatttggtc tcaattctgg tctttccatt 29700 tactggctgt atgagctcag gcaagttgtt taatctcccc agtctcagtt tcctcatctt 29760 tatggtgaca atactactgc ttactttgct gagacagatg gactaaagat aatatatata 29820 aagtgcttgg tgtggttcct attatagctt tattcttaca acactttttt aaacattgtg 29880 ttgattaaac atttacttga ttaaatgttt ttctctccca ttagactata cattctttga 29940 gaccagcagt gcttaatgtt tgtagaattc aacttttaat gagtttgaat tttaacagag 30000 tacatttgct aagttaaaag agtgattata ttaactttaa ttgtggaatt tcacgtaatg 30060 tgaaattcaa agctttatat atattttcag cagagactct taaaattatg ttttcgaaat 30120 aacaatttta tctccccaag caaaatgcca cctgcacctt tacaaacact ttaccgcagt 30180 tatgtgaacc ttcattgctc acccttgctg cagatcatta aatgtatgca taaatagaat 30240 tattaatgtt taccaaggtc ttaaagtcct ggtaagaaaa tattgagtgg tcaatataga 30300 aaggtcacct gttagctcct gatgcttgaa aacagaggtg ggacctggtg agcctgtctt 30360 cagctgatgc agtgccagca catctgggct gggcagggcc tccagtgcct tctgcacagg 30420 ttcacgtggc caactctgct tggacagtgg gatctgctcc ctggcagctg ggacacctgc 30480 TTCACGTGGC CAACTCTGCT TGGACAGTGG GATCTGCTCC CTGGCAGCTG GGACACCTGC 30480 tccccactgt tgagccattc ctcttgccag ggctgtttga gtatcagcca ctacattcct 30540 TCCCCACTGT TGAGCCATTC CTCTTGCCAG GGCTGTTTGA GTATCAGCCA CTACATTCCT 30540 ggccccctcc ttttagagag agatctctga ttttaagagg caaggtttaa agtaagacat 30600 GGCCCCCTCC TTTTAGAGAG AGATCTCTGA TTTTAAGAGG CAAGGTTTAA AGTAAGACAT 30600 ttcatgtgta gtagcagata ttattaactt tgttgtgcac agtatgcctt ggatcttctt 30660 TTCATGTGTA GTAGCAGATA TTATTAACTT TGTTGTGCAC AGTATGCCTT GGATCTTCTT 30660 ttcttgggta gggtacagat aaaatgatgg ttctacgagg ggtaagtagg aaaaattagt 30720 TTCTTGGGTA GGGTACAGAT AAAATGATGG TTCTACGAGG GGTAAGTAGG AAAATTAGT 30720 ggttctggat tctctccatg ccttcactcc attatagtca cagtggcaag atggagtatt 30780 GGTTCTGGAT TCTCTCCATG CCTTCACTCC ATTA TAGTCA CAGTGGCAAG ATGGAGTATT 30780 agcaagacag gtcctgagtg tggttgcggt ctaatttaaa cttttttttt tagacttact 30840 AGCAAGACAG GTCC TGAGTG TGGTTGCGGT CTAATTTAAA CTTTTTTTTT TAGACTTACT 30840 ctctcgccca ggtcggagtg cagtggtgca gtcttggctc actgcatcac tgcatccccc 30900 CTCTCGCCCC AGGT CGAGTG CAGTGG TGCA GTCTTGGCTC ACTGCATCAC TGCCATCCCCC 30900 agccaggctc aagcaattct catttctcag cttcctgagt agctgggact acagtcatgt 30960 AGCCAGGC TC AAGCAAT TCT CATTTCTCA G CTT CCTGAGT AGC TGGGACT ACAGTCATGT 30960 gcaaccatgc ctggctaatt tttttatttt tagtagagac gggatttcgc catgttggcc 31020 GCAACCATGC CTGGCTAATT TTTTTATTTT TAGTAGAGAC GGGATTT CGC CATGTTGGCC 31020 aggctggtct tgaacccctg gtctcaagtg atctgcccgc ctcccaaagt gctggggtta 31080 AGGCTGGTCT TG AACCCCTG GTCTCAAGTG ATCTGCCCGC CTCCCAAAGT GCTGGGGTT A 31080 caggcatgag ccactgtgtc caacctaatt taaactttat ctttgtaagt gccagggtaa 31140 tagaggctac ccagtttgcc attgtgctca cccaaagatg aaagtgctaa ggttgagtgg 31200 aagaattccc aattattaag cccctcctct ccctgtgttg aaaattctcc atgaaaaccc 31260 atctctggag gtgtatcaat cagtgttatc actgccctaa gccatttctg tcaaatccag 31320 tctttccagc atattgcata ttccctgtct tgccagtgaa atgtggtaaa gaagaaaaaa 31380 gttgccacag aatgtaattt ttaaaaccat agccgggcac ggtggctcac gcctgtaatc 31440 ctaacacttt gggaggccga ggcgggtgga tcacctgagg tcaggagttc aagagcagcc 31500 tgggcaacat ggtgaaaccc tgtctttact aaaaataaaa aaggccaggt gctgtggcac 31560 gcacctgtaa tcccagctac ttgggaggct gagggaggag aattgcttga acccaggagg 31620 tggaggttgc ggtgagctga gatcgcgcca ctgcattcca gcctgggcta cagagtgtga 31680 cttcatctca aataataata ataaaaccat aaaattccta caaagtgtga atatatattt 31740 atgaatgatg tctgtgtgta tccgtataca tacatccatt catatttctg aaaagctttc 31800 aaggagaatt tttaaacata gcagatgcat taatttgcat acaaagaagc cagttacgga 31860 tcattcttat ctagttatcc aagtcaacag tgcctttttt tttttttttt tttgagacaa 31920 ggtcttgctc tgtcgcctag gctggagtgc aatggcacaa tcgcggttca ctgcattctc 31980 gaccacccag actccagtga gtctcctacc ttagcctcct gagtagctaa gaccaccagc 32040 tacacctggc taatttttat tttttaattt tttgtagaga tgtctcccca tgttgccctg 32100 gctggtctca aatgcctggg ctcaaatggt ccccctgcct cagccttctg aagtgctggg 32160 attacaggaa tgagccactg tgcccagctt aatgatgcta ttaaactaca cctgttaacc 32220 ttctacttgc tttctgtggt tgacaagtaa ttctctttgc atttcttttt actaattcag 32280 atgggcttct cttttcttac tccaaattat tgaaagttat ttatttgtgt aaaatattac 32340 atagacatac acacacacaa tatatgtgtg ccttatgcat tttacaacat catctgaaca 32400 aatatttatt aactacctat tgtgtttcag actctgtgct agacatgcgg gatttcgcag 32460 tgatcagaac agtatctgcc caacagtccc acaaagaaat attaagcaag aataaaatct 32520 gtagccacat acctgtgtac attttttgag agcttactat gctctgggca ctgtgctaac 32580 attttcaaca ttatttcatg tagcactctc aacaactgtg tgagataagt tttatttatt 32640 gttaccattt ttccaaggag gaaaccgaaa cacagaaagg ttaagttact tgcctgaggt 32700 aacttagcat tgatagaatt aggatttgaa cctgggtagt atgactctta ggtccccatg 32760 agctggcaca caggttgcca atccatcagt taaccttccc atcactgggt cagagagcca 32820 aggagcagaa gggggataga acttccccag taccaaacta aggggctgag ctgggatcgc 32880 aaccacaggt tctgttccaa agctggggct ctttcgacaa ggccatgccc gccttgtctc 32940 ttatgtatgc tttttcactt gaggctgaat tttaggtctg agtcaactgc attttgaaaa 33000 aaatcatctg cttgaagacc ctgatgatgt aacaataaag gccattggaa aaccggcaat 33060 ctgtgaatct tcatatagat gaaatatcca ggttagaatt tatcatccta atctggatat 33120 ttttgagagt gaaaggagta ctgttataga tgctggggca tcaggagcca gccaggacgt 33180 ccaggcaagc cggggcttgt ggtcactctg tggacacgca gcatggtcat taatgcccac 33240 acaatgtgtg tattgttatt tgtagattcc attactcttt ggaatataaa cctctaagca 33300 agctaaaata actctactat gctttaatta tgcattgttg gtaaccataa caataaatcc 33360 ttggccaatt ccaatccaag gacataagca tttagagtca tttttaaagg agtgctccga 33420 tggtttagca gtcatgtaaa tgcatttata tgtaacagaa gacaaaaaca ataattaaga 33480 taaacataaa actcatccct tttgcctaca gaggcaactc tatttttaga actacaaatt 33540 tgggctggat gtggtgactc ttacctgtaa taccagcact ttgggagacc gaggtaggag 33600 gattgcttga ggccaggagt ttgaaaccag cctggccaac gcagcgagac catatctata 33660 caaaagaaaa gttaaaaact tagctgggca tggtggcatg tgtctgtagt cccagctact 33720 gaggaggctg aggcaggaca accacttgag cccaggagtt caaggctgca gtgagccatg 33780 attgtaccag tgcactctag cctgggcaac atagcaagac attatcttaa agaaaaattg 33840 tatgcatagc ctgagtacat gatttgaata tatttaggaa atagataatg ggataaaata 33900 ttcaaaagtg aaattatgca ggaaaattcc aggcatgtga ttgtcatata catagttgaa 33960 catttctatt ctctatgtag tttagacttt aattaaacag aataatctga ctttaatctc 34020 tcttgatact ataaaagtaa cttgaatttg ttcatttttt gttttttaat catgaaaact 34080 aatgcccaaa gtacctgtct aactcaaaat aattgtttct tttttcacac aaagccaatt 34140 gaatagattg gacaattgca tacttaagat ttaatatatt tgctcatcat attatgggtt 34200 ttaaaaaatc atttatcttg gagctaagta catttcacta tggtttttta tgttcattgt 34260 cgtaaaatat tatctttaat gtttcactag gttcactaat gttaggctga gggtgtttat 34320 tgttgtatca gatatccttt tgtgttttat ttaaatctat atttactagt tttaaatagt 34380 ttgccttatt gctaaggcta gtggaaaata gttaaagtgt gagaatgtgg tttttccctg 34440 tagtgttctt ttgagtagtt gacagctgtg aaaacattta gcaggcttcc ctccttttca 34500 gcgttggtta agctctgctt ctaaaggagt attcctagca gagcttgcca cagtatatcc 34560 aaaacattgc tgtattacag acgcaacatg gtttaaaagg aaaaattgag tgtgggtttc 34620 aagttgacac atgataaaag tagccctgaa gtgttcaatt agaaactgaa ggcctggatc 34680 tgataacttc atgggcagtg cccctgctct tttgtcttac gaccaatagg attttgactg 34740 ctcaggaaat ttgtatgcat gtacccagat atagacctag gcacctaaat gcacacgtgg 34800 atatatacga atatatggtg gacagtgcag atccaagcta ctcctgctaa cacatttctg 34860 gaaaacattc cctttatggg aggacttaac aaaaaaatta gtagctagtt ttactttggt 34920 ttacatatat tgtttttcta atatagaatt ctcattagtt ggttttatga atcatatcac 34980 caaatgaaaa tttgaaaagt atgtaacagt tttctattga gcgttctctg aatattcttg 35040 ccttcagtcc tgctgggagc cttttctcag atcaccagtg caagcctaca gtcatacatt 35100 gtttgtagtt gatacaacct caatgatata agttttttga tgattttaaa atatgagtaa 35160 ttaatcattt ataacctata ggttaaaatg aattataagt ggatgaaggt gggaacccaa 35220 atgaatcaaa gctatttaag tttggtattg acattgtgca tcttcccatt atttattata 35280 ccaaaaaaat cactttattt cttaaaaata gtacaacacg gccaggtgca gtggctcacg 35340 cctgtaatcc cagcactttg ggaggccgag gtgggtggat cacgaggtca ggagatcaag 35400 accatcctgg ctaacacagt gaaaccccgt ctctactaga agtacagaaa attagccggg 35460 cacggtggca ggcacctgta atctcagcac ttttgggaag ccaaggtggg cggatcatga 35520 ggtcaggaga tcaagaccat cctggctagc acggtgaaac cccatctcta ctaaaaatac 35580 aaaaaaaaaa aaaattagcc aggcatggca gcgtgcacct gtagtcccag ctacttggga 35640 ggctgaggca ggagaatggc gtgaacccgg gaggtggagc ttgcagtgag ctgagatcgc 35700 accactgcac tccagcctgg gtgacagagc gagactccat ctttaaaaaa aaaatagtcc 35760 aacacaagaa ggaaagaagg tacagaaatg gaaagttaag aatgaatcag tggagtaaaa 35820 catttctgag attggtgcat tttccttttg gaaatgtcat tcaaaccata tgaaattaaa 35880 tttggaaata atgttctgca gtctcttgaa tgggatttaa aatatgacaa tgcagaaaac 35940 acatatgggg atgaaaaccc ataggacatg tctcaaggag gacagacacc ctggggaaga 36000 acacttgtga gggtaccagg ctctcagaca caaggaaaga ggaatctgag ggattgctgg 36060 gaaggattct catcttatga ccccagctgt gggtagattt ggctggggag cccattctca 36120 ttgcaggggg cggtggcctc agccctgtgc atgctgactc ttcagtgctg tgctctggct 36180 gaggcgtgcc tctagtgagg gtgaccctgg gcaccgagca tattcagtca ccatcagcca 36240 aagctggttg taggagggct ggcggtggtc ttgggaatgt tgcttgaatc catcttgggg 36300 tgaaaatcag agtattggct catttctatt ttttatgatt attgttatta ttattattat 36360 tattgagaca ggatcttgcc ctgttgccca ggctggagtg cagtggcacg atcacagctc 36420 actgcaggtc acagctccca ggctcaagca atcctcccac ctcagcttcc cgagtagcta 36480 agacttcagg tgtgtgccac ctggcctggc tcatttttta atttattttt gtagagatga 36540 ggtctcccca tgttgtccag gctggtctca aactcctggg ttcaggtgat ccttctgcct 36600 cattcttccc aagtgctggg gttacaggca tgagccacca tgcccagcct ctccttcctg 36660 cctgcctgct tccctccctc cctcccttcc tccgttgctc ttttattttc tttccttttt 36720 tttcctgttg ctcaggctgg agtgcagtag tgcaatcttg gctcactgca gccttaacct 36780 cccaggctca aaccatcctc ccacctcagc ctcccaacta gctacagcta caggcataca 36840 ccaccacgcc tggctaattt tttatttttt gtagagccaa ggtcttacta tgttgcgcag 36900 gctgctctca tttctaatac actgagctca actgcttgct gggtgaggcc tgcatgagga 36960 ctttactcat gggctcatag aagccctttt aagctgctta gtgtccttag agtctccaga 37020 ggcatcccta acccagaatc ttttgactgt cctctggaga gaaaggcagt aggtctgtac 37080 caagaccaag gggcttgaag gcttggacgc tctgcttcca cctgcctctg gagtggccag 37140 aagtaatgcc tgttctcatg accagccagt ttgcctcact gctccggtgc caaaggccaa 37200 gttttgattg gaatgatgtc tgtatattct gaataaccag gaattttgtt tttctgattt 37260 acagatcttc aggcttttca tcagcatgat ttgccatctc tgtctttgtc atctagggtc 37320 tgtaattatc ctcaggtttt ctgccaatac ctgcattttc aaccagttag aaaaagaaga 37380 tgtaattatc ctcaggtttt ctgccaatac ctgcattttc aaccagttag aaaaagaaga 37380 gctaaaacag gtagaagcct gaaaccaagg ggcatatgca tttctgaaac attttcaaga 37440 gctaaaacag gtagaagcct gaaaccaagg ggcatatgca tttctgaaac attttcaaga 37440 aattgtctca attttatatt tctgtttata attataattt caatgtccag tggaggaagt 37500 aattgtctca attttatatt tctgtttata attataattt caatgtccag tggaggaagt 37500 tgtgagactc aatgtcagta gctaccactt actattctgt gcgcctcggt tcagattgtt 37560 tgtgagactc aatgtcagta gctaccactt actattctgt gcgcctcggt tcagattgtt 37560 tggggcagtg gtgttttatg ttgtgggtcc aaacccatga gtcatctggg aagttaattt 37620 tggggcagtg gtgttttatg ttgtgggtcc aaacccatga gtcatctggg aagttaattt 37620 attaggtcac aatgagcgtt ttaaaaaatg aaccataggc ccggcgcagt ggctcacgcc 37680 attaggtcac aatgagcgtt ttaaaaaatg aaccataggc ccggcgcagt ggctcacgcc 37680 tgtaatccca gcactttggg aggctgaggc aggtggagca tgaggtcagg agttgcagac 37740 tgtaatccca gcactttggg aggctgaggc aggtggagca tgaggtcagg agttgcagac 37740 cagcctgacc aacatggtga aaccccgtct ccactaaaaa tacaaaaatt aactgggagt 37800 cagcctgacc aacatggtga aaccccgtct ccactaaaaa tacaaaaatt aactgggagt 37800 ggtggcggcg cctgtaatcc caacttttcg ggaggctgag gcaggagaat cgcttgaacc 37860 ggtggcggcg cctgtaatcc caacttttcg ggaggctgag gcaggagaat cgcttgaacc 37860 caggaggcgg aggttgcagt gaatccagac tgcgccattg cactccagcc tgggcgacag 37920 caggaggcgg aggttgcagt gaatccagac tgcgccattg cactccagcc tgggcgacag 37920 aacaagactc tgtctcaaaa aaaaaaaaaa aaaaaaaaga actgtaacat aataggaaat 37980 aacaagactc tgtctcaaaa aaaaaaaaaa aaaaaaaaga actgtaacat aataggaaat 37980 attcaattct gagcccatca cacattttgt cttatgaaac ttttggtttt ttatatgtgg 38040 tgtatgtatg tatatgtggt gtatgtatgt atatgtggtg tatgtatgta tatgtggtgt 38100 atgtatgtac atgtgtatgt atgtatatgt ggtgtatgtg tatatgtggt gtatgtatat 38160 gtggtgtatg tatgtatatg tggtgtatgt atgtatatat ggtgtatgta tgcatgtgca 38220 agctctcact ataaatacac ttcttcctgt gggttgcagt aaaaaaaaat ttgaaaactg 38280 ttaccttaag actatttatt ttaaaaattt taataggtac atagtgtata tatttatggg 38340 gtacatgaaa tgttttgata taggtataca atgtgtaata atcacatcaa ggtaaatagg 38400 gtatctctga cctcaagcat ttgtcatttc tttgtgtttc aaacattcca gttatactct 38460 tgtagttatt tttaaatata taataaatta ttgttgactg tagttatcct gttgtactat 38520 caaatactag aacttacttc tttctatctc actggatttt tgtactcatt aaccatcccc 38580 agtttccttc ctctacccca ctacccttcc cagcccctga taaccaatat tctagtctct 38640 gtccccatga tttcaattgt tttaactttc agctcccaca agtgagtgag aacagttgaa 38700 gtttgtcttt ctgtgcctga cttatttcac ttaacataat gtcctccagt tccatccata 38760 ttgttgcaaa tggcaggatc tcattctctt ttatgactga atagtacttc attgtgtata 38820 tgtaccacaa tttcttgatc catttagcta ccgatggccc cttaggttgc tcccaaatct 38880 ttgctattgt gaatactgct gcaataaaca tgggagtgca gatatctctt caatacactg 38940 ccttcctttc ttttgggtat atacctagca ttgggattgc tgggtcatat ggtagttcta 39000 gttttagttt tttgaggaac ctctatgctg ttctccatag cggttgtact aaattacatt 39060 cccattaaca gtgtacgatg gtttcctttt ttccatatcc tcgctagcat ttgttattgc 39120 ctatctttta gataaaagtc attttaggcc gggtgcagtg gctcacgcct gtaatcccag 39180 cactttggga ggctgatgcg ggtggatcac gaggtcagga gatcgagacc atcctggcca 39240 acatggtgaa accccgtctc tactaaaaaa atacaaaaac ttagctgggc gtggtggcgg 39300 gtgcctgtag tcccagctac ttgggaggct gaggcaggag aatggtgtga acacaggagg 39360 cagagcttgc agtgagctga gattgcacca ctgcactcca gcctgggcga cagagcaaga 39420 ctctgtctca aaaaaaaaaa aaaaaaaagt cattttaatg gggtgtgata tcttaccgta 39480 gttttgattt gcatttctgt ggtgatcagt gatgttgagc accttttcat atgcctgttt 39540 gccatttgta tgtcttcttc tgagaaatat ctgttcagat cttttgccca gtttttaaat 39600 tagattatta aatttttttt ttcctattga gttgtttgag cttcttatat atcctggtta 39660 ttaatccttt ctcagatgga tggtttgcaa atatttttcc ccattctgtg ggctgtctct 39720 tcactttgtt gattgttttc ctttgctgtg cagaagcttt ttaacttgac atgattccat 39780 ttgtctattt ttgcttcggt cgcctgtgct tttggggcct tagggcaatt tctgatccag 39840 ggagagggct ttctttaagg ggtgtggggc atgaataagc caccagcaat gtcagggtat 39900 cttctttcct gtggctgaga aattccctaa tttaaactta ggattctgtt tgccatgaac 39960 atgctattaa aatagtggca ccctgggaca ctgtggggtt cagtgggctt ttgtgagcta 40020 tcctgaggtc ctaaccatca tttgtgacat tttttccatg gggaaaaatt cattttcagt 40080 tccaaagaag caacttattg gtgaattatt ttgagcagga tcctgtcata atcaggcact 40140 tccaaagaag caacttattg gtgaattatt ttgagcagga tcctgtcata atcaggcact 40140 gcttttatat tttttctgta ttcagtagta aatctacttg taccttaagc agtcatcgca 40200 gcttttatat tttttctgta ttcagtagta aatctacttg taccttaagc agtcatcgca 40200 ctcctttagc cccttttaat ttctaatgag gagtggtgtg ttataggagt ttggggtggc 40260 ctcctttagc cccttttaat ttctaatgag gagtggtgtg ttataggagt ttggggtggc 40260 ctgggggaag aagggttgct gaaggatgga gctcctgaaa gggagggtta atgaggatgt 40320 ctgggggaag aagggttgct gaaggatgga gctcctgaaa gggagggtta atgaggatgt 40320 gtgtccaaag ggtgaatgtc tgtgctcctc accagggaga agggcatctt gcatttacaa 40380 gtgtccaaag ggtgaatgtc tgtgctcctc accagggaga agggcatctt gcatttacaa 40380 tgatttataa ttcacctggg tcaggttcct ctgactatag gagccaattg gttacagaaa 40440 tgatttataa ttcacctggg tcaggttcct ctgactatag gagccaattg gttacagaaa 40440 tctttttttt tttttttttt tttgagatgg agtctcactc tggcaccccg gctggagtgc 40500 tctttttttt tttttttttt tttgagatgg agtctcactc tggcaccccg gctggagtgc 40500 agtggcgtga tctcggctca ctgcaacctc tgcctcttgg gttcaagcaa ttctctgcct 40560 agtggcgtga tctcggctca ctgcaacctc tgcctcttgg gttcaagcaa ttctctgcct 40560 cagcctcctg aatagctggg attgcaggtg cccgctacca cacctggcta attttttttt 40620 cagcctcctg aatagctggg attgcaggtg cccgctacca cacctggcta attttttttt 40620 tttttttttg tatttttagt agagacaggg tttcaccatc ttggccaggt tggtattaaa 40680 tttttttttg tatttttagt agagacaggg tttcaccatc ttggccaggt tggtattaaa 40680 ctcctgacct cgtgatccac ccaccttggc ctcccaaagt gctgggacta caggcgtgag 40740 ctcctgacct cgtgatccac ccaccttggc ctcccaaagt gctgggacta caggcgtgag 40740 ccaccacgcc tggctggtta cagaaatctt aatataagga gctcgttaca aaggcaagca 40800 taccctagat ctgcctgcta gccctgcgat agctacttct cagtaccttc tgtgccagac 40860 actcttctag gcccagggat acagcaggga gagaggtggg cagggcccct tcccctctgg 40920 agaggctgct gctgggccct ccgcacgctc tggcctccca tgctttggga atgcttgtca 40980 ctgccaagct gtgcagtgca tcaccatcca tggtgctcat gggcctgagg accactctgc 41040 ccacacctaa ggggggctgg gtggccagtg tttgaaaggc ttataagaaa gatcacaaga 41100 agtttcttct tgagaaatgc tacagtcttt aaaaacaaaa caaacaaaaa actacttaga 41160 attttgatag ccccagctca agaaaacaat aggtaggaca caagcttgtt cataggcagc 41220 agagaggtcc aaagcaatgt gcctcaaatc aagacgtgtg gctctgaaaa accttcttag 41280 ggaaggctcc cgccccacct agagcttccc aattccatag tatctaaggc cttgcaaaat 41340 ctacatctgt cacacgcccc cccggccccg ccccccatca caactccata gtgggaagaa 41400 cttatttcag tgcctatttt tcttgaggtt tataaaaaca aaatgtgtga agggaatgta 41460 tgctgattct cttgggacta attaaacaaa cagattcaga gaaagaaagc agagaagatt 41520 tgctgattct cttgggacta attaaacaaa cagattcaga gaaagaaagc agagaagatt 41520 taggacagtt aaaatgtaaa aaatgtaatt ttggggataa cagccgtcaa tttttccact 41580 taggacagtt aaaatgtaaa aaatgtaatt ttggggataa cagccgtcaa tttttccact 41580 cgagtgtttt tccaaagggc agagggcaag aagcatctta agtcctctcg aagctgagac 41640 cgagtgtttt tccaaagggc agagggcaag aagcatctta agtcctctcg aagctgagac 41640 acatttcaac tggagatatt agaagcaaat ttggtgccca agacaggtgc cagattttaa 41700 acatttcaac tggagatatt agaagcaaat ttggtgccca agacaggtgc cagattttaa 41700 ttacagttgg catttctgag catgcttttg gcagaggtgg ccatcttgtt ggaaaagcca 41760 ttacagttgg catttctgag catgcttttg gcagaggtgg ccatcttgtt ggaaaagcca 41760 tagattttga cactagcagt atgggtgccc agttgagaac gattgcaaat aatttccctc 41820 tagattttga cactagcagt atgggtgccc agttgagaac gattgcaaat aatttccctc 41820 ctattttcat cttgttggtt gtgagagaaa acttcagtgg ctaatattta aacaagatat 41880 ctattttcat cttgttggtt gtgagagaaa acttcagtgg ctaatattta aacaagatat 41880 tctctgttca gaggaaacag ttcattggaa ttgtcttggt ggcttggtgt gttggctctg 41940 tctctgttca gaggaaacag ttcattggaa ttgtcttggt ggcttggtgt gttggctctg 41940 ggggtaataa tttctatctt aaagtcttta aaaccagtta aaacaataaa ataggatatt 42000 ggggtaataa tttctatctt aaagtcttta aaaccagtta aaacaataaa ataggatatt 42000 tcattatgag aaatgaagtt ggcatgcacc aggttttcca ttaagttcca ggtcacagac 42060 tcattatgag aaatgaagtt ggcatgcacc aggttttcca ttaagttcca ggtcacagac 42060 aaataaagaa agtatgttcg ggggggaaaa atccttttta aaaataatta gggagctgtc 42120 aaataaagaa agtatgttcg ggggggaaaa atccttttta aaaataatta gggagctgtc 42120 tgttttgttc accagatgga aaatccgtag ttgtgggtgg tctgtgttgg agagggcatc 42180 tcacaattgg caggactatg cctttaactt ttcaagtcct cggacaaaaa tctcaacgcc 42240 atggactatt cagaaggctc cgaatgttca tcagtccttc caggtctgct tttctgaatt 42300 ttaccagttt tctaacaaga gtgtgtaagc aaaatgatta tggaatgccg aaataaatag 42360 tccattgttc gttgaattct gaaacagcat tcctgctagc cctggttttt tgtgactttt 42420 cttgctgtca ataccttatg atgataatct gattagtgga gctgcctgcg ttactattta 42480 aaggcccctt tttcttacat ttctgtgttc cagagtagat agggcagcca gatgtgctaa 42540 cagcataaga aatcatcata agcttgttat ttattgaaga tggatattgc actgttatca 42600 gtgtttaggg tcttttctac acttgctttg attctggtca aattaattca ataagaaaat 42660 gtacttttct tactcctttc ttgaactgat aatagaatgt tagctaccta gtgtcactag 42720 tatttttttt atctacacat caaggtatca tttttttcta gctcagtctt atgttttaca 42780 tggctttaca tctgcaagtt ggactttgaa atgaatattt tatgtttgaa cataaggata 42840 tatatatttt tatatttcat tcatacattt aacccatccc caaaattttc ttaaaaaact 42900 taataattac aggagatgta aattaataaa atcatctagg atgctaaatg cataaacttt 42960 caattgagtg aaaaaaagcc ctggaacgta aatcattttt ctcggacagc caatgtgttt 43020 ggctttggat gtacatcaga ttctgcctgt gcatagtaaa aatggcaatg atttattagt 43080 tccatctccc cccagcacag atgctgcaca gtctgtccct tttctgggca ctaaacagtg 43140 ggttttaaaa aacataattt cactcaagtt ttccgcttgc attctgtgtt gtactattaa 43200 tagcaaccct ttgttattca tgccagctcg cttctagcat aagacaatga tgtggggaaa 43260 cggttttaac taaaatacta ctaattggca cccatttttc atatactggt gcaggtgcag 43320 ttctgtgagg aatagaagca cctctaaaaa ttatgtcatg aagatagaaa agctaaacag 43380 ttacacagtt aagtaggtgt acctttgcaa tgtcattgtt tctttgaaaa atcttactta 43440 tttgccgata agttgaaggg tggggaaaat ttggaatatt gatggaggac ttatgtgtta 43500 aatgttctat gagtccacca ggcgctgtgg ctcacgcctg taatcccagc actttgggag 43560 gccaaatcac gcagactgct tgagccggga gcatgagacc aacctgggca acagagcgaa 43620 accccatctc tacaaatgat acaaaaatta gccacatgta gtggtgggca cctgtagtcc 43680 cagctactca ggaggctgag atgggaggat cacttgagcc cagaagttgg agtctgcagt 43740 aaacagtgat catgtgccac tgcactccag cctgggcaac agagcaacac tttgtctcaa 43800 aagaaaaaaa aaaaagttat atgatgcttc atgaaaaaaa aagaaagttt tcagcctgag 43860 caatatagca agacctcatc cctacaaaaa attttttaaa aatcagactg gtaggccaag 43920 cgcagtggct cacacctgta atcccagcac tttgggaggc cgaggcgggc ggatcacgag 43980 gtcaggagat agagaccatc atggctaaca cggtgaaacc ccatctctac taaaaataca 44040 aaaaattagc tgggcgtggt ggcgggcgcc tatagtccca gctactcggg aggctgaggc 44100 aggagaatgg cgtgaaccca ggaggtggag cttgcagtga gcagagattg cgccactgca 44160 ctccagcctg ggcgacagag ccagactccg tctcaaaaaa aaaatcagag tggtatggtg 44220 gcgcacacct gtagtcccag ctactgggaa ggctaaggca ggaggattgc ttgagcccag 44280 gcgcacacct gtagtcccag ctactgggaa ggctaaggca ggaggattgc ttgagcccag 44280 gagtttaagg ctacggtgaa ctatggttgt cccaccgtac tccagcctgg gtgacagaat 44340 gagtttaagg ctacggtgaa ctatggttgt cccaccgtac tccagcctgg gtgacagaat 44340 gagaccctgt ctcaaaaaaa gaagttttca agaggggaag aaataagaag agtgttagaa 44400 gagaccctgt ctcaaaaaaa gaagttttca agaggggaag aaataagaag agtgttagaa 44400 acctcagttg cccatctact ttcctctcct tgcccagggt gacagaaact gttgccaaat 44460 acctcagttg cccatctact ttcctctcct tgcccagggt gacagaaact gttgccaaat 44460 tctcactggc agcaacttga gttttgattt ggctcaggta catgtgctct tctagagagt 44520 tctcactggc agcaacttga gttttgattt ggctcaggta catgtgctct tctagagagt 44520 aaggtcagca cagaggctgc tggagggtgc tggaggctgc ctccatggac tctagcacca 44580 aaggtcagca cagaggctgc tggagggtgc tggaggctgc ctccatggac tctagcacca 44580 cctttcctcg tcctgtagct gaaatccttg gagtcattct tgcctccact agacccttgg 44640 cctttcctcg tcctgtagct gaaatccttg gagtcattct tgcctccact agacccttgg 44640 attctgtgca tctccacatg gctcccagcc attgtggctc tttttttctg ggactctcct 44700 attctgtgca tctccacatg gctcccagcc attgtggctc tttttttctg ggactctcct 44700 ctctcctgtc catcttccag aagtagcttt ctgaacacag acgtcattgc gctcctctcc 44760 ctctcctgtc catcttccag aagtagcttt ctgaacacag acgtcattgc gctcctctcc 44760 acaagcagca gtcactctca acactgagga atgaagacct tacagtgttc tttacaccct 44820 acaagcagca gtcactctca acactgagga atgaagacct tacagtgttc tttacaccct 44820 aacccaagat taccttccca gaatcatctt cctaccccat gtctctgacc ttccaatcac 44880 aacccaagat taccttccca gaatcatctt cctaccccat gtctctgacc ttccaatcac 44880 attaaaattc tcagggtggt gagatactcc agctcctttc tgtctctgcc atggctcatg 44940 ctgctgcttc tccctacaga gccctcctct catcttcctt gagatcttgc tgaccaaata 45000 tcttctcttc tgatgactta gcagtcaccc acactgccgt gggccagcag acacccttcc 45060 tctgccttcc agtagctttg cccaatgtga tctctgctct ttgtaccctg tgctgtctcc 45120 catgttagat ccaaccacct aggtcatagc gctactgctg gtacctgatt gcaggcccag 45180 catctgtctt ggtgccaagc acatgacaag aactgaataa ataggccagg tgaggtggct 45240 cacgcctgta atcccagcac ttggggaagc caaggcgagt ggcctgacgt caggagttct 45300 agactagact ggccaacatg gctaaacccc atctctgcta aaaatacaaa aattagctgg 45360 gcgtagtggc gcatgcctgt aatcccagct acttgggagg ctgaggtggg agaatcgctt 45420 gaacccagga gacggaggtt gtagtgagcc aagatcacac cactgcactc cagcctgggt 45480 gacacagcga gaccccgtct caaaaaagaa ttgaataaat attttttgag tgaaatggaa 45540 ttctgggttg tggctttcca tcagttaaga ttcttggttg tggccaggtg cagtggctca 45600 cgcctgtaat cccaacactt tgggagggag aggtgggtgg atcatgaggt caggagtttg 45660 agaccagcct ggccaacatg gtgaaacccc gtctctacta aaaatacaaa aaaattagcc 45720 aggcctggtg gcacgtgcct gtaatcccag ctacttggga ggctgaggca ggagaattgc 45780 ttgaactcgg gaggcagagg ttgcagtgag ccgagatcat gccattgcac tccagcctgg 45840 gcgacaaagc aagactccat cttgggggga aaaaaagatt cttggttgta agcaacagac 45900 accacatctg gctttggcaa gagggggttt tcctggaagg atatggacac attcacaccg 45960 tcggtggaaa gcaactgtag gcatgccgac atgaatcctt ctccattgtg tccctgctca 46020 gccacgagaa cagtgtgctg ccagggtgct cagtctgtaa ctctgctcag gatgcaggct 46080 ccagggaggg agtgtccaac tgggctaata gggcccaccc tttggctctg ggagagctgg 46140 gctccttgat gataatccca gcacactgtc agctgtgggt aggatgaggc aagttcctca 46200 gggggtgaag tgtgttttgg gatagtcaac agccagtgac tgttcaccac aggactaaac 46260 cttccatttc cagtgctcca gtgccccaac tggcctgtct ggtgtgattt gccaagtgtt 46320 ctgagcaagc agggaagttg tccattaaaa tacagatatt ggccagacat ggctcacacc 46380 tgtaatccca gcactttggg aggctgaagc gggtggatca cttgcactca ggagtttgag 46440 accagcctgg gcaatatagt gagaccctat ctcaattaaa aaaataaaaa gcagatatcc 46500 tgggaaggga taacactata gcccctgcta gccagcattg atttgccaaa agtaggacct 46560 gtgggtgcag gatccagagt agccatggat ggatgaggtg cgaaagggtc ccgaatgttc 46620 cagcaatgct acccaaatgg caagtggtgt cgtggcctct ttcacctcca tactcaggga 46680 gaagccactc agaggcctcc acataacagg caggctctcg cctgggtgtc ggaggtgagg 46740 atgctactga agcttattgc ttcttaagaa tcaatcaaat caaaacaatg gatgcccttc 46800 tcccaagtgg aagggctgtg ctggccacct tgtatgactg acacaaagtt gggaagagtt 46860 gcagtcatga gctggggaga cacaccctaa aagcagatga gctggtggcg gtaaataaca 46920 ctcccatgac gacaagtggg ctttgcacgt gtgtggtagc agggcatggc cctgggggag 46980 gtggggctac ctgaggaaaa tcacaggagg caggaatgta tactgtttgt tgggaaaaga 47040 gtggggctac ctgaggaaaa tcacaggagg caggaatgta tactgtttgt tgggaaaaga 47040 caagccaatc tgaagggcaa gggtttgtag aggggagtgg tggagagtga ggttggaagg 47100 caagccaatc tgaagggcaa gggtttgtag aggggagtgg tggagagtga ggttggaagg 47100 gtgtttgggt ccagttgtag acagccttga attctaggca aagacttcag atgttgtttt 47160 gtgtttgggt ccagttgtag acagccttga attctaggca aagacttcag atgttgtttt 47160 ctaagcacta ggagcacctg tggatgtttc agaatgggag tatcatgatg gatgtactat 47220 ctaagcacta ggagcacctg tggatgtttc agaatgggag tatcatgatg gatgtactat 47220 tttttaaaga gcaggaataa gcagcaggcc actgctaagg aatctgtgtc atgaggctgt 47280 tttttaaaga gcaggaataa gcagcaggcc actgctaagg aatctgtgtc atgaggctgt 47280 tagcctgggg acaaggagga ccggccaaaa aaaaaaaagt cctgtatata agggggagca 47340 tagcctgggg acaaggagga ccggccaaaa aaaaaaaagt cctgtatata agggggagca 47340 cccagcacac ttaggttcca gacctccatt cctagcaact cacctgtaac tcctggcagg 47400 cccagcacac ttaggttcca gacctccatt cctagcaact cacctgtaac tcctggcagg 47400 ggtctctgcc tccctctact catccaaaga tcatggtgct tgttcaccag ccttatgatg 47460 ggtctctgcc tccctctact catccaaaga tcatggtgct tgttcaccag ccttatgatg 47460 atccaaagaa atagtgatct ttcatgttta aataaaccta cttaattacg tgcatctgtt 47520 atccaaagaa atagtgatct ttcatgttta aataaaccta cttaattacg tgcatctgtt 47520 tggaaattca gacacaccac atgagaagct actgggactt ttttctcctt ttcttggctt 47580 tggaaattca gacacaccac atgagaagct actgggactt ttttctcctt ttcttggctt 47580 ctcatttgtt ctctagcttt ttcttttttt ttcatcctgg ccagttatta accttctcct 47640 ctcatttgtt ctctagcttt ttcttttttt ttcatcctgg ccagttatta accttctcct 47640 ccacttttga gtctcataat tgtccagatc tcccaggacg ctgacagtgt ggttttggtt 47700 cacttgttcc ttctcctgct caaatttcaa ttgattcaag ggtgatatgc tgggtatgat 47760 gggctggaca tgcccatttt cattcaactc atccataact aggatataag tgaggataca 47820 gatttgggtg ttatggccaa aagatcttca aatacaatga ctttatcaag gataatttct 47880 tcccccctca cttgaaacat ggttaggtgg tgcagggctg ataacgatgg ctcagtgagg 47940 tctcaggctc ctttagctat ggttctgcca acctcagcac atggcacaca tcttccccaa 48000 ggcggctgct tggtttgctc ccaccacgtg gggtaggggg acacatgcct tcctttatgg 48060 ggacactata tttctgctca ccctgaccac atcccatcaa tcagaacttt gacccatgaa 48120 tacactcaga tgcaggagag gctgggaaat gttttcctta gctgggcatc catggacctc 48180 tccctgggtc catcctagag agtggagtag cagcatttgc accccagacc tgaggggtag 48240 ctgaggctgg gctggcagtg gggagtgagg ccaggagctt ggatgtgcag gctggcatgt 48300 cccacacata tggggcaaag aggaggggct ggctggggct cgtgtcctcc tccgtaccct 48360 catcccaagc cccagtgcgg tagggtgggc tgctccaacc caaatcttct tctcccaagt 48420 cttctctttc ccagcatatt atgctgcttc catttcacca tagtttcact cactatcaag 48480 gtctccaaag caatcccatc atctacatat tacttgattg ttgtgtacag ggtattgggt 48540 gaaaatgaag gtgggcagag cacatagcct ccaacctcaa agccagtaat gtgccagatg 48600 aaaggtaagt cacagtcttt agattcagta gcattttaac aaaactaaaa tgtcaggccg 48660 gatgccgtgg ctcacgcctg taatcccaac actctgggag gctgaggcgg gctgatcacg 48720 aggtcaggag atctagacca tcctggctaa cacggtgaaa ccctgtctct actaaaaata 48780 caaaaaatta gctgggcatg gtggcgggcg cctgtagtcc cagctactag ggaggctgag 48840 gcaggagaat tgcttgaacc cgggaggcag aggttgcagt gagccgagat cgcgctccga 48900 agtccattct ttctgtgcag tggctgatct atcagaagaa gaaccccctt ccccaccagg 48960 cctaagaacc catgggagtg atggcccagc actatggtgt tgcatttctt tctttttttt 49020 ttttttagac ggagtcttgc tgtgtcatcc agggtggagt gcagtggcgt gatctcggct 49080 cactgcaatc tctgcttcct aggttcaagt gattctcctg cctcaacctc ccaagtagct 49140 gggattacag gcacctgcca tcatgcccgg ctaatttttg tatttttagt agagacgggg 49200 ttttaccatg ttggccaggc tggtcttgaa ctcctgacct cagatgatct gtctgcctcg 49260 gcctcccaaa gtgctgggat tataggcgtg agccactgcg cctggcctgg tgttgcattt 49320 cgttactggc aagatgatgt atgctctttc acgtgcccac atgcaaaggt gtgactcatg 49380 gaatgaagac agctcagcaa cctctagttg ctgggacaga aatgcaagtc agtgggctct 49440 ggcctttctc ccaggctgtg ttgccacctt ggtggctgga ggggcgctac tagggggtgt 49500 gcttggcaga gcctggggtg gcgtcctctt tcatcccact aaataacaca ttggtggctt 49560 gatctgccat gaagctgaca atcagagcaa gtatttgaag cctgggaagc tgtgtgggcc 49620 ttgcttgtgg gacctggtgt cagctgatga cgtggccctg tctgggtgcc cgtgagtcag 49680 atggcaggga tggcctgaca tcatgaacac tagctccacc tacgtcacat tggacttgac 49740 tctttacaga gcacctagga agagatattg atgagtgaac cattgcccag tcatcgagtt 49800 aattatttcc ccccagaaag gttaacatac tgaggacaga gagggcatgt gtaggcggca 49860 gggcgcactt ttcatttagt ctggttcgca ggtgcggtca agacgattcc ttattgtctt 49920 ggagccagcc cttttaagcc ttcctgcaca gctctgtgac agtctcgcgg ggagacctgt 49980 gaggccactt tccttttttc cagacccacc cccacctttg agattatcac tcttcctgct 50040 ttgttttctg aggctttttg gctttttagt ttatttgtaa ttagagcaga aaaggtacag 50100 taatcggata gaagggttgg gagtgataga atgagccagg atgcggtgga gaaatggggt 50160 ggagaacccc cagaccttag ggtggcgtgt ttgtcaggga gtagaccagg gtgcagttct 50220 aggcagacca ctagatggca ctcacacatt ggtgttcagt cgggtccacg gcactttcca 50280 ctttgggctt ctgtaaaatc tccccatgaa ctagccaaag ctttgtatct gtcatcccgc 50340 catttgtccc tctaacctct cttgatcgtc tacagatagg tatatatata tgttcctctc 50400 tagttcattt ttctcttaac atttagtgac cttaaaaacc tggcagaaat accttcaaag 50460 cctcccacct tgttagctgc attgccgcat ttttactgtt ctcctaagcc aatgtttaca 50520 tggcagtaaa tccatgttta gagtaggtaa tagagaaacc tgtttctctc tcagtcactc 50580 tgagggtatg tttgataaag aaatggaaat tgagaggatg agttattaaa aggattaata 50640 ggtaacaaat ttttttaaaa tttccattca aaattatggg ttggtaaatt gtaagatact 50700 taaaattaat gagttgtttt atcaaatcat tttctgttgt tagagaattt tcagcaagct 50760 gagattcaaa tgcaaaatgt cttattgcat actgctggct ctaggaagga ggagggacag 50820 acaggaaatc atggttgatg ttatctttac caaggtgcat gtggtgtgaa agcaaaaccc 50880 catcttaaat tcatcatccg ttctgatgtt gcacagaaat gaaaattaat ttctatatgt 50940 caactaaaat ggattctgct cctgcctggt tcaattctgt cttagctgcg tggacatctt 51000 ggggagctgc tctgatggta catgcagacg tgcttcatct gtgcacttct tagctgagat 51060 aacagtgctc cgagccccac tgttaaggtt aacctgtata ctgcagaagg acaacagatg 51120 gaaattctgg tttgcagcca ctgcagaaaa ataaaaacat gttccaaagg gaaaaggatc 51180 gaaattctgg tttgcagcca ctgcagaaaa ataaaaacat gttccaaagg gaaaaggatc 51180 ttgtatttct gcgtttgaac ctttgatccg cattcacatc tgaagggagg gctaaacctt 51240 ttgtatttct gcgtttgaac ctttgatccg cattcacatc tgaagggagg gctaaacctt 51240 ttggtggaga cacatggctt tgcatctgtg aaaacacagc tgtatagagc aaaacaaaac 51300 ttggtggaga cacatggctt tgcatctgtg aaaacacagc tgtatagagc aaaacaaaac 51300 tagtggaaaa tacaatggtt ttgctgtgtc aggctcaccg ctaatgcttg aattctatat 51360 tagtggaaaa tacaatggtt ttgctgtgtc aggctcaccg ctaatgcttg aattctatat 51360 aaatattgtc ttttgatagt tctttcaaaa atctttcttg actttccata taattgtcca 51420 aaatattgtc ttttgatagt tctttcaaaa atctttcttg actttccata taattgtcca 51420 ataatgttat tcttttaaat tcagatttgg ggctttttag gattaaggca caaatgtctc 51480 ataatgttat tcttttaaat tcagatttgg ggctttttag gattaaggca caaatgtctc 51480 ttttctttta aaaaggcagt aaaataaaca gattgtagtt agatcataat aacatctaaa 51540 ttttctttta aaaaggcagt aaaataaaca gattgtagtt agatcataat aacatctaaa 51540 ttttgaccta attagagggg gagatgggga aaaaaactct ttaaaagaag attaaggtaa 51600 ttttgaccta attagagggg gagatgggga aaaaaactct ttaaaagaag attaaggtaa 51600 agttctaact ttatccttga ttctgaattt tttttctttt ttggaaatat tgcatacatt 51660 agttctaact ttatccttga ttctgaattt tttttctttt ttggaaatat tgcatacatt 51660 ttatactgca agttacaatg tgaagggcct ttgaagcact ctaaagagac ttagctgctt 51720 ttatactgca agttacaatg tgaagggcct ttgaagcact ctaaagagac ttagctgctt 51720 tgttttgagc aattaaggtt tgttattgag tgaattaagc agctctgctg tggctcatct 51780 tgttttgagc aattaaggtt tgttattgag tgaattaagc agctctgctg tggctcatct 51780 gtaaatagtg gtcttcagcg agtttcttga tccgcttttc ccaggaacag aagccaagtc 51840 ggcttcctta ggagctgctc gccctgagtc tccatgtttc ctcgcagtcc ggggcagaac 51900 ccaagcccgt gccttcctcg ccctgccttc ggagtttcct gctattgacc tggcggggcg 51960 cgccagccct ccctgcccgc tggtgccgcg gtccctctcg gctgcagcag tggggcacag 52020 ggacgtcatg gacaaaagac gaggatgcca gttcgatgtg cgagcagaag gccgcctgcg 52080 tgcgctctgc agagcaaccg gcccggtttc ataaggggcc gccggcaggg gactgccatg 52140 gcgccccctc ctaccctttt tggcaccagc agcttcatcc gcaggcccac tcggggtgaa 52200 tagtgtggga gctctcaaga gaaagaattt taggtttgtc gatagtaatc accaccttca 52260 gcgctcctcc cactgctgga gagaccgcgg actgggtgac aagtcttcca ggctcccaga 52320 aagtttctgc tgcaaagcgc agggtccgcc tcccacttcc aggagggacc aggagggaac 52380 catcgcgaaa ccccgaggct gccggggact ctgactgagg cctttcctcc tgtgtccctc 52440 tcccccacgc cggcctcccc tcctttcaac tctctgcgct ttgaaaccgc ggagttgggg 52500 tagctaggcc ctccagcccg gcgacaggcc tcggccacag gcagcggggc ggggtgtgga 52560 caggaacaag gaggtggggt ggggaggaag cctcgggcag agcaggttct ctgcggagtt 52620 tcggcagccg ggtttatcca ttttgattaa agaaaagtaa aaaaaatccc ccaaaaccaa 52680 acaacaaaaa ccctctccca acccccaacc cgcctccaca gctgcatttt gttcctggga 52740 ctgttggtgt caccagagcc tcttttttta tgcaatccta gactccattg caaaggactt 52800 gtaaagccta ttaatttttc aggcaaaaca gatcaagcct gcctttgatg ctgctgtttc 52860 ctaatttagt attcaaaggt aaagaatcct actaccgtct cttcactttt tacataatta 52920 aaagagattt ttcccccagg ggcaaagaac tagttgctta gtattttttg aagtgaaagg 52980 tctaacttta tatcacatct gtttttatgc tctttcctaa acataaccaa tatttactct 53040 ttgctggtac ttagttgcat gctattcacc cttgcatttt aatttctgac agagagggga 53100 ttagtattcc tcttctaaat catcctctaa gctatgtgtg tgtgtgtgta tatatatata 53160 tgtatatata tgcgtgtgtg tgtgtatttc tcctctaaca aactgagagt gcaatttaaa 53220 aaactgtaga atcagtgatt gccctaactt gtgcatgact cttgatttca cacaggagca 53280 ttcaaaagtt ccgttaagtg ggctttcaca ggtttccatc tgctgtgttt aatagcaaat 53340 tgcgaccctt caaaccactt caaggccttt tgtgttctta tctgtgcatt tcaatacttg 53400 ctggggaagt gccttcagtt tcactacata tgcaaaaagc caggcttgaa ttttttgaca 53460 cccccttttc atttgttggt ggagtcctgt tttctattgt ttgcttgttg atgttggcaa 53520 aagtcattta tttaatgatg ccaggaatac tatgccctat tctttattgg caggaaatcc 53580 taaaacatcc gggagtagtt taaaaattaa tgcatgccgt tgaaacttac atattggcat 53640 agttttattt tcccatcata aggtagatga tattttaaag gtattatata gtaatagtca 53700 ccagaagcca attaaaacca ctaggattct atcaaaaacc tgtttggcgg cataaataat 53760 tgatatgtgg aaataaagca gcattctctg ggcaggggcc tcctgtacag acctgggtgg 53820 cgttagcagt ctctcttctg tgaaccaggg ggcgatttct gggcataata tcacctggat 53880 ttaaaaaact taataataag gccacacacg catttttaaa aaaacttccc ttgcctgtgg 53940 agacagagag gagatagcga tgttattttc aggtgtttga gcttcttaga gaaaaagaaa 54000 caaataccag actccttagg agctaggtgc gaatggaata aattgaggca ctccagcgac 54060 acatccagct tgttttaaat gatgtgatta aaaaggacat ctgagaacaa atttatgaag 54120 ttgttatggc aactgaaata cttggattga aatgtgtgta tctaaaacat gaaagattgc 54180 aggaagccgg agaaacaagc aaagcattgt gtttgggaaa ggcaattgaa cagtggtagc 54240 caaatattat tcctctgtcg gtcatccttc agggtaactg cctcggtttc cacttttaaa 54300 aacgtgtggt gattgatgct ttcatggact ctgttgcccc aagtccccaa gccttgtacc 54360 agcgtttgct cattggtcct ttgacctgtt gagaacacaa tctctgtatt gtgttcaatg 54420 cagggatgca tttagagttg cccctgcatc tacccattaa gtttatactc ctgtgttctc 54480 atattaacgt ctgtgtttgc gaatagtctc tctagtcttt gattcactaa ctcccccaat 54540 tctaacccac caatggccac caacaaacag ccacactcaa aggactcaac atatttaaat 54600 attccctaga attctttgct ctgttctaaa agctatgtat cccgtctaca taatttgatt 54660 ctattgtaat ttctcatggt tttaaaattt tccttaccag tacttgtaag agtattgaat 54720 aattgtttct ttcaatcaca ggcaaggata tttaaaatta acatgtggaa atgtgtgttg 54780 cagtgtgttg aaaaatcagt acttgaagtc acatctagaa tgcttccgat tttttccatc 54840 attagatttc agatgcagtg tccaaaaaaa ttgtttttaa aaaatgttgc tacaaatttc 54900 ttgcaatata ttttgggttt ttcactttaa atgaaaaaat agttaagtgt caattttagt 54960 gattttagta gaccatccca ttaaaaatat gctattttaa ttgaaaaaaa tttttgacaa 55020 aacacattta caccgaaaaa ttcaaacaaa ataatttctt ctttaacaat attgattgtt 55080 aaatgttgat taccaagtta tttggtctcc ttaattttaa gcataatact ctttactttg 55140 aatagtctta atccctaaag caaagcaaag taaaagagaa aaaaaagtag aagaatttcc 55200 agaaaattgt aattccctta aaccatgaat tagaactttt tactcttact gtagttactc 55260 tagttttttt agttagacat gagggtacaa tcaaacagct tcttttttgt aacagattgt 55320 tttcctactt cttgaaaaac tgagatcaaa acatttttct attctgttgc aaaattaact 55380 gttgttattc tgtcagacga gttattcact ctaaatgtct ttttaaacct gtatatttat 55440 ttttttatcc ctttttgcat atgcacatgg aagtgattcc attttaatat aactttttta 55500 ttttgaaaac ccataggttg aagatgatat aatcagagta ataataagga tgctccaagt 55560 atattaaaca taagaatttg gggtctttgt tacggaaata gatttccttt ttcctttctt 55620 tagtcttact gttttaagga ttttttttaa agtatgattt tgatgcttcc aaagtggttg 55680 ctattttatt tcaggtgggt catttgatag tttgttgatg ttttcaaggt ctctaaacaa 55740 agtctgatgt gttaacagtc ttattaacga tgttaaaata gaattaacag cttgtggatg 55800 cctccagagc acaatagcct gtcagcctcc cctgccccct tataaattgt gcccctgctg 55860 tcatcccttt gtttcaggcc caatcagcaa actaaacttt tgtgaagaga gaaggcagtt 55920 caagtgtttc tgccgcagct gaagggagga gctaatgtgg aatttattgg gagaccaatc 55980 agtgctggag ggaagaatga ccccaaattg gaagaaggac tttgttgacc cccttggctc 56040 cctgaccttt ccttgctatt ttgtcccttt aaagaggttt actttggttg tcgtttactt 56100 tgtgatgctt ttagtaaaag ccccagaagt tttaaaagtt ttggatttaa gatccctctc 56160 agcacagatt taattgtgcc gtgtgatttt gagtaacttg agtagatgtt aacactcaga 56220 attatttttt aaaaatctga cacaagctat taataatttt ttaaaggagt agaatttcat 56280 tgattctctt gagccaaaat agattctaaa tctcttctag tatctctttc ctgtagttac 56340 tttcatagag aatcagatta gttactgatg ttagactatt ggttctattt cgttgaattt 56400 atgctttaaa aaatagattt gtgtattcca aatctttaaa actttttttt tttttttttt 56460 tggtcaggca gtaattatag tgagtcagaa acaaatattt ttagaggaac cctttttttt 56520 tattagtatt actagtaaca aatttgacaa attgtcacat tttaaagtac agatttatgg 56580 gatagcctga caaatggata ccacttttcc tttgctactg tccaatttaa taatttataa 56640 agtgtggcat atttgtaaaa tagttaccca ttctcactga gcattacacc agttttggta 56700 agggatactg agacattgtt aagaattaag tattttttag attgcttgag cactttcctt 56760 gttcccttat cagcattctt ctcccttgta atgtggtttg ttgcttatat ttactttgca 56820 accggccact gttcaaggac ttcagacagt gctgaggtcc gagatggtca ctgctggtcc 56880 aaagaacaaa catctcaggc tagatgcagg acactaaata tgttcttcaa gccaaggtta 56940 aataaaatta ctcaaatgta gccaaatacg gggaatgaga tgcagttaat gaattgtggc 57000 tttgaaatat gatcatttgc caaggaatat gctgtgatga aaacggagca gtgtaaactc 57060 agcacttctc ttagctcgta gtgagctgct caggtaaacc ccgaggggaa tactttcttc 57120 tactgaataa accctaaaga taggcatcca ttgattgaac agtgtctgaa actggagcaa 57180 acagggcgcc gtttgatact agccccatgt gtgggtataa ataatgtttt ccagttggca 57240 tccttgagtt gtgctgcttg aatagggcca ctttacgtgg ctgaagaaga atgtctctgc 57300 agtagttctc cctgagcagt aactgcttgg tacctttcac ccacagcacc cattgctgcc 57360 catccccatc ccatgtctag tgtgccctca cattccactg ctggtcagca gaaccatctt 57420 atcagaaagt gtcaccaagc ccttttttgg tgtgcccgct gaatgagcac tccaggctgt 57480 ggagttcggg acatgccttg gtttgtgggg accatgctgc ctgcctgtcg agaccaagca 57540 tcgatactgt gtgtctacct gatgaaagtg tccagtatgt gtctgcatga cttggggaca 57600 ctaagaaaac caaagggatt agcaacaaag agagcttgtc acctttgtgc ggaagccagc 57660 tggcatctca cagggacaac ctacaacctg agctgctgcg tcctcactaa atctgggccc 57720 ctagggaccc cgttttactc ctgctctcct ggagcttatt acgggcctgg ctaccaaagg 57780 gaaagagggg aaaatagacc aggagcctta tgctagaacc atttattttg tttcacgtga 57840 tgcagacaga gataaaactg caaatttaat gaaactttaa caatcagtac aatgtttctc 57900 cttaagaact ttgtaaatag catttatctt tcaagagttc tttctctctt tttgtgatta 57960 ttttataaac ttaaaggaaa aagagaaaaa gtcagtggtt ccagcatttg ctttagtctg 58020 tgacttaaat ggattataac tcttgaccgc tgacatttac caagataaat cagtggtcat 58080 Threonine - alanine - threonine - alanine - asparagine - glycine - isoleucine - asparagine - serine - leucine - threonine - alanine - aspartic acid - histidine - phenylalanine - glutamine - aspartic acid - asparagine - serine - valine - glycine - serine 58080 agatgtggag cttgatgtct cttcggctct gggaccaatc cccttggaca aaagttttcc 58140 Arginine - methionine - glycine - alanine - aspartic acid - valine - serine - serine - glycine - leucine - glycine - proline - isoleucine - proline - leucine - glycine - asparagine - lysine - phenylalanine - proline 58140 tgtgttctta gtattctgaa ctggctacag caactttaag gaaaataaag gttacaaaaa 58200 Cysteine - valine - serine - valine - isoleucine - serine - glutamic acid - threonine - glycine - tyrosine - serine - asparagine - leucine - lysine - glutamic acid - asparagine - lysine - valine - threonine - lysine 58200 aagttctgac aatttgtttg cttttacatt ttcaaatttg tgaaatgtag agataatttt 58260 Lysine - serine - aspartic acid - asparagine - phenylalanine - valine - alanine - leucine - tyrosine - isoleucine - serine - asparagine - phenylalanine - valine - glutamic acid - methionine - valine - arginine - aspartic acid - asparagine - phenylalanine 58260 gttttcaaat ctttgtaatt ccctgaagca aatactttca agccagttgc aaaatgctgc 58320 Valine - phenylalanine - serine - asparagine - leucine - valine - isoleucine - proline - glutamic acid - alanine - asparagine - threonine - phenylalanine - glutamine - alanine - glutamine - leucine - alanine - asparagine - alanine - alanine 58320 tttagaaata attcatataa acatgcttct ctatttaatc acaaggggag atgtggagaa 58380 Leucine - arginine - asparagine - isoleucine - histidine - tyrosine - alanine - histidine - alanine - serine - tyrosine - isoleucine - threonine - lysine - glycine - glutamic acid - methionine - glycine - glutamic acid 58380 tggatgtttt attttttcag tagtttttgc tctataaaaa tattaaattg ctattatgat 58440 Tryptophan - aspartic acid - valine - phenylalanine - isoleucine - phenylalanine - serine - valine - phenylalanine - alanine - serine - tyrosine - lysine - isoleucine - alanine - tyrosine - aspartic acid 58440 tactaaagat acctttgcgt agtactttat aatttccatg tacttttata aacactatta 58500 Threonine - alanine - lysine - aspartic acid - threonine - phenylalanine - alanine - valine - threonine - phenylalanine - isoleucine - phenylalanine - histidine - threonine - phenylalanine - tyrosine - asparagine - histidine - leucine - threonine 58500 catataatct ttgaatgagt tctgcaggaa gatattattg tcctggtttt gtaggtgaag 58560 Histidine - tyrosine - asparagine - serine - glutamic acid - serine - leucine - glutamine - glutamic acid - aspartic acid - isoleucine - isoleucine - valine - proline - glycine - phenylalanine - valine - glycine - glutamic acid 58560 aaacaataaa tgcacacatt cataacgtgc gacgtagtgc cagctttgta ttgccagaaa 58620 Lysine - glutamine - asparagine - alanine - histidine - histidine - histidine - asparagine - valine - alanine - aspartic acid - valine - serine - alanine - glutamine - leucine - valine - leucine - alanine - glutamine - glutamic acid 58620 atcattttaa gaaatagatt tggtatgtaa aatggctaga aaacacaaca caaatttgga 58680 Isoleucine - histidine - phenylalanine - lysine - glutamic acid - asparagine - aspartic acid - tryptophan - tyrosine - valine - asparagine - glycine - leucine - lysine - asparagine - histidine - histidine - histidine - asparagine - phenylalanine - glycine 58680 acgaagaagc tgttttgatt ccacatgatc tctataaaat atgaccaaaa aaaccgggaa 58740 aataaaatca tctgatcatg tgcaagtcct tttttagtac acaattaggt gaatcttgtt 58800 tttatgattg cttagttttc atgtttataa aataccataa tatgccccat aattatagca 58860 taataataaa ataggaaaat aacatcttcc ttttaatttt taagacaaat tataatcaca 58920 atattatagc agaagaatgt ttttcaatta aaaagtggtt tggttaaaat atgatttttt 58980 ttcttctaaa aatcccaaaa tggccaaaat atatttaaaa atatatttgt tatatatatt 59040 tttatttaaa aataattttt tctcccactt gttaaagatt aagtggcata agtatttgtt 59100 ttggggcaat tcactgccga cattcagaaa cagaactgta ttattgggca caggaaaatc 59160 tgattagtgt ttctaagaat tagtatagca cagcaagaca gggaagcagt ccaaagtgct 59220 aaaggttaaa agcaatttat aaaaactagg aaaagacctt acttgcttaa aaaaaaaatt 59280 tcttaataat taaaattgtc taagtgatat tcttgaatat tttttgtaat agtttcatat 59340 tattttcata aaaatacttt ggttgaggca gcagcagagg aggaagaatt aatataaaga 59400 aaacaaattt tcctattttg caagatccaa aatcttatga atatagtttt tagtgaaaaa 59460 aattattaaa cttttacaca gtaaccctaa gaaaagtctc cacatatggt cccaaaggtt 59520 aaaaaaaaaa acccaaaact cctctgggga aataatcatt tcatgaaaag cagtctattt 59580 ggaatttcac tggagacact ctgagaatcg aatcgagctt aatttctgtc tttgtgataa 59640 tcaaatatac attttgcatt ctagaagttt agaggaggta ttatatgtcc atggttttct 59700 ttaaacaact ctaacgtcaa aatggtcaaa ctgttggact gattcatgtt agataaaaca 59760 gatgggattc tcagatcgtg caccgttcag ccgtttgatg gaattcgcgc caccctgacc 59820 agcgagccca gatggcgctt aaatctgtca ctctttaacg accctactgt tttttaaaaa 59880 tatttttata tttgagctca ttcagttaat ctgagagatt aatcttttcc aagatatgtc 59940 tggggagggc tttactgagg gtttcacttg gaggagctca gggagagtaa gaagaatgat 60000 gaggcctctt ggacacagga ctacttgcta gaaggaacct ggggaagaag ggctggtggt 60060 ccttgggctt tggaatattt ttgaggcaac aaggcccaga agtgcacgtg aagagtggta 60120 gtgacggggg ttaagggaag aggggatgga tggggagggc gtggtgcagg gaggtggagg 60180 tgaacactga catcgctgag acagtacagc ccctgccaag aagcttagat ctcagtaagg 60240 ggccaagtgc tgactgtgac cacacctgag tggctttagg cagcctgacc ttcactgcat 60300 ggggatgact cggtgggtgt gccccaggag cagagcccaa gatgggctcc aatgtgataa 60360 gcctctggga tggctctgga aagaacctgt tctgcaccac aactctcaca aatctcatgg 60420 ggtttttttc cctgtaggtt ttttcccccc tttcttaaat tgttatttcg agccagtcag 60480 cctactgaaa tgcttcagtc atttggggtg taagatgttg gccagctccc gaatctctaa 60540 ccttggatac atttgcagag tggcttggaa ttgctttcta ttcctgctgg ggtaagaaag 60600 ggaaaaatga atgatttttc attatgcaca catggtattt tttcccgcat ttgaatcaga 60660 tgtagcaggt gtcaaaatat ggtacagaat ttacagtatt aatgtgaaaa tttctactgt 60720 cccaaaaagc caccagagat ggacaaaccc acctatgtta acaaaagtaa aaatgccctg 60780 ctgtctttaa agatgttcct ggtggtattt gggctgctct gtgctagttt ttctggggat 60840 ctgtctttaa agatgttcct ggtggtattt gggctgctct gtgctagttt ttctggggat 60840 cacgtgatgc tgcctatact tgggggtctt gtggaaagca gtggatgcct ctcaccctgc 60900 cacgtgatgc tgcctatact tgggggtctt gtggaaagca gtggatgcct ctcaccctgc 60900 atagaagacg ggtccaaatg tcttgtgccc acctgagtgt ctacacactg aagacacgtt 60960 atagaagacg ggtccaaatg tcttgtgccc acctgagtgt ctacacactg aagacacgtt 60960 agaggtctgc tttagagagg ttttcctcct aaattaagga tctatgactt tcacatagac 61020 agaggtctgc tttagagagg ttttcctcct aaattaagga tctatgactt tcacatagac 61020 ccaaaagtga gcagagatgc aagttgtgtt gagaggttct tttggaagat gtggggcttt 61080 ccaaaagtga gcagagatgc aagttgtgtt gagaggttct tttggaagat gtggggcttt 61080 gggggtcttg gggactgggg gactgtcaga agtcactgct ctagtagaat gtggtgggaa 61140 gggggtcttg gggactgggg gactgtcaga agtcactgct ctagtagaat gtggtgggaa 61140 cttataacaa cagattctcc acagaaatgc cacacgtaac catcccttga ggatggagcc 61200 cttataacaa cagattctcc acagaaatgc cacacgtaac catcccttga ggatggagcc 61200 acgtatgctg taaacagagt ggaagaacat caagctcgta acttgtcatt ttcatcataa 61260 acgtatgctg taaacagagt ggaagaacat caagctcgta acttgtcatt ttcatcataa 61260 atctgctgta agcttgcaac agccacggtt tccccctccc cacccacata cacataggga 61320 atctgctgta agcttgcaac agccacggtt tccccctccc cacccacata cacataggga 61320 cattatggaa aggattgcag tatagtccta cttaaagaat cttgcttttt tttttttttt 61380 cattatggaa aggattgcag tatagtccta cttaaagaat cttgcttttt tttttttttt 61380 ttttttttag aaaataaaca attgctagaa gagtggagag accacttgca tctaactctg 61440 ttttttttag aaaataaaca attgctagaa gagtggagag accacttgca tctaactctg 61440 tggcaggttt aggattctgt gtaatcagta agacagagag ggaatgtgtc aactggcctc 61500 aatttcctag tagttttggg tgtttacaga gtacaattga acattttaag ggaagcaact 61560 tgtggtgcct gcattgaaaa tcattttgat gaagttatgg aatacatttg atattatcta 61620 aaaaacaaat ctcctacttt tttgactcag atatttctag tagtcaaaat aaacattcgc 61680 tcttaaattt tactagtcaa ctcatttcac attcacctga ttccctctaa aaacaaaaat 61740 ctttctacta ttccaattgg caaatggtct gttttggaac aaaaggatag atgcatttga 61800 agaattctct ggtcacctga gaaatgaagg tttgcccatc attttttatt aatgcttgcc 61860 aagtaattag gtatgaaaag atgaggagca aaaataaatc ttcatcttca atagaacatt 61920 agaggctggg catggtggct cacacctgta attccagcac tttaggaggc cgaggtggga 61980 ggattgattg aggtcagaag ttcgagaaca gcctgggcaa catggtgaaa ccctgtctct 62040 acataaaata caacaaatag ccatacatgg tggcacatgc ctgtagtccc agctacgtgg 62100 gaggctgggg tgggaggatc gtttgagccc aggaggcaga ggttgcagtg atccaagatc 62160 atgccactgc actccagcct gggcgacaga gtgagccctt gtcttaaaaa aaaaaaatct 62220 ttaatagaac actggcggcc gggcatggta gctcacgcct gtaatcccag cactgggagg 62280 ccaaggcagg tggatcacct gaggccagga gttcgagacc agcctgacca acatggtgaa 62340 accccatctc tattaaaaat acgaaaatta gctgggtgag atggcagttg cctgtgaatc 62400 ccagctactc gggaggctga ggcaggagaa tcacttgaac ctgggaggcg gaggttgcag 62460 tgagctgaga ttatgccact gcactccagc ctgggagaca gagcgaaact ccatctcaaa 62520 aaaaaaaaaa agaaaaaaag aatattggct atttgagcaa gatgaatgta ttttggtaga 62580 tagaagaaaa atgagaacta tcgctcatca ctgctgatgt ctaaatcccc tccctgggca 62640 tgaggatggt acaatgacac agcaacccca gcctgccggg cttctgggat gccaagctgg 62700 gggtgtcctg gccaggtcca aggtgctgga agccaatcca gaaaatagga tgaagggata 62760 aaatcacact ccaaggccgt gatatgattg aatcaaggtg ttaatctagg tgaagagtgc 62820 agaagccaga ttcactaggg gctccagggc aggacaagct ggttcagcgg ggagaaggca 62880 ggaggcagat gcccaagaag tgtgggcaga tctaggggtg actttatagg ccctcagctt 62940 tttttttttt ttgagatgga gttttgctct gtcaccaggc tggagtgcag tggcgcgatc 63000 tcggctcact gcaacctctg cctcccaggt tcaagcgatt ctcctgcctc agcttcccaa 63060 gtagctggaa ctacaggcat gtgccaccat gcccagctaa tttttttgta tttttagtat 63120 agaagggatt ttaccacgtt gaccaggatg ctctcgagct ctcaacctca tgatctgccc 63180 gcctcagcct cctaaagtgc tgggattaca ggtgtgagct gccgcgcccg gccaggtcct 63240 tagctttgaa tggggagtga ggcattaggg gcatggagac acgtgaagaa catgtagctt 63300 gtcctttctt ccttccagag ctttctctag ccatttgcaa aaattccaca agcacattat 63360 gcagtcattt ttctaggaac tggagagagg ggagaggtgg atgggagggg agggtgtcat 63420 aagccttgaa aaagttagat attccaaaga accttagctt ttcccaaata ggaaatggag 63480 agttgggtgg tagtttcaaa tcattatttc attttcttta cctctgaaaa gatttcttag 63540 tttactgaga gcctaccttc aattgcatgg atgttgtaca ttttcttttt catttcccat 63600 gaagatccct atttttagct tttaagagct gttactcaca actgagatac aaacagtgtt 63660 ctgcacccca cacagcatat atctcatgat tagtgacagg gttcctagaa ctgcaagaca 63720 agtttgggct tactcaatct gttaagtttc cttataggca tattttttca ttctattgtt 63780 ttagcaaggg gcctattaga aatttccatg tgtataaatg acactttaaa gagctgggtg 63840 caggctgggc gccatggctt acgcctgtaa tcccagcatt ttggaaggcc gaggcaggca 63900 gatcatttga agtcaggtgt tcgagaccag cctggccaac atagtgaaac cccgtctcta 63960 ttaaaaaaaa aaaaatgaaa attagccagg catggtggca tgcacctgta gtcccagcta 64020 ctcaagaggc tgaggtatga gaattgcttg aacctgggag gcggaggttg cagtgagcta 64080 agatcgtgcc actgcactcc agcctgggtg acagagcaaa actccatctt aaaaaaaaaa 64140 gagctaggtg cagtggtgca tgcctatagt cccagctact tgggagactg aggtgggagg 64200 atcacttgag cccaggagtt tgaggctata gtgagctatg attgcgtctg tgaatagcca 64260 ctgcggtcca gcctgagcaa agtagcgaga ccctgtctct aaaaaagtaa aaattaaatt 64320 taagaagttc ttcaccatcc aagttatgtt acgtggaatt tcccgtggca gtttcctgaa 64380 acataacttg tttatgtttc ctgaaacaaa acttacctgc agtttctaga cacgtgctct 64440 aaaaacaggc atctactggc ctgttttagg atttagaaat ctctgctctg cccttctttc 64500 aactatccaa atgaaaagaa tgttttaaca tttcatattt gtgaaagttt cctcaattac 64560 tgactttttt cagttttaca atttgttttt ggccaaaaca acagcaacaa caaaaatacg 64620 atttaactgc aaaattgacc ccctgatctt agctacacct tccgtggtcc ttttctacca 64680 ttcagcttga ccctccaccc atgggaggtc tctgctcatt gggaatactg ggccatgaaa 64740 catctcaata ttttccatgg tctttacttt atgcttgctc ctcttctgtt cttcatacag 64800 caagccagag aatcagatca tgtcacttcc ctgcatatct ttcaaaacat cccattgaac 64860 ttagaataca atccctcctc cttactgtgg ccaccaaagc cctgcattgc tggctcctgc 64920 tgacctctcc agcaacatct tatgccactc tctcccttgt ccgctgccct tatgacacac 64980 tgacctctcc agcaacatct tatgccactc tctcccttgt ccgctgccct tatgacacac 64980 cggtctcctt tcagtgcctg cggtatacag cgagctagtt cctgcctcag ggttgttgca 65040 cggtctcctt tcagtgcctg cggtatacag cgagctagtt cctgcctcag ggttgttgca 65040 ctagctggtt ctgctgccag attctttctg tagctggctc cttgtcctct tgaggcccta 65100 ctagctggtt ctgctgccag attctttctg tagctggctc cttgtcctct tgaggcccta 65100 aagccttccc taccatccta ttttaggact cttatcctca accccctgcc atgattctct 65160 aagccttccc taccatccta ttttaggact cttatcctca accccctgcc atgattctct 65160 ctcacagcat cctgtttctt tccttcatgg catttatcac agtctgtcaa tttttgtttt 65220 ctcacagcat cctgtttctt tccttcatgg catttatcac agtctgtcaa tttttgtttt 65220 ttgttttctc gtataatgtt tgtcttccct actgcaggcc aaagagtttg agctgagcag 65280 ttgttttctc gtataatgtt tgtcttccct actgcaggcc aaagagtttg agctgagcag 65280 ggagttgcta ctgaaagtgt ttgatcaggg gagggtatga ttgcacttta aggagattga 65340 ggagttgcta ctgaaagtgt ttgatcaggg gagggtatga ttgcacttta aggagattga 65340 tctatagcca tggacaaaat agatttaagt gcttggtgaa ggcaagggag agactgaaac 65400 tctatagcca tggacaaaat agatttaagt gcttggtgaa ggcaagggag agactgaaac 65400 agaaaagcta agaatagaaa actactgtag gctgggcacg atggctcaca cctgtaaccc 65460 agaaaagcta agaatagaaa actactgtag gctgggcacg atggctcaca cctgtaaccc 65460 cagcactttg ggaggccgag gtgggcagat cacttgaggt caggagtttg agaccggcct 65520 cagcactttg ggaggccgag gtgggcagat cacttgaggt caggagtttg agaccggcct 65520 ggccaaaatg gtgaaacctc acctctacta aaaatacaaa aattagccag gtgtgatggt 65580 ggccaaaatg gtgaaacctc acctctacta aaaatacaaa aattagccag gtgtgatggt 65580 gcgtgcctgt aatcccagct actcgggaga ctgaggcagg agaatcactt gaacctaaga 65640 ggcagaggtt gcagtgagcc aagatcacgc cactgtactc cagcttgggt gacacagcaa 65700 gactctttct caaaaaaaaa aaaaaaataa taaaagctac tgcaaaagtc ctcctagtag 65760 gtgacaggga ctcaaactgg taggtgagaa gggatgtgtg agagactgcc cttgtgcctg 65820 gactcacgtc cctgacttgt gcacctccct cccaagcaca cagtcccctt ctcacatcct 65880 cgccctcact ccacccttca ccgtctcctt ttgggtgtca cctctgaggc aggcactaca 65940 ggccttctgg gtgattcttg agtgatgggt atgtctcaca tctttttgtt gatgatacag 66000 ttcctttata cagagctgta ttgttattag ctgtcgtccc tcttgttaga gcccttctca 66060 aggaagaata aatacactgt aggtgactag gtgactcctg gaatcttttc ttcgggaatt 66120 tctttttttt tttctttttt ttttttttga gacagagtcc cactctgtta cccaggctgg 66180 agtgccagag tgcagtgttg cagtcatagc tcactgcgac cacaaactcc tgggctcaag 66240 tgatcctccc acttcagcct ccagagtagc tgagacaaca ggcatgcact actgtgccta 66300 gctgcctatc agtttttctt tactgggtaa gtataaatat ctaaacatcc acaagacatg 66360 gaattgagag tctcttttct ctgtcaccta gtgtatttca ggtctataat aaatattata 66420 cagggctggg tgtggtggct cacaccgata attccagcac tttgggaggc ttcagcaagt 66480 ggatcgcttg agcccaagag tttgagacca gcctgggcaa catggtgaaa ccccatctct 66540 accaaaaaaa aaaaaaagtt agccaggcgt ggtggtgtgt gcctatagtc ccagcttctt 66600 gggaggtgtg aggatcactt gagcccagga ggttgaggct gtagtgagct gtgattgtac 66660 caccacactc cagcctgggc aacagagcaa gaccctgtct caaaaaatat atattaaaca 66720 gcaacaataa cattttacct catttttctc cactgttcat tttgtttgaa aatctgtgtg 66780 gcaaaactct atgcccttaa cccttaaagt tgccattagc cagatgatgg caggggaagg 66840 gcgcatggga tcttcagcag tccttcatcc atgcgcccaa tgggtattga ggagccacca 66900 tgttcagggc actggggaga tccatccatg aacagaacca aaatctgccc ccctagagct 66960 tacaattcag caggctgatg gtaacaacag agataaataa gttggtcatg gagaatttta 67020 gaaagtgtcc agtgtcatgg tgcaacagaa aaggtgcaac aggggccagg catggtagct 67080 tagcctgtaa tcccagcact ttgagaggct gaggtaggca gatcacctga ggtcaggagt 67140 tcaagaccag cctggccaac atggtgaaac cctgtttcta ttaaaaatac aaaaattagc 67200 cgggcatggt ggcgcacacc tgtaatccca gctactcaga aggctgaggc aggagaatca 67260 cttgaacccg ggaggcagag gttgcagtaa ggtgagatag caccactgca ctctagcctg 67320 ggcgacagag cgagactccg tctcaaaaaa aaaaaaagaa atggtgcgac aggggcaaag 67380 gaaatgaggg gaggcaggat gctattttaa atagggggta aggataagcc ccattgagaa 67440 ggtaatatct gagcaaaagt ttgaaggaga ggaaggactt agccttgaat acttttgagg 67500 aagaatggtc taggtagagc agccctgtgt gtccgtggtt agcagggagg gctgtgggct 67560 ggtgtgaagt gagaggggtc cacagcaatg gaggtgagac cagaggggga accagaggcc 67620 agatcgctgg aaggatgttg gctttactcc aggtggaatg aggagccatt gcagagtttt 67680 taaacagatc cctccagaag ttatgttgag aataccatgt aggagacgag ggttgaagtg 67740 gggagacccg tcaggagata gttgctgtga gacaggtgag agctgctggt gtctcaaact 67800 gagtgagatg tacctgggat atgtttcgaa gcagaatcaa tgagatttcc tggtgggttg 67860 agtgtttctc agttggaaat aaaagaggag tcaaggatga ctgtcaaggt ttttggtttg 67920 tgtgactgga gggaaagagt ggctttcaaa atgggtttgg caataaatgg agcacgcttt 67980 tgttggggta ggggtggagg tcaagagttc agctttggac atgttgaatt taagatagct 68040 actagaattc gttttttttt agctactaga attctttttt tttttttttt tttttttttt 68100 tgagatggag tcttgctctg tcacccaggc tggagtgcaa tggtgtggtc ttggctcacc 68160 gcaacctccg cctcgcaggt tcaagcgatt ctcctgcctt agtctcctga gtagctggga 68220 ttacaggcac gtgccaccat gcctggctaa tttttgtatt tttaataaag acaggggttc 68280 accatgttgg ccaggctggt ctcgaactcc tgacctcgtg atctgcccat ctcggcctcc 68340 caaagtgctg ggattacagg cgtgagccac catgcccagc tgatagctac tagaattcta 68400 agtggaggta tgaagtaggc agttttgggg agagaggctg ggtctggaga tgtacgtgtg 68460 ggagctgtta gcagagaaat ggtatttaaa gtctgggtgg ggtgacttag gaatggacac 68520 agagctgact gagcagtcac accccgtttc cccacgaaga cctcagccca aggcaaccac 68580 taatctattt tctgtcttaa tgttggacac tccaacatta agggatgcgt aagaagaagt 68640 ggagggggaa ccagcaaagg agatggagaa gtcatgagca gtaagatagg atgattacca 68700 aggagtatat gtcctgggaa ccaagtgaaa acagtacagc aagttgggag ggggtgattg 68760 caccagatgt tgctgccact gggtcagata attaggaaac caagaatcga ctattgaaag 68820 ctggatgcaa tggtgcttgt ctataattcc agccactcag gagactgaga tgcaaggatg 68880 gcatgagccc aggagttcga ggctgcagag aacaatgatc gctcctgtga atagctgctg 68940 cattccagtc tgggcaacat agccagacca gaccctgtct cttaaaaaaa caaaaattgg 69000 gctattggat ttagccacgt gggggtcatg ggtgaccttg acaagagcag atttggtaga 69060 agcatggaat ggatttttcc taatattcct cctcttcagg ggcagacaat atttaaatga 69120 agcatggaat ggatttttcc taatattcct cctcttcagg ggcagacaat atttaaatga 69120 aaccgtattt ggagcactca tcaggcttta atattgatgc tgcatcttct actgtcacat 69180 aaccgtattt ggagcactca tcaggcttta atattgatgc tgcatcttct actgtcacat 69180 acataaaggg gaccaggacc actgcattac agatgtgact atttctacct tatatatgta 69240 acataaaggg gaccaggacc actgcattac agatgtgact atttctacct tatatatgta 69240 tgccctatat gtgtatatac tttatgtgtc tatatcagct ttattgagat ataatttaca 69300 tgccctatat gtgtatatac tttatgtgtc tatatcagct ttattgagat ataatttaca 69300 tctcatacag ttcacctatt taaagtatac agttcaatga atttaatggt ttttagtcta 69360 tctcatacag ttcacctatt taaagtatac agttcaatga atttaatggt ttttagtcta 69360 ttcagtgtta tgcaaccatc accatgatta attttatttc tatcactgtt tcttcagccc 69420 ttcagtgtta tgcaaccatc accatgatta attttatttc tatcactgtt tcttcagccc 69420 caaaagaaac cctgttccca ttagcagtca ctcctccttt tcccccaaaa ccctcagtcc 69480 caaaagaaac cctgttccca ttagcagtca ctcctccttt tcccccaaaa ccctcagtcc 69480 taggcaacca ctagtctact ttctgtcttt ctatatttgc cgattctgga ctcttcatat 69540 taggcaacca ctagtctact ttctgtcttt ctatatttgc cgattctgga ctcttcatat 69540 acatggaatc atacgatacg tggtcttttg tgactggctt ctttcactta gtataatgtt 69600 acatggaatc atacgatacg tggtcttttg tgactggctt ctttcactta gtataatgtt 69600 ttcaagcttc atccaagctg tagcatgtgg cagtacttca ttccttttta gggccaaata 69660 ttcaagcttc atccaagctg tagcatgtgg cagtacttca ttccttttta gggccaaata 69660 atattccact gtatgtacta atatatacca cattttattt attcattgat tagttgttgg 69720 atattccact gtatgtacta atatatacca cattttattt attcattgat tagttgttgg 69720 aaatttaggt tgtttctact ttttagctat tatgaataat gctgctatga acatttctgt 69780 acacattgtt gtatgggtat atttcttctg gggttatact taggaatgga attagaatgc 69840 agtagcatga tctcagctca ctgcaacctc cacctcccgg gttcaagttg ttcttgtgcc 69900 tcagcccccc aagtagctgg gatcacaggc actgtgcctc cacgcctggc taattttttg 69960 tatttttagt agagatagag tttcattatg ttggccatgc tggtctcgaa ctcctaccct 70020 caggtgatct gcccgcctcg gcctcccatg gtgctgggat tacaggcgtg agccaccacc 70080 cctggccaca gactggcttc cttccttccc tccgtccgtc cttccttcct tccttccttc 70140 cttccttcct tccttccttc cttccttcct ttcctccctt tctttttctt tttctatttc 70200 tctttctctc tctctttctt ccctcctctc ccctcccctc ctgtcctctt ctctctttct 70260 gacagagttt cactcttgtc acccaggctg gagtgcaatg gcgcaatctc ggctcactgc 70320 agcctccacc tcatttgttc tcatgtctta gccactcgag tagctgggat tacaggcacc 70380 tgccaccagg cctggctaat tgttttttta gtagagacag gatttcgcca tgttggccag 70440 gctggtctca aactcctgac ctcaagtcac ctcagcctcc caaagtgctg ggattacagg 70500 catgagccac cacacccagc cttcttcatt cttttttttt ttgaggtgga gttttgctct 70560 tgttgcccag gctggagtgc aatggcacaa tctcgactta ctgcaacctg cacctcccag 70620 gttcaagcga ttctcctgcc tcagcctccc gagtagctgg cattacaggc atgccccacc 70680 actgggtgaa aaatgcccag ctaattttgt atttttagta gagatggggt ttctccatgt 70740 tggtcaggct ggtctcgaac tctcgacctc aggtgatctg cccacctcag cctcccagag 70800 tgctgggatt acaggcatga gccactgcgc tcggcccatc attcttaata atattcccaa 70860 atggtgtata gataaatact gccacaattt caggactaga ggttaaaaaa aaaaaaagga 70920 caaagttgga cagtagagct gagagcacaa attgtcccct ttttttccca ggtccttagg 70980 ttcgtagacc tgtgtcagcc ataattactg ctgaatgtga ctcctgtgta tacggtcatt 71040 tttatttttt ctgttcctgg aggtaatgca ataagtcacc tcaaaaaggg cagagaatga 71100 gggagagaat ctacctgcat tggcgtattg aaaagatcag attatttcag aatccagatc 71160 attgtttact tttagctgca taaaaatatc caagtagatt ttcttataac ctctccaaga 71220 taaagatgcc agaaacggca gccaaataca ttgactttga gagaggaaaa aaaaaaagct 71280 tgtaattgcc gatcactcct tagatttact tctggggagc atcaattaaa aatctcaaaa 71340 cacgtggctt tttcagtaaa attatattaa tatggaggaa gagtttttat taaatatgag 71400 tgcacgtgtg tctgtgtctg tgtgtgtttt aaacgtttcc ataatgagtt cccaagtgct 71460 tttttggcag tagttattaa agcttgtgtg tttgcgattt atctggcaaa atgcctttag 71520 tgaagaaagg atatagtgtt ctcttggttc tttgcccaaa gtaattttgg agaaatctgt 71580 gactgtattt agttcatgtc attcattgga gacaagtcat tgtaattaca tttgatcaca 71640 atcagtttac agttgtttta tcatgttttc atgctgaata tctactcaga catcatagca 71700 aaatttaatg gctactgttt tatttgtaca taaaaagatt tatagccctt ttttgcagta 71760 cacatcgggt ttcaatcatc actgctacag agtgagagga aagctatgga taaagaacta 71820 tcagaaatcc ttgactacat aaaattttat ttcaggtcac cttgccataa aagatattga 71880 tcagaaatcc ttgactacat aaaattttat ttcaggtcac cttgccataa aagatattga 71880 atgtattttg tggcttttcc tcctgacatg ctaaccgttg tagaagagtt gcaatatttg 71940 atgtattttg tggcttttcc tcctgacatg ctaaccgttg tagaagagtt gcaatatttg 71940 gaggaaaccg tatgaaatgt ggaacttaat tcacttaaga tggtttacat ttgcgcgtgt 72000 gaggaaaccg tatgaaatgt ggaacttaat tcacttaaga tggtttacat ttgcgcgtgt 72000 tttccactgg aaatagccat gccagcctta tgaatggccc ctgggtaatg cagagtaaaa 72060 tttccactgg aaatagccat gccagcctta tgaatggccc ctgggtaatg cagagtaaaa 72060 ggcaatggca ttaccgatat tgttgagttt taaaaggaag ttgtcttctt aaccatacgg 72120 ggcaatggca ttaccgatat tgttgagttt taaaaggaag ttgtcttctt aaccatacgg 72120 ctgaacattt tgtcattgtt gtttcagaag gtctgctcta acatctgcac tgcggcagag 72180 ctgaacattt tgtcattgtt gtttcagaag gtctgctcta acatctgcac tgcggcagag 72180 ttgtgttaca gggtgttatt tacagctgcc aaatgctgga ggctctcatg tatcagacag 72240 ttgtgttaca gggtgttatt tacagctgcc aaatgctgga ggctctcatg tatcagacag 72240 tctctgattt gctaattgtg gaagccttga ttcaccagtg aatttgggta aattcactag 72300 tctctgattt gctaattgtg gaagccttga ttcaccagtg aatttgggta aattcactag 72300 ttatctccta agtgtcactt ctcaccatgc cttatttttc ccaaaaggtt gattccaagc 72360 ttatctccta agtgtcactt ctcaccatgc cttatttttc ccaaaaggtt gattccaagc 72360 tctgctgaaa tctgctcttg gtctatatgt ttgagtctac attgttgaag aagctccctt 72420 tctgctgaaa tctgctcttg gtctatatgt ttgagtctac attgttgaag aagctccctt 72420 tagaaaagtg taattaatgg gaaactttga catccagctg agaaaattag tttgcatcaa 72480 tagaaaagtg taattaatgg gaaactttga catccagctg agaaaattag tttgcatcaa 72480 aaaaggcaag atagaggatt aaagaaattt tagtcctaat ctaattaaca ccaattaaaa 72540 gatgcagatg aatttacacc aacaaattgt taattatgca ccaatttgcc ttttttctgc 72600 aaccaaactg aaatcaggaa agtattatta tcttttttgt ttgaggagac ttacattgtg 72660 ggtttgttac aatcatcttt tatataatta aaaggtaggc agaaagagcg tgaaatattt 72720 cagtctcctg acattttaat ttaggttaac tctttaaaaa tacaaagtac agaaaaaaaa 72780 aattccagaa agtacaggat aattctatta gcatttattc actgactaga tatttattga 72840 ggacctagta tgtgctggac acttaggtaa ccattgtttc tctcactcaa aatattttaa 72900 cttgtttgta tttctaatat attgctcatt agaaaaattg tatacgccag gcgtggaggc 72960 tcatgtctgt aatcccagca ttttgggagg ccgaggcggg cagatcacga ggtcaggaga 73020 ttgagatcat cctggctaac acggtgaaac cctgtctcta ctaaaaatac aaaaaattag 73080 ccgggtgtgg tggcgggcgc ctgtggtccc agctacttgg gaggctgagg caggagaatg 73140 gcgtgaaccc gggaggtgga ggttgcagta agccaagatt gcgccacagc actccagcct 73200 gggcgacaga gcgagactcc gactcaaaaa aaaaaaaaaa gaaaagaaaa agaaaaatag 73260 tataagctaa ttgtaccaaa attgtataag ctaattaaat tctgccctgg atacatgtca 73320 gaaaaggaga atcgaaattt tgggcctcaa gaaatgctct taatgtccca gaactcctct 73380 ttaatgtgac cttttagggt attcagcgtc catcttcctc acccggggaa tctgtgtttg 73440 tttcccagtg ttactcagat ctccactgtg ctgggacttt ctttttcctt gtgaatcact 73500 cttacagctc ctcagatggt gatctgagta attatgtaga gggtggtgaa gcacagacct 73560 tttagatctg caaatgacac atctgtccaa atgtcatgta atgagtaacc acaatccact 73620 ttatataagc catatttctt ctctttccaa aatgggaagg agagggtggg gggggaaatg 73680 tctggtagaa atgggaagaa aagcatttct atggattcct ctagccatta attacataga 73740 aaatgccaca gctgccctca gatttccagg tgcacgttat taaacgtcat tcaaaggaaa 73800 tgtttggcca actagatcct gatgtgagaa acagcatcct aaagagctat gtatgttact 73860 gtctaatgca tgtacttagg cattaaataa tattaaatga ccatattacc ttcttggggc 73920 aatagttaat ataaacattt aggatgctca gtttgtactt ttcagtattt gtgctggaaa 73980 cttttgcatt cacaactttg aaaactggct cgtgttttca tgggaagtgt cagtttaaag 74040 ttggcctact aattaatggg ggaatttcaa tatggcagtg cctatgacag caaatgaaaa 74100 gcattcttta taaacatcgt cttcctccac acattcacag gttagtatct gacacccatt 74160 tttaatttct agagcaccat ttgtacttct ctgtggtttt gcctgcaagt cttcgatgat 74220 gcagtccacc cttccataaa aacgtactcc aattttccta gattctactt tcattgtttt 74280 gtgaatcaag ttagaaggta cataacttag cctgatgaga tggcattcat aaggttagga 74340 tagcagagtt gcctaactaa tcttcctgtt gcaatgaaat aaaagaatga tatctaagtt 74400 gaccaacctt gtaaaaggta attttacaag aggcctccaa aaataacgga ctaggactgg 74460 ccagagaaga aaataaaccc ctcatcctca atccagaacc cagagtttgt catcactagt 74520 cgaaaagctt gtttgtttta tacctaagtg agaccttgag ttatccaatg tgcttccatc 74580 acagaatggc ccaccagggt agaaggattc tgagtgtatt ccctgatgct gttgccatgt 74640 acagaatggc ccaccagggt agaaggattc tgagtgtatt ccctgatgct gttgccatgt 74640 gcctatggaa tacctatgct ccatatttag ccctctttct gtgtagtagc agagacgtgg 74700 gcctatggaa tacctatgct ccatatttag ccctctttct gtgtagtagc agagacgtgg 74700 atggctgctg aaactaaggg cccagtcttt ggccttagga cagaccactg ggtttctccc 74760 atggctgctg aaactaaggg cccagtcttt ggccttagga cagaccactg ggtttctccc 74760 actgcctaaa accttagtag cattagcatc agacatggac ccaataaaca aactacctag 74820 actgcctaaa accttagtag cattagcatc agacatggac ccaataaaca aactacctag 74820 ttgataagcg ccgcattttt catgatgttt tactgtggcc tatccacaca ctctttccta 74880 ttgataagcg ccgcattttt catgatgttt tactgtggcc tatccacaca ctctttccta 74880 ttccatgaaa ggaagaaaaa gataatttga gttgtcctat aaattaaaat ttttcctttg 74940 ttccatgaaa ggaagaaaaa gataatttga gttgtcctat aaattaaaat ttttcctttg 74940 tgattatttt ggaccagcgc cataacattc agaagattat gagtggagtc tttagtctgt 75000 tgattatttt ggaccagcgc cataacattc agaagattat gagtggagtc tttagtctgt 75000 gaacaaagtc ctaagaagga agtagtgctg taacttactt catctaaaga agcatcatcg 75060 gaacaaagtc ctaagaagga agtagtgctg taacttactt catctaaaga agcatcatcg 75060 ctgaatgtaa agagtgattc tggattggat cttaagccgg aaaaataaat ggagataaag 75120 ctgaatgtaa agagtgattc tggattggat cttaagccgg aaaaataaat ggagataaag 75120 tacgttattg ggataatttg tgatatttaa ataaggcctg tagaatagat actagtatcg 75180 tacgttattg ggataatttg tgatatttaa ataaggcctg tagaatagat actagtatcg 75180 tacccatgtt acattttctg atttggataa ttgcaataaa gttgtgtagg agagtgtcct 75240 tacccatgtt acattttctg atttggataa ttgcaataaa gttgtgtagg agagtgtcct 75240 tggtttgggg aaccatttaa gagtataggg gcacaatgaa tataacttac agatggttca 75300 cggaaaaaat acaggtgcat actatataaa aataaattta tatctgcata tattatgcat 75360 atctacctat ctatatctgt atttatacat atggagagag agagaccaag agagacagag 75420 agagcagtaa tgataaagaa atgtggcaga atgttagtgc taacagttgg taagactggg 75480 taaggagaat tcaggaatta tttagctatc cttgcatttt tatttttatt ttttttgaga 75540 cagagtctcc cactgtcacc caggctagag ttcagtggca cggtctcggc tcattgcaac 75600 ctccaccttc cgggttcaag cgattctcct gcctcagcct cctgagtagc tggggttaca 75660 ggcatgtgcc acgacgcctg gttacttttt gtatttttag cagagacggg gtttcaccag 75720 ttggccaggc tggtctcaaa ttcctgacct caagtgatcc acctgtctcg gccccccaca 75780 gtgccaggat tacaggtgtg agccaccact ccctgcctcc ttgcatcttt tctgttagtt 75840 tcagattaca gtcatgtgtc acttaacaat gagaatacgt cttgagaaat gcatccttag 75900 gcaatttcat cgttgtgcta ccatcacaga atgtatttcc acaaacctag atgttacaac 75960 ctaccgcaca cctaggctgt gtggtatggc ttattgcccc tctgacataa acctacacaa 76020 catgtagcta ctgaatactg taggcaattg taacacagtg gtaaggattt cttttctttt 76080 cttttttttt ttggagacag agtctcgctc tgtcgcccag gctggaatgc agtggcgcag 76140 tctcggctca ctgcgtcctc cacctcctgg gttcaagtga ttctcctgcc tcagcctcct 76200 gagtagctgg gactacatgc gcctgccacc atgcctgact aatgtttgta tttttagtag 76260 agatggggtt ccaacatatt ggccaggctg gtttcgaact ccagaccttg tggtctgccc 76320 accttggcct cccaaagtgc tgggataaca ggagtgagcc accgcgccca gccaggggta 76380 aggatttcta tatctaaaca tatatgaaca gaaaaagggt acagtaaaaa tactgcataa 76440 aagatttaaa atggaacact cacagaaggc tcttaccatg aatggagctt gcaggacctc 76500 atgttgttct gggtgagtca gtgagtgagc agtgagtgaa tatgaaggcc taggacatta 76560 ctgtcactac tgtagacttt gtaaacacta tatacttaga ctatactaca tttattaaaa 76620 aaaacaaaaa acttttcttc agtagtaaat taaccttagc ttactataac attttttttt 76680 tcttttgaaa cggagtctca ttctgtcacc caggctggag tacagtggcg tgatcttggc 76740 tcactgcaat ctcagcctcc cgagttcaag tgattcttct gtctcagcct cctgagtagc 76800 tggcactaca ggcacctgtc accacgcccg gctaattttt gtattttcag tagagacggg 76860 ggtttcgcca tgttggccag gctggtctcg aactcctgac ctcaggtgat tcacccacct 76920 cgccctccca aagtgctggg attacaggtg tgagtcacca cacctggccc aactttttac 76980 tttataaact ttatatatat atatatgttt tgttttgttt tgtttgtttt tgagatggag 77040 tcttgctctg tcacccaggc tggagtgcag tggcacaatc tcagctcact gcaacctctg 77100 cctcccacgt tcaagcaatt ctcctgcctc agcctcccaa gtggctgggc ttgcaggtgc 77160 ccccccctac accaggctaa tttttgtgtt tttagtagag acatggtttc accattttgg 77220 ccaggctggt ctcgaactcc tgaccttgtg atccgctcac ctcagcctcc caaagtgcta 77280 ggattacagg tttgagccat cacgcctagc ctgtaaactt tgtttatatt tttaacttta 77340 tgactctttt atagtaacag taagtttaaa acacacattg tacaaaaata tttcctttat 77400 tgactctttt atagtaacag taagtttaaa acacacattg tacaaaaata tttcctttat 77400 ctcttattct atatgctttt ttatattttt aatttttatt ttttatactt tttgaacttt 77460 ctcttattct atatgctttt ttatattttt aatttttatt ttttatactt tttgaacttt 77460 ttgttaaaag caaaaacaca aacacataca ttagcctagg cctacacagg gtcaggatca 77520 ttgttaaaag caaaaacaca aacacataca ttagcctagg cctacacagg gtcaggatca 77520 tcaatattac tgtcttccac ctctacagct tgtcccagtg gaaggtcttt aggggcaata 77580 tcaatattac tgtcttccac ctctacagct tgtcccagtg gaaggtcttt aggggcaata 77580 acacgcacag agctgttgtc tcctgtgata acaatgcctt cttctggaat gcctcctcaa 77640 acacgcacag agctgttgtc tcctgtgata acaatgcctt cttctggaat gcctcctcaa 77640 ggacccacat gagtctattt tacagttaac tttttttttt tttaataagg aggcgtatag 77700 ggacccacat gagtctattt tacagttaac tttttttttt tttaataagg aggcgtatag 77700 tctaaaatga agataaaatg taaagcataa tataaaacca gtaacattta ttattatcaa 77760 tctaaaatga agataaaatg taaagcataa tataaaacca gtaacattta ttattatcaa 77760 gtattatgta ctgtgcatgg ttgtattgct atatatatat aaaatttttt ttttttttga 77820 gtattatgta ctgtgcatgg ttgtattgct atatatatat aaaatttttt ttttttttga 77820 gacagagtct cactctgtca cccaggctgg agagcagtgg cacaatctcg gctcactgca 77880 gacagagtct cactctgtca cccaggctgg agagcagtgg cacaatctcg gctcactgca 77880 agctccacct cccaggttca tgccattctc ctgcctcagc ctcctgagta gctgggatta 77940 agctccacct cccaggttca tgccattctc ctgcctcagc ctcctgagta gctgggatta 77940 caggcgcatg ccaccatgcc tggctaattt tttttttttt tttttgtatt tttaatagag 78000 caggcgcatg ccaccatgcc tggctaattt tttttttttt tttttgtatt tttaatagag 78000 acgacgtttc accatgttga ccaggctggt cttgaactcc tgaccttatg atctgcccac 78060 ctcggcctcc caaagtgcta tgattacaga tgtgagccac cgggcctggc atgtttgatt 78120 tttttttttt ttttttttga gacagattct ctttctgttg cctaggctgg agggcagtgg 78180 cgcgatctgc aacctctgtc tctcaggttt gagcaattct cgtgcctcag cctcccaaat 78240 agctgggatt acaggtgcgt gccaccacac ccagtttatt tttgtatttt tagccaccgg 78300 gcccggccca gtataatctt atgggagcac tgtggtatat ttggtctgtt gttgacctga 78360 aagttatgtg gcgcatgact gtatctaaaa acaaacaaac aaaaaattta aatagaaaag 78420 ctggattccg accacatgaa tgaaccttct tatgattctg gaaggaaggc aggttctact 78480 aagacagggt tcttaggcta ctaccactac ctcacccata aaggatcagc cttcttaacc 78540 cagccagtca caagggattc tgaggatgaa tgttcaccag ggcagattgg ttatgttcaa 78600 cagcttcatg tatttctgca aaatctacca gtccgatgat gcagtgaaca taatgatccc 78660 ctgtgaaata ataacaagat aaaatgttcc caggcccatt tgtttcccac tgctaccacc 78720 ccataagact gaaccatggg ttggagtgaa cacttgaacc aaaggttact gtattgtatg 78780 ttcctgttca cacaggactt cacaaaaacc ttcttttata aagaaaacaa ccagccaata 78840 aaacagcatt agagtacatt atatatgtga ccaattgttt gataaaatct gatgtaagcc 78900 ttcaggcgta attggagttc ttagcaaaat actttggttt ttaatatttc tatgaccctc 78960 taagggggac ttagcaatta ctcttaaatt ttcaaagtgg tgcatggagt tttagctgtg 79020 tgacttccat taaagtccat aggagtcttg ttgctaaaac atcacacaac tcacgaaaat 79080 gtgccccttc ttgtgaaatt gttagaagta atgggtatgg aagtatagca cagcatttct 79140 ggtacattta caagctttct gtacttgttt caaactttat acctttatga aaactcaata 79200 gttctacaaa aattgaaggg gggaaaaagc ctattatgtg tattccagaa gtgaagggta 79260 gaaaaaatac agtagaacta ataaaaaatt aaaataattc ttcaagaaaa gccaatgttt 79320 caacatattt ataatcttta tttcaaaatt aatatatcct ctcctgggtt tttaaagtaa 79380 gccttaaaat cgaacttaaa aaaatgaact tgactgacaa agtttcacta taaccacata 79440 caatgatttt gggtttcagc aacctcctct actgaaaccg atataattta agcatcatcg 79500 gagcaaaact tcactccttt agatggttgg tggtgtgagg ggtttgttcc cagtggataa 79560 gaggatggcc ttaaggtttg ctgagtcctt ctctctcaag aacctcttag aggccgggca 79620 cggtggctca agcctgtaat cccagcactt tgggaggcca aggtgggtgg atcacttgag 79680 gtcaggagtt cgagaccagc ctggccaaca tggtgaaacc ccgcctctac taaaaataca 79740 aaaattagct gggtttggtg gtgtgtgcct gtaatctcag ctactcgggt ggctgaggaa 79800 cgagaatcgc ttgaacccgg gaggtggagg ttgcagtgag cggagattgc gccattgcac 79860 tccagcctgg gcgacagaac aagaccctgt ttcaaaaaaa agaaaaagaa tctcatagac 79920 atttttctcc tgcgtccctg tgataagagt gctgtgattc ctgctataca gtcaaacaga 79980 ttcagagccc acctttgact ccaggaccaa atttccccct ttattctctt ttgtgctatg 80040 ctggagaaga cagtgtggca gttcccggga atttgtgtgg agctgtttca gggacactct 80100 gagctaagaa aagggtgaga gccctgggag gagaccttct ttcaatccag gctacatgct 80160 gagctaagaa aagggtgaga gccctgggag gagaccttct ttcaatccag gctacatgct 80160 ctgagttttt ccagggcata tatagctgct ccatttggac tatgctccta gactgacatg 80220 ctgagttttt ccagggcata tatagctgct ccatttggac tatgctccta gactgacatg 80220 ggcctgactc gcaggagccg gcagctccta gtgctgagct caccggctcg gcccagcacc 80280 ggcctgactc gcaggagccg gcagctccta gtgctgagct caccggctcg gcccagcacc 80280 agcaccacct cccctctgag catctggtgg ccttgctgat aacacagtga cacagccatt 80340 agcaccacct cccctctgag catctggtgg ccttgctgat aacacagtga cacagccatt 80340 tccttctata gcagaaacat ctttggagtt ctgtttccac agtcctttga gagacactct 80400 tccttctata gcagaaacat ctttggagtt ctgtttccac agtcctttga gagacactct 80400 cgaggccagg agctggcctc agacataaaa gtatttgttt caacagagag agagagagag 80460 cgaggccagg agctggcctc agacataaaa gtatttgttt caacagagag agagagagag 80460 agagagagac tcagtgactg ctttttgagt gaattgttgt tggctcagtt tcagtaatgc 80520 agagagagac tcagtgactg ctttttgagt gaattgttgt tggctcagtt tcagtaatgc 80520 aatgttttgc tttagattaa tttactaagc agtactaaag tatcatttat atacattatt 80580 aatgttttgc tttagattaa tttactaagc agtactaaag tatcatttat atacattatt 80580 gaatcttggg atgattctgt aagcattact ttctcctgaa atctccactt ctgtctggat 80640 gaatcttggg atgattctgt aagcattact ttctcctgaa atctccactt ctgtctggat 80640 agtggatttt aggataggtt tttaaaatgg aaaacttgac gcagattcaa attttcaaca 80700 agtggatttt aggataggtt tttaaaatgg aaaacttgac gcagattcaa attttcaaca 80700 cagcattgaa cgaatagcat cagttgggga cgagctatgc caccaattta aagtgtctca 80760 cagcattgaa cgaatagcat cagttgggga cgagctatgc caccaattta aagtgtctca 80760 gccggggtca gaattaggaa ctgtctaaac agggaatttc agaagggcag gggaggatat 80820 caaggcacaa atgggaggaa ataagccatt gttcctttag aaagcttagt aacctcgata 80880 atgagaggct atgaaatagt tctttttata actgtaactt cttaacttcc cagccctaat 80940 agctccaagg gtcacttgca gagagctccg gggctacacc tgccttgcat agattagggc 81000 ccgctgcagc tgtcttcatt tgcaagacta aggtgtagct ccaagaggct gtaggtctca 81060 gcatggagca ctcctcctca atctcaatag gctggtgttg cggattaaga tggccattgc 81120 aagccaggct cagaaattgc aagatgaggc aggtaacgat ttcctgggtt ttatgtatca 81180 ggccttatga ggtatgaagc gggaaacctt ttatatgcct gccttgaatt agccatacct 81240 tttacaatca gctctgttga agagcagctg cctggactat ctagcatacc catgggccag 81300 gcatgccttt ggcacacaca caggtttgta aagagcttgt gaatctatag gatgatctct 81360 tggtgaggac agaagggacc catagctagt cagcatcgga ctgagggctg gatccacttt 81420 ttcctgtaat gtgcgtttgt ttcacagaaa atacaattag gataattctt gtgtgttctc 81480 tacaggaagg aagtagtata ctgcttgtga atggtaggct aaatgaggga accattaaag 81540 tacaggaagg aagtagtata ctgcttgtga atggtaggct aaatgaggga accattaaag 81540 ggtttttaat agacttttga accaaaagac ataagggact atacttacat tgccttaact 81600 ggtttttaat agacttttga accaaaagac ataagggact atacttacat tgccttaact 81600 gagaaatgtt ttgcagtgtg agaagactta atcaaatctt cattacatac ctggtcaggg 81660 gagaaatgtt ttgcagtgtg agaagactta atcaaatctt cattacatac ctggtcaggg 81660 accaggctga atgtttcttt aagcaattgg ataaaatgca cattgaagac ttgttagtta 81720 accaggctga atgtttcttt aagcaattgg ataaaatgca cattgaagac ttgttagtta 81720 tgtgagaata tgtctgacat gcttatcatc tctcctgcgg gtgcctgagt atcagatact 81780 tgtgagaata tgtctgacat gcttatcatc tctcctgcgg gtgcctgagt atcagatact 81780 tggtttactg tgttgggttg aatgcttctg ttctaatttc taatgaactt taccaccaga 81840 tggtttactg tgttgggttg aatgcttctg ttctaatttc taatgaactt taccaccaga 81840 ctacttgatc atgactaagc ccaggaacaa tgcaaatgga attgccccct gatggggagg 81900 ctacttgatc atgactaagc ccaggaacaa tgcaaatgga attgccccct gatggggagg 81900 aatgggccca gatggccctt aaaaggtgat aattaggaaa tatagcatat ctgagtagct 81960 aatgggccca gatggccctt aaaaggtgat aattaggaaa tatagcatat ctgagtagct 81960 agaaagagag aaacttgtta aagacagtca atctgtttga agcaacggtc tttcccttaa 82020 agaaagagag aaacttgtta aagacagtca atctgtttga agcaacggtc tttcccttaa 82020 ttatatgcat tttcatttag agttgcttta aagtaaaatt tttcccagga aaaattttta 82080 ttatatgcat tttcatttag agttgcttta aagtaaaatt tttcccagga aaaattttta 82080 ttagtagtga aaacctagaa accacacaca ttttttgcta ctctgtttca gattattgaa 82140 ttagtagtga aaacctagaa accacacaca ttttttgcta ctctgtttca gattattgaa 82140 aattccttcc ttcatacatt tttttttggt ttccagatct gtgtaacaga ctcgctgtgt 82200 gtgttttagg ggccgtttta ggttatttta gctcttatgc taaaacagtg ggggagcgat 82260 taaaagtgag aaaaatatta gtatatattt gtaaactgag cttttaaaat taaaatagtt 82320 gcaggatatt acagtctgtg atttcatata ctttgtaaga ttgccttaaa tgtattgcct 82380 tcttggcagc ataatgccag atttttctcc aatatgtcat taaaataatt ttacatataa 82440 caagttgaac tttttaaaga cttatatttt aatctgaatt agcatattta ctaattatat 82500 tacttatttg gcttttcttt agtccgattc tttttttttt tttagtgtac aaacttggca 82560 ttactcttta agaataacaa gtgaactgga aatgagaggg tggtgacatg aagagagaga 82620 gaagaaatac attccttata tctcttccag ctcctgcatg gaatggagga tctgacagtg 82680 gccgttcctg ggtattaaag ccagtgcaaa agatagagcg tccaaaaggc aacttcttcc 82740 ttatttgtga agtcagaaag taaaatttac attgaagtgc taagctacat tattctgaat 82800 attttggcag tcatatttgg gcagatgagt tgctttcctg aaattactca gcaccgtcat 82860 tcatcctgac aaaagctagt ggtacagatg cacctttggg actgaggcca aggtgaaacc 82920 tcatcctgac aaaagctagt ggtacagatg cacctttggg actgaggcca aggtgaaacc 82920 tgcgttttcc aggtctgccc actcagtttt ggaaggactc tcaataaaac cggcctcaca 82980 tgcgttttcc aggtctgccc actcagtttt ggaaggactc tcaataaaac cggcctcaca 82980 ccccaagcat cacccctcct gtccttagta agtacactct ggctggaatc tagggtccca 83040 ccccaagcat cacccctcct gtccttagta agtacactct ggctggaatc tagggtccca 83040 gcatctagcc cttgggactt cacataaatt catcaagtat cttcctccac tcaaaaatct 83100 gcatctagcc cttgggactt cacataaatt catcaagtat cttcctccac tcaaaaatct 83100 cttacagaat ctttgacatt ttaagcttca ttactgcaaa gtggatatag ttcattctca 83160 cttacagaat ctttgacatt ttaagcttca ttactgcaaa gtggatatag ttcattctca 83160 gcttcaggat tccagaagca tacataccag gtcagaatgg agaggcatca cgctttcctt 83220 gcttcaggat tccagaagca tacataccag gtcagaatgg agaggcatca cgctttcctt 83220 cagcaagctt caggggagac aggatttagg gtgggctaca catataccag ttgtaggatg 83280 cagcaagctt caggggagac aggatttagg gtgggctaca catataccag ttgtaggatg 83280 ttttgaacta tattcttgca atcacagaca ggttgtccca gagaaaataa tctggctcta 83340 ttttgaacta tattcttgca atcacagaca ggttgtccca gagaaaataa tctggctcta 83340 acttctcttg ttatcagtga aaagcaaaaa ttacttagtg caggtatgct taaaggaaga 83400 acttctcttg ttatcagtga aaagcaaaaa ttacttagtg caggtatgct taaaggaaga 83400 aaaaggcaag agaaaagtga gaggtaaaaa ttattaccat tttaattcca tgattctcag 83460 aaaaggcaag agaaaagtga gaggtaaaaa ttattaccat tttaattcca tgattctcag 83460 caatgaagga gcatgattga aatcccatgt gcccagtcat atctgtaaat gagagtatga 83520 caatgaagga gcatgattga aatcccatgt gcccagtcat atctgtaaat gagagtatga 83520 tttttgaaga ctgtattcaa ttgtttcaac ttgagcaatt gcaagtaaga cttaggactg 83580 tgtttaccag ttactgttat ttactattac tttctaaatg tgacctgact tgaaaccttt 83640 atagatgcaa tttttatcca tcacagaaca agctctaaag aggagagcag cttcaacttt 83700 ttcaagtatt ttttttaaaa ggatttaact ttgaccaaca ccaaacttgc atcttgttaa 83760 ggtaactaat aactgactgc tataacaata tttagaattt tgtttaacag caactgtcaa 83820 aaaaacaagt taaaatagcc caggagatgg atggcgatta actctgcgtt ctttttagac 83880 tgtgttatta attagctctc tagactcaaa taggttacat cctctgagca gtaaattgca 83940 tgccatttgc aagctctgtg gatgaaattt acatgtggcc taaatgctat ccacattgtt 84000 ttgttaagta gcattttatc tctctggact ggtcccttgc ctcctccccc agccccagtg 84060 gaaagtttta ttcacttttc ttgcgggaag ggaaagtcgc atgtagtttg gtcatttggc 84120 gccatcttct tttttattgt taaatccact aaaggcagtc atgcctggca gtaccaggcg 84180 ggctccttcc ggggttctgg tcggctaagc attcagagtt ctttgctttt tatttactag 84240 aaatgttttc agcttctgtc tttgctttaa gatctgatta aaaggaacct taaaaaagga 84300 acatagattt tggtgtatgt ttttcctctc tacttctctt ccctgtgccc cccaaaccca 84360 atcaccaact tcccccatcc ccagcaaaca cgggcttttt tgtgtattta tttaaaagca 84420 ttttaggaag caccgtaata ccgcacagct tggaggttaa gatagaattg gatgtttccc 84480 tcaccatcag ttgatggaat catcagtgtt ccggttctat ttatcttcat gatttgagaa 84540 tgtttctgaa cagctgccct tccgaggatg ctggtattta gacagacctt cctgtcattt 84600 atctgactca caggcacaac tcttttgaaa gaattaatat caaacttaca tatgaaaatc 84660 taccatttag aaatggattc tttttcagta cctacttttc aattcaaccg gggagatccc 84720 atctatttta aaaaatcatt aatcacatac ttctgacatt ttccctttca gtttcctcag 84780 ggggaaaaaa gttatttgaa taacaaacaa acaaacaaaa acccctcatc gcaaatagct 84840 gtcagcctcc tatgtgaaca aaagcgctct ctcacactga gacctaccgt gttttgttta 84900 gttagaggca gcagggtggt agggacacta tgcagaaagg cttgggttaa tgagcacttt 84960 ctggattgag tgtctctcct gattaaccct tggattactt gctttgagtt gctaaccctc 85020 tctcactgcc cactcaagaa tttcccacat ggtagcctca tccatagatg cccttcccta 85080 gatgcactga tggtagacag cataaaaatg tgtaagcact gtagctttcc aatgaactga 85140 aacaaaaagc ttaagcatta ctttttattt ccttgatcta aagaacatgt tttgggggat 85200 attttcttaa tcaatatcta agttatctag agttataaat actataaata aattagttac 85260 aaactaattt ttagacacaa gtcttttgtt caaaccaact cttacataga aactctaaaa 85320 atttgggggg gaaggggaca gttggggatc agaatcactc cccattatct ctctgttggt 85380 gctccccaaa ggatcttcag ggaacgacgt ttaaaaacca ttttgtatca tgaaatggga 85440 cccgaaagac attctttcta gtgactttgt gttagattga gtcatatgaa attgccactg 85500 tgtgacccat tttgacctaa aaaaaaaacc aaaaaacaaa aaacaaaaaa ccagcactag 85560 ggtaccagag tatttttagt gtaatcactg tacgtgataa gaaaatggag aaatgagaaa 85620 gcttgaaact tagactataa gctcctcata agctaatagc gacttcctcc cttcttccgc 85680 agtcacctgg caagtccctt ctatgtgctg ctggttctag ggatacagag ataaaaggcg 85740 tagtcactgc ctccaaatca tcacagtgca aaggacaaat aaatacagaa ggacccatgg 85800 gatgaagaaa cagcccaggg tggtgggaac acagagcgag agtttctagg atcttgaaag 85860 ccatgttgca cggagctccc ggtccctgag ttttcagact gatctttctg tggactgtgc 85920 ctaagtaata acagtttcag aacgtagaaa ctcagtgtac atccttcata gccaggatgg 85980 atatgatagc ctggaagtga acagtaaatc cagtgatttg ttttcttcac tcattctttc 86040 attttaagaa agtgcttaga atccacaagt tgtgcttaat gccagcagac gcattacata 86100 tgccagccct gccccttagt cacaattact ccccacactg acctcccatt gcatttaaaa 86160 attgtcgctt tattgagaca tagttcagct ctcataaagt tcacatttgt tttttttgtt 86220 tttttttttt tgtttgtttt tttgagacag tcttgctcag tcacccaggc tggagtgcag 86280 tggcatgatc tcggctcact gcaagctccg cctcccgggt tcacgccatt ccccggcctc 86340 agcctcccta gtagctgaga ctacaggcac ccgccatcac gcccggttaa tttttttgaa 86400 cttttagtag agacggggtt tcaccatgtt agccaggatg gtcttgatct cctgacctta 86460 tgatccaccc gcctcaccct cccaaagtgc tgggattaca ggtgtgagct accgcctcca 86520 gccataaagt tcacattttt aaaggacaca cgtcagtggt tttcagtctc ttcacaaagt 86580 tgtgtaatga tcatcactat ctaattctag aacattttcc ttaccctaaa agaaacccca 86640 tacccattgg ggttcactcc ctatgcaccc ctcttccttt ctcctgacaa ccactaatct 86700 actttctaaa tctatagatt tgtctgttcg ggacatttcc tataaatggg actatataaa 86760 tatgcccttt tatctctggc ttccttcact ctgcataatg ttttcaaggt ttgtccatgt 86820 tgtagcatgc attagtactt cattcctttt tctggcccag taatgttcaa ttgtaggaat 86880 ataccccatt atccattcca atgatggata ttttttgggg gatatccact ttatggccat 86940 tataatttca ctatgaacat ttgtgtacaa gtttttggtt ggacatacgt tttcattttt 87000 gttgggtata tagctaggag tggaattgct gggtcttatg gtaactatgt ttaacatttt 87060 gaggaactgg gatgtatagc gttttgtgcc tgtgttatga aactaattat gaagcatctt 87120 atattcatta tcattagatc tgtctcctcc attaggtttt aagtttcttg agacagggat 87180 tatccttctt catctttgta tcccccacag tccagcactc accaaggcac aaagtaggtt 87240 cttaggaaac tggtcagaat aaaactgacc aatgcaacat cctccttccc ttaaaatcac 87300 cagcatacct gggaacacag gctttctctt tctctcttcc ctgtagcaag tgacagctta 87360 tcaggaatgg aggcagctcg gtattatcca tcaagccttc cagctgggtg cacaccctcc 87420 ttatctgctg tgggaccttc accagcatcc tgggatttcc tcaggctgcc tcatccccac 87480 aacggcagtg atgtgaccca ctctggttta tgcagatacg ttaagagaga ttattttaag 87540 taccactgaa acccaagcag aggcaaggct ggtggaaacc ccagtatttg ggggcacact 87600 ccctgtttga catggttcag gaatgactca tacatctact ttaaggggga tttggacaag 87660 gaggcactaa gatacctatc caggctttga cttgggagca ttgtgcagga ggtttaagaa 87720 agattatttt aagactatct cagaaggctg gagcagcatc tttctgattt cttctgctac 87780 ctttcactcg gagcctgtga tttctaacat gtatttcagg ttggatttaa gagagagctg 87840 gcccctactg aattggccac ctttgatgtt acagtactga ggaggggagg gacagatgga 87900 acaaaagtcg taattgccca aatttacaat aggttcccag cagcagctcg cacaaaagaa 87960 atgttaatgc aataaactgt aaatggaagg aagctaagca ggatttatgg ctggcgcaaa 88020 cctccatcaa ggcactggaa cctctttcct tcatctgtga ggcatttcag tggtcacagt 88080 agaattgggc tcatttgaaa atcagtggct gctctgtgtc ttccctccct tgtgtgtcct 88140 gcacccccta cacaaaccaa ttcctgtaaa aacctgggga atctgataga acaattggag 88200 aaaaacatca gtcattgagg gtcaagactg caaacccggc agagccaaga gtgcacacac 88260 atttgctgtg cacatgtatg tgttacagct gcaaccaaag cgtgcgtttt ccttccccaa 88320 gtctgtgttg gggacttccg cagacacagc agatggccag agtgtttcca cgtttcagcc 88380 tcgaagtgag agggcagcat gtcattcacc agcctggcac ttacccatcc atcagggcca 88440 tcgaagtgag agggcagcat gtcattcacc agcctggcac ttacccatcc atcagggcca 88440 cctctataaa tggctgtttt tcacatttgg ttccagcagg ttccatctcc attccggctc 88500 cctctataaa tggctgtttt tcacatttgg ttccagcagg ttccatctcc attccggctc 88500 acacatacct ttacctgtgt tgtatcatgt ggggtgctcc ttgtctcctt gtccagattg 88560 acacatacct ttacctgtgt tgtatcatgt ggggtgctcc ttgtctcctt gtccagattg 88560 ctggcccgct tgcttcatta attaaccgag gagcacgtct aaatggaaaa ttgctttctg 88620 ctggcccgct tgcttcatta attaaccgag gagcacgtct aaatggaaaa ttgctttctg 88620 agatctgtgt gtcctggctc catgctgtga ggggctccct gtaattaaac acgttttaag 88680 agatctgtgt gtcctggctc catgctgtga ggggctccct gtaattaaac acgttttaag 88680 agttgctgtc ttgcgcatgt gccctggaca tcggtgcttt ccccactggc tgggtagttc 88740 agttgctgtc ttgcgcatgt gccctggaca tcggtgcttt ccccactggc tgggtagttc 88740 tgtttcagac cttagaccta aatgataata acagttacaa ggaaacttgc atatcagttg 88800 tgtttcagac cttagaccta aatgataata acagttacaa ggaaacttgc atatcagttg 88800 attaagtgtg gggaattttt ttccttttga aagttggggg aaagaactcc tcagtgggga 88860 attaagtgtg gggaattttt ttccttttga aagttggggg aaagaactcc tcagtgggga 88860 agtgattttt ctgccctgta gggatgggaa aatggttaat catcatgggg gcctcaacag 88920 agtgattttt ctgccctgta gggatgggaa aatggttaat catcatgggg gcctcaacag 88920 acctcagcat gttcgcgaag tccttggcct caggttcctc atccatccca ccgcgagagc 88980 acctcagcat gttcgcgaag tccttggcct caggttcctc atccatccca ccgcgagagc 88980 gatacggctt gggtaagccg cgaatcccct cactgggctc caattattcc agaattctaa 89040 gatacggctt gggtaagccg cgaatcccct cactgggctc caattattcc agaattctaa 89040 ctttaaaaat aaaatattca tatttaaaag ttcctgctac cgtttcaaag aagcttccca 89100 agaagctctt cagagctggg gcatggagga acctgtcact gggtcccttt ctgctggggg 89160 atcagttcct tttataagta ggaaaagatt tctccatacc ccttgcaaag ttcatggctg 89220 acacccctat aacaaaagac aggttaacaa gagcaaagca cacaaatata tttaatacaa 89280 gttttatgtg acacaggagg cttctgaaat gaagcctcaa agacccaggg aaaaccgcgt 89340 ttttatggac agtcaagcag aaatatgatg agagaacaaa agggtttgat gtagcgtaat 89400 aaacttagca agcccgttca gattcttctt ggtgttcctg tgtgacattt tttttttcct 89460 ttttgttttg agacggagtc tccctctgtc gcccaggctg gagtgcagtg gcgtgatctt 89520 ggctcactgc aacctccgcc tcccgagttc aagagattct cctgcctcag cctcccaagt 89580 agctgggatt gcaggtacgc accaccacac ctggctaatt tttgtatttt tagtagagag 89640 agatttcacc atgttggcca ggctggtctc gaactcctga ccttagatga tcccctgcct 89700 tggcctccca aagtgctggg attacaggaa tgagccaccg tccccagtgt gacatttctt 89760 ttctctgggt acagggcagg acacctgtca catgagggtc ttcaggggag gagggagaag 89820 ttctctgggt acagggcagg acacctgtca catgagggtc ttcaggggag gagggagaag 89820 gtcagagggt gaccttccta cttctgcagt tttctcgatt tctttcagtt tacagtactc 89880 gtcagagggt gaccttccta cttctgcagt tttctcgatt tctttcagtt tacagtactc 89880 agtaggacaa ggtgcatatt tggggatatc gtgttctgag ccccaacagt aacaaagaac 89940 agtaggacaa ggtgcatatt tggggatatc gtgttctgag ccccaacagt aacaaagaac 89940 aaagaaagcc tttaggcagt ttttgctgag actctcaaat aggaatttag agtgatggtc 90000 aaagaaagcc tttaggcagt ttttgctgag actctcaaat aggaatttag agtgatggtc 90000 cttgagtcag agaggccact agagaaagtt tgactttcca tgtagaatag caagctctcc 90060 cttgagtcag agaggccact agagaaagtt tgactttcca tgtagaatag caagctctcc 90060 tctggcaagt cttttccaga caaggctcta taattttttg ctaataaatg tgtggacaaa 90120 tctggcaagt cttttccaga caaggctcta taattttttg ctaataaatg tgtggacaaa 90120 gttatgggat gtatagaagt ctgacaccaa aaccatgtca gagacttttt tttttttttt 90180 gttatgggat gtatagaagt ctgacaccaa aaccatgtca gagacttttt tttttttttt 90180 aaaccgagtc ttactctgtt gcctaggctg gagtgcagtg gcatggtcat ggctcactgc 90240 aaaccgagtc ttactctgtt gcctaggctg gagtgcagtg gcatggtcat ggctcactgc 90240 agcctcaacc ttctaggccc aagagttcct cccacctcag cctcctgaat agctgggacc 90300 agcctcaacc ttctaggccc aagagttcct cccacctcag cctcctgaat agctgggacc 90300 ataggtatat gccaccacac ccagctaatt tttagttttt ttatagagac agggtctccc 90360 ataggtatat gccaccacac ccagctaatt tttagttttt ttatagagac agggtctccc 90360 tgtgttgccc aggctggtct caaactcctg ggctccagca atccttccaa agtgcagggt 90420 tgtgttgccc aggctggtct caaactcctg ggctccagca atccttccaa agtgcagggt 90420 tatagacata agccactgca cgtgtcctgt aacaaagatt gttagtaaaa gttattgaaa 90480 tttaaatcta tgattcaagt cctgatacat aagacacatg gcacatggta aaaattgatt 90540 tgattacatc ctaggaggcc tagacttggg aaagaaggaa aaaaaaaagt atgaggctgg 90600 gcatggtagc ttgtgcctat aatctcagca ctttgggagg ctaaggcagg gggattgctt 90660 gaggccagga gttcaagacc agcctgggca acatagcaag accccatctc taccaaaaca 90720 aaacaaaaca aaacaaccac aaaaaaaacc cacacaaatt aaatttaaaa agaaaaagat 90780 gaaaagaaag atgagaagat aaggaaatat ctgtagacag tttaagtctc tatttttttc 90840 tacaaatgta cagttatgta ttgtaacata ctgcatcttt tgaccctgtc ttctctgttt 90900 caacagagct tatctttttc tatgtatttc tgctgctact gcttttccaa accacagcca 90960 ttgtcctgta atgcccttat gcatttggcc aagccctctt agcttcttta ttattgtggg 91020 gtcttagttg atatttgggg tgtatgacat tacaaaacac atgggatgta gatgtggatt 91080 gggagagccc ctgaattgta gcactctgct ttcctcatct gtaaaatggg tataatcaaa 91140 gtcttggaag ggtaattgta aaggggtgtg cagacagtgg ggttactgtt atttgcaagg 91200 atccgcccat agttttatct gcaaatgtca ctagcttttg atttactcac tctcgcttgc 91260 gtcttctttc gtgctgatgt tggctctgct ttatcccaca ggctttttat tattattatt 91320 acttttgaga caaggtcgct gtctgtcacc caggctggag tgcagtggtg caatcttggc 91380 tcactgcaac ctctgcctcc caggttcaag tgattctccc cactcagtag ctgggattac 91440 aggcacgtgc caccacaccc agctaatttt tgtatttttt tggtagagat ggggtttcac 91500 catgttggcc aggctgagtg atcctcccgc cttggcttcc caaagtgctg ggattacagg 91560 catgagcccc tgcacccagc ctttctttct ttcttttttt ttttttttga gacagggtct 91620 cactgtgttg tgcagtggct cgatcttggc tcactgcaac ctccgcctcc tgggttcaag 91680 cagatctcat gtctcagcct ctcgagcagc tgggagtaca ggcacccgcc accacaccca 91740 gctaattttt atattttttg gtagagatgg ggtttcacca tgttggccag gctggtctcg 91800 aactcctgac ctcaagtgat tcatttgtct cagtctccca aagtgctggg atttcaggcg 91860 tgagccaccc caccgggcct cccaacaggc ttttaaagca ccagggcagc agggcactgg 91920 gccaacaggt atatttggga atgtgcattt ggtttcatgt gtaataggaa agagattaat 91980 taaggttgtt agtgtggcct aatactaaat ttcaggaagc ctcacaggac acatgaaaca 92040 aatctttaca acacatatgt gttttcatgc tgctgaacat tttgaagacg ttttgccatg 92100 ctggagtttg tttgttttgt ttgtgttttt gtttgaggtg ccgtcttatt ctgtctctca 92160 ggctggagtg cagagttcag tggcacaatc ttggctcact gcaacctcca cctcccaggt 92220 tcaagctatt cttctgcctc agcctcccga gtagtgcctg gctgctgttt ttttttttta 92280 attgttaaat tagatgggct cagtattacc gtgccttgat ggagtttcta gttgagggat 92340 cagtgtcatt tctgggtccc catcagaatt acgttcaagc tataagtagt tatgcaaagt 92400 cgtaggttaa atgatttggg aataaaatgt catttgacgt gacagaaatg aacccttgga 92460 ggcccacatt tccactggga cttgctctca gttccagttc tcatgttaag taattttatt 92520 aactgaggca aggaggggca tgcggtgaca agggaaatat tttttgctaa cattttaatc 92580 ctaatcacaa aatttattgt gtgtggcttt atatgcattt gtagctataa agccctggaa 92640 gatgatgaaa gtaaaaatta gaatgtgtgg tttttctgcc caggtgttat ttaaagcatt 92700 gtatgaattg tcaaaaagat taattgttgt tgcttgcagc taaacagatg ataaatttta 92760 aaattatatt attactgtgt gtcagcataa aagctctgtg tcccaagtgt gtcacctttt 92820 tctgttgctg ttgacaaggt tcacatttta tagcacaccg ccagaaggtg ttgacatata 92880 ttcagattac tgattaaatg tttttaaaac gtgccactca gaaactaaca ctgatgttct 92940 atgaaagagt catttttcgg agtaagattt tttttttccc ttcatcatta acatggagga 93000 aatgttaaac actgacattt taaaaaatga acctgggact ggcgcagtgg ctcatgcctg 93060 taatcccagc accttaggag gccgaggtga gcggatcacc tgaggtcggg agttcgagac 93120 cagcctgacc aacatggaga aaccccgtct ctattaaaaa tacaaaatta gctgggtgtg 93180 gtggtgggca cctgtaatcc cagctactcg agaggctgag gcaggagact ccctcgaacc 93240 cgggaggtgg aggttgtggt aagctgagat cgtgccattg cactccagcc tgggcaataa 93300 gagcgaaact ccggtctcta aataaataaa taaataaaaa cgtggctggg cgtggtggct 93360 cacgcctgta atctcagcac tttgggaggc tgaggcaggc agatcacgag gtcaggagtt 93420 cgagaccaac ctggccaata tggcgaaacc ccatctttac gaaaaatacc aaaaattagg 93480 tgggcatggt ggcacgcgcc tgtaatccca gctactcaga aggctgaggc aggaaaattg 93540 cttgaacccg ggaggcggag gttgcagtga gccgagatcg cgccaccgta ctccagcctg 93600 ggtgacagac caagactccg tctcaaaaaa aaaaataaaa ataggccagg cgcagtggct 93660 cacgcctgta atcccagcac tttgggaggc caaggtgggc agatcacaag gtcaggagtt 93720 cgagaccatc ctggctaaca aggtgaaacc ccgtctctac taaaaataca aaaaaattag 93780 ctgggcgtgg tggcgggtgc ctgtagtccc agctactcga ggctgaggca ggagaatggc 93840 gtgaaccacg gaggcggagc ttgcagtgag ccgagattgc accactgcac tccagcctgg 93900 gcgacagagc gagactccat ctaaaaaagt aaaataaaat aaaatgaaaa taataaataa 93960 ataaataaac ctaatgttac tgcactttcc catttcaaat tgcagccagc aatgaaaaca 94020 ggcattagca ttcccatttc ttctcttctc cttcactgga agggcaacaa caaacacacc 94080 cacggtgtaa aaacagcacc aaggccgggt gcagtggctc acacctgtaa tctcagcact 94140 ttgggaggcc gaggcatgcg gatcacctga ggtcgggagt tcgagactag cctgaccaac 94200 atggagaaac cccatctcta cttaaaatta ggcgagcatg gtggcgcgcg cctatgatcc 94260 cagctactca ggaggctgag gcaggagaat cacttgaacc tggaggcgga ggttgcggtg 94320 agcggagatc gcaccattgc actccagcct gggcaacaag agaaaaactc catctcaaaa 94380 caaaacaaaa caaaacaaca gcaccaagga aggcgctaaa tggctacttg taaacaaccc 94440 aggttggaca ctggacccac gacaaggagg caagttccgt cctgtggtgg tgtttgtgat 94500 gggaagtctg gggctttgac tccctcaagt ttccaggcta acacacacaa ctcttcccag 94560 cagctgaggg gccccgaagc cttgatgtcc ctttcacgtt gaaggacctc actggtcttg 94620 gctgtttgcg ttttgtggat ggagtgggcc agtcttcaga gaactcaaat gtgtatttgc 94680 tgcagggagt gataagtaaa cataatgcct caagaatcaa catttgaatg cttttctttt 94740 tggcgttaag gtataatggg acactggcaa tatatgctgt ctctctgaac acagcatgaa 94800 agaacttgaa aacaaatgat ttagtattca ttacggaaaa gttggatttt tttccctctc 94860 tgattgtcaa gatttcttcg tcgatgtcag ttttccagtg tgaatttttc ctctccactc 94920 agaggaagtt gacgttggtt cagtgatgtt acagaggatg ctgtaagaca gccaggagcc 94980 caggccggtt gtcacttcat cccagcatgg tgcaaagtta gagatgttgc agtgacagcc 95040 ttgggtgccc tcaggggcaa ccaagtctct aaataatctc tgtgaccaaa agtacatagg 95100 aggttgtaaa tatcccacaa caaaaataaa gatagctgcc atttcccaga acttaatgca 95160 tatcaggaaa tgttacctct ttatatatat tgtctcattt aaatccagta agatagcagc 95220 agctgtggga ggatgtggtt tttgtttttg tttttgtttt gagacagagt cttgctctgt 95280 cacccaggct ggagtgcagt ggcgggatct cggctcactg caagctccgc ctcccgggtt 95340 cacactgttc tcctgcctca gcctcccaaa gtagctggga ctacaggcgc ctgccaccac 95400 gcctggctaa tttttttgta tttttaatag agacggggtt tcacatgtta gccaggatgg 95460 tctcgatctc ctgacctcgt gatctgcctg cctcggcctc ccaaagtgct gggattacag 95520 gcgtgagcca ccgcgcccgg cctgaggatg tggttttatt cccatttaac aaaggcttga 95580 gtatcacttc ttatttattt atttatttta ttattattat ttttgagacg aagtctcact 95640 ctgtcaccca ctctggagag caggggtgtg atctcagctc actgcaacct ctgcctccca 95700 agttctagtg attctcctgc ctcagcttcc cgaatagctg ggagtacagg tgtgcaccac 95760 cttgcacggc taatttttgt attagtagag atggggattc accatgttgg ccaggctggt 95820 ctcaaactcc tgacctcagg tgatctgccc atcttggcct cccaaagtgc tgaggttaca 95880 tactttatta acaaaggagt gaagtgatac cccctctccc ctgtaagtag cagccaacat 95940 tcaaacccag gtgtgtcaga ttttagagcc acactgacat tcagcagcta ctgactgatt 96000 gtctactatg tgttaggcat tgttttcatc acacctttta ttttttgttt tgttttttag 96060 ggacagagtt ttgctttgtg gcccaggtgt agtgcagtgg catgatgttt gctcactgca 96120 gcctcgagct cctggcctca agtgatcctc ctgttttggc ctcccaaaat gctaaggtgc 96180 tgggatcaca ggtatgcgcc actccgctca gcccacatat ttttaagtct gctttttaac 96240 ctaatttact tattttgggg gtcttaaaaa attggctcac agccgggcgc ggtggctcat 96300 gcctgtaatc ccagcacttt gggaggccaa ggcgggtgga tcacgtgagg ccaggagttc 96360 aagaccagcc tgatcaacat ggtgaaaccc tgtctctact aaaaaataca aaaattagcc 96420 gggcgtggtg tcgcatgcct gtaatccgag ctacttagga ggctgaggca ggagagtcgc 96480 ttgaacctgg gaggcggagg ttgcagtgag ctgagatcac accatggcac tccagcctgg 96540 gcaacaagag tgaaattccg tctcaaaaaa aaaaaaaaaa aacctattta aaaatgaagg 96600 ctcatgcaga atgctctgca agttcaaaat aaaacagagc tctgagcgcc tagaggcttt 96660 ttgccttaag gggcgaggag gaaaaagatt aagatgctcc ttggaaaata gccctttttg 96720 ttgccttaag gggcgaggag gaaaaagatt aagatgctcc ttggaaaata gccctttttg 96720 gctccatctc ctttactgtg gtcacctgca catgccttta gttcttctgg atttatatct 96780 gctccatctc ctttactgtg gtcacctgca catgccttta gttcttctgg atttatatct 96780 aagcacgttg ctctgctaca taagaagctg acaacatcag ttaccttcct ggaggtgaag 96840 aagcacgttg ctctgctaca taagaagctg acaacatcag ttaccttcct ggaggtgaag 96840 gagagacaaa ggggctgagt gggtgagagg aatactttcc accatgaact ctttggtacc 96900 gagagacaaa ggggctgagt gggtgagagg aatactttcc accatgaact ctttggtacc 96900 tttcggaacc acgtgaatac agtatctttt caaaagaaaa gggtgctata ttcattgtcc 96960 tttcggaacc acgtgaatac agtatctttt caaaagaaaa gggtgctata ttcattgtcc 96960 tagactatta agcactttaa aaggcacttc aggtaatcta gttgaagtgc aaatttcact 97020 tagactatta agcactttaa aaggcacttc aggtaatcta gttgaagtgc aaatttcact 97020 gaacccagac tattatgcaa acaggactag ctctgtattt attcttttat gttgtggctt 97080 gaacccagac tattatgcaa acaggactag ctctgtattt attcttttat gttgtggctt 97080 gccatccaat ttgtgtttcc ggggctgggc acgaggtggc tctagcctgt aatccccgca 97140 gccatccaat ttgtgtttcc ggggctgggc acgaggtggc tctagcctgt aatccccgca 97140 gtttgggagg ccaaagcagg cgatcagttg gggccaccat ctccattgtc cctttttaaa 97200 gtttgggagg ccaaagcagg cgatcagttg gggccaccat ctccattgtc cctttttaaa 97200 tttaagtgac catagcttct tagtatgata tcaggttttt gttttcttcc caccttgtct 97260 tttaagtgac catagcttct tagtatgata tcaggttttt gttttcttcc caccttgtct 97260 ttgtttttta gtgtgtggat gcctttgaaa ttagtttttt tgtttgcttg gttttttttt 97320 ttgtttttta gtgtgtggat gcctttgaaa ttagtttttt tgtttgcttg gttttttttt 97320 gagatggagt ctcactctgt cgccaggctg gagtgcagtg gcgcgatctc ggctcactgc 97380 aacctctgcc tgccaggttc aagcgattct cctgcctcag cctcccaagt agctgggact 97440 ataggcacat gccaccatgc ccagctaatt ttttgtattt ttagtagaga cggtatttca 97500 ccgtgttagc cagagtagtc cgatctcctg acctcgtgat ccgcccacct tggcctccca 97560 tagcactggg attacacata tgagccactg tgcctggcca aaattagttc attttttata 97620 catgtccttt acacaggaga tgtattcaga aagattatct atattttttc ctttaaacaa 97680 tgcatttgtt tgtttgtttt atttgtaagc tcatctcctg ggtttagagc agaatgaaaa 97740 atttgacctt aatcttctct tatccactgg ttggaagctc ctacttttat gtcaacagac 97800 ataaaaaagc ttatacatat aagataatgt atagcatgca taaagattta tagaatgcta 97860 tgtctcataa atttcacagt tataaaacaa aacctaagct tatgcatgtg gagatattcc 97920 tgtgtttgca gttgtcagtg acttctagag aatgtggtaa aggaaaaaag cacctgtgga 97980 agttatttat gtaaaagttg cttatgtaaa gactcaagcc attaacaaga gaaggagact 98040 tctttatatt gttaatgaca ttcagatctt tcttctgaat gagggttgtc actctgaatg 98100 tctttatatt gttaatgaca ttcagatctt tcttctgaat gagggttgtc actctgaatg 98100 agtgttgtca ttcatttcct ccaagccttc agatatcttt ttcctttatc tgacacattt 98160 agtgttgtca ttcatttcct ccaagccttc agatatcttt ttcctttatc tgacacattt 98160 caggaaagag taagggaaag atttatttat attactttga tggtttcatt ttctttgtgg 98220 caggaaagag taagggaaag atttatttat attactttga tggtttcatt ttctttgtgg 98220 aagtccagtg acccagatat agagatcttc ttggaatata ttactttgat ggttactttc 98280 aagtccagtg acccagatat agagatcttc ttggaatata ttactttgat ggttactttc 98280 tctttttgga agtccagtga cccagttata cggatgtcct cagaacattt gaagtctagt 98340 tctttttgga agtccagtga cccagttata cggatgtcct cagaacattt gaagtctagt 98340 tctgcattat aaaatagccc agttgccata ggttgatttc ctgtaaacct aaatcatgat 98400 tctgcattat aaaatagccc agttgccata ggttgatttc ctgtaaacct aaatcatgat 98400 tgcaaactaa tttggatgct aattaggact catataatga ttatatcaat ttataacttg 98460 tgcaaactaa tttggatgct aattaggact catataatga ttatatcaat ttataacttg 98460 tctaacaaca atgcatttct ttctttgcta ggttaagtat ttacactatg acctgtgaac 98520 tctaacaaca atgcatttct ttctttgcta ggttaagtat ttacactatg acctgtgaac 98520 agggaaaccg gcctgtgttg agtactttct atgagccagg tgctcgactg tgtacaaact 98580 agggaaaccg gcctgtgttg agtactttct atgagccagg tgctcgactg tgtacaaact 98580 tagaagaacc aggtgcacac gccaggtttg ggggccgttt gacagtcagc attcaataaa 98640 tagaagaacc aggtgcacac gccaggtttg ggggccgttt gacagtcagc attcaataaa 98640 tgtgttgttt aagttgatat aattaattta aaaggaagta gcaggtgatg gtagtacact 98700 tgtgttgttt aagttgatat aattaattta aaaggaagta gcaggtgatg gtagtacact 98700 tcattgtcct agaatattaa atactttgtt gtcagcagac ggacttcttt catatgcaca 98760 ggtcatcttg agcacggatc ctaagcctag tctttaatcc ttcctagcat cctacatgag 98820 ccaaacttag acaaaaaaca aggcaactct tggttatcta gggcaatttg aaatctcagc 98880 tacctccact gtgtccagct caccgcagtt atgcatctat ttacaacatt atgtttgcaa 98940 cattatgttt aagcaaacca caatgaaagc attaaacata gaataactat gatccagcaa 99000 ttttacttcc aagcatatac cgaaaagtgt tcaaagcagg gactcagata ctgtgttcct 99060 gggtacatgg gtacacccat gtttatagca gcattattct tgatagccaa aagatggaag 99120 caacccaggt gtccatcgac aggtgaatgg gtaactacaa tgtggtgtac acatgcacac 99180 acacacacac acgcaagaat attattcaac cttaagaaag aatgaaattc taccagtcac 99240 ggtggctcac atctgtaatc ccagcacttt gggaggctac ggtgggcgga tcacctgagg 99300 tcaggagttt aagaccatcc tgaccaacat ggtgaaaccc tgtctctact aaaagtatga 99360 aattagccag gtgtggtggc acatgcctgt catcccagct acttgggagg ctgaggcagg 99420 agaatcgctt ggacctggaa ggtggaggtt gcagtgagcc aagattgtgc ctttgtactc 99480 agaatcgctt ggacctggaa ggtggaggtt gcagtgagcc aagattgtgc ctttgtactc 99480 catcctggag agcaaaattc catctcaaaa aaaaaaagca tgaaattctg atacctgcta 99540 catcctggag agcaaaattc catctcaaaa aaaaaaagca tgaaattctg atacctgcta 99540 taacacagat gaaccttgaa gatatgctga gtgaaataag ccatatgcaa aaggacaaat 99600 taacacagat gaaccttgaa gatatgctga gtgaaataag ccatatgcaa aaggacaaat 99600 atgtgattgt acttatatga ggtacctaga gtagtcaaat tcacagcaca gaaatgcgaa 99660 atgtgattgt acttatatga ggtacctaga gtagtcaaat tcacagcaca gaaatgcgaa 99660 tggtggttac ccggggctgg ggggaaggag ggaatgggga attggtgtta gtgggcttac 99720 tggtggttac ccggggctgg ggggaaggag ggaatgggga attggtgtta gtgggcttac 99720 agtttcagta gggggtgatg gaaaagttct agagatacat agtggtaatg gatgcacaat 99780 agtttcagta gggggtgatg gaaaagttct agagatacat agtggtaatg gatgcacaat 99780 attgtgaaag gacaaaacta tacctaaaaa tggttcaaat gctaaattgt atgttatgta 99840 attgtgaaag gacaaaacta tacctaaaaa tggttcaaat gctaaattgt atgttatgta 99840 tattttgccc caattttttt ttttgttttt ttttgtttta aagacagagt tttcctcttg 99900 tattttgccc caattttttt ttttgttttt ttttgtttta aagacagagt tttcctcttg 99900 ctgcccaggc tggagtgcaa tggcatgatc tcggctcact gcaacctccg cctcctgggc 99960 ctgcccaggc tggagtgcaa tggcatgatc tcggctcact gcaacctccg cctcctgggc 99960 tcaagcaatt ctcctgcctc agccttccaa gtagctggga ttacaggctc gtgccgtcat 100020 tcaagcaatt ctcctgcctc agccttccaa gtagctggga ttacaggctc gtgccgtcat 100020 gccccgccaa tttttgtatt tttagtagag acagggtttc accatcttgg ccaggctggt 100080 gccccgccaa tttttgtatt tttagtagag acagggtttc accatcttgg ccaggctggt 100080 tttgaactcc taacctcgtg atccacccac tttggcctcc caaagtgctg gcattacagg 100140 cgtgagctac cgtgcctgac gtattttgcc ccaattttta aaagcaacat aataaagaca 100200 aagccactgg gaaggatata aaatgcccct gagccaaaca ctgtatagcc aaaagagcaa 100260 aactacaaaa actgcaccaa gcttctttgt aaattcattc ttccaccctc caggacactc 100320 ttgcctggtt tgcatcctga gagtctcctg gggttccagt tatttcaagg tcacagtcag 100380 gtaggcacgg tggctgtgct atggtggaaa tggcttttgg attcctcgta attttggatt 100440 atcggtgccc tgtttttctg agaccaaaac ttatcaatag agtattacgg aaggtgatat 100500 gtagtaagtg cacctttctt gggcattcag tctaaattca atagtaaagg gaaaatcaca 100560 tatagtcaag tccaaattgc tcatgatgtt acagtagatc tacatgtccc taacttgatt 100620 acctattttc cctccagcac tggagtgcat ctctgcttct tcagccgagc tccagaagga 100680 gaagatggct gcatttagag tcagactgga ggaaaaggag cattttgata ttttatttta 100740 tttttattta ttttttagag acaggttctt gtgttgtcat ccagactgga atgtagtggt 100800 tgcagctcac tgcagccttg acctcctggg ctgaagccat cctcaagcct cagcctcctg 100860 agtagccagg actacaggtg ccgtgccacc aagcccagcg aatttttgta tttttttgta 100920 gagacggatc ctgctgtgtt gtccaggctt gaaatttgat tttaaacata ttaggcatca 100980 cactaagccc ccaagacaag atagaagctt acagaacctt gactatctgg aatgactttc 101040 acagagtgat ttcatctttc ttcttaccct caatgagtaa cgtgaaggct gagcaggtca 101100 tactgttcaa attatttatc ttctttagac ttttttcttc caaacaccag tagttgaatt 101160 tgagtacttt aagtgctagt attatgcaat ttcattttat tttattttat tttatttttg 101220 agacagagtc ttactctgtt gcccagactg gagtgtagtg gtgcgatctc ggctcactgc 101280 aacctccgcc tcccgggttc aagcaattct cctgcctcag cctcccaggt agctgtgttt 101340 acagtcatgc gccaccacgc ccggctaatt tttttgtgtg tctttagtag agacgggatt 101400 tcaccgtgtt ggccaggctg atctcgaact cctgacctcg tgatctgccc ccctcggcct 101460 cccaaagtgc tgggattaca ggcttgagcc attgcaccca gcctggatta atgatttcga 101520 atgaacattt acaatgatat gttttataat aaaacacatt aaaaagtttg aaacactagt 101580 tactagaaac agttctctct gtaatgacag ttagtcatgt tgttgctgca gtgacagttg 101640 acttggtgtg agggtattga taaagcctgg tgatttgtac acttgcccat attatgcttt 101700 tcccaggcta gctatgcaaa tctcatttcc cttacatttg aaagtcattt cagaattgtt 101760 tcattagggt tgaggcaagc acaaatcatg aatgcctgtg tcttctacca ccaaacagat 101820 tctgttggac ccacaactga tatgctttca gccaggttcc atttagcatt caccctggac 101880 ttagggactc tacaagagtg ctgggtattt agttttgttt tgttttgctt tttgagacag 101940 agtcttgctc tgtcacccag gctggattga agtggcgcaa tctcagctca ctgcaacttc 102000 caccccctgg gttcattctc ctgcctcagc ctcccaagta gcagggatta caggtgccca 102060 ctaccacacc cggctaattt ttttgtattt ttggtaaaga ctgggtttca ccatgttggc 102120 caggctggtc ttgaactcct gacctcaggt gatccacccg ccttggcctc ccaaagtgct 102180 aggattacag gcgtgagccc tcacacccgg cctgatgctg ggtgttttgt acagtggtcc 102240 caccttactg tgtagaaatt ctcagtcatc cacctaattt gtttatttcc cattcatata 102300 cttcatttat tcactcactc acttggccat ttatgcatat ttattgattt tgttaaaaag 102360 atacctgaat gagtggaaca ctgagatgat atagtggaat ggacggatag tggttagccc 102420 cttggaccca tgagggccag gttctgaaat tccagctctg aaattctacc agctggttga 102480 ccttgggcaa attacttagc ttctctgtgc ctcatttctt catttataaa atggagataa 102540 taggttaaga gggttagatg agacattcat tcatagtact tggcacacag taagagccca 102600 gtaaatacta gctgttgtta tggattgaat tgtgttgcct ccccacaaac tcatatgttg 102660 aagtcctaac cacaggtact cagaatgtga ttgtatttgg aaacatggcc attgtagatg 102720 taattattta agaagagtga ttaggctggg catggtggct cacacctgta atcccaacaa 102780 ctttgggagc ctgaggtggg cggatgatga gggcaggagt tcgagatcag cctggccaat 102840 atggtgaaac cctgtctcta ctaaaaatac aaaaattagc cgggcgtggt ggcaggcacc 102900 tgtagtccca gctactcggg aggctgaggc aggagaactg cttgaaccag gggggcggag 102960 gttccagtga accgagataa cgccactgca ctccagcctg ggcggcagag cgagactctg 103020 tctaaaaaaa aagaaagaaa aaaaaggagt gattaggttg ggcactaatc caatatgatt 103080 gctgtcctta aaaaaagaga aaatttgggc acagagatag acatgcatag agggacaatg 103140 gccatccaca agccaaggag agggtcctgg aacaaatcct ttcttaccag tcctcagaag 103200 gagccagccc tgctgacacc tcagtttcag acttccagcc tccacagctg tgagacaata 103260 aatgtctgtc agtgaagcca ctcagtctgt ggttctctgt cacagcagcc ctggccgaag 103320 acacagctgg cagcaccgtc ctccgctact gtgtttctca ttgacccgag aacctgcctt 103380 tgtccatgtc ctggccaatg ggagatacat tttctccttc tcagcttgta ttcctgtcct 103440 tttctttctt tttttacaaa atagcataca aaaaagttcc tagcaagaag aggtgctcca 103500 gaagttccct ggctgtttca ttcctgtgtc tcttcctaaa aagctgcagt tgaaaagata 103560 gaatttgttt tctcttagca cattcttctc actttctaag cttgattttt ctttctttga 103620 aagagtgtaa ccccttcaaa attaactcct agagattgct cacctccctc ccaaactttc 103680 gatgaggagc gtttattttg tagactctta tgtgtgaatt ctttcaataa ataggggatg 103740 cagagaagca gctttgtgga gtgaagcagt aggaaagagc ctgttgggga gagagcttcg 103800 ccactgcatt tcagtagctg gtttcgcccc tacctttttt ttttttcttt ctttcttttt 103860 tttttttttt tttgagacgg agtctcgctc tgtcgcccag tctggagtgc agtggcgcta 103920 tctgggctca ctgcaagctc cgcctcccag gttcacgcca ttctcctgcc tcagcctctc 103980 cgagtagctg gggctacagg cgcccgccac caagcccggc taattttttg tattttcagt 104040 agagatgggg tttcaccgtg gtctcgatct gctgacctcg tgatctgccc gcctcggcct 104100 cccaaagtgc tgggattaca agcgtgagcc accgtgcccg gcccaccctt acctattttt 104160 aaatggtgac ttgtgttttt atttgtttgt ttttaaagac acttgaaatt ggacagtgtt 104220 tctccatcct ctccagaggc ccagagcaat gtcgcaagga tgcattctgg acgaggcagt 104280 cacggggccg cacatggcat ttgttcttct cagggaccgt ctgctgtgtt tctctgcccc 104340 agtggagctg ggggaataat aaagctctta ttatttatac tgcacagcat ttttttcagt 104400 ggtccttgtc attgagactt atgaagacaa tgtaggacaa gagggaactt gtgagttttt 104460 atgcaggtag ggaagaaaag taaaaggggc tatatttcca gggatattgc tttttcccaa 104520 acccctgcac ttccccagtc cttttcttta tcttcctttc ccaaatgaga ggtgtccttc 104580 ttctccggta tcagccccct agtttcttcc ctgtcctcgg ggaatcttgg tacaccattt 104640 agcccctgtc ttggtttttg actctgtccc tttgtggaat tcttacccat agcttacagt 104700 gatgtttcac atacatctct gccaaagatg aagtggaaaa taacctttgc tttgtaagca 104760 catctccttc aagctatcga caagtctcct ggcttctctt agtttctaac cccttttttt 104820 tttttttttt ttttttgaga cagagttttg ctgttgttgc ccaggatgga gtgcagtggc 104880 gtgatctcgg ctcactgcaa cctccgtctc ctaggttcaa gtgattatca tgcctcagcc 104940 tcctgagtag ctgggattac aggcgtctgc caccacaccc agctgatttt tgtactttta 105000 gtagagaggg gttttgccag gctagtcttg agctcctggc ctcaagtgat ctgactgcct 105060 tggcctccca aagtgccagg attacaggtg tgagccactg ctcacagtta tctatttgtc 105120 ttagtccatt taggctgcta taacaaaata ccataaactg ggtgtcttat aagcaacaga 105180 aatttatttc ccacacttct ggagacttgg aggtccaaga tcaggcagtg gcaggttcag 105240 tgtctggtgc taaggaagca cctgcgcctc ccggtccaca gacgcacacc ttctcactgc 105300 gtcctcatat gacagaaggg gtgagggagt ccgtcttttt tttttttttt ttttttttga 105360 gacggagttt cgctctcgtt gcccaggcta gagtgcaatg gcgcaatctc ggcacgcaac 105420 ctccacttcc cgggttcaag cgattctcct gcctcagcct cccaagtagc tgggattaca 105480 ggcatgcacc accacgcccg gctaattttg tatttttagt agagatggcg tttctccatg 105540 ttggtcaggc tggtcttgaa ctcccgacct caggtgatct gcccacctca gcctcccaaa 105600 gtgctgggat tacaggcgtg agccaccgca cgcagctttt tttatttttt attttttaat 105660 atttttgaga cagagtcttg ctgtgtcacc aggctggagt ccagtggtgc catcttggct 105720 cacggcaacc ttcacctcct gggttcaagc gattctcctg cctcagcctc cagagtagct 105780 gggattacag gcgcccacca tcacgcccgg ctaatttttt gtatttttag tagagacagg 105840 atttcaccat cttggccagg ctggtctaga actcctgacc tcgtgatcca cccgccttgg 105900 cctcccaaag tgctgggatt acaggcgtga gggagtctgt cttttataaa ggtgctgatt 105960 tcatttatga gagctcagtc ctcgtgacta atcaccaacc aaaggcctca cctccacatt 106020 tgatcacact gggaatgaag acttcaacat actttgcagg ggacagaaac attctgtcta 106080 tgatcacact gggaatgaag acttcaacat actttgcagg ggacagaaac attctgtcta 106080 tagtaccatt aactttctgc tatggaagtt gaagacagtt ctcttttgtc gtattatacc 106140 tagtaccatt aactttctgc tatggaagtt gaagacagtt ctcttttgtc gtattatacc 106140 cctgatacac acttcagtgt ttatactgtg ccaaggaaag cagtgttcac agctgagcca 106200 cctgatacac acttcagtgt ttatactgtg ccaaggaaag cagtgttcac agctgagcca 106200 tacggtacac tacaattttc ctttctcata taactttttg tccccttgga gttagcaatt 106260 tacggtacac tacaattttc ctttctcata taactttttg tccccttgga gttagcaatt 106260 gacttttttt tttttttttt ttttgagaca gggtctcgct ctatagcgca ggctagaatg 106320 gacttttttt tttttttttt ttttgagaca gggtctcgct ctatagcgca ggctagaatg 106320 cagtggcgtg atctcggttc actgtgacct ctgcctcctg ggctcaagcg atcctcccgc 106380 cagtggcgtg atctcggttc actgtgacct ctgcctcctg ggctcaagcg atcctcccgc 106380 ctcagcctcc caagtagctg ggaatacagg tgcgtgccac catacccagc taatttttgt 106440 ctcagcctcc caagtagctg ggaatacagg tgcgtgccac catacccagc taatttttgt 106440 ttttttgagt tttttttttg gtagagacag agttttgcca cattgcccag gttggtctcc 106500 ttttttgagt tttttttttg gtagagacag agttttgcca cattgcccag gttggtctcc 106500 aagtctggag ctcaagaatc tccctgcctt ggcctccaaa agtgctggga ttacaagtgt 106560 aagtctggag ctcaagaatc tccctgcctt ggcctccaaa agtgctggga ttacaagtgt 106560 gagctaccac acctggccag tgattgactc ttttgctaaa actaattttt tcttccaact 106620 gagctaccac acctggccag tgattgactc ttttgctaaa actaattttt tcttccaact 106620 ctttgacaaa gtcgtcaata ttaagcaata tacccttact acttttgatt ttgtttttaa 106680 ctttgacaaa gtcgtcaata ttaagcaata tacccttact acttttgatt ttgtttttaa 106680 aggaattcct tctggagtct tctatcctga acttgtcgct ttctaggcca gctgcatttc 106740 tgtgtttcta aaacttccat tgtttataaa acaccataat cctggcgttt gctctgtttg 106800 tactctgtat ttccagtatc tggaaaccat gtcttctttc ttggcttatt tcctcattca 106860 gggctgtgtg tcttctagta gttatgtgaa aagagggttc gtgaaagtta aatgtttttg 106920 aggctatatg tctgaagagg tctttggtgt tcttcagact tcatggaaaa cattttccct 106980 aagaatgtta aaggcattgc tccctttttt gcttctaatg ctgccattgc tttaaaatgg 107040 tgccattttg attcttgatc ctttttctgt aaattttttt ctctcttgaa aatctctttc 107100 ttgaagatct ctcttgaaga tctctttctc cctagtcctc tcaaactttg cattgatgta 107160 ctttgatggg atcatttttg ttaattatgt aggtgttcca taggcctttc agtctgaaaa 107220 tttatgtgtt ttagttctag gagctttctt tgtatttctc tatttctttt tttttttttt 107280 ttttttttta agacagagtc ttggtctgtc gcccaggctg gagtgcagtg gtgtgatctt 107340 ggctcactgc aacctccgcc tcccaggtcc aggtgattct cctgcctcag cctcctgagt 107400 ggctcactgc aacctccgcc tcccaggtcc aggtgattct cctgcctcag cctcctgagt 107400 agctgggatt acaggcatct gccaccatgc ctggctaatt tttgtatttc tagtagagac 107460 agctgggatt acaggcatct gccaccatgc ctggctaatt tttgtatttc tagtagagac 107460 ggggtttcac catgttagct gggctgatct tgaactcctg gcctcaggtg atccgccctc 107520 ggggtttcac catgttagct gggctgatct tgaactcctg gcctcaggtg atccgccctc 107520 cttggccttc caaagtgctg ggattacagg catgagccac catgtctggc ctcaaatttt 107580 cttggccttc caaagtgctg ggattacagg catgagccac catgtctggc ctcaaatttt 107580 tatttttgat agctttttca tgttctctaa tgtttctgat tgattgattg attgagaaag 107640 tatttttgat agctttttca tgttctctaa tgtttctgat tgattgattg attgagaaag 107640 ggtcttgctc tgttgcccag gctggagtgc agtggtgcaa tcatggctca ttgcagcctc 107700 ggtcttgctc tgttgcccag gctggagtgc agtggtgcaa tcatggctca ttgcagcctc 107700 aacctcctgg gctcaaatga tcctcctgcc tcagcctccc aagtagctgg gactacaggc 107760 aacctcctgg gctcaaatga tcctcctgcc tcagcctccc aagtagctgg gactacaggc 107760 acaagacagt atacctggct aatttttaaa tatatttttt tgtagagaca gggtttcgct 107820 acaagacagt atacctggct aatttttaaa tatatttttt tgtagagaca gggtttcgct 107820 atgtgcccag gttggtctca aactcctggg ctcaagcaat ttgctttcag cttcctaaaa 107880 atgtgcccag gttggtctca aactcctggg ctcaagcaat ttgctttcag cttcctaaaa 107880 tgctaggatt acaggtgtga gtacatggca cctggccgtg tttgtttgtt tgtttttgag 107940 tgctaggatt acaggtgtga gtacatggca cctggccgtg tttgtttgtt tgtttttgag 107940 acagggtctt gctctgtcac ccaggctgga gtgcagtggt acaatctcgg ctcattgcag 108000 acagggtctt gctctgtcac ccaggctgga gtgcagtggt acaatctcgg ctcattgcag 108000 ccttgacctc ctgggctcaa gcaatcctct acctcagcct accatgtttc ttttataaat 108060 gatattttat tcctgttttt gcatgtatta taatatctta tttctctgtt tgttctgttt 108120 tttgctctgt tctgctcttt gcattgtccg tttctcttga gtgctcttcc tggttgtttg 108180 tttcggtttt tatctttcaa gttgataact tttgtcgaag gtctggtggt gtttgactgt 108240 cagtccatat ttaaaaatga ggaactggcc tggtgtggtg gctcacgcct gtaatcccag 108300 cactttggga ggccgaggtg ggcggatcac ctgaggtcag ctgtttgaga ccagcctggc 108360 caacatggtg aaaccccatc tctactaaaa atacaaaaat tagcttggtg tggtggcaca 108420 cgcctgtaat cccagctact caggaggctg aggcaggaga actgcttgaa ccggggaggt 108480 agaggttgca gtgagtggag actgcgccac tgcacttcct cctgggtgac agagtgagac 108540 tccgtctcaa aaataaataa ataaataaat aataaaaata aaaatgagga acttaaggac 108600 caggagcagt ggctcctgcc tctaatccca gcactttggg aggccgaggt gggcggatca 108660 cttgaggtca ggagttcgag accagcctgg ccaaaatggt gaaaccgcgt ctctactaaa 108720 aacacaaaaa attagccagg catggtggcg ggtgcctgta atctcagcta cttgggaggc 108780 tgaggcacaa gaatcacttg agcccaggag gcagaggttg cagtgagccg agatcatacc 108840 actatactct agcctgggca acagagcgag actctgtctc taaataaata aataaataaa 108900 taaaaaggag gaacgtaacc atcaatagag agttatttgt gtatggaact gatgggcctc 108960 actgtaaggt tatcagctgg gggcccacca gcttcaccag aggatcacca catatcagtg 109020 tctgtaggct tgttctcctg ggctcttctg tttttcagag aggtatcctg tgctctccag 109080 cttcggggga ttagacctgg cttccagcat tctagaagcc aaggctagga ggcaactggg 109140 gaattctcat tattccatac ctggactttc actcagcccg tttcagcttt tgactaactc 109200 ctccccttca ctgtgttttg cctccacaaa gtgaaagctt tttggggttc catttctcca 109260 gggatttact tcctttgccc ttttggaatg ggacaggtta aggtctaagt gctctttata 109320 gagattttct ttttttcttt ttcttttctt tttttttttt ttgagacagt ctcactcgga 109380 ctggagtgca gtggtgtgat ctcaactcac tgcagcctgg acctctaggg ctcaggtgat 10944...

Claims

1. A method for generating microglia from stem cells, comprising the steps of: (a) targeted insertion of a nucleotide sequence encoding a transcriptional regulator protein into a first safe harbor site in a genome; and (b) targeted insertion of the coding sequence for transcription factor PU.1 (SEQ ID NO: 1) into a second safe harbor site in the genome, wherein the gene is operably linked to an inducible promoter that is regulated by the transcription factor protein; expression of PU.1 (SEQ ID NO: 2); and (c) culturing the stem cells obtained from steps (a) and (b) with exposure to at least one growth factor or small molecule, which recapitulates signaling during at least one of the stages of embryonic development of microglia, or the stages of proliferation, differentiation, or polarization of adult microglia.

2. 2. The method of claim 1, wherein at least one of the growth factors or small molecules is selected from the group consisting of activin A (SEQ ID NO: 7), BMP4 (SEQ ID NO: 8), FGF (SEQ ID NO: 9), VEGF-A (SEQ ID NO: 10), LY294002, CHIR99021, SCF (SEQ ID NO: 11), IL-3 (SEQ ID NO: 12), IL-6 (SEQ ID NO: 13), CSF1 (SEQ ID NO: 14), IL-34 (SEQ ID NO: 15), CSF2 (SEQ ID NO: 16), CD200 (SEQ ID NO: 17), CX3CL1 (SEQ ID NO: 18), TGFβ1 (SEQ ID NO: 19), and IDE1.

3. The method of claim 1 or 2, wherein at least one of the growth factors is CSF1 (SEQ ID NO: 14) or IL-34 (SEQ ID NO: 15).

4. The method of any one of the preceding claims, wherein at least one of the small molecules is CHIR99021, LY294002, or IDE1.

5. 2. The method of claim 1, wherein the first and second safe harbor sites on the genome are different.

6. 10. The method according to any one of the preceding claims, further comprising the insertion of the coding sequence of the gene for the transcription factor CEBPB (SEQ ID NO: 3) and its expression.

7. 2. The method according to claim 1, further comprising the insertion of the coding sequence of the gene for the transcription factor RUNX1 (SEQ ID NO: 4) and its expression.

8. 10. The method according to any one of the preceding claims, further comprising the insertion of the coding sequence of the gene for the transcription factor IRF8 (SEQ ID NO: 5) and its expression.

9. 10. The method according to any one of the preceding claims, further comprising the insertion of the coding sequence of the gene for the transcription factor SALL1 (SEQ ID NO: 6) and its expression.

10. 2. The method of any one of the preceding claims, wherein the transcriptional regulator protein is reverse tetracycline transactivator (rtTA) (SEQ ID NO: 20) and its activity is regulated by doxycycline or tetracycline.

11. 2. The method of claim 1, wherein the inducible promoter comprises a Tet responsive element (TRE) (SEQ ID NO: 21).

12. 20. The method of claim 1, wherein the first and second safe harbor sites on the genome are selected from the group consisting of the hROSA26 locus (SEQ ID NO: 22), the AAVS1 locus (SEQ ID NO: 23), the CLYBL gene (SEQ ID NO: 24), the CCR5 gene (SEQ ID NO. 25), the HPRT gene (SEQ ID NO. 26), or a gene having site ID 325 (SEQ ID NO: 27) on chromosome 8, a gene having site ID 227 (SEQ ID NO: 28) on chromosome 1, a gene having site ID 229 (SEQ ID NO: 29) on chromosome 2, a gene having site ID 255 (SEQ ID NO: 30) on chromosome 5, a gene having site ID 259 (SEQ ID NO: 31) on chromosome 14, a gene having site ID 263 (SEQ ID NO: 32) on chromosome X, a gene having site ID 264 (SEQ ID NO: 33) on chromosome 2, a gene having site ID 265 (SEQ ID NO: 34) on chromosome 3, a gene having site ID 266 (SEQ ID NO: 35) on chromosome 4, a gene having site ID 267 (SEQ ID NO: 36) on chromosome 5, a gene having site ID 269 (SEQ ID NO: 37) on chromosome 6, a gene having site ID 268 (SEQ ID NO: 38) on chromosome 7, a gene having site ID 269 (SEQ ID NO: 39) on chromosome 8, a gene having site ID 270 (SEQ ID NO: 39) on chromosome 9, a gene having site ID 271 (SEQ ID NO: 39) on chromosome 10, a gene having site ID 272 gene with site ID 231 (SEQ ID NO: 34) on chromosome 4, gene with site ID 315 (SEQ ID NO: 35) on chromosome 5, gene with site ID 307 (SEQ ID NO: 36) on chromosome 16, gene with site ID 285 (SEQ ID NO: 37) on chromosome 6, gene with site ID 233 (SEQ ID NO: 38) on chromosome 6, gene with site ID 311 (SEQ ID NO: 39) on chromosome 134, gene with site ID 301 (SEQ ID NO: 40) on chromosome 7, gene with site ID 293 (SEQ ID NO: 41) on chromosome 8, gene with site ID 319 (SEQ ID NO: 42) on chromosome 11, gene with site ID 329 (SEQ ID NO: 43), and a gene with site ID 313 (SEQ ID NO: 44) on the X chromosome.

13. 2. The method of any one of the preceding claims, wherein the stem cells are pluripotent stem cells, induced pluripotent stem cells (iPSCs), neural progenitor cells, hematopoietic stem cells, or embryonic stem cells (ESCs).

14. 2. The method of any one of the preceding claims, wherein the stem cells are human or mouse stem cells.

15. Microglia obtainable by the method according to any one of claims 1 to 14, preferably expressing at least one microglial surface protein selected from the group consisting of: ITGAM (CD11B) (SEQ ID NO: 45), ITGAX (CD11C) (SEQ ID NO: 46), CD14 (SEQ ID NO: 47), CD16 (SEQ ID NO: 48), ENTPD1 (CD39) (SEQ ID NO: 49), PTPRC (CD45) (SEQ ID NO: 50), CD68 (SEQ ID NO: 51), CSF1R (CD115) (SEQ ID NO: 52), CD163 (SEQ ID NO: 53), CX3CR1 (SEQ ID NO: 54), TREM2 (SEQ ID NO: 55), P2RY12 (SEQ ID NO: 56), TMEM119 (SEQ ID NO: 57), and HLA-DR (SEQ ID NO: 58).

16. 16. The microglia of claim 15 for use in therapy.

17. 17. Use of microglia according to claim 15 or 16 for the in vitro diagnosis of a disease.

18. 18. Use of microglia according to claim 17, wherein the disease is selected from the group consisting of: diseases of the central nervous system, preferably neurodegenerative diseases; more preferably Alzheimer's disease, Parkinson's disease, frontotemporal dementia or amyotrophic lateral sclerosis; neuroinflammatory or autoimmune diseases, preferably multiple sclerosis, autoantibody-mediated encephalitis or infectious diseases, neurovascular diseases; preferably stroke, vasculitis; traumatic brain injury and cancer.

19. 17. Use of microglia according to claim 15 or 16 for in vitro culture with cerebral organoids.

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