Abstract:
Methods are disclosed in which the expression of a specific gene, or combinations of genes, is controlled spatially and temporally to develop intra- and interspecies chimeras. A transgenic EC/ES/P/iPS cell line is created which conditionally expresses a suicide or compromiser gene configured to compromise all cell lineages except that corresponding to a target tissue/organ. The EC/ES/P/iPS cell line is injected into donor embryos having a specific target gene deficiency or embryos genetically engineered to be complementary compromised in lineages corresponding to the target tissue/organ cell lineages of the EC/ES/P/iPS line. One or more stimuli is provided to the embryo to activate compromiser genes for ablation of non-target tissues/organs of the EC/ES/P/iPS line and target tissues/organs of the host embryo, resulting in a chimeric animal having target tissues/organs derived from the genotype of the transgenic cell line and all remaining tissues/organs derived from the donor embryo.

Description:
REFERENCE TO RELATED APPLICATION 
       [0001]    This application claims priority to co-pending PCT application PCT/U.S.08/056,204, filed on Mar. 7, 2009, the disclosure of which is incorporated herein by reference, which claims priority to U.S. provisional application No. 60/690,169, filed on Mar. 9, 2007, entitled “A Novel Method for Conditional and Inducible Transgene Expression to Specifically and Precisely Direct the Development of Embryonic Cells, Embryonic Stem Cells and Precursor Cells”, the disclosure of which is incorporated herein by reference. 
     
    
     BACKGROUND 
       [0002]    The present disclosure relates to methods to direct the development of embryonic cells, embryonic stem, precursor and induced pluripotent stem (EC/ES/P/iPS) cells to any cell type, tissue or organ system in vitro or in vivo in an exclusive manner, particularly for the creation of chimeras. 
         [0003]    The human and mouse genome sequences together created an unprecedented opportunity to develop new, genetically engineered animal models to expedite the development of new treatment modalities to address and relieve human pain and suffering due to diseases. The differentiation program of EC/ES/P/iPS cells is one of the central questions in biology. Furthermore, isolation of tissue-specific stem cells presents a potentially powerful opportunity to develop effective therapeutics to facilitate repair of damaged or diseased organs. The best hope for more rapid discovery of effective prevention and treatment of cancer, cardiovascular disease, diabetes and other catastrophic human diseases, is via enhanced animal models of human health and disease. 
         [0004]    Transplantation of organs is a well-known and accepted life-saving procedure for many of these human diseases, such as end-stage kidney, liver, heart and lung diseases. From both a medical and an economic point of view, organ transplantation is often preferable to alternative forms of therapy. But, the insufficient number of donor organs limits the application of this technique and can lead to unnecessary loss of life when other procedures prove ineffectual. Experimental techniques, such as xenotransplantation, have become increasingly more important to develop new methods of creating organ availability. 
         [0005]    In past years several kinds of EC/ES/P/iPS cells have been isolated and their differentiation potential has been tested both in vivo and in vitro. However, none of these early studies addressed the “true” physiological fate of such stem cells and progenitor cells as a part of normal development. Several years ago, a novel cell-mapping system was developed which is based on expressing Cre or Flp recombinase in a stem cell or progenitor cell population. See, Dymecki and Tomasiewicz, Dev. Biol. 201:57-65 (1998); Gu et al., Development 129:2447-2457 (2002); and Zinyk et al., Curr. Biol. 8:665-668 (1998). Cre-mediated excision of the “floxed” sequences (i.e., loxP-flanked termination sequences) or Flp-mediated excision of the FRT-flanked sequences in the reporter constructs was shown to result in the permanent expression of the reporter in all the descendant cells. Since Cre or Flp can be introduced into these cells transgenically by using stem cell (or progenitor cell) specific promoter and/or enhancer elements in mice, this strategy permits analysis of the fate of these precursor cells throughout the cells&#39; life in complex organ systems in vivo. A good example of the power of this new recombination-based fate-mapping system is the fate determination of Flk1 +  cells in mice and proof that Flk1 +  cells also exhibit a differentiation potential for the other mesodermal lineages than endothelial cells. See, Motoike et al., Genesis 28:75-81 (2003). 
         [0006]    Matsumura et al. (2004) reported a new transgenic mouse model with a lineage-specific cell disruption system to express DT which was silent and harmless without the co-expression of Cre recombinase. This mouse provided a model for a variety of studies addressing the consequences of specific cell-type ablations produced by activation of DT expression when it was bred with lineage/cell-specific Cre-expressing mice. See, e.g., Brockschnieder et al., Genesis 44:322-327 (2006) and Kisanuki et al., Developmental Biology 230,230-242 (2001). However, these conditional gene targeting systems have a number of limitations, as they are either spatially controllable or temporally controllable—but not both. 
         [0007]    A mutant ligand binding domain of the human estrogen receptor has also been fused to the Cre recombinase by Metzger and Chambon (2001). In transgenic mouse lines produced with this modification, the nuclear localization of the Cre recombinase leads to action that is tamoxifen dependent. These mice have been used to generate cell/organ specific spatio-temporally controlled somatic mutations. The system has been also used in enriching for desired cell types in stem cell differentiation studies. 
         [0008]    Two predominant methods have been developed for introducing ES cells into pre-implantation-stage embryos: the so-called injection chimeras and aggregation chimeras. The injection of embryonic cells directly into the cavity of blastocysts is one of the fundamental methods for generating chimeras. ES cells can also be injected into blastocysts, which is probably the most common method for introducing genetic alterations performed in ES cells into mouse by producing germ-line-transmitting chimeras (Bradley et al., Nature 309:255-256 (1984)). Chimeras can also be created by aggregation of embryonic cells with morula-stage embryos. Although ES cells are typically established from the blastocyst stage, they are still capable of integrating one day earlier into the eight-cell-stage embryos. By taking advantage of this property, a relatively simple way of introducing ES cells back into embryonic environment has been developed (Nagy and Rossant, Gene Targeting: A Practical Approach, pp. 177-206 Oxford University Press (1999). Thus, ES cells can also be aggregated with morula-stage embryos to generate chimeras. 
       SUMMARY 
       [0009]    According to the present method, a novel combination of known genetic tools are used to provide genetically engineered cell, embryo or animal models in which embryonic cells, embryonic stem, precursor and induced pluripotent stem (EC/ES/P/iPS) cells can be precisely directed into desired cell types in intra- or interspecies chimeric composition with differently altered cells in vitro or in vivo. Using this method the expression of a specific gene, or combinations of genes, can be controlled spatially and temporally to develop intra- and interspecies chimeras. 
         [0010]    In a preferred embodiment, the method comprises three steps. The first step is to make a transgenic EC/ES/P/iPS cell line which conditionally expresses a suicide or cell progression/existence compromiser gene. Suitable suicide/compromiser genes include Diphtheria Toxin A (DT A), Herpes Simplex Virus-Thymidine Kinase (HSV-TK) or hypoxanthine phosphoribosyltransferase (hprt), although other such genes are contemplated. In the context of the present method, the suicide/compromiser gene is operable to kill target cells or place the target cells at a disadvantage once it is expressed. The time and the type of target cells, i.e., when and where the compromiser gene expression occurs, are controlled by using genetic tools. In certain embodiments, suitable genetic tools include the Cre/loxP, Flp-FRT, and the Tet-inducible recombination systems. In this step, the location of the compromiser gene expression is determined by the gene lineage corresponding to target tissue or organ cells to be derived from the transgenic cell line. Specifically, the compromiser gene is configured to compromise all lineages except that corresponding to the target tissue/organ. 
         [0011]    The second step is to aggregate/inject these EC/ES/P/iPS cells into donor embryos. The embryos may have specific gene deficiencies (i.e., knock-out embryos) corresponding to the target lineage. Alternatively, these embryos may be genetically engineered to be complementary compromised in lineages where the EC/ES/P/iPS cells component would be expected to colonize—i.e., the lineage corresponding to the target tissue/organ. The embryo will be a host for the introduced EC/ES/P/iPS cells, establishing the part of the organism where its cells are not compromised. The EC/ES/P/iPS cell contribution may not or may be withdrawn by specific compromiser expression. The complementing part in the organism will be derived exclusively from the introduced EC/ES/P/iPS cells. 
         [0012]    The last step of the present embodiment is to apply one or more stimuli to activate the compromiser gene(s) for ablation of undesired tissues/organs of the EC/ES/P/iPS cells and of the host embryo. The stimuli may include exposure of the embryos to a recombination control, such as a particular drug. In a specific example, a suitable drug is a tetracycline. 
         [0013]    The present method provides a genetic engineering system for whole organism- or cell-based approaches which can specifically and precisely direct the development of EC/ES/P/iPS cells to desired cell types, tissues or organ systems in vitro or in vivo in an exclusive manner. Using this method, the expression of a specific gene, or combinations of genes, can be controlled spatially and temporally to develop intra- and interspecies in vivo or in vitro chimeric conditions. In these chimeras, a specific cell type, tissue and/or organ system will come exclusively from one component (genotype) and the other cells, tissues and organs are originated from the other component (genotype). For example, this method allows the establishment of a human vasculature (blood vessels) and hematopoietic (blood) system in non-human species such as the mouse or the pig. The method will also enable new approaches to increase the precision of gene therapy methods by differentiating EC/ES/P/iPS cells to specific cell lineages. 
         [0014]    According to an alternative embodiment, the method may use genetically modified early cleavage stage embryos or morula embryos (embryonic cells) instead of genetically modified EC/ES/P/iPS cells, in combination with counterpart early cleavage stage or morula embryos instead of blastocysts. These complementary genetically modified cells can then be physically aggregated to produce a viable embryo chimera which can then be transferred to a recipient animal host for gestation and production of live offspring (Nagy et al., Manipulating the Mouse Embryo: A Laboratory Manual, 3d Ed. (2003). A further variation of this method can be to make EC/ES/P/iPS embryonic cell aggregates. 
     
    
     
       DESCRIPTION OF THE FIGURES 
         [0015]      FIG. 1  is diagram showing the steps of one embodiment of the methods disclosed herein. 
           [0016]      FIG. 2  depicts the construction of the LoxP-tet-O-DT-A-pA-loxP [SEQUENCE NO. 1] plasmid used in one embodiment of the method. 
           [0017]      FIG. 3  depicts the construction of the HSC-SCL-Cre-ER T -pA plasmid [SEQUENCE NO. 2] used in one embodiment of the method. 
           [0018]      FIG. 4  depicts the construction of the Endothelial-SCL-Cre-ER T -pA plasmid [SEQUENCE NO. 3] used in one embodiment of the method. 
       
    
    
     DETAILED DESCRIPTION OF THE EMBODIMENTS 
       [0019]    Specific language is used to describe several embodiments of this invention to promote an understanding of the invention and its principles. It must be understand that no specific limitation of the scope of this invention is intended by using this specific language. Any alteration and further modification of the described methods or devices, and any application of the principle of this invention are also intended that normally occur to one skilled in this art. 
         [0020]    The methods disclosed herein provide genetically engineered animal models that will be extremely helpful to provide new treatment modalities to address human diseases. These animal models may provide a foundation for producing transplantable human organs or tissues, or make such organs and tissues available for drug testing, for instance. In this model, the development of embryonic, embryonic stem, precursor and induced pluripotent stem (EC/ES/P/iPS) cells in an in vitro and in vivo chimeric organism can be precisely directed to any cell type, tissue or organ system in an exclusive manner. In one example, this method allows the establishment of a human vascular endothelium (blood vessels) and hematopoietic (blood) system in non-human species such as the mouse or the pig. 
         [0021]    The present method first makes use of cell depletion due to compromiser genes. Examples of suitable compromiser genes include: diphtheria toxin A (DT A), as demonstrated by Ivanova et al., in the article “In vivo genetic ablation by Cre-mediated expression of diphtheria toxin fragment A”, Genesis 43:129-135 (2005), the disclosure of which is incorporated herein by reference; or Herpes Simplex Virus-Thymidine Kinase (HSV-TK). The present method further makes use of certain genetic tools such as: Cre/LoxP as disclosed by Sauer et al., in U.S. Pat. No. 4,959,317, the disclosure of which is incorporated herein by reference; or Flp/FRT, as described by Wahl et al., in U.S. Pat. No. 5,654,182, the disclosure of which is also incorporated herein by reference. These tools further include recombination systems, such as the recombination system demonstrated by Nagy in the article “Cre recombinase: the universal reagent for genome tailoring”, Genesis 26:99-109 (2000), the disclosure of which is incorporated herein by reference. 
         [0022]    In a final step of the method, inducible gene expression system are implemented, such as the tetracycline inducible system described by Bujard et al., in U.S. Pat. No. 5,814,618, the disclosure of which is incorporated herein by reference; or by Belteki et al., in the article “Conditional and inducible transgene expression in mice through the combinatorial use of Cre-mediated recombination and tetracycline induction”, Nucleic Acids Research 33, No. 5 (2005), the disclosure of which is also incorporated herein by reference. Using a combination of these tools, the present method contemplates precisely spatially and temporally controlling the expression of cell-specific genes (compromiser) during the development or differentiation processes. 
         [0023]    By way of example the method disclosed herein allows the establishment of a human vasculature (blood vessels) and hematopoietic (blood) system in a non-human species such as the mouse or the pig. First, a novel mouse embryonic stem cell (ESC) line will be created which combines all the required genetic tools and inducible systems. In this ESC line, tetracycline inducible compromiser genes are flanked by recombinase attachment sites, such as loxP sites, so that recombinase will delete the compromiser in the lineage of its specificity of expression. A novel transgenic mice line will be produced which is specific gene deficient or in which the inducible compromiser has exactly complementing specificity of expression. This can be achieved by making the reverse tetracycline transactivator recombinase excision conditional, as described by Gossen et al., in the article “Transcriptional activation by tetracyclines in mammalian cells”, Science 23 Jun. 1995 268:1766-1769 (1995), the disclosure of which is incorporated herein by reference. 
         [0024]    Chimeras will be formed between these ESC and embryos and the chimeras will be incubated or will be transferred to pseudo-pregnant recipients, such as in a manner described by Voncken in “Genetic modification of the mouse: Transgenic mouse—methods and protocols”, Methods in Molecular Biology, Volume 209 (2003), the disclosure of which is incorporated herein by reference. By administering inducible drugs to the recipient mice, such as doxycycline (a derivative of tetracycline), at specific times in development of the embryo, the expression of recombinase and compromiser genes in the chimeric embryos/fetuses will be regulated. This method will be used to establish chimeras in which, by way of non-limiting example, there is a vascular endothelium and hematopoietic system from one genotype (i.e., from the donor ESCs) with all other tissues from another genotype (i.e., from the recipient), as depicted in the diagram of  FIG. 1 . 
       EXAMPLES 
       [0025]    The following examples will serve to illustrate the application of the methods described herein. 
       Example 1 
     Spatial and Temporal Regulation of Endothelial and Hematopoietic-Specific Gene Expression and its Application in Mouse Esc-Mouse Chimeras 
       [0026]    FLK1 is a receptor tyrosine kinase and the main signaling receptor for Vascular Endothelial Growth Factor-A (VAGF-A) during embryonic development and adult neovascularization. (Millauer et al., Cell 72:835-846 (1993), Nature 367:576-579 (1994); Goede et al., Lab Invest. 78:1385-1394 (1998)). Analysis of FLK1 knock-out mice by Shalaby et al., (Nature 376:62-66 (1995), Cell 89:981-990 (1997)) revealed a central role of FLK1 in hematopoietic and endothelial development. Licht and co-workers created a novel transgenic mouse line of FLK1-Cre and then cross-bred with the LacZ report mouse line. (Licht et al., Development Dynamics 229:312-318 (2003)). They detected strong, reproducible LacZ staining primarily in the endothelium of blood vessels, but also in circulating blood cells. An almost complete vascular staining was found at mid-gestation and persisted in all organ systems examined in adult mice. 
         [0027]    The stem cell leukemia gene (SCL) encodes a basic helix-loop-helix transcription factor with a pivotal role in both hematopoiesis and endothelial development. During mouse development, SCL is first expressed in extra-embryonic mesoderm, and is required for the generation of all hematopoietic lineages and normal yolk sac angiogenesis. SCL deficient embryos lacked yolk sac hematopoiesis and large vitelline vessels although endothelial capillary spaces were present in SCL-l-yolk sac, as demonstrated by Lorraine, et al. (Proc. Natl. Acad. Sci. USA, VOL. 92, pp. 7075-7079), and substantiated by Shivdasani et al. (Nature (London) 373:432-434 (1995)). To address that the lineage relationship between embryonic and adult hematopoietic stem cells (HSC) in the mouse exists, Joachim et al. (Blood 1 April, Vol. 105, No. 7 (2005)) generated transgenic mice which expressed the tamoxifen inducible Cre-ER T  recombinase under the control of the stem-cell enhancer of SCL locus (HSC-SCL-Cre-ER T -pA) (Sanchez, et al. Development 126:3891-3904 (1999), Development 128:4815-4827 (2001); Gottgens, et al., EMBO J 21:3039-3050 (2002)). and proved that tamoxifen-dependent recombination occurred in more than 90% of adult long-term HSCs. This experiment was a clear demonstration of successful inducible genetic manipulation of HSCs in vivo. 
         [0028]    The FLK1 and SCL play crucial roles in the establishment of hematopoietic and endothelial cell lineages in mice. Changwon et al. (Development and Disease 131:2749-2762 (2004)) have previously used an in vitro differentiation model of embryonic stem (ES) cells and demonstrated that hematopoietic and endothelial cells develop via sequentially generated FLK1 +  and SCL +  cells. 
         [0029]    Where the Cre recombinase expression specificity is determined by the endothelial and blood precursor specific promoters, cells derived from the ESC component of the chimeras and differentiated into all non-endothelium and non-hematopoietic (i.e., non-target) lineages will be eliminated by inducing the expression of compromiser genes. At the same time, cells derived from the donor ESC line that developed into target endothelium and hematopoietic lineages will not express the compromiser genes and therefore will survive. Reciprocally, the cells derived from embryo component of the chimeras and differentiated into endothelium and hematopoietic lineages will be eliminated by inducing the expression of compromiser genes. Conversely, cells derived from the embryo component and developed into all non-endothelium and non-hematopoietic lineages will not express the compromiser genes and therefore will survive. As a result, in these chimeras the ESC and embryo components will complement each other; the endothelium and hematopoietic cells will be built from the ESC component, while the embryo component will provide the remaining cells/structure of the chimera. 
         [0030]    Applying the present method to this example, a new mouse ESC line will be created which contains LoxP-tet-O-DT-A-pA-loxP ( FIG. 2  and SEQUENCE NO. 1), Rosa26-rtTA-IRES-EGFP-pA (Enhanced Green Fluorescent Protein, as disclosed in U.S. Pat. No. 5,625,048, the disclosure of which is incorporated herein by reference), FLK1-Cre-pA and HSC-SCL-Cre-ER T -pA ( FIG. 3  and SEQUENCE NO. 2). Mouse SCL−/− recipient blastocysts will be created by breeding SCL−/+mice or mouse recipient blastocysts will be created which contain tet-O-DT-A-pA, Rosa26-LoxP-STOP-LoxP-rtTA-IRES-EGFP-pA, FLK1-Cre-pA and HSC-SCL-Cre-ER T -pA. The new ESC line will then be injected into recipient blastocysts and embryo transfer performed according to suitable techniques, such as that described by Voncken. 
         [0031]    A Tet-On and Cre-LoxP system will be combined to regulate specific genes&#39; expression by introducing a recombination control drug, such as tetracycline, into the host embryos. In the stem cells system, when endothelial/hematopoietic cell-specific promoters of FLK1 and SCL express, Cre recombinase will be expressed followed by excision of LoxP recognition sites which contain DT-A. Meanwhile, the lineages other than the target endothelial and hematopoietic lineage will express DT-A which kills the cells. In the recipient blastocysts system, SCL−/− blastocysts are hematopoietic and endothelial cells deficient which will be rescued by stem cells because in the blastocysts, this gene regulatory program is working in an opposite way relative to that in stem cell line. When FLK1 and SCL are expressed, Cre recombinase is expressed followed by excision of STOP gene which stops expression of rtTA. After this stop is removed, the tet-O system is activated and DT-A will be expressed. The result is that the recipient blastocysts will be hematopoietic and endothelial deficient and will be “rescued” by the cells coming from donor stem cell system. 
         [0032]    By phenotyping the resulting chimeras to confirm different genotypes of the vascular endothelium and hematopoietic system vs. other tissues, it will be possible to identify if the endothelial and hematopoietic cells differentiated from the ESC line rescued the target lineage of the recipient blastocysts. 
         [0033]    Alternatively, a stem cell line will be made with constructs of SCL-Cre and Rosa 26-loxP-TK-loxP. By injecting this cell line into SCL −/− embryos, the hematopoietic and endothelial system in the SCL −/− embryos will be replaced with the corresponding system from the stem cell line. 
       Example 2 
     Spatial and Temporal Regulation of Endothelial and Hematopoietic-Specific Gene Expression and its Application in Human ESC-Mouse Chimeras 
       [0034]    The highly conserved basic helix-loop-helix (bHLH) transcription factor SCL has been shown in mice and zebrafish to play a crucial role in patterning of mesoderm into blood and endothelial lineages by regulating the development of the hemangioblast. See, for instance, Labastie et al., Blood 92:3624-3635 (1998) and Lorraine et al., EMBO J. 15:4123-4129 (1996), Proc. Natl. Acad. Sci. USA Vol. 92, pp. 7075-7079 (1995). To address the role SCL plays in normal human developmental hematopoiesis, Elias&#39;s work (Elias, et. al, Blood 106:860-870 (2005)) provide insight into the role that key hematopoietic genes may play in human embryonic development. Elias&#39; data revealed that SCL was the first and most dramatically up-regulated gene coinciding with emergence of primitive hematopoiesis and was expressed abundantly in all hematopoietic colonies. 
         [0035]    The SCL gene is expressed in a subset of blood cells, endothelial cells, and specific regions of the brain and spinal cord. This pattern of expression is highly conserved throughout vertebrate evolution from zebrafish to mammals. Systematic analysis of the murine SCL locus has identified a series of independent enhancers, each of which directs reporter gene expression to a subdomain of the normal SCL expression pattern. Of particular interest is a 3′enhancer that directs expression to blood and endothelial progenitors throughout ontogeny. See, Sanchez, et al., Development 126:3891-3904 (1999). Joachim, et al. (Blood 104:1769-1777 (2004)) generated endothelial-SCL-Cre-ER T  mice using inducible Cre recombinase driven by the 5-endothelial enhancer of the SCL locus. By intercrossing with Cre reporter mice, Joachim found Cre-mediated recombination in almost all endothelial cells of the developing vasculature. 
         [0036]    Combining all this information, mouse-human chimeras can be made using the methods described in Example 1. A new human ESC line will be created which contains LoxP-tet-O-DT-A-pA-loxP ( FIG. 2  and SEQUENCE NO. 1), Rosa26-rtTA-IRES-EGFP-pA and SCL-Cre-pA ( FIG. 3  and SEQUENCE NO. 3). Meanwhile, mouse SCL−/− recipient blastocysts will be created, or alternatively recipient blastocysts will be created which contain tet-O-DT-A-pA, Rosa26-LoxP-STOP-LoxP-rtTA-IRES-EGFP-pA, and SCL-Cre-pA. The new ESC line will be injected into recipient blastocysts and embryo transfer will be performed. 
         [0037]    The site-specific recombination systems will be activated at a pre-determined time in the development of the embryo by administration of a recombination control, such as the drug doxycycline. Expression of the suicide/compromiser genes in the ESC line and the donor embryo will result in reciprocal ablation of the non-target cells in the ESC line and the target cells in the donor embryo. The ESC line will thus provide the target cells, in this case vascular endothelium and hematopoietic tissues, for the developing chimeric mouse. The resulting chimeras can be phenotyped to confirm different genotypes of the vascular endothelium and hematopoietic system vs. other tissues. In these chimeras, the endothelial and hematopoietic cells will be human genome background while all the other tissues and organs will be mouse genome background. 
       Example 3 
     Spatial and Temporal Regulation of Endothelial and Hematopoietic-Specific Gene Expression and its Application in Human ESC-Pig Chimeras 
       [0038]    The chronic shortage of human organs, tissues and cells for transplantation has inspired research on the possibility of using animal donor tissue instead of human donor tissue. Transplantation over a species barrier is associated with rejections which are difficult to control. Therefore, it is has been proposed that successful pig to human xenotransplantation requires donor pigs to be genetically modified. See, Prather et al. Theriogenology 59:115-123 (2003); and Kolber-Simonds et al. PNAS 101:7335-7340 (2004). Vascular endothelium is the most immediate barrier between the xenogeneic donor organ and host immune and non-immune defense systems. Thus, these cells are the prime targets for such genetic modifications. 
         [0039]    Godwin et al. (Xenotransplantation 13(6):514-521 (2006)) cloned and characterized the regulatory elements of the pig intercellular adhesion molecule-2 (ICAM-2) gene. They observed that a 0.90-kb pig ICAM-2 promoter fragment had strong activity in pig endothelial cells but not in non-endothelial cells. Deletion analysis revealed that the majority of promoter activity was specified by a 0.48-kb sub-fragment with significant homology to the human ICAM-2 promoter. Significant enhancer activity was identified within the first intron of the pig ICAM-2 gene. 
         [0040]    The Tie2 promoter and intron/enhancer element has been previously shown to drive reporter genes in vitro and in vivo. Inclusion of a Tie2 intronic enhancer element in conjunction with the Tie2 promoter in Tie2-βgal transgenic mice has resulted in expression in embryonic and adult endothelium as expected, as reported by Schlaeger et al. (Proc. Nat. Acad. Sci. USA 94:3058-3063 (1997)). This same type of promoter-element transgene design was used to generate Tie2-Cre and Tie2-GFP transgenic mice, and Tie2-GFP transgenic Zebrafish (Constien et al. Genesis 30:36-44 (2001); Motoike et al. Genesis 28:75-81 (2000)). Hao et al. (Transgenic Research DI 10.1007/s11248-00609020-8 (2006)) have generated transgenic Yucatan pigs that express the eNOS cDNA under the Tie2 endothelial-specific promoter and Tie2 intron/enhancer element and have demonstrated a similar expression profile in the endothelial compartment in the Tie2-eNOS transgenic swine by immunohistochemistry. 
         [0041]    So far, there is no specific gene known which will regulate the differentiation of hematopoietic stem cells from embryonic stem cells in pig. But, it is known that the pattern of SCL gene expression is highly conserved throughout vertebrate evolution from zebrafish to mammals. Thus a promoter of SCL gene can be used to regulate the hematopoietic development in swine. 
         [0042]    Consequently, pig-human chimeras can be made using the methods described in Example 1. A new human ESC line will be created which contains LoxP-tet-O-DT-A-pA-loxP, Rosa26-rtTA-IRES-EGFP-pA, SCL-Cre-pA and ICAM-Cre-pA/Tie2-Cre-pA. Concurrently, pig SCL−/− recipient blastocysts will be created or alternatively recipient blastocysts will be created which contain tet-O-DT-A-pA, Rosa26-LoxP-STOP-LoxP-rtTA-IRES-EGFP-pA, SCL-Cre-pA and ICAM-Cre-pA/Tie2-Cre-pA. The new ESC line will be injected into recipient blastocysts and embryo transfer will be performed. 
         [0043]    The site-specific recombination systems will be activated at a pre-determined time in the development of the embryo by administration of a recombination control, such as the drug doxycycline. Expression of the suicide/compromiser genes in the ESC line and the donor embryo will result in reciprocal ablation of the non-target cells in the ESC line and the target cells in the donor embryo. The ESC line will thus provide the target cells, in this case vascular endothelium and hematopoietic tissues, for the developing chimeric pig. Finally, the resulting chimeras will be phenotyped to confirm different genotypes of the vascular endothelium and hematopoietic system vs. other tissues. In these chimeras, the endothelial and hematopoietic cells will be human genome background while all the other tissues and organs will be pig genome background. 
       Example 4 
     Spatial and Temporal Regulation of any Organ/Tissue-Specific Gene Expression and its Application in Chimeras 
       [0044]    Based on the method described above, chimeras of any species can be for which EC/ES/P/iPS cells are available and for which the specific promoter/enhancer required to genetically control the chimeric characteristics is known. These chimeras can be created at various stages of embryonic development. In the present example this process can be used at a point in development in the formation of the initial three (triploblastic) tissue layers, namely the endoderm, ectoderm and mesoderm. In this example, inducing chimerism in one of these tissue lineages will result in all subsequent cells, tissues and organs that are derived from a different genotype. 
         [0045]    For example, using this method, a pig with a human endoderm lineage can be made. In one specific embodiment, when a specific promoter/enhancer for endoderm is observed which might be called END, the new ESC line of any kind of background would be created which contains LoxP-tet-O-DT-A-pA-loxP, Rosa26-rtTA-IRES-EGFP-pA and END-Cre-pA. Meanwhile, END−/− recipient blastocysts would be created or alternatively blastocysts of any kind of background would be created which contain tet-O-DT-A-pA, Rosa26-LoxP-STOP-LoxP-rtTA-IRES-EGFP-pA, and END-Cre-pA. The new ESC line would be injected into recipient blastocysts and embryo transfer performed. 
         [0046]    The site-specific recombination systems will be activated at a pre-determined time in the development of the embryo by administration of a recombination control, such as the drug doxycycline. Expression of the suicide/compromiser genes in the ESC line and the donor embryo will result in reciprocal ablation of the non-target cells in the ESC line and the target cells in the donor embryo. The ESC line will thus provide the target cells for the developing chimeric animal. Finally, the resulting chimeras would be phenotyped to confirm different genotypes of all the tissues/organs coming from endoderm layers vs. other tissues/organs. In these chimeras, the cells coming from endoderm layer will be one genome background and all the other tissues and organs will be the other genome background. 
       Example 5 
     Spatial and Temporal Regulation of Specific Gene Expression and its Application in Embryonic Cell Derived Chimeras In Vitro 
       [0047]    Examples 1-4 described above contemplate spatial and temporal regulation of specific gene expression in vivo. In the present example, this method will be used in vitro as well. As in the prior examples, a new ESC line or ECs will be created which contains three transgenes: (1) loxP-tet-O-DT-A-pA-loxP, (2) Rosa26-rtTA-IRES-EGFP-pA, (3) FLK1-Cre-pA/HSC-SCL-Cre-ERT-pA. Instead of blastocysts injection, chimeras will be made by ES cell-diploid/tetraploid embryo aggregation and injection. 
         [0048]    The new ESC line will be created to contain LoxP-tet-O-DT-A-pA-loxP, Rosa26-rtTA-IRES-EGFP-pA and END-Cre-pA. Meanwhile, END−/− recipient diploid embryos would be created or alternatively embryos of any kind of background would be created which contain tet-O-DT-A-pA, Rosa26-LoxP-STOP-LoxP-rtTA-IRES-EGFP-pA, and END-Cre-pA. ESC line will be aggregated with recipient embryos and cultured in vitro. Before embryo transfer, inducible drugs will be administered which will result in embryo chimeras having endoderm lineage that comes from the ESC line while the ectoderm and mesoderm lineages come from the recipient blastocysts. 
         [0049]    The resulting chimeras would be phenotyped in vitro to confirm different genotypes of all the tissues/organs coming from endoderm layers vs. other tissues/organs. In these chimeras, the cells coming from endoderm layer will be one genome background and all the other tissues and organs will be the other genome background. 
         [0050]    While the invention has been illustrated and described in detail in the drawings and foregoing description, the same should be considered as illustrative and not restrictive in character. It is understood that only the preferred embodiments have been presented and that all changes, modifications and further applications that come within the spirit of the invention are desired to be protected. 
         [0000]    
       
         
               
               
             
               
               
             
               
               
               
             
               
               
             
               
               
             
               
               
               
             
               
               
             
               
               
             
               
               
               
             
           
               
                 SEQUENCE NO. 1 
                   
               
             
          
           
               
                 Restriction analysis on pMC-loxp-tight-DTa-(R).seq 
                   
               
               
                 Methylation: dam-No dsm-No 
               
               
                 Enzymes with &gt;3 sites are not shown 
               
               
                   
               
               
                 Screened with 51 enzymes, 64 sites found 
               
               
                 AstII GACGT/C 1: 5117 
               
               
                 Acc651 G/GTACC 1: 532 
               
               
                 ApaI GGGCC/C 1: 466 
               
               
                 ApaLI G/TGCAC 3: 220, 3616, 4862 
               
               
                 BamHI G/GATCC1: 1682 
               
               
                 BglI GCCNNNN/NCCG 3: 294, 461, 4315 
               
               
                 BglII A/GATCT 1: 468 
               
               
                 BsaBI GATNN/NNATC 1: 1876 
               
               
                 BssHII G/CGCGC 1: 1357 
               
               
                 ClaI AT/CGAT 1: 475 
               
               
                 EcoICRI GAG/CTC 2: 2552, 2909 
               
               
                 EcoRI G/AATTC 3: 445, 719, 2517 
               
               
                 EcoRV GAT/ATC 1: 482 
               
               
                 HindIII A/AGCTT 2: 486, 2944 
               
               
                 HpaI GTT/AAC 1: 1775 
               
               
                 KpnI GGTAC/C 1: 536 
               
               
                 loxp 2: 514, 2867 
               
               
                 MscI TGG/CCA 2: 1042, 1983 
               
               
                 NcoI C/CATGG 2: 1392, 2504 
               
               
                 NdeI CA/TATG 1: 227 
               
               
                 NheI G/CTAGC 1: 2845 
               
               
                 NotI GC/GGCCGC 1: 2892 
               
               
                 PmeI GTTT/AAAC 1: 2902 
               
               
                 PstI CTGACA/G 3: 816, 1013, 2940 
               
               
                 PvuI CGAT/CG 2: 322, 4565 
               
               
                 PvuII CAG/CTG 3: 351, 1066, 3127 
               
               
                 SacI GAGCT/C 2: 2554, 2911 
               
               
                 ScaI AGT/ACT1: 4675 
               
               
                 SmaI CCC/GGG 2: 458, 2927 
               
               
                 SpeI A/CTAGT 1: 2913 
               
               
                 StuI AGG/CCT 3: 591, 662, 2567 
               
               
                 XbaI T/CTAGA 2: 538, 1883 
               
               
                 XbaI &lt;Methy&gt; T/CTAGATC 1: 538 
               
               
                 XhoI C/TCGAG 2: 450, 2919 
               
               
                 XmaI C/CCGGG 2: 456, 2925 
               
               
                 XmnI GAANN/NNTTC 2: 2881, 4794 
               
               
                   
               
               
                 Non Cut Enzymes 
               
               
                 Acc65I&lt;Methy&gt;AflII      ApaI&lt;Methy&gt; BstEII     BstXI       ClaI&lt;Methy&gt; 
               
               
                 I-PpoI      I-SceI     MscI&lt;Mety&gt; NruI        NruI&lt;Methy&gt; SacII 
               
               
                 SalI 
               
               
                   
               
             
          
           
               
                 ORIGIN 
                   
                   
               
               
                     1 
                 CTGCCTCGCG CGTTTCGGTG ATGACGGTGA AAACCTCTGA CACATGCAGC TCCCGGAGAC 
               
               
                    61 
                 GGTCACAGCT TGTCTGTAAG CGGAGCCGGG AGCAGACAAG CCCGTCAGGG CGCGTCAGCG 
               
               
                   121 
                 GGTGTTGGCG GGTGTCGGGG CGCAGCCATG ACCCAGTCAC GTAGCGATAG CGGAGTGTAC 
               
               
                   181 
                 TGGCTTAACT ATGCGGCATC AGAGCAGATT GTACTGAGAG TGCACCATAT GCGGTGTGAA 
               
               
                   241 
                 ATACCGCACA GATGCGTAAG GAGAAAATAC CGCATCAGGC GCCATTCGCC ATTCAGGCTA 
               
               
                   301 
                 CGCAACTGTT GGGAAGGGCG ATCGGTGCGG GCCTCTTCGC TATTACGCCA GCTGGCGAAG 
               
               
                   361 
                 GGGGGATGTG CTGCAAGGCG ATTAAGTTGG GTAACGCCAG GGTTTTCCCA GTCACGACGT 
               
               
                   421 
                 TGTAAAACGA CGGCCAGGGC CAGTGAATTC TCGAGCCCGG GGGGCCCAGA TCTATCGATG 
               
               
                   481 
                 ATATCAAGCT TGGTACTATA ACTTCGTATA GTATACATTA TACGAAGTTA TGGTACCTCT 
               
               
                   541 
                 AGATCGACAG TGTGGTTTTG CAAGAGGAAG CAAAAAGCCT CTCCACCCAG GCCTGGAATG 
               
               
                   601 
                 TTTCCACCCA ATGTCGAGCA GTGTGGTTTT GCAAGAGGAA GCAAAAAGCC TCTCCACCCA 
               
               
                   661 
                 GGCCTGGAAT GTTTCCACCC AATGTCGAGC AAACCCCGCC CAGCGTCTTG TCATTGGCGA 
               
               
                   721 
                 ATTCGAACAC GCAGATGCAG TCGGGGCGGC GCGGTCCCAG GTCCACTTCG CATATTAAGG 
               
               
                   781 
                 TGACGCGTGT GGCCTCGAAC ACCGAGCGAC CCTGCAGCCA ATATGGGATC GGCCATTGAA 
               
               
                   841 
                 CAAGATGGAT TGCACGCAGG TTCTCCGGCC GCTTGGGTGG AGAGGCTATT CGGCTATGAC 
               
               
                   901 
                 TGGGCACAAC AGACAATCGG CTGCTCTGAT GCCGCCGTGT TCCGGCTGTC AGCGCAGGGG 
               
               
                   961 
                 CGCCCGGTTC TTTTTGTCAA GACCGACCTG TCCGGTGCCC TGAATGAACT GCAGGACGAG 
               
               
                  1021 
                 GCAGCGCGGC TATCGTGGCT GGCCACGACG GGCGTTCCTT GCGCAGCTGT GCTCGACGTT 
               
               
                  1081 
                 GTCACTGAAG CGGGAAGGGA CTGGCTGCTA TTGGGCGAAG TGCCGGGGCA GGATCTCCTG 
               
               
                  1141 
                 TCATCTCACC TTGCTCCTGC CGAGAAAGTA TCCATCATGG CTGATGCAAT GCGGCGGCTG 
               
               
                  1201 
                 CATACGCTTG ATCCGGCTAC CTGCCCATTC GACCACCAAG CGAAACATCG CATCGAGCGA 
               
               
                  1261 
                 GCACGTACTC GGATGGAAGC CGGTCTTGTC GATCAGGATG ATCTGGACGA AGAGCATCAG 
               
               
                  1321 
                 GGGCTCGCGC CAGCCGAACT GTTCGCCAGG CTCAAGGCGC GCATGCCCGA CGGCGAGGAT 
               
               
                  1381 
                 CTCGTCGTGA CCCATGGCGA TGCCTGCTTG CCGAATATCA TGGTGGAAAA TGGCCGCTTT 
               
               
                  1441 
                 TCTGGATTCA TCGACTGTGG CCGGCTGGGT GTGGCGGACC GCTATCAGGA CATAGCGTTG 
               
               
                  1501 
                 GCTACCCGTG ATATTGCTGA AGAGCTTGGC GGCGAATGGG CTGACCGCTT CCTCGTGCTT 
               
               
                  1561 
                 TACGGTATCG CCGCTCCCGA TTCGCAGCGC ATCGCCTTCT ATCGCCTTCT TGACGAGTTC 
               
               
                  1621 
                 TTCTGAGGGG ATCGGCAATA AAAAGACAGA ATAAAACGCA CGGGTGTTGG GTCGTTTGTT 
               
               
                  1681 
                 CGGATCCGTC GAGGCAGTGA AAAAAATGCT TTATTTGTGA AATTTGTGAT GCTATTGCTT 
               
               
                  1741 
                 TATTTGTAAC CATTATAAGC TGCAATAAAC AAGTTAACAA CAACAATTGC ATTCATTTTA 
               
               
                  1801 
                 TGTTTCAGGT TCAGGGGGAG GTGTGGGAGG TTTTTTAAAG CAAGTAAAAC CTCTACAAAT 
               
               
                  1861 
                 GTGGTATGGC TGATTATGAT CCTCTAGACT CACACCACAG AAGTAAGGTT TCCTTCACAA 
               
               
                  1921 
                 AGAGATCGCC TGACACGATT TCCTGCACAG GCTTGAGCCA TATACTCATA CATCGCATCT 
               
               
                  1981 
                 TGGCCACGTT TTCCACGGGT TTCAAAATTA ATCTCAAGTT CTACGCTTAA CGCTTTCGCC 
               
               
                  2041 
                 TGTTCCCAGT TATTAATATA TTCAACGCTA GAACTCCCCT CAGCGAAGGG AAGGCTGAGC 
               
               
                  2101 
                 ACTACACGCG AAGCACCATC ACCGAACCTT TTGATAAACT CTTCCGTTCC GACTTGCTCC 
               
               
                  2161 
                 ATCAACGGTT CAGTGAGACT TAAACCTAAC TCTTTCTTAA TAGTTTCGGC ATTATCCACT 
               
               
                  2221 
                 TTTAGTGCGA GAACCTTCGT CAGTCCTGGA TACGTCACTT TGACCACGCC TCCAGCTTTT 
               
               
                  2281 
                 CCAGAGAGCG GGTTTTCATT ATCTACAGAG TATCCCGCAG CGTCGTATTT ATTGTCGGTA 
               
               
                  2341 
                 CTATAAAACC CTTTCCAATC ATCGTCATAA TTTCCTTGTG TACCAGATTT TGGCTTTTGT 
               
               
                  2401 
                 ATACCTTTTT GAATGGAATC TACATAACCA GGTTTAGTCC CGTGGTACGA AGAAAAGTTT 
               
               
                  2461 
                 TCCATCACAA AAGATTTAGA AGAATCAACA ACATCATCAG GGTCCATGGT GGCGGCGAAT 
               
               
                  2521 
                 TCTCCAGGCG ATCTGACGGT TCACTAAACG AGCTCTGCTT ATATAGGCCT CCCACCGTAC 
               
               
                  2581 
                 ACGCCTACCT CGACATACGT TCTCTATCAC TGATAGGGAG TAAACTCGAC ATACGTTCTC 
               
               
                  2641 
                 TATCACTGAT AGGGATAAAC TCGACATACG TTCTCTATCA CTGATAGGGA GTAAACTCGA 
               
               
                  2701 
                 CATACGTTCT CTATCACTGA TAGGGAGTAA ACTCGACATA CGTTCTCTAT CACTGATAGG 
               
               
                  2761 
                 GAGTAAACTC GACATCGTTC TCTATCACTG ATAGGGAGTA AACTCGACAT ACGTTCTCTA 
               
               
                  2821 
                 TCACTGATAG GGAGTAAACT CGACGCTAGC ATAACTTCGT ATAGCATACA TTATACGAAG 
               
               
                  2881 
                 TTATTCTAGC GCGGCCGCGT TTAAACGAGC TCACTAGTCT CGAGCCCGGG ATCGACTGCA 
               
               
                  2941 
                 GCCAAGCTTG GCGTAATCAT GGTCATAGCT GTTTCCTGTG TGAAATTGTT ATCCGCTCAC 
               
               
                  3001 
                 AATTCCACAC AACATACGAG CCGGAAGCAT AAAGTGTAAA GCCTGGGGTG CCTAATGAGT 
               
               
                  3061 
                 GAGGTAACTC ACATTAATTG CGTTGCGCTC ACTGCCCGCT TTCCAGTCGG GAAACCTGTC 
               
               
                  3121 
                 GTGCCAGCTG CATTAATGAA TCGGCCAACG CGCGGGGAGA GGCGGTTTGC GTATTGGCGC 
               
               
                  3181 
                 TCTTCCGCTT CCTCGCTCAC TGACTCGCTG CGCTCGGTCG TTCGGCTGCG GCGAGCGGTA 
               
               
                  3241 
                 TCAGCTCACT CAAAGGCGGT AATACGGTTA TCCACAGAAT CAGGGGATAA CGCAGGAAAG 
               
               
                  3301 
                 AACATGTGAG CAAAAGGCCA GCAAAAGGCC AGGAACCGTA AAAAGGCCGC GTTGCTGGCG 
               
               
                  3361 
                 TTTTTCCATA GGCTCCGCCC CCCTGACGAG CATCACAAAA ATCGACGCTC AAGTCAGAGG 
               
               
                  3421 
                 TGGCGAAACC CGACAGGACT ATAAAGATAC CAGGCGTTTC CCCCTGGAAG CTCCCTCGTG 
               
               
                  3481 
                 CGCTCTCCTG TTCCGACCCT GCCGCTTACC GGATACCTGT CCGCCTTTCT CCCTTCGGGA 
               
               
                  3541 
                 AGCGTGGCGC TTTCTCAATG CTCACGCTGT AGGTATCTCA GTTCGGTGTA GGTCGTTCGC 
               
               
                  3601 
                 TCCAAGCTGG GCTGTGTGCA CGAACCCCCC GTTCAGCCCG ACCGCTGCGC CTTATCCGGT 
               
               
                  3661 
                 AACTATCGTC TTGAGTCCAA CCCGGTAAGA CACGACTTAT CGCCACTGGC AGCAGCCACT 
               
               
                  3721 
                 GGTAACAGGA TTAGCAGAGC GAGGTATGTA GGCGGTGCTA CAGAGTTCTT GAAGTGGTGG 
               
               
                  3781 
                 CCTAACTACG GCTACACTAG AAGGACAGTA TTTGGTATCT GCGCTCTGCT GAAGCCAGTT 
               
               
                  3841 
                 ACCTTCGGAA AAAGAGTTGG TAGCTCTTGA TCCGGCAAAC AAACCACCGC TGGTAGCGGT 
               
               
                  3901 
                 GGTTTTTTTG TTTGCAAGCA GCAGATTACG CGCAGAAAAA AAGGATCTCA AGAAGATCCT 
               
               
                  3961 
                 TTGATCTTTT CTACGGGGTC TGACGCTCAG TGGAACGAAA ACTCACGTTA AGGGATTTTG 
               
               
                  4021 
                 GTCATGAGAT TATCAAAAAG GATCTTCACC TAGATCCTTT TAAATTAAAA ATGAAGTTTT 
               
               
                  4081 
                 AAATCAATCT AAAGTATATA TGAGTAAACT TGGTCTGACA GTTACCAATG CTTAATCAGT 
               
               
                  4141 
                 GAGGCACCTA TCTCAGCGAT CTGTCTATTT CGTTCATCCA TAGTTGCCTG ACTCCCCGTC 
               
               
                  4201 
                 GTGTAGATAA CTACGATACG GGAGGGCTTA CCATCTGGCC CCAGTGCTGC AATGATACCG 
               
               
                  4261 
                 CGAGACCCAC GCTCACCGGC TCCAGATTTA TCAGCAATAA ACCAGCCAGC CGGAAGGGCC 
               
               
                  4321 
                 GAGCGCAGAA GTGGTCCTGC AACTTTATCC GCCTCCATCC AGTCTATTAA TTGTTGCCGG 
               
               
                  4381 
                 GAAGCTAGAG TAAGTAGTTC GCCAGTTAAT AGTTTGCGCA ACGTTGTTGC CATTGCTACA 
               
               
                  4441 
                 GGCATCGTGG TGTCACGCTC GTCGTTTGGT ATGGCTTCAT TCAGCTCCGG TTCCCAACGA 
               
               
                  4501 
                 TCAAGGCGAG TTACATGATC CCCCATGTTG TGCAAAAAAG CGGTTAGCTC CTTCGGTCCT 
               
               
                  4561 
                 CCGATCGTTG TCAGAAGTAA GTTGGCCGCA GTGTTATCAC TCATGGTTAT GGCAGCACTG 
               
               
                  4621 
                 CATAATTCTC TTACTGTCAT GCCATCCGTA AGATGCTTTT CTGTGACTGG TGAGTACTCA 
               
               
                  4681 
                 ACCAAGTCAT TCTGAGAATA GTGTATGCGG CGACCGAGTT GCTCTTGCCC GGCGTCAATA 
               
               
                  4741 
                 CGGGATAATA CCGCGCCACA TAGCAGAACT TTAAAAGTGC TCATCATTGG AAAACGTTCT 
               
               
                  4801 
                 TCGGGGCGAA AACTCTCAAG GATCTTACCG CTGTTGAGAT CCAGTTCGAT GTAACCCACT 
               
               
                  4861 
                 CGTGCACCCA ACTGATCTTC AGCATCTTTT ACTTTCACCA GCGTTTCTGG GTGAGCAAAA 
               
               
                  4921 
                 ACAGGAAGGC AAAATGCCGC AAAAAAGGGA ATAAGGGCGA CACGGAAATG TTGAATACTC 
               
               
                  4981 
                 ATACTCTTCC TTTTTCAATA TTATTGAAGC ATTTATCAGG GTTATTGTCT CATGAGCGGA 
               
               
                  5041 
                 TACATATTTG AATGTATTTA GAAAAATAAA CAAATAGGGG TTCCGCGCAC ATTTCCCCGA 
               
               
                  5101 
                 AAAGTGCCAC CTGACGTCTA AGAAACCATT ATTATCATGA CATTAACCTA TAAAAATAGG 
               
               
                  5161 
                 CGTATCACGA GGCCCTTTCG TCTTCAAGAA// 
               
               
                   
               
             
          
           
               
                 SEQUENCE NO. 2 
                   
               
             
          
           
               
                 HSC-CRE-ERt 
                   
               
               
                 LOCUS Untitled 13033 bp DNA linear SYN 03-JAN.-2008 
               
               
                 DEFINITION. 
               
               
                 ACCESSION. 
               
               
                 KEYWORDS. 
               
               
                 FEATURES Location/Qualifiers 
               
               
                 BASE COUNT 3087 a 31210 c 3427 g 3399 t 
               
               
                   
               
             
          
           
               
                 ORIGIN 
                   
                   
               
               
                     1 
                 ACGACTGGAG AGATGGCTCA CTGGTTAAGA GCACTGACTA TTCTTCCAGA GGTCCTGAGT 
               
               
                    61 
                 TCAATTCCCA ACAACCACAT GGTGGCTCAG AACCATCTGT AATGGGATCT GATGCCCTCT 
               
               
                   121 
                 CCTAGTGTGT CTGAAGGCAG CCACAGTGTG TGTGTGTGTG TGTGTGTGTG TGTATACATA 
               
               
                   181 
                 TACATATATA TGTATATATA TAATTTTTGC ATATTAAATC TATAAAAAAA AAACCCAGTG 
               
               
                   241 
                 AGATCCGAGT TCTGTGTATT GAGAATACCA AGGTGTATGG TGTGTGTGTG TGGGGGGGAA 
               
               
                   301 
                 GAGGACACTT CATTGGAATA ATTCAAGGAA GAGCTTTCTT TATATTTTCT CCATCAGGAG 
               
               
                   361 
                 GGGAGCCCAG ATTCTAGTGA CTTCTGGAGC ACTTTCCCAA GTCTTAAGAG TCCAGCTGAG 
               
               
                   421 
                 CAGAATGGGG TGGAGTGTGA AGGGTAGTAG GACCAGAATC CAGGATTAGC TTCAGTCCTT 
               
               
                   481 
                 GACTCCCTTT CTTATGATAG GGTAGCTACT TGCAGAATAC AACGGTGGGT TTGCTTAGTG 
               
               
                   541 
                 TAGGCTGCTT TCCTCTTGGC CGGGAATATT TCTGACATCC TTGGTTGAAT AGAGCAGAGT 
               
               
                   601 
                 TCTTGCAGCT TCCACACCCT ACTTCACCAC CATAGTCTTT CTGGGTGTAT ATTTGCAGCG 
               
               
                   661 
                 CATGTGTGTA GCAGTAGATC GGGAGAGGGT TCCTATAGCA CTGGACAGAT TCCCCGCCAA 
               
               
                   721 
                 AACCAAAAGG GGGGCGGGAA GGACACGCTT GCTCGGGGGA TTAGTTCCCT CCCCTTCCCC 
               
               
                   781 
                 TGTGGCCTAA GAAGGAGGGA CTGGGTGATC TTTCTCTTCT CTGTGCATTT CCTTCCTCCT 
               
               
                   841 
                 TTTTCCCGTC GATTTTTGTC TCTCTGCCTG TATTCCTTTT CTCCCAAGGT TTCTGCCATC 
               
               
                   901 
                 TTTCTCCAGC ACAATTCCTA CCCTTGGACA CTGTGCCTTC CGGGCTTGTC CCACCCTTTT 
               
               
                   961 
                 CTTCCAATCT AGAGACACCC CCACATTGCT CCAGCTCCAG GCCTGTGGGC CTTCACGCCA 
               
               
                  1021 
                 GCAGGGTTGG GGTGTGCGTC CACGTGGTGC TGAGTTTGTC CTGTCCGCTT TTCAGGTTTC 
               
               
                  1081 
                 AGTGCGTGAT CTCCTCTCTG CCCCTTACCC TGTTACAGGA TGACGGAGCG GCCGCCGAGC 
               
               
                  1141 
                 GAGGCGGCAC GCAGTGACCC GCAACTAGAG GGACAGGACG CGGCCGAGGC CCGCATGGCC 
               
               
                  1201 
                 CCCCCGCACC TAGTCCTGCT CAACGGCGTC GCCAAGGAGA CGAGCCGCGC AGCCCCGGCT 
               
               
                  1261 
                 GAGCCCCCCG TCATCGAGCT AGGAGCGCGC AGCGGCGCGG GGGGCGGCCC TGCCAGTGGG 
               
               
                  1321 
                 GGCGGTGCCG CGAGGGACTT AAAGGGCCGC GACGCAGTAG CAGCCGAAGC TCGCCTTCGG 
               
               
                  1381 
                 GTGCCCACCA CCGAGCTGTG CAGACCTCCC GGACCCGCCC CGGCGCCCGC GCCCGCTTCG 
               
               
                  1441 
                 GTTCCTGCAG AGCTGCCTGG AGACGGCCGC ATGGTGCAGC TGAGCCCGCC CGCGCTGGCA 
               
               
                  1501 
                 GCCCCTGCCG GCCCCGGCCG AGCGCTGCTC TATAGCCTTA GCCAGCCGCT CGCCTCACTA 
               
               
                  1561 
                 GGCAGGTGAG CATCCCGGTC CCCTGCGGCG TTCTGGGTGC AGGCGAGGGT CGAGAGGAGG 
               
               
                  1621 
                 GGGTGGTGGC TTAAGATTCC AAGAGGAACG AGCCCAGAGA CCAGAGTCTC TCCCGCAACC 
               
               
                  1681 
                 CTCCCGCTAG TGGGAAAGGG GTCCCCTGTG AGACAGACTG TCAGGAAGGA CCGGTGGTCA 
               
               
                  1741 
                 GGGGACGACA GTTGTGTAGA AACCGGGGGT GGTCGCCTGC ACTGTTGAGG GTGCGGGTCT 
               
               
                  1801 
                 GTGGGTGAGT GTAAAAAGCT GCAGAGGTTG CTGACTACTG TTGAGTAGGC GGGATTCTTT 
               
               
                  1861 
                 AATATGAGTT CTGGGCCAGT GTCTGAATGC CCCTCTGCAG CAGAGGTGAG GTTCGCCACA 
               
               
                  1921 
                 AAGGGTGAAC TCTTCAGGAA GCTGCCGCGG TGGGTGGACA GGCTGGAGAG AAAGATCTAA 
               
               
                  1981 
                 GGCCGTTGCT GAGGGCAGCT CTTCTCAGCC TCTGCTAGGA TGCAGTGAGC GACACTGTCA 
               
               
                  2041 
                 TCCGCTCCTA ATCCTTCTGT CCCTTACCTG CGTGGTTGGT CTCCTTGCTG GGCCCTGTGG 
               
               
                  2101 
                 TGAGGGAAGC TGAATGGCCA GCAGAGTGTA GGACAGGCGG TAGGAAAGAA TTATAGGACA 
               
               
                  2161 
                 ACACGATGGT AGAGCAGTAG GGAGCGCTGT CAAGGGTTGG TGAGTGGGAG GTGGGGGGTG 
               
               
                  2221 
                 GTGCCGATCT GTGATCAGAG AGTGATGGTC GGTGAGGTCT GAGGGGACAA TGTGAGACCC 
               
               
                  2281 
                 TTTGTGGTGT GGGAGTTCTC TACTAGCACT TCCATCCCTC ACGTGTTGTC CTGTGTAGGT 
               
               
                  2341 
                 ACTTGTCTCT GAGCAAAGGT CTACCAGGAT TGAAGGAGAT TTTGTGTGTG TGTGTGTGTG 
               
               
                  2401 
                 TGTGTGTGTG TGTGTGTGTG TGTGTGTGTA CTTCAGCACA GGAATACGCC GCCTTGCCCC 
               
               
                  2461 
                 TCCCATTTAT GTATTGTTCC ATATATTCAC CCTCTTCGCT TCTGTGAATG CATGCATACT 
               
               
                  2521 
                 CAATTCAATC TGCATTTTAA GTGTGCAGGA GCAGGGGGTG CCTTAGCAGG AGGGGACTGA 
               
               
                  2581 
                 AGACACACAG GGAGAATCCA TCTAAGGAGT CTTTTTGTCT TTAACCTCAT TGTGATCTAC 
               
               
                  2641 
                 CTTCTCTTTC CATAGTGGGT TCTTTGGGGA ACCGGATGCC TTCCCCATGT TCACCAACAA 
               
               
                  2701 
                 CAACCGGGTG AAGAGGAGGC CCTCCCCATA AATTCCACCA TGTCCAATTT ACTGACCGTA 
               
               
                  2761 
                 CACCAAAATT TGCCTGCATT ACCGGTCGAT GCAACGAGTG ATGAGGTTCG CAAGAACCTG 
               
               
                  2821 
                 ATGGACATGT TCAGGGATCG CCAGGCGTTT TCTGAGCATA CCTGGAAAAT GCTTCTGTCC 
               
               
                  2881 
                 GTTTGCCGGT CGTGGGCGGC ATGGTGCAAG TTGAATAACC GGAAATGGTT TCCCGCAGAA 
               
               
                  2941 
                 CCTGAAGATG TTCGCGATTA TCTTCTATAT CTTCAGGCGC GCGGTCTGGC AGTAAAAACT 
               
               
                  3001 
                 ATCCAGCAAC ATTTGGGCCA GCTAAACATG CTTCATCGTC GGTCCGGGCT GCCACGACCA 
               
               
                  3061 
                 AGTGACAGCA ATGCTGTTTC ACTGGTTATG CGGCGGATCC GAAAAGAAAA CGTTGATGCC 
               
               
                  3121 
                 GGTGAACGTG CAAAACAGGC TCTAGCGTTC GAACGCACTG ATTTCGACCA GGTTCGTTCA 
               
               
                  3181 
                 CTCATGGAAA ATAGCGATCG CTGCCAGGAT ATACGTAATC TGGCATTTCT GGGGATTGCT 
               
               
                  3241 
                 TATAACACCC TGTTACGTAT AGCCGAAATT GCCAGGATCA GGGTTAAAGA TATCTCACGT 
               
               
                  3301 
                 ACTGACGGTG GGAGAATGTT AATCCATATT GGCAGAACGA AAACGCTGGT TAGCACCGCA 
               
               
                  3361 
                 GGTGTAGAGA AGGCACTTAG CCTGGGGGTA ACTAAACTGG TCGAGCGATG GATTTCCGTC 
               
               
                  3421 
                 TCTGGTGTAG CTGATGATCC GAATAACTAC CTGTTTTGCC GGGTCAGAAA AAATGGTGTT 
               
               
                  3481 
                 GCCGCGCCAT CTGCCACCAG CCAGCTATCA ACTCGCGCCC TGGAAGGGAT TTTTGAAGCA 
               
               
                  3541 
                 ACTCATCGAT TGATTTACGG CGCTAAGGAT GACTCTGGTC AGAGATACCT GGCCTGGTCT 
               
               
                  3601 
                 GGACACAGTG CCCGTGTCGG AGCCGCGCGA GATATGGCCC GCGCTGGAGT TTCAATACCG 
               
               
                  3661 
                 GAGATCATGC AAGCTGGTGG CTGGACCAAT GTAAATATTG TCATGAACTA TATCCGTAAC 
               
               
                  3721 
                 CTGGATAGTG AAACAGGGGC AATGGTGCGC CTGCTGGAAG ATGGCGATCT CGAGCCATCT 
               
               
                  3781 
                 GCTGGAGACA TGAGAGCTGC CAACCTTTGG CCAAGCCCGC TCATGATCAA ACGCTCTAAG 
               
               
                  3841 
                 AAGAACAGCC TGGCCTTGTC CCTGACGGCC GACCAGATGG TCAGTGCCTT GTTGGATGCT 
               
               
                  3901 
                 GAGCCCCCCA TACTCTATTC CGAGTATGAT CCTACCAGAC CCTTCAGTGA AGCTTCGATG 
               
               
                  3961 
                 ATGGGCTTAC TGACCAACCT GGCAGACAGG GAGCTGGTTC ACATGATCAA CTGGGCGAAG 
               
               
                  4021 
                 AGGGTGCCAG GCTTTGTGGA TTTGACCCTC CATGATCAGG TCCACCTTCT AGAATGTGCC 
               
               
                  4081 
                 TGGCTAGAGA TCCTGATGAT TGGTCTCGTC TGGCGCTCCA TGGAGCACCC AGGGAAGCTA 
               
               
                  4141 
                 CTGTTTGCTC CTAACTTGCT CTTGGACAGG AACCAGGGAA AATGTGTAGA GGGCATGGTG 
               
               
                  4201 
                 GAGATCTTCG ACATGCTGCT GGCTACATCA TCTCGGTTCC GCATGATGAA TCTGCAGGGA 
               
               
                  4261 
                 GAGGAGTTTG TGTGCCTCAA ATCTATTATT TTGCTTAATT CTGGAGTGTA CACATTTCTG 
               
               
                  4321 
                 TCCAGCACCC TGAAGTCTCT GGAAGAGAAG GACCATATCC ACCGAGTCCT GGACAAGATC 
               
               
                  4381 
                 ACAGACACTT TGATCCACCT GATGGCCAAG GCAGGCCTGA CCCTGCAGCA GCAGCACCAG 
               
               
                  4441 
                 CGGCTGGCCC AGCTCCTCCT CATCCTCTCC CACATCAGGC ACATGAGTAA CAAAGGCATG 
               
               
                  4501 
                 GAGCATCTGT ACAGCATGAA GTGCAAGAAC GTGGTGCCCC TCTATGACCT GCTGCTGGAG 
               
               
                  4561 
                 ATGCTGGACG CCCACCGCCT ACATGCGCCC ACTAGCCGTG GAGGGGCATC CGTGGAGGAG 
               
               
                  4621 
                 ACGGACCAAA GCCACTTGGC CACTGCGGGC TCTACTTCAT CGCATTCCTT GCAAAAGTAT 
               
               
                  4681 
                 TACATCACGG GGGAGGCAGA GGGTTTCCCT GCCACAGTCT GAGAGCTCCC TGGCGGAATT 
               
               
                  4741 
                 CGGATCTTAT TAAAGCAGAA CTTGTTTATT GCAGCTTATA ATGGTTACAA ATAAAGCAAT 
               
               
                  4801 
                 AGCATCACAA ATTTCACAAA TAAAGCATTT TTTTCACTGC ATTCTAGTTG TGGTTTGTCC 
               
               
                  4861 
                 AAACTCATCA ATGTATCTTA TCATGTCTGG TCGAGATCTA AGGAAGACCC TGAATTCTGT 
               
               
                  4921 
                 TCTCATACTC CATACCCCAT ATCTTTCTTC CTCTGTGTCT TCCTTGCCCT TAAAGAAATT 
               
               
                  4981 
                 GCAGCATTCC AAGAACAATA TCTGTACAAA GGGGGAAATG TAAGCATGAG AAAACATTAA 
               
               
                  5041 
                 AAAAAAAAAA CAGTGATGAA CATAACCACA GAGAGAATCC CACCCTTCAA GAATAATTCA 
               
               
                  5101 
                 TGTTTATTTG TGGTGGCAAA TAACAAAATG GTACAACCTT TATCCTTTTC CAGAAACAAA 
               
               
                  5161 
                 AACCAAGGGC ACAGCAACTA GAGTGAGCTG ACAGCTATTT TGGCCTTTTT GGTGGGTCTA 
               
               
                  5221 
                 GCCGTACTTG GGATCCCAGT GGTACATGAC CCTCTGCCGA AGGCTTGCCT CAGTCTGTGT 
               
               
                  5281 
                 ACATAGCACG CCATGTCTGT GGGCAAGCCC AGCACTTTGC GTCAGTGTCG TACTGTATGT 
               
               
                  5341 
                 AATGAACTGT GTTGGTCTCT GTGTTTTTTT TTTCTGAAGA AGAGGAGTAA CTACTCCGGG 
               
               
                  5401 
                 TACCTTGATA TTTGTACAGC CTATAGGCCA ACACTGCGGG CGTGTGACTC TTTATTGAAA 
               
               
                  5461 
                 AACAAAAACA AAAAAATACC AGTGTGGTGA TGATAGTGTG TGTATATATA TATAAGGTTA 
               
               
                  5521 
                 TATGGGGAAG ATTTCTAAAT AAAAGTTTTA CAAAGGGGCC TGGACTTTGT ACTTGGACTT 
               
               
                  5581 
                 TGCCCCCTAG AGTCTGAGAA TGGGAACATC AAGGGGAAAG GCTGACAGCT TTTAGGAAGT 
               
               
                  5641 
                 AGGATCTAGC TTCCAGTCTC AGCCTGTCGG GGAGGAAGGA GGCTACCCTA TGGGGGGGTT 
               
               
                  5701 
                 TCCTTTTCCC CCCTTCTGCA AGGCTCCAAG GGCTTCAGTA TCCTGTCCTT GTGTTTGCAG 
               
               
                  5761 
                 CCCTAGACAG CCTAGACCTC TCTGTGTAGG GTCAGCTTTC TCCTTGTTAG ATCACTTTCC 
               
               
                  5821 
                 CAAGTTGGGA CCATTGCTCC CAGTGAGAGC TTAGGACAGA AAAATGTAGC TGTTATCCAC 
               
               
                  5881 
                 CATTGGTGTC CATAGATTTC CTGATGACTC AGTGGGGGTT GCATCTTTTA CACTTGACTT 
               
               
                  5941 
                 TTTTTTTAAA GGTTAAAAAA TATTTTATGT ATAGAGATGT TTTATGTGTA TAGGTACAGT 
               
               
                  6001 
                 GCCCACAGAT GCCAGAAAAG GGAGTCGGAT TCCCTGGAAC TGGAGTTGCA AACCGTTGTG 
               
               
                  6061 
                 AGTTGCCCTG TAGGTGCTGG AGTTTCATGA ATAGAATTTG GGAAAGAGAC TGGGTCTTGG 
               
               
                  6121 
                 GGAGGCCATT ATGCATGGAC GTTTGGTCTC CTGGGAGTTT GTAAGCTGGG CATCTTCTGT 
               
               
                  6181 
                 CTTCTCATTT AACAAGCATT TGCTGAGCTC CTGCTCTGGG CAGACACTGT TCTGTTGGGG 
               
               
                  6241 
                 AGGGTTCAGC ATTGAATGAA ACAAGCATGG ATGCTCTCCA CTGCACCTTA CATTTTAGCA 
               
               
                  6301 
                 GGGGGATGTT GAATGCAGAA ACACATACAA GTAGAGTTAA ATAGTTAGAA AGCAAATTAG 
               
               
                  6361 
                 TATTAACCCA CAGTGAGTTT TATTCAGGCC AGCCTGGGCT ACAGTCTCAA AAACCAAAGC 
               
               
                  6421 
                 CAAGAAAGGT GGTAAGGAAC AAAAGTGGGC AGATCAACAG GGATAGTTCA GGAAGGCCC 
               
               
                  6481 
                 TAGGGTGCCA TCTTTTTCAT TCAGGATCAG ATGATTCCTG GTGTCAGAGA CAGTTTTGTC 
               
               
                  6541 
                 CCAGGGACAG GTTGGGTCTT TCTATCTACA TGCCCTGAGA TGGCTTTTTT CTTTCTTCTT 
               
               
                  6601 
                 CTCTGGACCT CAGTACTCAA CCCCAAATCT ACAGACATGG ACTAGCTCAG ATTCAACAAT 
               
               
                  6661 
                 TGGGAGGGAA TTCAATAGTC TCACCGTTAA TTCCCAGCTG GCCTGTCTCT AGTCTCAGCT 
               
               
                  6721 
                 GTGTTTTGTC CTCTTAGCTT CTATCCATCT ACAGGGAGAG GGTAGGATTC AGCCTGAGTG 
               
               
                  6781 
                 TCAATATCTG ATCCAGCTAC TGGGAAGCTC CTCAGATATG CCTCTCTTTG GCCTAGGACA 
               
               
                  6841 
                 AGGATGGTAG GATTTGGCCT TGGGGAGGGG AGAAAAATGG ATATTTAGGC TTATAGACCT 
               
               
                  6901 
                 GAGGAACTAT CATGATAGGA GAGAAAGAAA GAGGACAGAG AAGGAAGAAT GTGTTTGGG 
               
               
                  6961 
                 GTGGAGGAAG TGGCCAGTAT GCTCAGTACA ACTGAGGGGC CATGCACGGA AAGGCTGAGT 
               
               
                  7021 
                 TAACTGGTTT GAGGCAGCTG GTGACTGGAA AGAGCTGCAG AGAGGAGTGA ATAGAGGTAG 
               
               
                  7081 
                 TGACCTGAGG ACTCAGAGAT GTCACTTCCC ATCTTGTAAG ATTTTCCTCA GGAGAAATGA 
               
               
                  7141 
                 AGCTTTCCAT GTAATGGTGA CAAAGAGAGC CCGAGGATTC TGATCACTCC CGGAGTTCAT 
               
               
                  7201 
                 CGATGGGGCA GAGACCCAGA GAGAAAATGT CTTCTCAAGC CTTGTATCTC AGAGTGGTGT 
               
               
                  7261 
                 GTAGGCAGGC CCATTCTCCC TGTCCCAAGA AAATGTTGTC TCTGAAGCCC AGAATCCCTG 
               
               
                  7321 
                 ACTCCACAAG GGAAGAAAAG TGCCCTGAGG CCTGGCCTGA GGTGTTTTGC TGATCTGTTC 
               
               
                  7381 
                 CCCTTTATTT CTTACCACTC CATTTGTGTG TGTGTGTGTG TGTGTGTGTG TGTTTGCTTA 
               
               
                  7441 
                 TTTGTTTTTC TGAAACAGGG TCTCATGTGG CCTCAAACCC ACTAAGTTGC TGAGGCTGAC 
               
               
                  7501 
                 TTTGAACTTC CGATCTTCCT GCCTCTGTCT CCAGAGTGCT GGGATTACAG GTGTGCACTA 
               
               
                  7561 
                 GAATACCAGG TTTATTCAGT GACAGGAGCT AAATCCAGAG CTTTGTGCAT ATTAGGCAAG 
               
               
                  7621 
                 CACTCTACAA CCAGACTGCA TCCCCACCCC ACGCCTCACT CTTTTGTGCC TACCGTACTA 
               
               
                  7681 
                 GCTTTCTTCC TTTTTGTTTT AGACTGTTTT ATTGGTTTTT GACTCCCAGA TGTTGAATTT 
               
               
                  7741 
                 TGGTTTATTT TTCACATAAC AGCCCATCTT CCTCTTTGCC CACTCTCATT TGGTTGAATT 
               
               
                  7801 
                 GTCCCTGAAG TCCAGGAAGT TTTCCTGACT CCATGGGACT GGGTGCCTCC TTTGCATCCC 
               
               
                  7861 
                 CATGGGACCC CAGGTATGCT GGCCCTTCCT GCCCTAACAT TTGCTTATTT AGTTGCTTCT 
               
               
                  7921 
                 TCACTGAAAC ACAAACCCCT CAGAGCTGAA ACCAAGTCTG ATTAAGCCCT CTGCACCAGC 
               
               
                  7981 
                 ACCTTAGGGT ACAGACACTC GGTTCTTTCC CCACTGGCCA TGAACAGCCC TTCTCCTCCC 
               
               
                  8041 
                 ACTGGCTCTC TATTTTCTCT CTGGGCCTGG CGTCTGACCT GGCATCTGGC AAGGACCTGA 
               
               
                  8101 
                 AAGGCTGGTA TAGAGTGGTG AAGACCAGGC ATGGAGGCTA TGGATCCAGT CAGCTGTCTG 
               
               
                  8161 
                 GCCTCCTCAC GCCGGTCCCT ACCTGCTTCC TTTTTAATAA AATAAGTGTG TGTTCCTCAG 
               
               
                  8221 
                 AAGCTGTCAC TGTGTCATTA GCTTCCTCGC ACCCCCTACC CGGACACACC CCCCTGCCCA 
               
               
                  8281 
                 TGTAAACCTG TTACCTATTC ACAGAGCTTA ATTGTCATGA ATCTAAGTAA AGGGTTACCC 
               
               
                  8341 
                 AGGGGAGGTG ACACAAAGCC CTGAGTTGGA AGGGGCTTGA GCAAGGTGAA GTAGGTGTGA 
               
               
                  8401 
                 ATTCAGGGCG ACACCCAAGG TTAGAGATCC AGACCACATA GGAAGGTCAG GAAATAGAAG 
               
               
                  8461 
                 AGGAGGCCAG TAGACAGCTA GAGTTCATAG AGAAAATGGC TTTACTTTCC TTATGGGCAA 
               
               
                  8521 
                 GAGGGCTACA CAAATTTAGG CCCAGGACAG GTGGTGGTAG TGAAGAGCTT GCTGGCTGGA 
               
               
                  8581 
                 GGACTGGCTC TGTGGATGAC CATGGGGACA GTGAGGAAGG ACAGTTGGTG TGGAACAGTT 
               
               
                  8641 
                 GGTGAAGGGA GTAACTGGGG CCTGGGTGGA AGTGAGAAGA AAAGAGCAGC CAGGCTCTGG 
               
               
                  8701 
                 AGGAGCTTGG CCTGGTCAGA ATCACTTGGG GCTTAAGGGC TTAAGTATTG CTACTGGGTG 
               
               
                  8761 
                 TGCTGGCTTG TGACTTTGAG TGAGTCACTA TCATTCTGAG GTTTGGTTTC TTTATCTGTT 
               
               
                  8821 
                 AAACAGAGAT GTTAACAGTC ATCTTCCAGG ACTGTCATGG GACTTCAGCA TAATATATGC 
               
               
                  8881 
                 AAAGTATCTG TGTTTCATTA AAAAATGATT CTATAGAAAG AGCTACGGAA ATATCTATAA 
               
               
                  8941 
                 GAAAGCATTC TTTTTCCAAG AAACAGGACC AGGAGGGATG GGACTGTCCT AACAGAAGAG 
               
               
                  9001 
                 ACGAGGGAAG GACATGAGTG TGAGGGAATA TTAATCCCTC ACTCAACAGC AGGACTTTTG 
               
               
                  9061 
                 TGTGCCTGTC TTATGTCAGG AAAGGAGGGG TAGCCAGTCT TGACCACCCA TTTTGACTTC 
               
               
                  9121 
                 AGAGGCTGGA GAGCAGAGTG GAAGCTGGGA ATAGGAAGGA ATCCTAGAGG CAAGTGCTAT 
               
               
                  9181 
                 GGGAGGAGCT TAGTGGTGTG GTGTGGGCAG CCTAGCTCTG ACAGTAAAGT CCCTGAGCAA 
               
               
                  9241 
                 GTTGTGCTGA ACTGAACTGT CCTGAGGGGC AAGGTTGGGA GGTATCTGGG AGATTTCACA 
               
               
                  9301 
                 TTCTGTCTTG AGCATTACCT AGTTTTCAGT GGTGGAGCGG GCTGGTCCAG GAATGCTGGC 
               
               
                  9361 
                 TTCCTCCTGG GCCCCATACT CTTGCCAAGG CTACCTGGGG TGAGGCAATG CTCCCCCACC 
               
               
                  9421 
                 TCACTTTGCC TTCCAGCTCC TACTTAAGCT CTCCCCACTG GTTTGCTCTG AGGCCTGCCC 
               
               
                  9481 
                 CTCCCCAGCT CCTGGGCTTT CTCTCCACAC AATAACAGGA TGTGATCTTC GAAGAGAGGA 
               
               
                  9541 
                 AGTGGGGGAG GACTGCTGTG CCGATAGCAG GGAAGGAGGG GGGCTTCTGA CTCTCCCCTC 
               
               
                  9601 
                 TCCAGCCCTC CTTTGCTCTG TAGGCCAGCC CCTGCAGCTC CTTGATCCCC CTAAGCCCTA 
               
               
                  9661 
                 CCTCAAGCTT CTATCTGAGA CAAGTAGGGA TGAAGGGTCT TTAGGCCCAT GTAGGACTGC 
               
               
                  9721 
                 TTGCCTATGG AGAGACATGC CTTGGCCACA CCGTCTTCAG GATCTACCTT CTGGAGAGAC 
               
               
                  9781 
                 TTGCTGGCCT AGCTTTAGAT GCTGGGTTGT TTTCTGCCCG GAGCTGCTGG AGTCTAAGGG 
               
               
                  9841 
                 TGGGCAGGTG GGTCATTCTG TAGGGCTCCA TCTGTCCAGT GCACTCCCAA GTCCACACGA 
               
               
                  9901 
                 GCATGATTCA GTGCAGGGAG TGCGTGATAG CATCAATCTA AAGGTCTATG TCAAATGCTG 
               
               
                  9961 
                 GTTTGGCTTG CACAGTGTGT GTCAGGCTGC AAAAATGGAC AGTGAAATCC AGAAAGACAA 
               
               
                 10021 
                 GGAGCATGAG GAAGGAGCAA GGCTAGGCTG GAACCCAGCA CTAGGTCATT GGGTTACCGC 
               
               
                 10081 
                 CTCTTCGAGC CAGGGATGTT CTTAGAACTT CCAAAGTTGA TGGGAAAGTT TTAGATCGAG 
               
               
                 10141 
                 TCGACCGATG CCCTTGAGAG CCTTCAACCC AGTCAGCTCC TTCCGGTGGG CGCGGGGCAT 
               
               
                 10201 
                 GACTATCGTC GCCGCACTTA TGACTGTCTT CTTTATCATG CAACTCGTAG GACAGGTGCC 
               
               
                 10261 
                 GGCAGCGCTC TTCCGCTTCC TCGCTCACTG ACTCGCTGCG CTCGGTCGTT CGGCTGCGGC 
               
               
                 10321 
                 GAGCGGTATC AGCTCACTCA AAGGCGGTAA TACGGTTATC CACAGAATCA GGGGATAACG 
               
               
                 10381 
                 CAGGAAAGAA CATGTGAGCA AAAGGCCAGC AAAAGGCCAG GAACCGTAAA AGGCCGCGT 
               
               
                 10441 
                 TGCTGGCGTT TTTCCATAGG CTCCGCCCCC CTGACGAGCA TCACAAAAAT CGACGCTCAA 
               
               
                 10501 
                 GTCAGAGGTG GCGAAACCCG ACAGGACTAT AAAGATACCA GGCGTTTCCC CCTGGAAGCT 
               
               
                 10561 
                 CCCTCGTGCG CTCTCCTGTT CCGACCCTGC CGCTTACCGG ATACCTGTCC GCCTTTCTCC 
               
               
                 10621 
                 CTTCGGGAAG CGTGGCGCTT TCTCATAGCT CACGCTGTAG GTATCTCAGT TCGGTGTAGG 
               
               
                 10681 
                 TCGTTCGCTC CAAGCTGGGC TGTGTGCACG AACCCCCCGT TCAGCCCGAC CGCTGCGCCT 
               
               
                 10741 
                 TATCCGGTAA CTATCGTCTT GAGTCCAACC CGGTAAGACA CGACTTATCG CCACTGGCAG 
               
               
                 10801 
                 CAGCCACTGG TAACAGGATT AGCAGAGCGA GGTATGTAGG CGGTGCTACA GAGTTCTTGA 
               
               
                 10861 
                 AGTGGTGGCC TAACTACGGC TACACTAGAA GAACAGTATT TGGTATCTGC GCTCTGCTGA 
               
               
                 10921 
                 AGCCAGTTAC CTTCGGAAAA AGAGTTGGTA GCTCTTGATC CGGCAAACAA ACCACCGCTG 
               
               
                 10981 
                 GTAGCGGTGG TTTTTTTGTT TGCAAGCAGC AGATTACGCG CAGAAAAAAA GGATCTCAAG 
               
               
                 11041 
                 AAGATCCTTT GATCTTTTCT ACGGGGTCTG ACGCTCAGTG GAACGAAAAC TCACGTTAAG 
               
               
                 11101 
                 GGATTTTGGT CATGAGATTA TCAAAAAGGA TCTTCACCTA GATCCTTTTA AATTAAAAAT 
               
               
                 11161 
                 GAAGTTTTAA ATCAATCTAA AGTATATATG AGTAAACTTG GTCTGACAGT TACCAATGCT 
               
               
                 11221 
                 TAATCAGTGA GGCACCTATC TCAGCGATCT GTCTATTTCG TTCATCCATA GTTGCCTGAC 
               
               
                 11281 
                 TCCCCGTCGT GTAGATAACT ACGATACGGG AGGGCTTACC ATCTGGCCCC AGTGCTGCAA 
               
               
                 11341 
                 TGATACCGCG AGACCCACGC TCACCGGCTC CAGATTTATC AGCAATAAAC CAGCCAGCCG 
               
               
                 11401 
                 GAAGGGCCGA GCGCAGAAGT GGTCCTGCAA CTTTATCCGC CTCCATCCAG TCTATTAATT 
               
               
                 11461 
                 GTTGCCGGGA AGCTAGAGTA AGTAGTTCGC CAGTTAATAG TTTGCGCAAC GTTGTTGCCA 
               
               
                 11521 
                 TTGCTACAGG CATCGTGGTG TCACGCTCGT CGTTTGGTAT GGCTTCATTC AGCTCCGGTT 
               
               
                 11581 
                 CCCAACGATC AAGGCGAGTT ACATGATCCC CCATGTTGTG CAAAAAAGCG GTTAGCTCCT 
               
               
                 11641 
                 TCGGTCCTCC GATCGTTGTC AGAAGTAAGT TGGCCGCAGT GTTATCACTC ATGGTTATGG 
               
               
                 11701 
                 CAGCACTGCA TAATTCTCTT ACTGTCATGC CATCCGTAAG ATGCTTTTCT GTGACTGGTG 
               
               
                 11761 
                 AGTACTCAAC CAAGTCATTC TGAGAATAGT GTATGCGGCG ACCGAGTTGC TCTTGCCCGG 
               
               
                 11821 
                 CGTCAATACG GGATAATACC GCGCCACATA GCAGAACTTT AAAAGTGCTC ATCATTGGAA 
               
               
                 11881 
                 AACGTTCTTC GGGGCGAAAA CTCTCAAGGA TCTTACCGCT GTTGAGATCC AGTTCGATGT 
               
               
                 11941 
                 AACCCACTCG TGCACCCAAC TGATCTTCAG CATCTTTTAC TTTCACCAGC GTTTCTGGGT 
               
               
                 12001 
                 GAGCAAAAAC AGGAAGGCAA AATGCCGCAA AAAAGGGAAT AAGGGCGACA GGAAATGTT 
               
               
                 12061 
                 GAATACTCAT ACTCTTCCTT TTTCAATATT ATTGAAGCAT TTATCAGGGT TATTGTCTCA 
               
               
                 12121 
                 TGAGCGGATA CATATTTGAA TGTATTTAGA AAAATAAACA AATAGGGGTT CCGCGCACAT 
               
               
                 12181 
                 TTCCCCGAAA AGTGCCACCT GACGCGCCCT GTAGCGGCGC ATTAAGCGCG GCGGGTGTGG 
               
               
                 12241 
                 TGGTTACGCG CAGCGTGACC GCTACACTTG CCAGCGCCCT AGCGCCCGCT CCTTTCGCTT 
               
               
                 12301 
                 TCTTCCCTTC CTTTCTCGCC ACGTTCGCCG GCTTTCCCCG TCAAGCTCTA AATCGGGGGC 
               
               
                 12361 
                 TCCCTTTAGG GTTCCGATTT AGTGCTTTAC GGCACCTCGA CCCCAAAAAA CTTGATTAGG 
               
               
                 12421 
                 GTGATGGTTC ACGTAGTGGG CCATCGCCCT GATAGACGGT TTTTCGCCCT TTGACGTTGG 
               
               
                 12481 
                 AGTCCACGTT CTTTAATAGT GGACTCTTGT TCCAAACTGG AACAACACTC AACCCTATCT 
               
               
                 12541 
                 CGGTCTATTC TTTTGATTTA TAAGGGATTT TGCCGATTTC GGCCTATTGG TTAAAAAATG 
               
               
                 12601 
                 AGCTGATTTA ACAAAAATTT AACGCGAATT TTAACAAAAT ATTAACGCTT ACAATTTGCC 
               
               
                 12661 
                 ATTCGCCATT CAGGCTGCGC AACTGTTGGG AAGGGCGATC GGTGCGGGCC TCTTCGCTAT 
               
               
                 12721 
                 TACGCCAGCC CAAGCTACCA TGATAAGTAA GTAATATTAA GGTACGTGGA GGTTTTACTT 
               
               
                 12781 
                 GCTTTAAAAA CCTCCCACAC CTCCCCCTGA ACCTGAAACA TAAAATGAAT GCAATTGTTG 
               
               
                 12841 
                 TTGTTAACTT GTTTATTGCA GCTTATAATG GTTACAAATA AAGCAATAGC ATCACAAATT 
               
               
                 12901 
                 TCACAAATAA AGCATTTTTT TCACTGCATT CTAGTTGTGG TTTGTCCAAA CTCATCAATG 
               
               
                 12961 
                 TATCTTATGG TACTGTAACT GAGCTAACAT AACCCGGGAG GTACCGAGCT CTTACGCGTG 
               
               
                 13021 
                 CTAGCTCGAG ATC 
               
               
                 // 
               
               
                   
               
             
          
           
               
                 SEQUENCE NO. 3 
                   
               
             
          
           
               
                 Endothelial-CRE-ERt 
                   
               
               
                 LOCUS Endo-pGL2-Promoter 12191 bp DNA circular SYN 12-DEC.-2007 
               
               
                 DEFINITION. 
               
               
                 ACCESSION. 
               
               
                 KEYWORDS. 
               
               
                 FEATURES Location/Qualifiers 
               
               
                 BASE COUNT 3021 a 3013 c 2707 g 3451 t 
               
               
                   
               
             
          
           
               
                 ORIGIN 
                   
                   
               
               
                     1 
                 CCCGGGAGGT ACCCTGGGCT ACACAGAGAT AGATGTCTTT TGCCACAGCT TCTCCTGGCA 
               
               
                    61 
                 ACCCAAAGCT ACCTGGCAGA GTCCAGTCTG CCTAACACCT ATGAATCTAT GAGATACCTT 
               
               
                   121 
                 AAAAAGCATA TCCTTCTTCT ATACATCTTT CCACTTCCTC CTCTTCTCCA CCCTATTCAT 
               
               
                   181 
                 CAGACAACTG TCTCAGTCAG TGGGGAACAT GAAGAGGGGA TATGGATGCT TGCTTTCACA 
               
               
                   241 
                 GGTGCCTCTG CATAAAGGGA GTTCTCAGTG AGCTGGAGCA GAGGCTATGG AGGAAGGAAG 
               
               
                   301 
                 CAGAGATGAC AGATTAAAGA CAGATGCAGA GACAAGGCCT TTATACAGGA AAGAGGAGCA 
               
               
                   361 
                 GATTCAGAGT TTGCCTGAGC CTAAGATAGA GACCGGAGAA ATGAAAGGCA GAGTGAGCAA 
               
               
                   421 
                 GATAAGAGAT GAAAGGAATA GACCCCGGGG TTCCTCCACT GCATCCTCAT TACAGATAGG 
               
               
                   481 
                 AAAACTGAAG GTCAGAGGAA GTGGTGGTTC TACTTCCTAC GGATGTATCC ATCACTCTTG 
               
               
                   541 
                 TAAATACACT GGGTCAGGTC CTCCCTTCTC CAGCACTTTC CTCTTGCCCT TGTGCACTAG 
               
               
                   601 
                 GACTGAGTAA CACAAGTGAC ACCCAGTGGG AGGCTCTTGG ACAAGTCAAC CAGGAAGAGG 
               
               
                   661 
                 GAGAGAGGAG ACAGTGTAGA CAAAATAGAT TGACAGGGAA GTTTTTCTGG ACTAGGTAAG 
               
               
                   721 
                 CTTGAAGAAG GGCACAGAGG GTGTAAACAA CTGTATTAAG GTATATGGTT TATGTGCAGT 
               
               
                   781 
                 AAGATATACC ATTTCAGCAT CCAGTTAGCT GAGGTTTGAC AACACTTTTA TAGTCATTAC 
               
               
                   841 
                 CACAATCAAT GTATAGAATA TGTCTTTGAT AGAAAGTTCC GTTGTGTCTC TTGTATCCAC 
               
               
                   901 
                 TCCATGTTGC ATTTTATCCT TCTGAACTCT GTCATTGTAG ATTCATTTTG CTTTTCTTCC 
               
               
                   961 
                 CCTAGGGTTT CATGTACATG TAGCTAGTTA CTATAGACAC TGTCTGATCG CTTTCACTCT 
               
               
                  1021 
                 GCATGAATTG GAGGTTTTCT ATGTTTTTAC ATGTATCTGA AGTTATTTTT ATTGCTCAGC 
               
               
                  1081 
                 CATATCCAGT TGCATGCCTG GACCAATATA TGTACCTATC TGTTGATATG CTTTGAAAGT 
               
               
                  1141 
                 ATTTCTACTT TTTTTTTGTT TGTTTTTTGT TTTGTTTTTG AGACTATGTG GCTCTGGCTG 
               
               
                  1201 
                 TCCTGGAACT CGAAATATAG GCCAGGCTGG TGTGGCACTC ACAGAGACTC ACCTGCTTCT 
               
               
                  1261 
                 GTCTCTCACA TTCTGGGATT AAAGGTGTGG ACCATTATGC CCTGCTTATT TATTTTTTGA 
               
               
                  1321 
                 TGCTATGGAT AAGTCTTTAT ATGCATATAT TCCTGTGGAT ATGTTTTTAT GTCTCTCAAT 
               
               
                  1381 
                 AAATATGTAT TATTAACCAT ATGATTAAAG GGGCTCAAGT GTGTGTATTT TTCTCCAATA 
               
               
                  1441 
                 TGGCTGAGTA GAGTTTGCAT TCCCTTCAGT GTACAAGAAT GCCAGTTGAC CTTTATTCCT 
               
               
                  1501 
                 ATCAACACTT GGTCTTGTCT GTCTTTCACG TTCTAGCCAT ATTGACAGAC TTGTCATGGT 
               
               
                  1561 
                 AGCTCTTTGT AGTTTGAATT TCTGTTTCTC TGGTGTAACT TTCTTTTCAT GAAAGTTTTG 
               
               
                  1621 
                 TTTTCTAAGT TCATAAGGAT TTTCAACTCA GACACATTAC AGTGATACTC ACTGTTCGGC 
               
               
                  1681 
                 TGAAGATTTT AAAGTAGTTC ACACAGGAAG AAATGTCATA TAGCCAACAG GGGTGGAGAG 
               
               
                  1741 
                 GACAATAGGC CATGTTGTTC TAGGCTACAC AGCAGTTAAA TGACAAGAGT GAGCCTGCTT 
               
               
                  1801 
                 TCTCACCTCC AAAGTAGCGT CACCAGGCGG CATGACACTG TCATTGTCTA CAGTCAGATG 
               
               
                  1861 
                 ACAGGTGGAC ACAAGGGCAG AAGAGGTACA CACAGAGAGA TGCTCAGTAC ATGCATGTGC 
               
               
                  1921 
                 AGGGCCTGGA GGCATATCTA CTGTCTTGAT GTGTGTCATA AACCTGGCCA CTGTCCTGAT 
               
               
                  1981 
                 GACCATCGGC AGCTATTTGC GACAGAGTTG GTGGTTGTGC GTGTATTGTC TTCTAATGGC 
               
               
                  2041 
                 TTGAACAAGT AAAACATTAA TGGCAGAATG CTCTCTCCTG AGGACAGAAA GCTTGGGAAC 
               
               
                  2101 
                 ACAAACTGGG GACACAGCTT TGGTCCTCTG TGTACTTCTA GAAGATGCAT AGGTTGCACA 
               
               
                  2161 
                 AGGAAGATAG GAGGCTAGAG AGCCCGCTGC CTTCTGCAGC TGCTCATTCA TTTTGCTTTG 
               
               
                  2221 
                 GATTTTTTCC TTTCATTTCT CTTTCTTTCT TTCTTTCTTT CTTCCTTTCT CACCAATGGT 
               
               
                  2281 
                 GCTCTAGTTC TTAAGCTGTG TGCTGCAGAC ATCATCCTGG AGGCTGGTGA AACACACCTG 
               
               
                  2341 
                 GCCTCTCTTC CAGAGGAGCC TAGGGTCCCC TTCCAGAACT GACTTCTCTA AGGACATGGC 
               
               
                  2401 
                 CCCTCCTTTG AAAGTCATAC ATTAGAGCAA AGCCCTTTCC ATCCCTGCAA ATGCTGATGG 
               
               
                  2461 
                 CAAGGCTGGG ATAAGAACAT GGAAATGATT TCATCTGTGG GGTTCTGGGC TCAGCCTTGC 
               
               
                  2521 
                 AAACTAGAAT GGCAGGGGCT CATTCCTAGT AAGGAACAGA GGCAAAATAT GGAGGACAGT 
               
               
                  2581 
                 TATATGGAAA TGAATTGGAG CAGGTTATGA CATCTCCTTA AATGGGCATA TTTACCATCA 
               
               
                  2641 
                 ATAAGTTTTA TAAAACCCAC TGTCAGGTAT GGGCAATTAT CACCTCCTCT TTACAGAGGA 
               
               
                  2701 
                 GGAAAATGGA AGAGGCTATC TTGCCTATGC TCATGCAGCC CAGTGAGAAA GCAGGAATGA 
               
               
                  2761 
                 GGGCTCAGAC ATGCTAGTCA ATGGTTCTGC TCTGCTGCCT GGAGGCACCA GAATGTCCCG 
               
               
                  2821 
                 GCTGGGAATT CTTTATTCAC AGCAAGTTGC TTAGATGTCT GAGCTATCTA CTAAGTGGAA 
               
               
                  2881 
                 GTCCCGACCT TCCCTACGTC TTTGAGCTGT TGTAAAATGA ACGGAATTGA CATTATGAAG 
               
               
                  2941 
                 TGTTTAGGTC TGGCACGATA CAAATTCGTT ATAAACCCAT CTGCCCACCA GAGTGCTGGC 
               
               
                  3001 
                 AGACCGAACT TCTCCAGGGG TGGAAGCTCA GAGATGGTAC AGCACCTGAA AACATTGCAA 
               
               
                  3061 
                 ACCCTGGACT CTGGAGGGCG GACAACGTAG GCCCTGGGAG TGGAGGAGCC TGTCCCCTGC 
               
               
                  3121 
                 TCTTGCCTAC CCGGGGCCAG ACTCCAGACT CCCTGGTTCC TCACCTCCCC GCCCCCTCAC 
               
               
                  3181 
                 CACCCCCACC GAGGCGCTCC GAATTTCCTG CCCGACCGAG GCCCGGCTCG GGCGGGTGGA 
               
               
                  3241 
                 GGAGGGCTGG CATTTCCTGG CCGCCGCGTC ACTGGCTCAG CGGTGCTCGG ACAAAGCGCT 
               
               
                  3301 
                 GACCGACAGG CACCAGAAGC TATTTCAGGC GGCGCCCAGC TTAGCGCGCA GTTTCCGTTT 
               
               
                  3361 
                 TTCCACCGTC GGAAACAGGG AACAGGGAGC TTGCAGACGT CACAAACCCC CAGCCTCAGG 
               
               
                  3421 
                 CGTGGGTCCA GGGACCAGGA GAGGCAAGGC CCATGTGTTA GAAACAGGGT AGAGGCAGAC 
               
               
                  3481 
                 GCTATCCCCG CACCTTCTAT CCAACCTTAC TCCTTAACTG TCCTTGGAAA CACCAGAGAA 
               
               
                  3541 
                 GGCCATTTCA CACCCAGGAA AATGATCCAG TCGTCGTTGG TCAAGCCAAA TGCATAACCT 
               
               
                  3601 
                 TTTCAAGCCC ATAAACCTCG AGACAGCCTT ACCCCATTCC CTCTCCTGAA TTAACTAACC 
               
               
                  3661 
                 TGCCCCCAGA CATCCTGGAT TCTTCGATTT TCATTATTCA ACGGCGTCGT AGTTCTTCCA 
               
               
                  3721 
                 AACTCAGTCT TAAATACCCT GTGCGAAACA TCTACCCCAC ACCTTCTCTT CCATCTCCTG 
               
               
                  3781 
                 GAAGGAGAAT TAGAACAAGC TCTAACCTCT TTTCTCTGGT CACAGAACAC TTAGCCTTCA 
               
               
                  3841 
                 CCTCCCAGCT CCCCACACCA ACACAGCCCC TACCGCCATT TCAACCCAAG GCTTTCCTTT 
               
               
                  3901 
                 CCTTTCCTTT CCTTTCCTTT CCTTTCCTTT CCTTTCCTTT CCTTTCCTTT CCTTTCCTTT 
               
               
                  3961 
                 CCTTTCCTTT CCTTTCCTTT CCTTTTTCCT TTTTTATTAG ATATTTTCTT TATTTACATT 
               
               
                  4021 
                 TCAAATGTTA TCCCTTTCCT AGTTTCCCCT CCGAAAGTCC CCTATCCCTT CCCCCTCCCC 
               
               
                  4081 
                 CTCCCCCTGC TCCCCAACCC ACCCACTCCT GCTTCCTGGC CCTGGCATTC CCCTATACTG 
               
               
                  4141 
                 GGGCATAGAG CCTTCACAGG ACCAAGGGCC TCTCCTCCCA TTGATGACCA ATTAGGCCAT 
               
               
                  4201 
                 ACTCATGGCT CTAACTGCAT ACTGCATATG CAGTTAGAGC CATGAGTCCC ACCATGTGTT 
               
               
                  4261 
                 TTCTTTGATT GGTGGTTTAG TCCCAGGGAG CTCTGGGGGC ACTGGTTAGT TCATATTGTT 
               
               
                  4321 
                 GTTCCTCCTA TGGGGCTGCA AACCCCTTCA GCTCCTTGGA TACTTTCTCT AGTTCCTTCA 
               
               
                  4381 
                 TTCCAAGGGT TTTCTAAAAA AGCAAATCCG ATCTTACATA GGACAGCAAG CCCTTATGTA 
               
               
                  4441 
                 AACACAGTGG TAAAAACAAA ACCCTCAATT CTTCCACCCA TACTGTACCA GTTTTCTGTT 
               
               
                  4501 
                 TCTTACATTA ACTTTCCCCC TTTCTGTGTC AGCCCTTGGT CCAGGACGCC TGGCTTTCCT 
               
               
                  4561 
                 GGGAAGCACA CCCAGTTAGC TCACATACAA TATAGTTAGC CCATATAACC AAGCGAAGGC 
               
               
                  4621 
                 AGCACAGCTG GACTTTCATC AAATGTCACA GAGGAACAGA CAGGTGTAAC TAATACTCCA 
               
               
                  4681 
                 TCTCCAGTAT TGGTCCTGAA ATCTAGGAGG GGCAGAACTC AAAACAGGTG CTACTCTTTG 
               
               
                  4741 
                 GAATCAGCCC TTGACTGAGT CTCAGTCTGT GACCGGGTTC AGAGCTACTG GAAGGTCAAT 
               
               
                  4801 
                 GCAGTTTGGG ATGCTTAGTG GGGTCTATGG AATGGAAATT GAACAAGAGA GTTCAGATAG 
               
               
                  4861 
                 GTGCTGGCGT TACTCTAGCT ATAAGTTTGA TCAAGTTACA TCTCTTTGCC CCCTACTTTC 
               
               
                  4921 
                 CTTTAACATT CTTAATTTCT GTATGGCAAC CAGACAAATG CCTATGATAT CTTATTAGCT 
               
               
                  4981 
                 CCCTACCACC CACTTTTTAT ATTATTTCTC ATGTATATGA AACTTTGGCA TTTAAAATAT 
               
               
                  5041 
                 TATTATTATT CATTTAGGCT TTTTGAGAAA GGGTTTCTCT GTGTAGTCTT GGCTGTCCTG 
               
               
                  5101 
                 AAACTCCATA TGTAGATTAG GCTGGCCTCT AACTCAAAGG ATCTGCCTGC CTGCCTCAAT 
               
               
                  5161 
                 GAGAGCTGGG ATTAAAGGTA TGTGCGTTTA TTCTTTGAGT ATTTTATAAA ATGAATTTTG 
               
               
                  5221 
                 ATCATATTCA CCTCCTATTA CCCCTCATCT CCTCCTATCC TGATGTCTTT TTTTTTTTTT 
               
               
                  5281 
                 TAAATCTACA GAGAGGCTAG AGAGATGGCT CAGTGGTTAA GAGCATTGGT TGCTCTTCTG 
               
               
                  5341 
                 GAGGACCTGG ATTTGATTTC CAGTGTCTAT ATGGTAGCTC ACAAACTCCT GTTTCAGAGG 
               
               
                  5401 
                 AGCTAATGTG TCTGCCTTCT TCTAGTCTCT GGATACAGTA CATAGACATT AATACAGGCA 
               
               
                  5461 
                 AAACCAGGGA CCACACCCTT AAAAATGATT CCCCTTCTCA AGAAGTGTTC AACTGTCAAT 
               
               
                  5521 
                 AGCTCTTCAG TTAGCGGTGA AGGCTCATGA ACACCCCCCC CCCACACACA CACACACACA 
               
               
                  5581 
                 TACACCTACC TTCTGGCTAG AATCTTGACT GGCTTGATCC TGAGCAGGCA ACCACAGCTG 
               
               
                  5641 
                 TGAGTTGGTC CTTTTCTGAC CAGAAGGTAG TCACTCTGGT CTTCCTTAAC TGCTCTTACT 
               
               
                  5701 
                 ATCTTTTTCT TCTTTTCTTC TCTTCTGCTT CAAGGCAGGG TTTCACTATG AACCCGTGGC 
               
               
                  5761 
                 TGGTGGCTGA CCTGGCTCTC TATACCAGGC TGTCTTTGAA TTCATGGGTG TCCACCTCCC 
               
               
                  5821 
                 TCTGCCTCCC AAACACCACC ATGTGTCCAC CATGGTCTCA CTCACGTAGC CCAAACTGGC 
               
               
                  5881 
                 CAAGAATTCT GCTATTTTTG CCCTTATCTT TTGGATTACA ACAGCCAGTT TCCACCATCA 
               
               
                  5941 
                 TAGAAAGAAA AACCCAGATA CTCCTCAGCC CATTGGGTTC CTACAATGTA CCTATGGGCT 
               
               
                  6001 
                 TCAATGTCAA ACTTCTTTCA AATCAGGCTT CTGTCCCTCA CTAGAAATTT AATATTGAGT 
               
               
                  6061 
                 TATTGACACA GCCCTGTCAC CCCCTCCCCC CACTGTTTTC TCACCTATAA ATTAGGAATA 
               
               
                  6121 
                 ATAAAAGCAC CAATGGGGAG AAGTTGGATG AAGGGGGAAA AGTATCACTA AAGCACACTA 
               
               
                  6181 
                 TTATTCAAAG GTGCCCTAAT GATATCCAAT TGTATGGTAT TTAAAAAATA AAAAATAAAA 
               
               
                  6241 
                 GCATTACGAA ACCGGGTGTG GTGGTGCATG CCTTTAATCC CAGCACTCCA GAGGCAGAGA 
               
               
                  6301 
                 CAGGTGGATC TGCATCTCAA TTAGTCAGCA ACCATAGTCC CGCCCCTAAC TCCGCCCATC 
               
               
                  6361 
                 CCGCCCCTAA CTCCGCCCAG TTCCGCCCAT TCTCCGCCCC ATGGCTGACT AATTTTTTTT 
               
               
                  6421 
                 ATTTATGCAG AGGCCGAGGC CGCCTCGGCC TCTGAGCTAT TCCAGAAGTA GTGAGGAGGC 
               
               
                  6481 
                 TTTTTTGGAG GCCTAGGCTT TTGCAAAAAG CTCGATCCTG AGAACTTCAG GGTGAGTTTG 
               
               
                  6541 
                 GGGACCCTTG ATTGTTCTTT CTTTTTCGCT ATTGTAAAAT TCATGTTATA TGGAGGGGGC 
               
               
                  6601 
                 AAAGTTTTCA GGGTGTTGTT TAGAATGGGA AGATGTCCCT TGTATCACCA TGGACCCTCA 
               
               
                  6661 
                 TGATAATTTT GTTTCTTTCA CTTTCTACTC TGTTGACAAC CATTGTCTCC TCTTATTTTC 
               
               
                  6721 
                 TTTTCATTTT CTGTAACTTT TTCGTTAAAC TTTAGCTTGC ATTTGTAACG AATTTTTAAA 
               
               
                  6781 
                 TTCACTTTTG TTTATTTGTC AGATTGTAAG TACTTTCTCT AATCACTTTT TTTTCAAGGC 
               
               
                  6841 
                 AATCAGGGTA TATTATATTG TACTTCAGCA CAGTTTTAGA GAACAATTGT TATAATTAAA 
               
               
                  6901 
                 TGATAAGGTA GAATATTTCT GCATATAAAT TCTGGCTGGC GTGGAAATAT TCTTATTGGT 
               
               
                  6961 
                 AGAAACAACT ACATCCTGGT CATCATCCTG CCTTTCTCTT TATGGTTACA ATGATATACA 
               
               
                  7021 
                 CTGTTTGAGA TGAGGATAAA ATACTCTGAG TCCAAACCGG GCCCCTCTGC TAACCATGTT 
               
               
                  7081 
                 CATGCCTTCT TCTTTTTCCT ACAGCTCCTG GGCAACGTGC TGGTTATTGT GCTGTCTCAT 
               
               
                  7141 
                 CATTTTGGCA AAGAATTGTA ATACGACTCA CTATAGGGCG AATTCCACCA TGTCCAATTT 
               
               
                  7201 
                 ACTGACCGTA CACCAAAATT TGCCTGCATT ACCGGTCGAT GCAACGAGTG ATGAGGTTCG 
               
               
                  7261 
                 CAAGAACCTG ATGGACATGT TCAGGGATCG CCAGGCGTTT TCTGAGCATA CCTGGAAAAT 
               
               
                  7321 
                 GCTTCTGTCC GTTTGCCGGT CGTGGGCGGC ATGGTGCAAG TTGAATAACC GGAAATGGTT 
               
               
                  7381 
                 TCCCGCAGAA CCTGAAGATG TTCGCGATTA TCTTCTATAT CTTCAGGCGC GCGGTCTGGC 
               
               
                  7441 
                 AGTAAAAACT ATCCAGCAAC ATTTGGGCCA GCTAAACATG CTTCATCGTC GGTCCGGGCT 
               
               
                  7501 
                 GCCACGACCA AGTGACAGCA ATGCTGTTTC ACTGGTTATG CGGCGGATCC GAAAAGAAAA 
               
               
                  7561 
                 CGTTGATGCC GGTGAACGTG CAAAACAGGC TCTAGCGTTC GAACGCACTG ATTTCGACCA 
               
               
                  7621 
                 GGTTCGTTCA CTCATGGAAA ATAGCGATCG CTGCCAGGAT ATACGTAATC TGGCATTTCT 
               
               
                  7681 
                 GGGGATTGCT TATAACACCC TGTTACGTAT AGCCGAAATT GCCAGGATCA GGGTTAAAGA 
               
               
                  7741 
                 TATCTCACGT ACTGACGGTG GGAGAATGTT AATCCATATT GGCAGAACGA AAACGCTGGT 
               
               
                  7801 
                 TAGCACCGCA GGTGTAGAGA AGGCACTTAG CCTGGGGGTA ACTAAACTGG TCGAGCGATG 
               
               
                  7861 
                 GATTTCCGTC TCTGGTGTAG CTGATGATCC GAATAACTAC CTGTTTTGCC GGGTCAGAAA 
               
               
                  7921 
                 AAATGGTGTT GCCGCGCCAT CTGCCACCAG CCAGCTATCA ACTCGCGCCC TGGAAGGGAT 
               
               
                  7981 
                 TTTTGAAGCA ACTCATCGAT TGATTTACGG CGCTAAGGAT GACTCTGGTC AGAGATACCT 
               
               
                  8041 
                 GGCCTGGTCT GGACACAGTG CCCGTGTCGG AGCCGCGCGA GATATGGCCC GCGCTGGAGT 
               
               
                  8101 
                 TTCAATACCG GAGATCATGC AAGCTGGTGG CTGGACCAAT GTAAATATTG TCATGAACTA 
               
               
                  8161 
                 TATCCGTAAC CTGGATAGTG AAACAGGGGC AATGGTGCGC CTGCTGGAAG ATGGCGATCT 
               
               
                  8221 
                 CGAGCCATCT GCTGGAGACA TGAGAGCTGC CAACCTTTGG CCAAGCCCGC TCATGATCAA 
               
               
                  8281 
                 ACGCTCTAAG AAGAACAGCC TGGCCTTGTC CCTGACGGCC GACCAGATGG TCAGTGCCTT 
               
               
                  8341 
                 GTTGGATGCT GAGCCCCCCA TACTCTATTC CGAGTATGAT CCTACCAGAC CCTTCAGTGA 
               
               
                  8401 
                 AGCTTCGATG ATGGGCTTAC TGACCAACCT GGCAGACAGG GAGCTGGTTC ACATGATCAA 
               
               
                  8461 
                 CTGGGCGAAG AGGGTGCCAG GCTTTGTGGA TTTGACCCTC CATGATCAGG TCCACCTTCT 
               
               
                  8521 
                 AGAATGTGCC TGGCTAGAGA TCCTGATGAT TGGTCTCGTC TGGCGCTCCA TGGAGCACCC 
               
               
                  8581 
                 AGGGAAGCTA CTGTTTGCTC CTAACTTGCT CTTGGACAGG AACCAGGGAA AATGTGTAGA 
               
               
                  8641 
                 GGGCATGGTG GAGATCTTCG ACATGCTGCT GGCTACATCA TCTCGGTTCC GCATGATGAA 
               
               
                  8701 
                 TCTGCAGGGA GAGGAGTTTG TGTGCCTCAA ATCTATTATT TTGCTTAATT CTGGAGTGTA 
               
               
                  8761 
                 CACATTTCTG TCCAGCACCC TGAAGTCTCT GGAAGAGAAG GACCATATCC ACCGAGTCCT 
               
               
                  8821 
                 GGACAAGATC ACAGACACTT TGATCCACCT GATGGCCAAG GCAGGCCTGA CCCTGCAGCA 
               
               
                  8881 
                 GCAGCACCAG CGGCTGGCCC AGCTCCTCCT CATCCTCTCC CACATCAGGC ACATGAGTAA 
               
               
                  8941 
                 CAAAGGCATG GAGCATCTGT ACAGCATGAA GTGCAAGAAC GTGGTGCCCC TCTATGACCT 
               
               
                  9001 
                 GCTGCTGGAG ATGCTGGACG CCCACCGCCT ACATGCGCCC ACTAGCCGTG GAGGGGCATC 
               
               
                  9061 
                 CGTGGAGGAG ACGGACCAAA GCCACTTGGC CACTGCGGGC TCTACTTCAT CGCATTCCTT 
               
               
                  9121 
                 GCAAAAGTAT TACATCACGG GGGAGGCAGA GGGTTTCCCT GCCACAGTCT GAGAGCTCCC 
               
               
                  9181 
                 TGGCGGAATT CGGATCTTAT TAAAGCAGAA CTTGTTTATT GCAGCTTATA ATGGTTACAA 
               
               
                  9241 
                 ATAAAGCAAT AGCATCACAA ATTTCACAAA TAAAGCATTT TTTTCACTGC ATTCTAGTTG 
               
               
                  9301 
                 TGGTTTGTCC AAACTCATCA ATGTATCTTA TCATGTCTGG TCGACCGATG CCCTTGAGAG 
               
               
                  9361 
                 CCTTCAACCC AGTCAGCTCC TTCCGGTGGG CGCGGGGCAT GACTATCGTC GCCGCACTTA 
               
               
                  9421 
                 TGACTGTCTT CTTTATCATG CAACTCGTAG GACAGGTGCC GGCAGCGCTC TTCCGCTTCC 
               
               
                  9481 
                 TCGCTCACTG ACTCGCTGCG CTCGGTCGTT CGGCTGCGGC GAGCGGTATC AGCTCACTCA 
               
               
                  9541 
                 AAGGCGGTAA TACGGTTATC CACAGAATCA GGGGATAACG CAGGAAAGAA CATGTGAGCA 
               
               
                  9601 
                 AAAGGCCAGC AAAAGGCCAG GAACCGTAAA AAGGCCGCGT TGCTGGCGTT TTTCCATAGG 
               
               
                  9661 
                 CTCCGCCCCC CTGACGAGCA TCACAAAAAT CGACGCTCAA GTCAGAGGTG GCGAAACCCG 
               
               
                  9721 
                 ACAGGACTAT AAAGATACCA GGCGTTTCCC CCTGGAAGCT CCCTCGTGCG CTCTCCTGTT 
               
               
                  9781 
                 CCGACCCTGC CGCTTACCGG ATACCTGTCC GCCTTTCTCC CTTCGGGAAG CGTGGCGCTT 
               
               
                  9841 
                 TCTCATAGCT CACGCTGTAG GTATCTCAGT TCGGTGTAGG TCGTTCGCTC CAAGCTGGGC 
               
               
                  9901 
                 TGTGTGCACG AACCCCCCGT TCAGCCCGAC CGCTGCGCCT TATCCGGTAA CTATCGTCTT 
               
               
                  9961 
                 GAGTCCAACC CGGTAAGACA CGACTTATCG CCACTGGCAG CAGCCACTGG TAACAGGATT 
               
               
                 10021 
                 AGCAGAGCGA GGTATGTAGG CGGTGCTACA GAGTTCTTGA AGTGGTGGCC TAACTACGGC 
               
               
                 10081 
                 TACACTAGAA GAACAGTATT TGGTATCTGC GCTCTGCTGA AGCCAGTTAC CTTCGGAAAA 
               
               
                 10141 
                 AGAGTTGGTA GCTCTTGATC CGGCAAACAA ACCACCGCTG GTAGCGGTGG TTTTTTTGTT 
               
               
                 10201 
                 TGCAAGCAGC AGATTACGCG CAGAAAAAAA GGATCTCAAG AAGATCCTTT GATCTTTTCT 
               
               
                 10261 
                 ACGGGGTCTG ACGCTCAGTG GAACGAAAAC TCACGTTAAG GGATTTTGGT CATGAGATTA 
               
               
                 10321 
                 TCAAAAAGGA TCTTCACCTA GATCCTTTTA AATTAAAAAT GAAGTTTTAA ATCAATCTAA 
               
               
                 10381 
                 AGTATATATG AGTAAACTTG GTCTGACAGT TACCAATGCT TAATCAGTGA GGCACCTATC 
               
               
                 10441 
                 TCAGCGATCT GTCTATTTCG TTCATCCATA GTTGCCTGAC TCCCCGTCGT GTAGATAACT 
               
               
                 10501 
                 ACGATACGGG AGGGCTTACC ATCTGGCCCC AGTGCTGCAA TGATACCGCG AGACCCACGC 
               
               
                 10561 
                 TCACCGGCTC CAGATTTATC AGCAATAAAC CAGCCAGCCG GAAGGGCCGA GCGCAGAAGT 
               
               
                 10621 
                 GGTCCTGCAA CTTTATCCGC CTCCATCCAG TCTATTAATT GTTGCCGGGA AGCTAGAGTA 
               
               
                 10681 
                 AGTAGTTCGC CAGTTAATAG TTTGCGCAAC GTTGTTGCCA TTGCTACAGG CATCGTGGTG 
               
               
                 10741 
                 TCACGCTCGT CGTTTGGTAT GGCTTCATTC AGCTCCGGTT CCCAACGATC AAGGCGAGTT 
               
               
                 10801 
                 ACATGATCCC CCATGTTGTG CAAAAAAGCG GTTAGCTCCT TCGGTCCTCC GATCGTTGTC 
               
               
                 10861 
                 AGAAGTAAGT TGGCCGCAGT GTTATCACTC ATGGTTATGG CAGCACTGCA TAATTCTCTT 
               
               
                 10921 
                 ACTGTCATGC CATCCGTAAG ATGCTTTTCT GTGACTGGTG AGTACTCAAC CAAGTCATTC 
               
               
                 10981 
                 TGAGAATAGT GTATGCGGCG ACCGAGTTGC TCTTGCCCGG CGTCAATACG GGATAATACC 
               
               
                 11041 
                 GCGCCACATA GCAGAACTTT AAAAGTGCTC ATCATTGGAA AACGTTCTTC GGGGCGAAAA 
               
               
                 11101 
                 CTCTCAAGGA TCTTACCGCT GTTGAGATCC AGTTCGATGT AACCCACTCG TGCACCCAAC 
               
               
                 11161 
                 TGATCTTCAG CATCTTTTAC TTTCACCAGC GTTTCTGGGT GAGCAAAAAC AGGAAGGCAA 
               
               
                 11221 
                 AATGCCGCAA AAAAGGGAAT AAGGGCGACA CGGAAATGTT GAATACTCAT ACTCTTCCTT 
               
               
                 11281 
                 TTTCAATATT ATTGAAGCAT TTATCAGGGT TATTGTCTCA TGAGCGGATA CATATTTGAA 
               
               
                 11341 
                 TGTATTTAGA AAAATAAACA AATAGGGGTT CCGCGCACAT TTCCCCGAAA AGTGCCACCT 
               
               
                 11401 
                 GACGCGCCCT GTAGCGGCGC ATTAAGCGCG GCGGGTGTGG TGGTTACGCG CAGCGTGACC 
               
               
                 11461 
                 GCTACACTTG CCAGCGCCCT AGCGCCCGCT CCTTTCGCTT TCTTCCCTTC CTTTCTCGCC 
               
               
                 11521 
                 ACGTTCGCCG GCTTTCCCCG TCAAGCTCTA AATCGGGGGC TCCCTTTAGG GTTCCGATTT 
               
               
                 11581 
                 AGTGCTTTAC GGCACCTCGA CCCCAAAAAA CTTGATTAGG GTGATGGTTC ACGTAGTGGG 
               
               
                 11641 
                 CCATCGCCCT GATAGACGGT TTTTCGCCCT TTGACGTTGG AGTCCACGTT CTTTAATAGT 
               
               
                 11701 
                 GGACTCTTGT TCCAAACTGG AACAACACTC AACCCTATCT CGGTCTATTC TTTTGATTTA 
               
               
                 11761 
                 TAAGGGATTT TGCCGATTTC GGCCTATTGG TTAAAAAATG AGCTGATTTA ACAAAAATTT 
               
               
                 11821 
                 AACGCGAATT TTAACAAAAT ATTAACGCTT ACAATTTGCC ATTCGCCATT CAGGCTGCGC 
               
               
                 11881 
                 AACTGTTGGG AAGGGCGATC GGTGCGGGCC TCTTCGCTAT TACGCCAGCC CAAGCTACCA 
               
               
                 11941 
                 TGATAAGTAA GTAATATTAA GGTACGTGGA GGTTTTACTT GCTTTAAAAA CCTCCCACAC 
               
               
                 12001 
                 CTCCCCCTGA ACCTGAAACA TAAAATGAAT GCAATTGTTG TTGTTAACTT GTTTATTGCA 
               
               
                 12061 
                 GCTTATAATG GTTACAAATA AAGCAATAGC ATCACAAATT TCACAAATAA AGCATTTTTT 
               
               
                 12121 
                 TCACTGCATT CTAGTTGTGG TTTGTCCAAA CTCATCAATG TATCTTATGG TACTGTAACT 
               
               
                 12181 
                 GAGCTAACAT AA 
               
               
                 //