Patent Publication Number: US-2020291433-A1

Title: Globin gene therapy for treating hemoglobinopathies

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     This application is a Continuation of International Patent Application No. PCT/US18/064256 filed on Dec. 6, 2018, which claims priority to U.S. Provisional Application No. 62/595,277 filed on Dec. 6, 2017, the content of each of which is incorporated by reference in its entirety, and to each of which priority is claimed. 
    
    
     SEQUENCE LISTING 
     The specification further incorporates by reference the Sequence Listing submitted herewith via EFS on Jun. 2, 2020. Pursuant to 37 C.F.R. § 1.52(e)(5), the Sequence Listing text file, identified as 0727341087_SL.txt, is 268,531 bytes and was created on May 31, 2020. The Sequence Listing, electronically filed herewith, does not extend beyond the scope of the specification and thus does not contain new matter. 
     INTRODUCTION 
     The presently disclosed subject matter provides expression cassettes and vectors comprising such expression cassettes that express a globin protein, e.g., a human β-globin protein. The presently disclosed subject matter further provides expression cassettes that comprise a globin gene or a functional portion thereof operably linked to a β-globin locus control region (LCR) comprising a plurality of Dnase I hypersensitive sites. The expression cassettes of the presently disclosed subject matter comprise one or more insulators that counteract the effect of enhancer elements. The insulators disclosed herein do not substantially adversely impact the titer of a vector that comprises the presently disclosed expression cassettes. The expression cassettes and vectors can be used for treating a hemoglobinopathy, e.g., β-thalassemia, and sickle cell anemia. 
     BACKGROUND 
     β-thalassemia and sickle cell anemia are severe congenital anemias that are caused by defective production of the β chain of hemoglobin. In β-thalassemia, the β chain deficit leads to the intracellular precipitation of excess α-globin chains, causing ineffective erythropoiesis and hemolytic anemia (Weatherall and Clegg (1981), Stamatoyannopoulos et al., (1994), Weatherall (2001), Steinberg (2001)). In the most severe forms found in homozygotes or compound heterozygotes, anemia is lethal within the first years of life in the absence of any treatment (Cooley and Lee ( 1925 )). Lifelong transfusion therapy is needed to correct anemia, suppress ineffective erythropoiesis and inhibit gastrointestinal iron absorption (Weatherall and Clegg (1981), Stamatoyannopoulos et al. (1994), Weatherall (2001), Steinberg (2001)). However, transfusion therapy itself leads to iron overload, which is lethal if untreated. The prevention and treatment of iron overload are the major goals of current patient management (Giardina (2001)). The only current curative treatment to cure β-thalassemia is to provide erythroid precursors harboring normal globin genes through allogeneic bone marrow transplantation (BMT) (Giardini and Lucarelli (1994), Boulad et al. (1998), Lucarelli et al. (1999), Tisdale and Sadelain (2001)). 
     In sickle cell anemia, the hemoglobin β chain is mutated at amino acid position 6 (Glu→Val), leading to the synthesis of β S  instead of the normal β A  chain (Steinberg (2001), Pauling et al. (1949)). The resulting hemoglobin, HbS, causes accelerated red cell destruction, erythroid hyperplasia and painful vaso-occlusive ‘crises’ (Steinberg (2001)). Vaso-occlusion can damage organs, eventually causing long-term disabilities (e.g. following stroke or bone necrosis), and sometimes sudden death. While a very serious disorder, the course of sickle cell disease is typically unpredictable (Steinberg (2001)). By increasing production of fetal hemoglobin (Swank and Stamatoyannopoulos (1998)) and suppressing hematopoiesis, hydroxyurea can produce a measurable clinical benefit (Platt et al. (1984)), Charache et al. (1992), Atweh and Loukopoulos (2001)). Since hydroxyurea is a cytotoxic agent, there is a great need for alternative, less toxic drugs to induce γ-globin gene expression (Perrine et al. (2005), Stamatoyannopoulos (2005)). As for β-thalassemia, allogeneic bone marrow transplantation (BMT) is at present the only curative therapy for sickle cell disease (Tisdale and Sadelain (2001), Vermylen et al. (1998), Luzzatto and Goodfellow (1989)). 
     BMT, however, is not available as a therapeutic option to most patients suffering from β-thalassemia or sickle cell disease, due to the lack of an HLA-matched bone marrow donor for most individuals. Furthermore, although potentially curative, allogeneic BMT is not devoid of complications. Safe transplantation requires the identification of a histo-compatible donor to minimize the risks of graft rejection and graft-versus-host disease (Tisdale and Sadelain (2001), Vermylen et al. (1998), Luzzatto and Goodfellow (1989)). Because of the greater risks associated with matched-unrelated or mismatched transplants, most patients have to settle for life-long transfusion therapy, which does not correct ineffective erythropoiesis and exacerbates systemic iron accumulation. Moreover, despite the considerable improvement in life expectancy in the last decades (Borgna-Pignatti et al. (2004), Telfer et al. (2009), Ladis et al. (2011)), the risk of some serious complications arising over the long term from viral infections, iron toxicity and liver cirrhosis, remain (Mancuso et al. (2006)). These medical risks, together with the socio-economic cost of chronic β-thalassemia, underscore the need for safe, effective and curative therapies. 
     The only means to cure rather than treat severe β-thalassemia is to provide the patient with healthy hematopoietic stem cells (HSCs). HSCs normally give rise to all blood cell types, including 20 billion RBCs per day in adults. HSCs can be harvested from a donor with wild-type β-globin genes to yield long-lived red blood cells (RBCs) with a normal content in hemoglobin. Alternatively, one may genetically correct the patient&#39;s own HSCs, which at once resolves the search for a donor and eliminates the risks of graft-versus-host disease and graft rejection associated with allogeneic BMT (Sadelain (1997), Sadelain et al. (2007)). Globin gene transfer aims to restore the capacity of the β-thalassemic subject&#39;s own blood-forming stem cells to generate RBCs with a sufficient hemoglobin content Sadelain et al. (2007), Persons and Tisdale (2004), Sadelain (2006)). The goal in patients with sickle cell anemia is to prevent sickling, which can be achieved by diluting the endogenous HbS with a non-sickling Hb that incorporates the vector-encoded globin chain. The patient&#39;s own HSCs are the cells that have to be genetically modified to ensure long-lasting therapeutic benefits and achieve a curative stem cell-based therapy. 
     The implementation of globin gene transfer for the treatment of severe β-thalassemia and sickle cell anemia requires the efficient introduction of a regulated human β- or β-like globin gene in HSCs. The β-globin gene (or β-like variant) must be expressed in erythroid-specific fashion and at high level, especially for the treatment of transfusion-dependent beta-zero thalassemias. 
     The globin vectors developed to date present shortcomings that may limit or even preclude their safe use in thalassemia and sickle cell patients. Some of the β-globin locus control region (LCR) components contained in the vectors, in particular Dnase I hypersensitive site-2 (HS2), may have non-erythroid activity, exposing patients to the risk of insertional oncogenesis as seen with non-specific expression vectors. Further, the use of large LCR segments can be detrimental to the production of high titer vectors and the efficient transduction of patients HSCs. Accordingly, there is a need for novel globin expression cassettes that allow for therapeutic expression of a globin gene (e.g., human β-globin gene) in erythroid-specific and differentiation stage-specific fashion with minimal risk of insertional oncogenesis, and that enable high level transduction, thus improving their safety when used in treating thalassemia and sickle cell patients. 
     SUMMARY OF THE INVENTION 
     The presently disclosed subject matter generally provides enhancer blocking insulators, and certain insulators additionally possess barrier insulator activity. The presently disclosed subject matter also provides expression cassettes comprising one or more insulators and allows for expression of a globin gene (e.g., a human β globin gene). Also provided are vectors comprising such expression cassettes, cells transduced with such expression cassettes or such vectors, and uses of such expression cassettes for treating hemoglobinopathies (e.g., β-thalassemia and sickle cell anemia). 
     In certain non-limiting embodiments, the presently disclosed subject matter provides an expression cassette comprising a globin gene or a functional portion thereof operably linked to a β-globin locus control region (LCR) that comprises a Dnase I hypersensitive site-2 (HS2) region, a Dnase I hypersensitive site-3 (HS3) region, and a Dnase I hypersensitive site-4 (HS4) region, wherein the HS4 region has a length of less than about 800 bp and does not comprise the nucleotide sequence set forth in SEQ ID NO: 50, which is provided below. 
     
       
         
           
               
            
               
                 [SEQ ID NO: 50] 
               
               
                 TGAGCCCCTTTTCCTCTAACTGAAAGAAGGAAAAAAAAAATGGAACCCAA 
               
               
                   
               
               
                 AATATTCTACATAGTTTCCATGTCACAGCCAGGGCTGGGCAGTCTCCTGT 
               
               
                   
               
               
                 TATTTCTTTTAAAA 
               
            
           
         
       
     
     In certain embodiments, the HS2 region does not comprise the nucleotide sequence set forth in SEQ ID NO: 52, which is provided below. 
     
       
         
           
               
            
               
                 [SEQ ID NO: 52] 
               
               
                 GTATATGTGTATATATATATATATATATTCAGGAAATAATATAT 
               
            
           
         
       
     
     In certain embodiments, the HS2 region has nucleotides 45-860 of SEQ ID NO: 9. In certain embodiments, the HS2 region has the nucleotide sequence set forth in SEQ ID NO: 33. 
     In certain embodiments, the expression cassette further comprises a Dnase I hypersensitive site-1 (HS1) region. 
     The presently disclosed subject matter also provides an expression cassette comprising a globin gene or a functional portion thereof operably linked to a β-globin locus control region (LCR) that comprises a HS1 region, a HS3 region, and a HS4 region and does not comprise a core sequence of HS2 region, wherein the HS4 region has a length of less than about 800 bp and does not comprise the nucleotide sequence set forth in SEQ ID NO: 50. In certain embodiments, the core sequence of HS2 has the nucleotide sequence set forth in SEQ ID NO:20 or SEQ ID NO:21. In certain embodiments, the β-globin LCR does not comprise a HS2 region that sustains the enhancer activity of HS2. 
     In various of these embodiments, the HS4 region has a length of between about 700 bp and about 800 bp. In certain embodiments, the HS4 region has a length of about 750 bp. In certain embodiments, the HS4 region does not comprise the nucleotide sequence set forth in SEQ ID NO: 
     51, which is provided below. 
     
       
         
           
               
            
               
                 [SEQ ID NO: 51] 
               
               
                 TTTAATCTAACAATTATGAACAGCAATGAGATAATATGTACAAAGTACCC 
               
               
                   
               
               
                 AGACCTATGTGGTAGAGCATCAAGGAAGCGCATTGCGGAGCAGTTTTTTG 
               
               
                   
               
               
                 TTTGTTTGTTTTTGTATTCTGTTTCGTGAGGCAAGGTTTCACTCTGCTGT 
               
               
                   
               
               
                 CCAGGCTGGAGTGCAGTGGCAAGATCATGTCTCACTGCAGCCTTGAC 
               
            
           
         
       
     
     In various of these embodiments, the HS3 region has a length of about 1300 bp. In certain embodiments, the HS3 region has the nucleotide sequence set forth in SEQ ID NO: 5. In certain embodiments, the HS3 region has the nucleotide sequence set forth in SEQ ID NO: 34. 
     In various of these embodiments, the HS1 region has a length of about 600 bp. In certain embodiments, the HS1 region has the nucleotide sequence set forth in SEQ ID NO: 3. 
     In certain embodiments, the β-globin LCR comprises or consists essentially of a HS2 region having nucleotides 45-860 of SEQ ID NO: 9, a HS3 region having the nucleotide sequence set forth in SEQ ID NO: 5, and a HS4 region having nucleotides 115-868 of SEQ ID NO: 6. 
     In certain embodiments, the β-globin LCR comprises or consists essentially of a HS2 region having the nucleotide sequence set forth in SEQ ID NO: 33, a HS3 region having the nucleotide sequence set forth in SEQ ID NO: 34, and a HS4 region having the nucleotide sequence set forth in SEQ ID NO: 35. 
     In certain embodiments, the β-globin LCR comprises or consists essentially of a HS1 region having the nucleotide sequence set forth in SEQ ID NO: 3, a HS3 region having the nucleotide sequence set forth in SEQ ID NO: 5, and a HS4 region having nucleotides 115-868 of SEQ ID NO: 6. 
     In certain embodiments, the β-globin LCR comprises or consists essentially of a HS1 region having the nucleotide sequence set forth in SEQ ID NO: 3, a HS3 region having the nucleotide sequence set forth in SEQ ID NO: 34, and a HS4 region having the nucleotide sequence set forth in SEQ ID NO: 35. 
     In certain embodiments, the β-globin LCR comprises or consists essentially of a HS1 region having the nucleotide sequence set forth in SEQ ID NO: 3, a HS2 region having nucleotides 45-860 of SEQ ID NO: 9, a HS3 region having the nucleotide sequence set forth in SEQ ID NO: 5, and a HS4 region having nucleotides 115-868 of SEQ ID NO: 6. 
     In certain embodiments, the β-globin LCR comprises or consists essentially of a HS1 region having the nucleotide sequence set forth in SEQ ID NO: 3, a HS2 region having the nucleotide sequence set forth in SEQ ID NO: 33, a HS3 region having the nucleotide sequence set forth in SEQ ID NO: 34, and a HS4 region having the nucleotide sequence set forth in SEQ ID NO: 35. 
     In various of these embodiments, the expression cassette further comprises at least one an insulator comprising the CTCF binding site sequence set forth in SEQ ID NO:18. In certain embodiments, the at least one insulator has the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, or SEQ ID NO: 31. In certain embodiments, the expression cassette comprises two insulators, each comprising the CTCF binding site sequence set forth in SEQ ID NO:18, for example, but not limited to, where one or both insulators comprise the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO: 24, SEQ ID NO:25, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, or SEQ ID NO: 31. 
     In certain embodiments, the expression cassette further comprises at least one erythroid-specific enhancer. In certain embodiments, the at least one erythroid-specific enhancer is positioned within the β-globin LCR. In certain embodiments, the at least one erythroid-specific enhancer is positioned between the HS1 region and the HS3 region of the β-globin LCR. In certain embodiments, the at least one erythroid-specific enhancer has the nucleotide sequence set forth in SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, or SEQ ID NO: 17. In certain embodiments, the at least one erythroid-specific enhancer has the nucleotide sequence set forth in SEQ ID NO: 15. In certain embodiments, the at least one erythroid-specific enhancer is between about 100 and about 200 bp in length. In certain embodiments, the expression cassette comprises one, two or three erythroid-specific enhancers. 
     The presently disclosed subject matter also provides an expression cassette comprising at least one erythroid-specific enhancer disclosed herein and a globin gene or a functional portion thereof, wherein the expression cassette does not comprise a β-globin locus control region (LCR). 
     The presently disclosed subject matter further provides an expression cassette comprising at least one erythroid-specific enhancer disclosed herein and a globin gene or a functional portion thereof that is operatively linked to a β-globin locus control region (LCR) that comprises a HS3 region, and does not comprise a core sequence of a HS1 region, a core sequence of a HS2 region, or a core sequence of a HS4 region. 
     In certain embodiments, the globin gene is selected from the group consisting of β-globin gene, γ-globin gene, and δ-globin gene. In certain non-limiting embodiments, the globin gene is human β-globin gene. In certain non-limiting embodiments, the human β-globin gene is a wild-type human β-globin gene. In certain non-limiting embodiments, the human β-globin gene is a non-wild-type human β-globin gene, including, but not limited to, a human β-globin gene comprising one or more deletions of intron sequences, a human β-globin gene encoding at least one (e.g., one or two) anti-sickling amino acid residue (e.g., a human β-globin gene encoding anti-sickling hemoglobin 1 (HB AS1), and a human β-globin gene encoding anti-sickling hemoglobin 2 (HB AS2)), and a human β-globin gene comprising one or more deletions of intron sequences and encoding at least one anti-sickling amino acid residue. In certain non-limiting embodiments, the human β-globin gene is a human β A -globin gene encoding a threonine to glutamine mutation at codon 87 (β A-T87Q ). In certain non-limiting embodiments, the human β-globin gene is a human β A -globin gene encoding a threonine to glutamine mutation at codon 87 (β A-T87Q ) and comprising a deletion in intron 2. In certain non-limiting embodiments, the human β-globin gene is a human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 (β A-E22A ). In certain non-limiting embodiments, the human β-globin gene is a human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 (β A-E22A ) and comprising a deletion in intron 2. In certain non-limiting embodiments, the human β-globin gene is a human β A -globin gene encoding an asparagine to lysine mutation at codon 80 (β −N80K ). In certain non-limiting embodiments, the human β-globin gene is a human β A -globin gene encoding an asparagine to lysine mutation at codon 80 (β −N80K ) and comprising a deletion in intron 2. In certain non-limiting embodiments, the human β-globin gene is a human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and an asparagine to lysine mutation at codon 80. In certain non-limiting embodiments, the human β-globin gene is a human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and an asparagine to lysine mutation at codon 80 and comprising a deletion in intron 2. In certain non-limiting embodiments, the human β-globin gene is a human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and a threonine to glutamine mutation at codon 87. In certain non-limiting embodiments, the human β-globin gene is a human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and a threonine to glutamine mutation at codon 87 and comprising a deletion in intron 2. In certain embodiments, the deletion in intro  2  is a deletion of about 370 bp. 
     In certain embodiments, the expression cassette further comprises a β-globin promoter. In certain embodiments, the β-globin promoter is positioned between the globin gene or functional portion thereof and β-globin LCR. In certain non-limiting embodiments, the β-globin promoter is a human β-globin promoter that is about 265 bp in length. In certain non-limiting embodiments, the β human β-globin promoter has the nucleotide sequence set forth in SEQ ID NO:11. 
     In certain embodiments, the expression cassette further comprises a human β-globin 3′ enhancer. In certain embodiments, the human β-globin 3′ enhancer is positioned in the upstream of the globin gene or functional portion thereof. In certain embodiments, the β-globin 3′ enhancer is between about 700 and about 900 bp in length, e.g., between about 800 and 900 bp in length. In certain embodiments, the human β-globin 3′ enhancer is about 880 bp in length. In certain embodiments, the human β-globin 3′ enhancer has the nucleotide sequence set forth in SEQ ID NO:12. 
     In certain embodiments, the expression cassette allows for expression of the globin gene or functional portion thereof in a mammal. In certain embodiments, the expression cassette allows for expression of a human β-globin gene. In certain embodiments, the expression of the globin gene or functional portion thereof is restricted to erythroid tissue. 
     The presently disclosed subject matter also provides recombinant vectors comprising the above-described expression cassettes. In certain embodiments, the recombinant vector is a retroviral vector. In certain embodiments, the retroviral vector is a lentivirus vector. In certain embodiments, the expression cassette comprised in the recombinant vector comprises one insulator. In certain embodiments, the expression cassette comprised in the recombinant vector comprises two insulators, e.g., two of the insulator comprising the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, or SEQ ID NO: 32. In certain embodiments, the recombinant vector further comprises a Woodchuck hepatitis post-regulatory element (WPRE) in the 3′ long terminal repeat (LTR) of the vector. In certain embodiments, the recombinant vector further comprises a bovine growth hormone polyadenylation signal in the 3′ long terminal repeat (LTR) of the vector. 
     In addition, the presently disclosed subject matter provides non-naturally occurring or engineered nucleases comprising the above-described expression cassettes. In certain embodiments, the nuclease is selected from the group consisting of a non-naturally occurring or engineered zinc-finger nuclease (ZFN), a non-naturally occurring or engineered meganuclease, and a non-naturally occurring or engineered transcription activator-like effector nuclease (TALEN). In certain embodiments, the nuclease comprises a DNA binding domain and a nuclease cleavage domain. In certain embodiments, the nuclease binds to a genomic safe harbor site. In certain embodiments, the nuclease generates a double strand break (DSB) at the genomic safe harbor site. In certain embodiments, the expression cassette comprised in the nuclease comprises two insulators, e.g., two of the insulator comprising the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, or SEQ ID NO: 32. In certain embodiments, the nuclease allows for targeted delivery of the expression cassette. The presently disclosed subject matter also provides polynucleotides encoding the above-described nucleases, and vectors comprising the polynucleotides. In certain embodiments, the vector is a lentiviral vector. 
     Furthermore, the presently disclosed subject matter provides non-naturally occurring or engineered CRISPR-Cas systems comprising the above-described expression cassettes. In certain embodiments, the CRISPR-Cas system comprises a CRISPR-Cas nuclease and single-guide RNA. In certain embodiments, the CRISPR-Cas system binds to a genomic safe harbor site. In certain embodiments, the CRISPR-Cas system generates a double strand break (DSB) at the genomic safe harbor site. In certain embodiments, the expression cassette comprised in the CRISPR-Cas system comprises two insulators, e.g., two of the insulator comprising the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, or SEQ ID NO: 32. In certain embodiments, the CRISPR-Cas allows for targeted delivery of the expression cassette. The presently disclosed subject matter also provides polynucleotides encoding the above-described CRISPR-Cas systems, and vectors comprising the polynucleotides. In certain non-limiting embodiments, the vector is a lentiviral vector. 
     In certain embodiments, the genomic safe harbor site is an extragenic genomic safe harbor site. In certain embodiments, the genomic safe harbor site is located on chromosome 1. In certain embodiments, the genomic safe harbor meets all of the following five criteria: (1) distance of at least 50 kb from the 5′ end of any gene (e.g., from the 5′ end of the gene), (ii) distance of at least 300 kb from any cancer-related gene, (iii) within an open/accessible chromatin structure (measured by DNA cleavage with natural or engineered nucleases), (iv) location outside a gene transcription unit and (v) location outside ultraconserved regions (UCRs), microRNA or long non-coding RNA of the human genome. 
     Additionally, the presently disclosed subject matter provides cells transduced with the above-described expression cassettes, cells transduced with the above-described recombinant vectors, cells transduced with the above-described nucleases, cells transduced with the above-described CRISPR-Cas systems. In addition, the presently disclosed subject matter provides cells transduced with the above-described vectors. In certain embodiments, the cell is selected from the group consisting of a hematopoietic stem cell, an embryonic stem cell, an induced pluripotent stem cell, and a hemogenic endothelium cell. In certain non-limiting embodiments, the hematopoietic stem cell is a CD34 +  hematopoietic stem cell. In certain embodiments, the cell is transduced ex vivo. 
     Also provided are pharmaceutical compositions comprising an effective amount of the above-described cells and a pharmaceutically acceptable carrier. The presently disclosed subject matter also provides pharmaceutical compositions for treating a hemoglobinopathy comprising an effective amount of the above-described cells and a pharmaceutically acceptable carrier. 
     Furthermore, the presently disclosed subject matter provides kits for treating a hemoglobinopathy comprising the above-described cells. In certain embodiments, the kits further comprise written instructions for using the cell for treating a subject having a hemoglobinopathy. 
     In addition, the presently disclosed subject matter provides methods of treating a hemoglobinopathy in a subject, comprising administering an effective amount of the above-described cells to the subject, thereby restoring the subject&#39;s ability to produce red blood cells containing normal hemoglobin. In certain embodiments, a therapeutically relevant level of hemoglobin is produced in the subject following administering the cell to the subject. In certain amendments, the method comprises administering an effective amount of the cell transduced with the above-described recombinant vector. In certain embodiments, the vector copy number of the recombinant vector in the cell that provides for the therapeutically relevant level of hemoglobin in the subject is about 0.5-2 vector copy number per cell. In certain embodiments, the method corrects ineffective erythropoiesis in the subject. In certain embodiments, the method does not incur the risk of graft-versus-host disease in the subject. In certain embodiments, the method does not comprise administering an immunosuppressive agent. In certain embodiments, the cell is selected from the group consisting of a hematopoietic stem cell, an embryonic stem cell, an induced pluripotent stem cell, and a hemogenic endothelium cell. In certain non-limiting embodiments, the subject is a human. In certain embodiments, the cell is from the subject. In certain non-limiting embodiments, the cell is from bone marrow of the subject. 
     In accordance with the presently disclosed subject matter, the hemoglobinopathy can be selected from the group consisting of hemoglobin C disease, hemoglobin sickle cell disease (SCD), sickle cell anemia, hereditary anemia, thalassemia, β-thalassemia, thalassemia major, thalassemia intermedia, α-thalassemia, and hemoglobin H disease. In certain non-limiting embodiments, the hemoglobinopathy is β-thalassemia. In certain non-limiting embodiments, the hemoglobinopathy is sickle cell anemia. 
    
    
     
       BRIEF DESCRIPTION OF THE FIGURES 
       The following Detailed Description, given by way of example, but not intended to limit the invention to specific embodiments described, may be understood in conjunction with the accompanying drawings. 
         FIG. 1  depicts a recombinant vector comprising an expression cassette in accordance with one non-limiting embodiment of the presently disclosed subject matter. “RRE” represents for “Rev Response Element”. cPPT represents for “Central Polypurine tract”. 
         FIG. 2  depicts a recombinant vector an expression cassette in accordance with one non-limiting embodiment of the presently disclosed subject matter. 
         FIG. 3  depicts a recombinant vector an expression cassette in accordance with one non-limiting embodiment of the presently disclosed subject matter. 
         FIG. 4  depicts a recombinant vector an expression cassette in accordance with one non-limiting embodiment of the presently disclosed subject matter. 
         FIGS. 5A-5C  represent the genotoxicity of insulator A1.  FIG. 5A  demonstrates the gammaretroviral vector genotoxicity assay used.  FIG. 5B  notices the increased survival of mice receiving 32D cells transduced with insulated gammaretroviral vector. Also notice the results obtained with cHS4 and with the uninsulated control.  FIG. 5C  shows that insulator A1 decreased the risk of genotoxicity. 
         FIG. 6  represents normalized β chain expression in thalassemic Hbb th3/+  mice 8 and 44 weeks post-treatment. 
         FIG. 7  represents the evaluation of enhancer activity in non-erythroid K562 cells. 
         FIG. 8  represents the erythroid-specific enhancers in accordance with certain embodiments of the presently disclosed subject matter (SEQ ID NOS 13-17, respectively, in order of appearance). 
         FIG. 9  represents the erythroid-specific enhancers in accordance with certain embodiments of the presently disclosed subject matter (SEQ ID NO: 26). 
         FIGS. 10A-10B  depict various recombinant vectors comprising the presently disclosed expression cassettes.  FIG. 10A  depicts recombinant vector #18, and #49-#55.  FIG. 10B  shows recombinant vector #18, #40, #49, #52-#55, and #57-#62. 
         FIG. 11  represents the titer of the recombinant vectors comprising the presently disclosed expression cassettes. 
         FIG. 12  represents the titer of the recombinant vectors comprising the presently disclosed expression cassettes. 
         FIG. 13  depicts recombinant vectors comprising expression cassettes in accordance with certain non-limiting embodiments of the presently disclosed subject matter. 
         FIG. 14  depicts recombinant vectors comprising expression cassettes in accordance with certain non-limiting embodiments of the presently disclosed subject matter. 
         FIG. 15  depicts recombinant vectors comprising expression cassettes in accordance with certain non-limiting embodiments of the presently disclosed subject matter. 
         FIG. 16  depicts the average Hb production for vectors (TNS9.B87.A1 SNS23.2.B87.A1, SNS26.B87.A1, and SNS27.2B.87.A1). 
         FIG. 17  depicts the titration assay relative to various vectors. 
         FIG. 18  depicts the average of ΔHb normalized per VCN for various vectors. 
         FIG. 19  depicts the average of total Hemoglobin (Hb) in thalassemic mouse peripheral blood at time points of 6-week, 5-month, and 9-month. 
         FIG. 20  depicts the average gain in Hb level (ΔHb) normalized to vector copy (VCN) at time points of 6-week, 5-month, and 9-month. ΔHb=Hb level−7.5 (baseline level in thalassemic mice). 
     
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     The presently disclosed subject matter generally provides expression cassettes that allow for expression of a globin gene (e.g., human β-globin gene). In certain non-limiting embodiments, the expression cassette comprises at least one insulator comprising the CTCF binding site sequence set forth in SEQ ID NO:18, for example, but not limited to, an insulator comprising the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO: 24, SEQ ID NO: 25, or SEQ ID NO: 27 and a globin gene or a functional portion thereof operably linked to a β-globin locus control region (LCR). The expression of the globin gene induced by the presently disclosed expression cassettes is erythroid-specific, differentiation stage-specific, high-level, and sustained. The presently disclosed subject matter also provides recombinant vectors, non-naturally occurring or engineered nucleases, and non-naturally occurring or engineered CRISPR-Cas systems comprising such expression cassettes, and cells transduced with such expression cassettes, recombinant vectors, nucleases and CRISPR-Cas systems. The presently disclosed expression cassettes and vectors comprising thereof provide for a safe gene transfer therapy as therapeutic transgene expression is achieved (e.g., a therapeutically relevant level of hemoglobin is produced) with a low vector copy number per cell (e.g., 0.5-2, 1-2, or even 0.5-1). In addition, the presently disclosed subject matter provides methods of using such transduced cells for treating a hemoglobinopathy (e.g., β-thalassemia and sickle cell anemia). 
     I. Definitions 
     Unless defined otherwise, all technical and scientific terms used herein have the meaning commonly understood by a person skilled in the art to which this invention belongs. The following references provide one of skill with a general definition of many of the terms used in this invention: Singleton et al., Dictionary of Microbiology and Molecular Biology (2nd ed. 1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988); The Glossary of Genetics, 5th Ed., R. Rieger et al. (eds.), Springer Verlag (1991); and Hale &amp; Marham, The Harper Collins Dictionary of Biology (1991). As used herein, the following terms have the meanings ascribed to them below, unless specified otherwise. 
     As used herein, the term “expression cassette” refers to a nucleic acid construct, generated recombinantly or synthetically, with a series of specified nucleic acid elements, which permit transcription of a particular nucleic acid in a target cell. The expression cassette can be incorporated into a plasmid, chromosome, mitochondrial DNA, plastid DNA, virus or nucleic acid region. The expression cassette portion can include a gene to be transcribed and elements that control the expression of the gene (e.g., a promoter). 
     As used herein, the term “β-globin locus control region (LCR)” refers to a polynucleotide composed of one or more Dnase I hypersensitive site (HS) regions, including a HS1 region, a HS2 region, a HS3 region, and a HS4 region. The structure of many LCRs of the β-globin genes have been published, e.g., human (Li et al.,  J. Biol. Chem.  (1985); 260:14,901; Li et al.,  Proc. Natl. Acad. Sci.  (1990) 87:8207); mouse (Shehee et al.,  J. Mol. Biol.  (1989); 205:41); rabbit (Margot et al., J. Mol. Biol. (1989); 205:15); and goat (Li, Q., et al.,  Genomics  (1991); 9:488), each of which are incorporated by reference herein. In certain embodiments, the β-globin LCR comprises a HS2 region (e.g., a β-globin LCR comprising a HS2 region, a HS3 region and a HS4 region; and a β-globin LCR comprising a HS1 region, a HS2 region, a HS3 region and a HS4 region). In certain embodiments, the β-globin LCR does not comprise a HS2 region (e.g., a β-globin LCR comprising or consisting essentially of a HS1 region, a HS3 region, a HS4 region). In certain embodiments, the β-globin LCR does not comprise a HS2 region or a HS1 region (e.g., a β-globin LCR comprising or consisting essentially of a HS3 region and a HS4 region). In certain embodiments, the β-globin LCR does not comprise a HS1 region, a HS2 region or a HS4 region (e.g., a β-globin LCR comprising or consisting essentially of a HS3 region). 
     As used herein, the term “recombinant” includes reference to a cell or vector, that has been modified by the introduction of a heterologous nucleic acid or that the cell is derived from a cell so modified. Thus, for example, recombinant cells express genes that are not found in identical form within the native (non-recombinant) form of the cell or express native genes that are otherwise abnormally expressed, under expressed or not expressed at all as a result of deliberate human intervention or may have reduced or eliminated expression of a native gene. 
     As used herein, the term “globin” refers to a family of heme-containing proteins that are involved in the binding and transport of oxygen. Subunits of vertebrate and invertebrate hemoglobins, vertebrate and invertebrate myoglobins or mutants thereof are included by the term globin. 
     As used herein, the term “wild-type” refers to the normal gene, virus, or organism found in nature without any mutation or modification. 
     The terms “polynucleotide”, “nucleotide”, “nucleotide sequence”, “nucleic acid” and “oligonucleotide” are used interchangeably. They refer to a polymeric form of nucleotides of any length, either deoxyribonucleotides or ribonucleotides, or analogs thereof. Polynucleotides may have any three-dimensional structure, and may perform any function, known or unknown. The following are non-limiting examples of polynucleotides: coding or non-coding regions of a gene or gene region, loci (locus) defined from linkage analysis, exons, introns, messenger RNA (mRNA), transfer RNA, ribosomal RNA, short interfering RNA (siRNA), short-hairpin RNA (shRNA), micro-RNA (miRNA), ribozymes, cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, isolated DNA of any sequence, isolated RNA of any sequence, nucleic acid probes, and primers. A polynucleotide can comprise one or more modified nucleotides, such as methylated nucleotides and nucleotide analogs. In particular embodiments, the presently disclosed subject matter provides polynucleotides encoding one or more globin genes or functional portions thereof. If present, modifications to the nucleotide structure may be imparted before or after assembly of the polymer. The sequence of nucleotides may be interrupted by non-nucleotide components. A polynucleotide may be further modified after polymerization, such as by conjugation with a labeling component. Such polynucleotides need not be about 100% identical with an endogenous nucleic acid sequence, but will typically exhibit substantial identity. Polynucleotides having “substantial identity” to an endogenous sequence are typically capable of hybridizing with at least one strand of a double-stranded nucleic acid molecule. By “hybridize” is meant pair to form a double-stranded molecule between complementary polynucleotide sequences (e.g., a gene described herein), or portions thereof, under various conditions of stringency. (See, e.g., Wahl, G. M. and S. L. Berger (1987) Methods Enzymol. 152:399; Kimmel, A. R. (1987) Methods Enzymol. 152:507). 
     For example, stringent salt concentration can ordinarily be less than about 750 mM NaCl and 75 mM trisodium citrate, e.g., less than about 500 mM NaCl and 50 mM trisodium citrate, or less than about 250 mM NaCl and 25 mM trisodium citrate. Low stringency hybridization can be obtained in the absence of organic solvent, e.g., formamide, while high stringency hybridization can be obtained in the presence of at least about 35% formamide, e.g., at least about 50% formamide. Stringent temperature conditions can ordinarily include temperatures of at least about 30° C., e.g., of at least about 37° C. or of at least about 42° C. Varying additional parameters, such as hybridization time, the concentration of detergent, e.g., sodium dodecyl sulfate (SDS), and the inclusion or exclusion of carrier DNA, are well known to those skilled in the art. Various levels of stringency are accomplished by combining these various conditions as needed. In a preferred: embodiment, hybridization will occur at 30° C. in 750 mM NaCl, 75 mM trisodium citrate, and 1% SDS. In certain embodiments, hybridization occurs at 37° C. in 500 mM NaCl, 50 mM trisodium citrate, 1% SDS, 35% formamide, and 100 μg/ml denatured salmon sperm DNA (ssDNA). In certain embodiments, hybridization occurs at 42° C. in 250 mM NaCl, 25 mM trisodium citrate, 1% SDS, 50% formamide, and 200 μg/ml ssDNA. Useful variations on these conditions can be readily apparent to those skilled in the art. 
     For most applications, washing steps that follow hybridization can also vary in stringency. Wash stringency conditions can be defined by salt concentration and by temperature. As above, wash stringency can be increased by decreasing salt concentration or by increasing temperature. For example, stringent salt concentration for the wash steps can be less than about 30 mM NaCl and 3 mM trisodium citrate, e.g., less than about 15 mM NaCl and 1.5 mM trisodium citrate. Stringent temperature conditions for the wash steps can ordinarily include a temperature of at least about 25° C., e.g., of at least about 42° C., or of at least about 68° C. In certain embodiments, wash steps occur at 25° C. in 30 mM NaCl, 3 mM trisodium citrate, and 0.1% SDS. In certain embodiments, wash steps occur at 42° C. in 15 mM NaCl, 1.5 mM trisodium citrate, and 0.1% SDS. In certain embodiments, wash steps occur at 68° C. in 15 mM NaCl, 1.5 mM trisodium citrate, and 0.1% SDS. Additional variations on these conditions will be readily apparent to those skilled in the art. Hybridization techniques are well known to those skilled in the art and are described, for example, in Benton and Davis (Science 196:180, 1977); Grunstein and Rogness (Proc. Natl. Acad. Sci., USA 72:3961, 1975); Ausubel et al. (Current Protocols in Molecular Biology, Wiley Interscience, New York, 2001); Berger and Kimmel (Guide to Molecular Cloning Techniques, 1987, Academic Press, New York); and Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, New York. 
     As used herein, the terms “polypeptide” and “protein” are used interchangeably to refer to a polymer of amino acid residues and to variants and synthetic analogues of the same. Thus, these terms apply to amino acid polymers in which one or more amino acid residues are synthetic non-naturally occurring amino acids, such as a chemical analogue of a corresponding naturally occurring amino acid, as well as to naturally-occurring amino acid polymers. Particular embodiments of the presently disclosed subject matter also include polypeptide “variants.” Polypeptide “variant” refers to polypeptides that are distinguished from a reference polypeptide by the addition, deletion, truncations, and/or substitution of at least one amino acid residue, and that retain a biological activity. In certain embodiments, a polypeptide variant is distinguished from a reference polypeptide by one or more substitutions, which may be conservative or non-conservative, as known in the art. In certain embodiments, a variant polypeptide includes an amino acid sequence having at least about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity or similarity to a corresponding sequence of a reference polypeptide. In certain embodiments, the amino acid additions or deletions occur at the C-terminal end and/or the N-terminal end of the reference polypeptide. In certain embodiments, the amino acid deletions include C-terminal truncations of about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 15, about 20, about 25, about 30, about 35, about 40, about 45, 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, about 100, about 105, about 110, about 115, about 120, about 125, about 130, about 135, about 140, about 145, about 150, about 155, about 160, about 165, about 170, or about 175 or more amino acids, including all intervening numbers of amino acids, e.g., 25, 26, 27, 29, 30 . . . 100, 101, 102, 103, 104, 105 . . . 170, 171, 172, 173, 174, etc. 
     As noted above, polypeptides of the presently disclosed subject matter may be altered in various ways including amino acid substitutions, deletions, truncations, and insertions. Methods for such manipulations are generally known in the art. For example, amino acid sequence variants of a reference polypeptide can be prepared by mutations in the DNA. Methods for mutagenesis and nucleotide sequence alterations are well known in the art. See, for example, Kunkel (1985, Proc. Natl. Acad. Sci. USA. 82: 488-492), Kunkel et al., (1987, Methods in Enzymol, 154: 367-382), U.S. Pat. No. 4,873,192, Watson, J. D. et al., Molecular Biology of the Gene, Fourth Edition, Benjamin/Cummings, Menlo Park, Calif., 1987) and the references cited therein. Guidance as to appropriate amino acid substitutions that do not affect biological activity of the protein of interest may be found in the model of Dayhoff et al., (1978) Atlas of Protein Sequence and Structure (Natl. Biomed. Res. Found., Washington, D.C.). 
     As used herein, the term “substantially identical” refers to a polypeptide or a polynucleotide exhibiting at least 50% identity to a reference amino acid sequence (for example, any one of the amino acid sequences described herein) or a nucleic acid sequence (for example, any one of the nucleic acid sequences described herein). Preferably, such a sequence is at least about 60%, e.g., about 80%, about 85%, about 90%, about 95%, or about 99% identical at the amino acid level or nucleic acid to the sequence used for comparison. 
     Sequence identity or homology is typically measured using sequence analysis software (for example, Sequence Analysis Software Package of the Genetics Computer Group, University of Wisconsin Biotechnology Center, 1710 University Avenue, Madison, Wis. 53705, BLAST, BESTFIT, GAP, or PILEUP/PRETTYBOX programs). Such software matches identical or similar sequences by assigning degrees of homology to various substitutions, deletions, and/or other modifications. In an exemplary approach to determining the degree of identity or homology, a BLAST program may be used, with a probability score between e-3 and e-100 indicating a closely related sequence. The percentage of identity between two sequences can also be determined with programs such as DNAMAN (Lynnon Biosoft, version 3.2). Using this program two sequences can be aligned using the optimal alignment algorithm (Smith and Waterman, 1981). After alignment of the two sequences the percentage identity can be calculated by dividing the number of identical nucleotides between the two sequences by the length of the aligned sequences minus the length of all gaps. 
     Terms that describe the orientation of polynucleotides include: 5′ (normally the end of the polynucleotide having a free phosphate group) and 3′ (normally the end of the polynucleotide having a free hydroxyl (OH) group). Polynucleotide sequences can be annotated in the 5′ to 3′ orientation or the 3′ to 5′ orientation. 
     As used herein, a “single guide RNA” or a “synthetic guide RNA” refers to the polynucleotide sequence comprising the guide sequence, the tracr sequence and the tracr mate sequence. The term “guide sequence” refers to the about 20 bp sequence within the guide RNA that specifies the target site and may be used interchangeably with the terms “guide” or “spacer”. The term “tracr mate sequence” may also be used interchangeably with the term “direct repeat(s)”. 
     The terms “non-naturally occurring” or “engineered” are used interchangeably and indicate the involvement of the hand of man. The terms, when referring to nucleic acid molecules or polypeptides mean that the nucleic acid molecule or the polypeptide is at least substantially free from at least one other component with which they are naturally associated in nature and as found in nature. 
     As used herein, the term “expression” refers to the process by which a polynucleotide is transcribed from a DNA template (such as into and mRNA or other RNA transcript) and/or the process by which a transcribed mRNA is subsequently translated into peptides, polypeptides, or proteins. Transcripts and encoded polypeptides may be collectively referred to as “gene product.” If the polynucleotide is derived from genomic DNA, expression may include splicing of the mRNA in a eukaryotic cell. 
     As used herein, the term “treating” or “treatment” refers to clinical intervention in an attempt to alter the disease course of the individual or cell being treated, and can be performed either for prophylaxis or during the course of clinical pathology. Therapeutic effects of treatment include, without limitation, preventing occurrence or recurrence of disease, alleviation of symptoms, diminishment of any direct or indirect pathological consequences of the disease, preventing metastases, decreasing the rate of disease progression, amelioration or palliation of the disease state, and remission or improved prognosis. By preventing progression of a disease or disorder, a treatment can prevent deterioration due to a disorder in an affected or diagnosed subject or a subject suspected of having the disorder, but also a treatment may prevent the onset of the disorder or a symptom of the disorder in a subject at risk for the disorder or suspected of having the disorder. 
     As used herein, the term “subject” or “individual” refers to a vertebrate, such as a human or non-human animal, for example, a mammal. Mammals include, but are not limited to, humans, primates, farm animals, sport animals, rodents and pets. Non-limiting examples of non-human animal subjects include rodents such as mice, rats, hamsters, and guinea pigs; rabbits; dogs; cats; sheep; pigs; goats; cattle; horses; and non-human primates such as apes and monkeys. The term “immunocompromised” as used herein refers to a subject who has an immunodeficiency. The subject is very vulnerable to opportunistic infections, infections caused by organisms that usually do not cause disease in a person with a healthy immune system, but can affect people with a poorly functioning or suppressed immune system. 
     As used herein, the term “isolated cell” refers to a cell that is separated from the molecular and/or cellular components that naturally accompany the cell. As used herein, the term “isolated” refers to material that is free, substantially free, or essentially free to varying degrees from components which normally accompany it as found in its native state. “Isolate” denotes a degree of separation from original source or surroundings. 
     As used herein, the term “cell population” refers to a group of at least two cells expressing similar or different phenotypes. In non-limiting examples, a cell population can include at least about 10, at least about 100, at least about 200, at least about 300, at least about 400, at least about 500, at least about 600, at least about 700, at least about 800, at least about 900, at least about 10 3  cells, at least about 10 4  cells, at least about 10 5  cells, at least about 10 6  cells, at least about 10 7  cells, or at least about 10 8  cells expressing similar or different phenotypes. 
     As used herein, the term “cleavage” refers to the breakage of the covalent backbone of a DNA molecule. Cleavage can be initiated by a variety of methods including, but not limited to, enzymatic or chemical hydrolysis of a phosphodiester bond. Both single-stranded cleavage and double-stranded cleavage are possible, and double-stranded cleavage can occur as a result of two distinct single-stranded cleavage events. DNA cleavage can result in the production of either blunt ends or staggered ends. In certain embodiments, fusion polypeptides are used for targeted double-stranded DNA cleavage. 
     As used herein, the term “cleavage half-domain” refers to a polypeptide sequence which, in conjunction with a second polypeptide (either identical or different) forms a complex having cleavage activity (preferably double-strand cleavage activity). The terms “first and second cleavage half-domains;” “+ and − cleavage half-domains” and “right and left cleavage half-domains” are used interchangeably to refer to pairs of cleavage half-domains that dimerize. 
     As used herein, the term “chromosome” refers to a chromatin complex comprising all or a portion of the genome of a cell. The genome of a cell is often characterized by its karyotype, which is the collection of all the chromosomes that comprise the genome of the cell. The genome of a cell can comprise one or more chromosomes. 
     As used herein, the term “gene” includes a DNA region encoding a gene product, as well as all DNA regions which regulate the production of the gene product, whether or not such regulatory sequences are adjacent to coding and/or transcribed sequences. Accordingly, a gene includes, but is not limited to, promoter sequences, terminators, translational regulatory sequences such as ribosome binding sites and internal ribosome entry sites, enhancers, silencers, insulators, boundary elements, replication origins, matrix attachment sites and locus control regions 
     The terms “operative linkage” and “operatively linked” (or “operably linked”) are used interchangeably with reference to a juxtaposition of two or more components (such as sequence elements), in which the components are arranged such that both components function normally and allow the possibility that at least one of the components can mediate a function that is exerted upon at least one of the other components. By way of illustration, a transcriptional regulatory sequence, such as a promoter, is operatively linked to a coding sequence if the transcriptional regulatory sequence controls the level of transcription of the coding sequence in response to the presence or absence of one or more transcriptional regulatory factors. A transcriptional regulatory sequence is generally operatively linked in cis with a coding sequence, but need not be directly adjacent to it. For example, an enhancer is a transcriptional regulatory sequence that is operatively linked to a coding sequence, even though they are not contiguous. 
     A “functional region” or “functional portion” of a protein, polypeptide or nucleic acid is a protein, polypeptide or nucleic acid whose sequence is not identical to the full-length protein, polypeptide or nucleic acid, yet retains the same function as the full-length protein, polypeptide or nucleic acid. A functional region can possess more, fewer, or the same number of residues as the corresponding native molecule, and/or can contain one or more amino acid or nucleotide substitutions. Methods for determining the function of a nucleic acid (e.g., coding function, ability to hybridize to another nucleic acid) are well-known in the art. Similarly, methods for determining protein function are well-known. For example, the DNA-binding function of a polypeptide can be determined, for example, by filter-binding, electrophoretic mobility-shift, or immunoprecipitation assays. DNA cleavage can be assayed by gel electrophoresis. The ability of a protein to interact with another protein can be determined, for example, by co-immunoprecipitation, two-hybrid assays or complementation, both genetic and biochemical. 
     As used herein, the term “promoter” refers to a recognition site of a polynucleotide (DNA or RNA) to which an RNA polymerase binds. The term “enhancer” refers to a segment of DNA which contains sequences capable of providing enhanced transcription and in some instances can function independent of their orientation relative to another control sequence. An enhancer can function cooperatively or additively with promoters and/or other enhancer elements. 
     As used herein, the term “vector” refers to any genetic element, such as a plasmid, phage, transposon, cosmid, chromosome, virus, virion, etc., which is capable of replication when associated with the proper control elements and which can transfer gene sequences into cells. Thus, the term includes cloning and expression vehicles, as well as viral vectors and plasmid vectors. 
     As used herein, the term “modulate” refers to altering positively or negatively. Exemplary modulations include an about 1%, about 2%, about 5%, about 10%, about 25%, about 50%, about 75%, or about 100% change. 
     As used herein, the term “increase” refers to alter positively by at least about 5%, including, but not limited to, alter positively by about 5%, by about 10%, by about 25%, by about 30%, by about 50%, by about 75%, or by about 100%. 
     As used herein, the term “reduce” refers to alter negatively by at least about 5% including, but not limited to, alter negatively by about 5%, by about 10%, by about 25%, by about 30%, by about 50%, by about 75%, or by about 100%. 
     As used herein, the term “about” or “approximately” means within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, “about” can mean within 3 or more than 3 standard deviations, per the practice in the art. Alternatively, “about” can mean a range of up to 20%, up to 10%, up to 5%, or up to 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, preferably within 5-fold, and more preferably within 2-fold, of a value. 
     II. Insulators 
     Several cases of vector-related malignant transformation have been reported in clinical settings, associated with the activation of cellular oncogenes by vector-encoded enhancers (Baum et al. (2006), Nienhuis et al. (2006), Ramezani et al. (2006)) and various vector modifications have been performed or proposed to reduce vector genotoxicity (Baum et al. (2006), Nienhuis et al. (2006), Ramezani et al. (2006)). A class of DNA elements known as chromatin insulators has been recognized as one approach to improve vector safety and performance (Emery (2011)). 
     Insulators are naturally occurring DNA elements that help from the functional boundaries between adjacent chromatin domains. Insulators bind proteins that modify chromatin and alter regional gene expression. The placement of insulators in the vectors described herein offer various potential benefits including, but not limited to, 1) shielding of the vector from positional effect variegation of expression by flanking chromosomes (i.e., barrier activity, which may decrease position effects and vector silencing); and 2) shielding flanking chromosomes from insertional trans-activation of endogenous gene expression by the vector (enhancer blocking). There are two basic classes of chromatin insulators: (a) barrier insulators that block the encroachment of silencing heterochromatin into adjoining regions of open chromatin that are transcriptionally permissive, and (b) enhancer blocking insulators that prevent enhancer-mediated transcriptional activation of adjoining regions. The sequences that mediate these activities are physically separable and mechanistically distinct (Recillas-Targa et al. (2002)). Chromatin insulators do not exhibit inherent transcriptional enhancing or repressing activities on their own. As such, they make ideal elements for reducing the interaction between gene transfer vectors and the target cell genome. Insulators can help to preserve the independent function of genes or transcription units embedded in a genome or genetic context in which their expression may otherwise be influenced by regulatory signals within the genome or genetic context (see, e.g., Burgess-Beusse et al. (2002)  Proc. Nat&#39;l Acad. Sci. USA,  99: 16433; and Zhan et al. (2001)  Hum. Genet.,  109: 471). 
     The problems created by insertional mutagenesis of viral vectors are widely known (Nienhuis (2013), Baum et al. (2006), Nienhuis et al. (2006)) as is the evidence that the risks of genotoxicity can be reduced by the use of chromatin insulators (Arumugam et al. (2007), Emery (2011), Evans-Galea et al. (2007), Rivella et al. (2000), Emery et al. (2000), Emery et al. (2002), Yannaki et al. (2002), Hino et al. (2004), Ramezani et al. (2003), Ramezani et al. (2008)). The presently disclosed subject matter provides novel insulators that are powerful enhancer blocking insulators, and certain insulators additionally possess barrier insulator activity. In vertebrates, the function of enhancer blocking insulators is mediated through the zinc-finger DNA-binding factor CTCF (Gaszner and Felsenfeld (2006), Wallace and Felsenfeld (2007)). In general, these elements are thought to function through physical loop structures, which are established by CTCF-mediated interactions between adjacent insulator elements or through CTCF-mediated tethering of the chromatin fiber to structural elements within the nucleus. The first characterized vertebrate chromatin insulator is located within the chicken β-globin locus control region. This element, which contains a DNase-I hypersensitive site-4 (cHS4), appears to constitute the 5′ boundary of the chicken β-globin locus (Prioleau et al. (1999) EMBO J. 18: 4035-4048). A 1.2-kb region containing the cHS4 element displays classic insulator activities, including the ability to block the interaction of globin gene promoters and enhancers in cell lines (Chung et al. (1993) Cell, 74: 505-514), and the ability to protect expression cassettes in  Drosophila  (Id.), transformed cell lines (Pikaart et al. (1998)  Genes Dev.  12: 2852-2862), and transgenic mammals (Wang et al. (1997)  Nat. Biotechnol.,  15: 239-243; Taboit-Dameron et al. (1999) Transgenic Res., 8: 223-235) from position effects. Much of this activity is contained in a 250-bp region. Within this stretch is a 49-bp cHS4 element (Chung et al. (1997)  Proc. Natl. Acad. Sci., USA,  94: 575-580) that interacts with the zinc finger DNA binding protein CTCF implicated in enhancer-blocking assays (Bell et al. (1999)  Cell,  98: 387-396). 
     Insulators, such as cHS4, can block the interaction between enhancers and promoters when placed between these elements (Evans-Galea et al. (2007), Chung et al. (1997), Bell et al. (1999), Ryu et al. (2007), Ryu et al. (2008)). Several studies have demonstrated the ability of the cHS4 insulator to reduce position-effect silencing of gammaretroviral vectors (Evans-Galea et al. (2007), Rivella et al. (2000), Emery et al. (2000), Emery et al. (2002), Yannaki et al. (2002), Hino et al. (2004), Ramezani et al. (2006), Yao et al. (2003), Nishino et al. (2006), Aker et al. (2007), Li and Emery (2008)), and lentiviral vectors (Bank et al. (2005), Arumugam et al. (2007), Puthenveetil et al. (2004), Evans-Galea et al. (2007), Ramezani et al. (2003), Aker et al. (2007), Ma et al. (2003), Chang et al. (2005), Pluta et al. (2005)). Those appropriately designed studies demonstrated that inclusion of the 1.2 kb version of the cHS4 insulator increased the likelihood and/or consistency of vector transgene expression in at least some settings (Arumugam et al. (2007), Emery (2011), Evans-Galea et al. (2007), Emery et al. (2002), Yannaki et al. (2002), Hino et al. (2004), Ramezani et al. (2006), Aker et al. (2007), Li and Emery (2008), Pluta et al. (2005). Jakobsson et al. (2004)). Nevertheless, the degree of protection afforded by the cHS4 insulator is far from complete. In addition, the inclusion of the 1.2 Kb cHS4 can adversely affect vector titers while the smallest cHS4 core has been proven ineffective (Aker et al. (2007), Jakobsson et al. (2004)). By contrast, the insulators of the presently disclosed subject matter do not affect adversely the titers of viral vectors, and are more powerful and effective than the cHS4 insulator. 
     The presently disclosed insulators are identified through genomic approaches, e.g., using genomic approaches to identify insulators that are powerful enhancer blockers as well as barrier insulators of the human genome. The presently disclosed insulators enhance the safety of gene therapy (e.g., stem cell gene therapy, globin gene therapy). For gene therapy of the hemoglobinopathies, powerful enhancers are required to achieve therapeutic levels of globin gene expression. Powerful insulators therefore represent one means to protect the genomic environment from the powerful enhancers of the integrating vectors. 
     The presently disclosed insulators possess powerful enhancer blocking activity. For example, and not by way of limitation, an insulator of the present disclosure can reduce the activity of an enhancer element by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98% or at least about 99%. In certain embodiments, the insulators possess barrier activity in addition to enhancer blocking activity. The presently disclosed insulators substantially decrease the risks of insertional mutagenesis and genotoxicity associated with viral vectors. Furthermore, when at least one presently disclosed insulator is incorporated into a vector, the insulator(s) does not adversely effect vector titers of the vector. In certain embodiments, the presently disclosed insulators (e.g., insulator A1) increase the in vivo expression of the globin gene or functional portion thereof. 
     In certain embodiments, the insulator comprises a Transcriptional repressor CTCF binding site, which has the nucleotide sequence set forth in SEQ ID NO: 18, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 18] 
               
               
                 CACCAGGTGGCGCT. 
               
            
           
         
       
     
     In certain embodiments, the insulator has the nucleotide sequence set forth in SEQ ID NO:1, which is provided below, or a sequence which is at least about 80%, about 85%, about 90%, about 95%, about 98, about 99% or about 100% % identical (homologous), to SEQ ID NO:1. SEQ ID NO: 1 is provided below. 
     
       
         
           
               
            
               
                 [SEQ ID NO: 1] 
               
               
                 TCCTTCCTTTCTAAATGACGAGAGAGACAGAAGAATTCTTCAAGGTTAGT 
               
               
                   
               
               
                 GTGTCCAGCATGCAACCTTTCCTTCCTGGATGAGCATCCCTGGAGTAGGA 
               
               
                   
               
               
                 GAGCCAGCCTGCCTCCTGCGCTGGCACAGAGCCCGGTTCCCTAGACAACT 
               
               
                   
               
               
                 GCCTCTCCAAATCTGATGTCCAGCGCCACCTGGTGTCCACATCAAGCAGA 
               
               
                   
               
               
                 CACAATTAATAGTCAACCTGTTCAGGAAAACTGTGAGGGGGAAAAAAAAG 
               
               
                   
               
               
                 AAAGAGGATTTATGAAGGGAAAAGAAAGTTTAGAGGATATGCCACGATTG 
               
               
                   
               
               
                 GCTAG 
               
            
           
         
       
     
     In certain embodiments, the insulator comprises or has the nucleotide sequence as set forth in SEQ ID NO:24, or a sequence which is at least about 80%, about 85%, about 90%, about 95%, about 98, about 99% or about 100% identical (homologous), to SEQ ID NO: 24. SEQ ID NO: 24 is provided below. 
     
       
         
           
               
            
               
                 [SEQ ID NO: 24] 
               
               
                 CCAATC GTGGCATATC CTCTAAACTT TCTTTTCCCT 
               
               
                   
               
               
                 TCATAAATCC TCTTTCTTTT TTTTCCCCCT CACAGTTTTC 
               
               
                   
               
               
                 CTGAACAGGT TGACTATTAA TTGTGTCTGC TTGATGTGGA 
               
               
                   
               
               
                 CACCAGGTGG CGCTGGACAT CAGATTTGGA GAGGCAGTTG 
               
               
                   
               
               
                 TCTAGGGAAC CGGGCTCTGT GCCAGCGCAG GAGGCAGGCT 
               
               
                   
               
               
                 GGCTCTCCTA TTCCAGGGAT GCTCATCCAG GAAGGAAAGG 
               
               
                   
               
               
                 TTGCATGCTG GACACACTAA CCTTGAAGAA TTCTTCTGTC 
               
               
                   
               
               
                 TCTCTCGTCA TTTAGAAAGG AAGGA. 
               
            
           
         
       
     
     In certain embodiments, the insulator comprises or has the nucleotide sequence as set forth in SEQ ID NO:25 (which is the reverse complement of SEQ ID NO: 1), or a sequence which is at least about 80%, about 85%, about 90%, about 95%, about 98, about 99% or about 100% identical (homologous), to SEQ ID NO: 25. SEQ ID NO: 25 is provided below. 
     
       
         
           
               
            
               
                 [SEQ ID NO: 25] 
               
               
                 CTAGCCAATCGTGGCATATCCTCTAAACTTTCTTTTCCCTTCATAAATCC 
               
               
                   
               
               
                 TCTTTCTTTTTTTTCCCCCTCACAGTTTTCCTGAACAGGTTGACTATTAA 
               
               
                   
               
               
                 TTGTGTCTGCTTGATGTGGACACCAGGTGGCGCTGGACATCAGATTTGGA 
               
               
                   
               
               
                 GAGGCAGTTGTCTAGGGAACCGGGCTCTGTGCCAGCGCAGGAGGCAGGCT 
               
               
                   
               
               
                 GGCTCTCCTACTCCAGGGATGCTCATCCAGGAAGGAAAGGTTGCATGCTG 
               
               
                   
               
               
                 GACACACTAACCTTGAAGAATTCTTCTGTCTCTCTCGTCATTTAGAAAGG 
               
               
                   
               
               
                 AAGGA 
               
            
           
         
       
     
     In certain embodiments, the insulator comprises or has the nucleotide sequence as set forth in SEQ ID NO: 27, or a sequence which is at least about 80%, about 85%, about 90%, about 95%, about 98, about 99% or about 100% identical (homologous), to SEQ ID NO: 27. SEQ ID NO: 27 is provided below. 
     
       
         
           
               
            
               
                 [SEQ ID NO: 27] 
               
               
                 ctggttctac tcattacatt ccaatcgtgg catatcctct 
               
               
                   
               
               
                 aaactttctt ttcccttcat aaatcctctt tctttttttt 
               
               
                   
               
               
                 ccccctcaca gttttcctga acaggttgac tattaattgt 
               
               
                   
               
               
                 gtctgcttga tgtggacacc aggtggcgct ggacatcaga 
               
               
                   
               
               
                 tttggagagg cagttgtcta gggaaccggg ctctgtgcca 
               
               
                   
               
               
                 gcgcaggagg caggctggct ctcctattcc agggatgctc 
               
               
                   
               
               
                 atccaggaag gaaaggttgc atgctggaca cactaacctt 
               
               
                   
               
               
                 gaagaattct tctgtctctc tcgtcattta gaaaggaagg 
               
            
           
         
       
     
     In certain embodiments, the insulator comprises or has the nucleotide sequence as set forth in SEQ ID NO: 28, or a sequence which is at least about 80%, about 85%, about 90%, about 95%, about 98, about 99% or about 100% identical (homologous), to SEQ ID NO: 28. SEQ ID NO: 28 is provided below. 
     
       
         
           
               
            
               
                 [SEQ ID NO: 28] 
               
               
                 CCAATC GTGGCATATC CTCTAAACTT TCTTTTCCCT 
               
               
                   
               
               
                 TCATAAATCC TCTTTCTTTT TTTTCCCCCT CACAGTTTTC 
               
               
                   
               
               
                 CTGAACAGGT TGACTATTAA TTGTGTCTGC TTGATGTGGA 
               
               
                   
               
               
                 CACCAGGTGG CGCTGGACAT CAGATTTGGA GAGGCAGTTG 
               
               
                   
               
               
                 TCTAGGGAAC CGGGCTCTGT GCCAGCGCAG GAGGCAGGCT 
               
               
                   
               
               
                 GGCTCTCCTA TTCCAGGGAT GCTCATCCAG GAAGGAAAGG 
               
               
                   
               
               
                 TTGCATGCTG GACACACTAA CCTTGAAGAA TTCTTCTGTC 
               
               
                   
               
               
                 TCTCTCGTCA TTTAGAAAGG AAGG 
               
            
           
         
       
     
     In certain embodiments, the insulator comprises or has the nucleotide sequence as set forth in hg18 coordinates 76229933 to 76230115 of chromosome 1. 
     In certain embodiments, the insulator comprises or has the nucleotide sequence between residues 68041 and 68160 (or SEQ ID NO: 29 provided below), or between residues and 68041 and 68210 (or SEQ ID NO: 30 provided below), or between residues 68041 and 68280 (or SEQ ID NO: 31 provided below), or between residues 68005 and 68305 (or SEQ ID NO: 24), of  Homo sapiens  chromosome 1 clone RP11-550H2, GenBank Accession No. AC092813.2, or a sequence at least about 80%, about 85%, about 90%, about 95%, about 98, about 99% or about 100% identical (homologous) thereto. 
     
       
         
           
               
            
               
                 (SEQ ID NO: 29) 
               
               
                 TCATAAATCC TCTTTCTTTT TTTTCCCCCT CACAGTTTTC 
               
               
                   
               
               
                 CTGAACAGGT TGACTATTAA TTGTGTCTGC TTGATGTGGA 
               
               
                   
               
               
                 CACCAGGTGG CGCTGGACAT CAGATTTGGA GAGGCAGTTG 
               
               
                   
               
               
                 (SEQ ID NO: 30) 
               
               
                 TCATAAATCC TCTTTCTTTT TTTTCCCCCT CACAGTTTTC 
               
               
                   
               
               
                 CTGAACAGGT TGACTATTAA TTGTGTCTGC TTGATGTGGA 
               
               
                   
               
               
                 CACCAGGTGG CGCTGGACAT CAGATTTGGA GAGGCAGTTG 
               
               
                   
               
               
                 TCTAGGGAAC CGGGCTCTGT GCCAGCGCAG GAGGCAGGCT 
               
               
                   
               
               
                 GGCTCTCCTA 
               
               
                   
               
               
                 (SEQ ID NO: 31) 
               
               
                 TCATAAATCC TCTTTCTTTT TTTTCCCCCT CACAGTTTTC 
               
               
                   
               
               
                 CTGAACAGGT TGACTATTAA TTGTGTCTGC TTGATGTGGA 
               
               
                   
               
               
                 CACCAGGTGG CGCTGGACAT CAGATTTGGA GAGGCAGTTG 
               
               
                   
               
               
                 TCTAGGGAAC CGGGCTCTGT GCCAGCGCAG GAGGCAGGCT 
               
               
                   
               
               
                 GGCTCTCCTA TTCCAGGGAT GCTCATCCAG GAAGGAAAGG 
               
               
                   
               
               
                 TTGCATGCTG GACACACTAA CCTTGAAGAA TTCTTCTGTC 
               
            
           
         
       
     
     III. Expression Cassettes 
     The presently disclosed subject matter provides expression cassettes comprising one or more insulators. In certain embodiments, an expression cassette comprises at least one insulator disclosed in Section II, and a globin gene or a functional portion thereof. In certain embodiments, an expression cassette comprises at least one insulator comprising or having the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, or SEQ ID NO: 31, and a globin gene or a functional portion thereof. 
     In certain embodiments, an expression cassette comprises a globin gene or a functional portion thereof, and at least one insulator disclosed in International Patent Publication No. WO2015/138852. WO2015/138852 is herein incorporated by reference in its entirety. In certain embodiments, an expression cassette comprises a globin gene or a functional portion thereof operably and at least one insulator comprising or having the nucleotide sequence set forth in SEQ ID NO: 32, or a fragment thereof (e.g., nucleotides 1 to 320, 10 to 320, 20 to 320, 21 to 320, 55 to 320, 55 to 300, 57 to 300, 57 to 296, 57 to 226, 57 to 200, or 57 to 176 of SEQ ID NO: 32). SEQ ID NO:32 is provided below. 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 32) 
                   
               
               
                 CTGGTTCTAC TCATTACATT CCAATCGTGG CATATCCTCT AAACTTTCTT TTCCCTTCAT  60 
                   
               
               
                   
               
               
                 AAATCCTCTT TCTTTTTTTT CCCCCTCACA GTTTTCCTGA ACAGGTTGAC TATTAATTGT 120 
               
               
                   
               
               
                 GTCTGCTTGA TGTGGACACC AGGTGGCGCT GGACATCAGA TTTGGAGAGG CAGTTGTCTA 180 
               
               
                   
               
               
                 GGGAACCGGG CTCTGTGCCA GCGCAGGAGG CAGGCTGGCT CTCCTATTCC AGGGATGCTC 240 
               
               
                   
               
               
                 ATCCAGGAAG GAAAGGTTGC ATGCTGGACA CACTAACCTT GAAGAATTCT TCTGTCTCTC 300 
               
               
                   
               
               
                 TCGTCATTTA GAAAGGAAGG                                             320 
               
            
           
         
       
     
     In certain embodiments, the globin gene or functional portion thereof is operably linked to a β-globin LCR, e.g., a β-globin LCR disclosed in section 3.1. 
     In certain embodiments, the expression cassette does not comprise a β-globin LCR. In certain embodiments, the expression cassette comprises at least one insulator (e.g., one disclosed in Section II and 3.6), and does not comprise a β-globin LCR. In certain embodiments, the expression cassette comprises at least one insulator comprising or having the nucleotide sequence set forth in SEQ ID NO: 1, and does not comprise a β-globin LCR. In certain embodiments, the expression cassette comprises an erythroid-specific enhancer (e.g., an erythroid-specific enhancer disclosed in section 3.5) and does not comprise a β-globin LCR. In certain embodiments, the expression cassette comprises an erythroid-specific enhancer having the nucleotide sequence set forth in SEQ ID NO:15, and does not comprise a β-globin LCR. In certain embodiments, the expression cassette comprises at least one insulator comprising or having the nucleotide sequence set forth in SEQ ID NO: 1, and an erythroid-specific enhancer having the nucleotide sequence set forth in SEQ ID NO:15, and does not comprise a β-globin LCR. 
     3.1. β-Globin LCR 
     The human β-globin gene cluster consists of five genes embedded within one of many olfactory receptor gene arrays (Bulger et al.,  PNAS  (1999); 96:5129-5134). The cluster spans over 80 kb on chromosome 11p15.4, and includes the five expressed β-like genes and cis-acting regulatory elements that direct their stage-specific expression during ontogeny (Forget (2001),  Molecular Mechanism of Beta Thalassemia . Steinberg M H et al., Eds. Disorders of Hemoglobin. Genetics, Pathophysiology and Clinical Management, Cambridge University Press, Cambridge). The genes are arranged in the order of their developmental expression (Stamatoyannopoulos et al., (2001) Hemoglobin Switching. In: Stamatoyannopoulos G, et al., Eds. Molecular Basis of Blood Disorders, W.B. Saunders, Philadelphia, Pa.), 5′-ε- G γ- A γ-ψη-δ-β-3′. The α-like globin gene cluster (5′-ξ2-ψξ1-ψα2-ψα1-α2-α1-θ-3′) is located very close to the telomere of the short arm of chromosome 16 and spans about 40 kb. The expression of genes encoded within these two independent clusters is limited to erythroid cells and balanced so that the output of the β-globin-like chains matches that of the α-chains. This fine-tuned balance is regulated at the transcriptional, posttranscriptional and posttranslational levels. 
     Developmental stage-specific expression is controlled by a number of proximal or distal cis-acting elements and the transcriptional factors that bind to them. In the case of the β-globin gene (HBB), the proximal regulatory elements comprise the β-globin promoter and two downstream enhancers, one located in the second intron of β-globin and the other approximately 800 bp downstream of the gene (Antoniou et al.,  EMBO J . (1988); 7:377-384; Trudel et al.,  Genes Dev . (1987); 1:954-961; Trudel et al.,  Mol. Cell. Biol . (1987); 7:4024-4029). The most prominent distal regulatory element is the β-globin LCR, located 50-60 kb upstream of the HBB and composed of several sub-regions with heightened sensitivity to DNaseI in erythroid cells (Forget (2001); Grosveld et al.,  Cell  (1987); 51:975-985; Talbot et al.,  Nature  (1989); 338:352). The most prominent property of the LCR is its strong, transcription-enhancing activity. An exemplary nucleotide sequence of the human β-globin region on chromosome 11 is set forth in SEQ ID NO:19 (GenBank Access No.: NG_000007.3), which is provided below: 
     
       
         
           
               
               
            
               
                 [SEQ ID NO: 19] 
                   
               
               
                 ggatcctcacatgagttcagtatataattgtaacagaataaaaaatcaattatgtattcaagttgctagtgtcttaagaggttcacatt 
                   
               
               
                   
               
               
                 tttatctaactgattatcacaaaaatacttcgagttacttttcattataattcctgactacacatgaagagactgacacgtaggtgcct 
               
               
                   
               
               
                 tacttaggtaggttaagtaatttatccaaaaccacacaatgtagaacctaagctgattcggccatagaaacacaatatgtggtataaat 
               
               
                   
               
               
                 gagacagagggatttctctccttcctatgctgtcagatgaatactgagatagaatatttagttcatctatcacacattaaacgggactt 
               
               
                   
               
               
                 tacatttctgtctgttgaagatttgggtgtggggataactcaaggtatcatatccaagggatggatgaaggcaggtgactctaacagaa 
               
               
                   
               
               
                 agggaaaggatgttggcaaggctatgttcatgaaagtatatgtaaaatccacattaagcttctttctgcatgcattggcaatgtttatg 
               
               
                   
               
               
                 aataatgtgtatgtaaaagtgtgctgtatattcaaaagtgtttcatgtgcctaggggtgtcaaatactttgagtttgtaagtatatact 
               
               
                   
               
               
                 tctctgtaatgtgtctgaatatctctatttacttgattctcaataagtaggtatcatagtgaacatctgacaaatgtttgaggaacaat 
               
               
                   
               
               
                 ttagtgtttacctattcaccaaaatttattaaatgcctaatctgtatcagatatacaattatctggcgaaatctgtaattcctaattta 
               
               
                   
               
               
                 aacagctgtgtagcctaattagggataaaggcatgcaaacccataatttgtgtaggttgaaatgagctatagaaaaatgcagtatattt 
               
               
                   
               
               
                 atcagaagtctttagggtcatgaaaaggaatggtcaactgacactgccagggactcatatgtaagagataactaatgtgaagtgacttt 
               
               
                   
               
               
                 aaaggagaaattagcagaagttttctttccatgtctcctcatcatgttacaataacggaagagattaaaacaacaaatacatttagaca 
               
               
                   
               
               
                 gcaatgtttatcctggttagatgttttaatctaaatctatcttggagtgttaaaatgcatttgctcacctactttaaaatataaatgaa 
               
               
                   
               
               
                 ggtaggaacctgtagatacaaaaagttggagaaaaaaagacaataaagatgacaaaaatctattaatccttgatagaaaatgagaagag 
               
               
                   
               
               
                 ataaaacactggtttacataaagaaaataagatggatagatagcagatccttataaaagtgataatttgagaaaaaaaatactccatat 
               
               
                   
               
               
                 tctgagtttcttcacataaaataatacaaatctgctgtggtaagttacaaagagatagattttttatcattatataaaagatattttaa 
               
               
                   
               
               
                 acagagttatacaacaaaggaacagactatgtcatatattctcacttatcactataaacatctcagaaaaatctgcaaaatcatttcat 
               
               
                   
               
               
                 agcattttaaatagttaggaataatgtagaaaactgaaacagttctaagtttcccacaaacttagagtctcaaatgttgcattacctaa 
               
               
                   
               
               
                 cttacctgcaaatattttatacaaatttgcacatgctactctagtcaaaaatatatgtacattatgggtattttctgtgtgtaacttgg 
               
               
                   
               
               
                 ttctagttgcttctttcagaaatagcctctatttttgatttacctgataaaatcacattcctctccaaagccttctaaatacttccaga 
               
               
                   
               
               
                 ctaactactttttagtacatctaagaagaaaagagttttgtctcttatccacctctgagtcaaaaagcagcatgtccatcaattggtac 
               
               
                   
               
               
                 atagttcccacagccccacttagctctggattggagttctacttggcattgtttgcaactacatggacgtaaaatgcatggattctctt 
               
               
                   
               
               
                 gaaaaaatgtttctgccatgatgttctctgaaagagactaaccttccctcgctttgcagagaaagactcgtgtaatccttgacaatgtc 
               
               
                   
               
               
                 atctcatctatttattcccatgtctacccatatgtgaccttcatgtctttgctctaagcccctacatcctcaatctacacactaggata 
               
               
                   
               
               
                 gtataaaagtaatagtaataatagtagtaatagtaataacaatacaatgattatggcttatactatacacaagacactgttgatatatt 
               
               
                   
               
               
                 atttcatttagtattcacagtaactctgtgcctcaagtactattgtaataccctttaagaggaggaaactgaggcacagggccctaaag 
               
               
                   
               
               
                 taatattccaagatgaagtggctactaactgacagagggcataattcaactcatgatatttggctctagaatacatgctctgaatcatt 
               
               
                   
               
               
                 atacaataataattcatgaggaaacattttttaaagcctaagttatttgctctgaaataagacataatttggggtgagaaagcttagat 
               
               
                   
               
               
                 tccatgaagtattacagcatttggtagtctttttgcactccaggtcttatttttactgcttaaacataataaaacatatggttcagtat 
               
               
                   
               
               
                 gcctttgattttacaataatattcctgttatttttggaagcacagggtgtgggataatgctaattactagtgattagtattgagaggtg 
               
               
                   
               
               
                 acagcgtgctggcagtcctcacagccctcgctcgctcttggcgcctcctctgcctgggctcccacattggtggcacttgaggagccctt 
               
               
                   
               
               
                 cagccggccgctgcactgtgggagcccttttctgggctggccaaggccagagccggctccctcagcttgccaggaggtgtggagggaca 
               
               
                   
               
               
                 gacgcgggcaggaaccgggctgtgcgccgtgcttgagggagttccgggtgggcatgggctccgaggaccccgcactcggagccgccagc 
               
               
                   
               
               
                 cggccccaccggccgcgggcagtgaggggcttagcacctgggccagcagctgctgtgctcaattcctcgccgggccttagctgccttcc 
               
               
                   
               
               
                 tgcggggcagggctcgggacctgcagcgcgccatgcctgagcctccccaccttcatgggctcctgtgcggcccgagcctcgccgacgag 
               
               
                   
               
               
                 cgccgccccctgctccagggcacccagtcccatcgaccacccaagggctgaagagtgcgggcgcacggcaggggactggcaggcagctc 
               
               
                   
               
               
                 cccctgcagcccaggtgcgggatccactgggtgaagccggctaggctcctgagtttgctggggatgcgaagaacccttatgtctagata 
               
               
                   
               
               
                 agggattgtaaatacaccaattggcactctgtatctagctcaaggtttgtaaacacaccaatcagcaccctgtgtctagctcagggttt 
               
               
                   
               
               
                 gtgaatgcaccaatcaacactctatctagctactctggtggggccttggagaacctttatgtctagctcagggattgtaaatacaccaa 
               
               
                   
               
               
                 tcggcagtctgtatctagctcaaggtttgtaaacacaccaatcagcaccctgtgtctagctcagggtttgtgaatgcaccaatcaacac 
               
               
                   
               
               
                 tctgtatctagctactctggtggggacgtggagaacctttatgtctagctcagggattgtaaatacaccactcggcagtctgtatctag 
               
               
                   
               
               
                 ctcaaggtttgtaaacacaccaatcagcaccctgtgtctagctcagggtttgtgaatgcaccaatcaacactctgtatctagctactct 
               
               
                   
               
               
                 ggtggggacttggagaacctttgtgtggacactctgtatctagctaatctggtggggacgtggagaacctttgtgtctagctcatggat 
               
               
                   
               
               
                 tgtaaatgcaccaatcagtgccctgtcaaaacagaccactgggctctaccaatcagcaggatgtgggtggggccagataagagaataaa 
               
               
                   
               
               
                 agcaggctgcccgagccagcagtggcaacccgctcgggtccccttccacactgtggaagctttgttctttcgctctttgcaataaatct 
               
               
                   
               
               
                 tgctgctgctcactgtttgggtctacactgcctttatgagctgtaacgctcaccgcgaaggtctgcagcttcactcttgaagccagcga 
               
               
                   
               
               
                 gaccacgaacccaccgggaggaacgaacaactccagaggcgccgccttaagagctggaacgttcactgtgaaggtctgcagcttcactc 
               
               
                   
               
               
                 ctgagccagcgagaccacgaacccatcagaaggaagaaactccgaacacatccaaacatcagaacgaacaaactccacacacgcagcct 
               
               
                   
               
               
                 ttaagaactgtaacactcaccacgagggtccccggcttcattcttgaagtcagtgaaaccaagaacccaccaattccggacacagtatg 
               
               
                   
               
               
                 tcagaaacaatatgagtcactaaatcaatatacttctcaacaatttccaacagcccttgcaattaacttggccatgtgactggttgtga 
               
               
                   
               
               
                 ctaaaataatgtggagataataatgtgttactccctaaggcagagtgcccttctatcattctctttcccttcctctatgtggcagaaag 
               
               
                   
               
               
                 taaaagattctgaaatgataaagtcaatcacaggaaggcacctggactcctggcccactgcttggaggagagcactcaggaccatgaac 
               
               
                   
               
               
                 atctgactgtgacgtagcaataaagaaacccacgtttcatatgaaactgcttaaaattaatggcacaagtcatgtttttgatgttgcac 
               
               
                   
               
               
                 atttgtctttatttgtggcttgttttgcttccacatcaatccactcaaggcctacattctgctataatgcaatttcaagttctttacag 
               
               
                   
               
               
                 gccgagaaaaatgaatctgaattcctgacctccaaaagtgatcaagatatttttagttcaggctccaaaattttctcattttcataggt 
               
               
                   
               
               
                 tttcctcgattgatcattattcatgatttgcaaggaatcattcaatgttttctaaatctattactgcatcctgacacatatgacatttt 
               
               
                   
               
               
                 aactatgttccagatttttgaatgaagagtgtaaattttaaatgttttcaccacaaaaaataagtatgtgaagtggtggatttgttaat 
               
               
                   
               
               
                 tagccttatttaaccatttaatattgtacacgtacaccaaagcatcatgttgtaccccatgaatacacacaattattatttgtcaattt 
               
               
                   
               
               
                 aaaatgaaataataaaaaataacaaaggcattagcctctgcattgcctttaccggtcatcctcacggtgactaacgcaaaaaacgttct 
               
               
                   
               
               
                 atttcatccttacaaacatccctatctttgatgcctctttgtctagatctctatcccctcctgttttctctacgttatttatatgggta 
               
               
                   
               
               
                 tcatcaccatcctggacaacatcaggacagatatccctcaccaagccaatgttcctctctatgttggctcaaatgtccttgaactttcc 
               
               
                   
               
               
                 tttcaccaccctttccacagtcaaaaggatattgtagtttaatgcctcagagttcagcttttaagcttctgacaaattattcttcctct 
               
               
                   
               
               
                 ttaggttctcctttatggaatcttctgtactgatggccatgtcctttaactactatgtagatatctgctactacctgtattatgcctct 
               
               
                   
               
               
                 acctttattagcagagttatctgtactgttggcatgacaatcatttgttaatatgacttgcctttcctttttctgctattcttgatcaa 
               
               
                   
               
               
                 atggctcctctttcttgctcctctcatttctcctgccttcacttggacgtgcttcacgtagtctgtgcttatgactggattaaaaattg 
               
               
                   
               
               
                 atatggacttatcctaatgttgttcgtcataatatgggttttatggtccattattatttcctatgcattgatctggagaaggcttcaat 
               
               
                   
               
               
                 ccttttactctttgtggaaaatatctgtaaaccttctggttcactctgctatagcaatttcagtttaggctagtaagcatgaggatgcc 
               
               
                   
               
               
                 tccttctctgatttttcccacagtctgttggtcacagaataacctgagtgattactgatgaaagagtgagaatgttattgatagtcaca 
               
               
                   
               
               
                 atgacaaaaaacaaacaactacagtcaaaatgtttctctttttattagtggattatatttcctgacctatatctggcaggactctttag 
               
               
                   
               
               
                 agaggtagctgaagctgctgttatgaccactagagggaagaagatacctgtggagctaatggtccaagatggtggagccccaagcaagg 
               
               
                   
               
               
                 aagttgttaaggagcccttttgattgaaggtgggtgcccccaccttacagggacaggacatctggatactcctcccagtttctccagtt 
               
               
                   
               
               
                 tccctttttcctaatatatctcctgataaaatgtctatactcacttccccatttctaataataaagcaaaggctagttagtaagacatc 
               
               
                   
               
               
                 accttgcattttgaaaatgccatagactttcaaaattatttcatacatcggtctttctttatttcaagagtccagaaatggcaacatta 
               
               
                   
               
               
                 cctttgattcaatgtaatggaaagagctctttcaagagacagagaaaagaataatttaatttctttccccacacctccttccctgtctc 
               
               
                   
               
               
                 ttaccctatcttccttccttctaccctccccatttctctctctcatttctcagaagtatattttgaaaggattcatagcagacagctaa 
               
               
                   
               
               
                 ggctggttttttctaagtgaagaagtgatattgagaaggtagggttgcatgagccctttcagttttttagtttatatacatctgtattg 
               
               
                   
               
               
                 ttagaatgttttataatataaataaaattatttctcagttatatactagctatgtaacctgtggatatttccttaagtattacaagcta 
               
               
                   
               
               
                 tacttaactcacttggaaaactcaaataaatacctgcttcatagttattaataaggattaagtgagataatgcccataagattcctatt 
               
               
                   
               
               
                 aataacagataaatacatacacacacacacacattgaaaggattcttactttgtgctaggaactataataagttcattgatgcattata 
               
               
                   
               
               
                 tcattaagttctaatttcaacactagaaggcaggtattatctaaatttcatactggatacctccaaactcataaagataattaaattgc 
               
               
                   
               
               
                 cttttgtcatatatttattcaaaagggtaaactcaaactatggcttgtctaattttatatatcaccctactgaacatgaccctattgtg 
               
               
                   
               
               
                 atattttataaaattattctcaagttattatgaggatgttgaaagacagagaggatggggtgctatgccccaaatcagcctcacaatta 
               
               
                   
               
               
                 agctaagcagctaagagtcttgcagggtagtgtagggaccacagggttaagggggcagtagaattatactcccactttagtttcatttc 
               
               
                   
               
               
                 aaacaatccatacacacacagccctgagcacttacaaattatactacgctctatactttttgtttaaatgtataaataagtggatgaaa 
               
               
                   
               
               
                 gaatagatagatagatagacagatagatgatagatagaataaatgcttgccttcatagctgtctccctaccttgttcaaaatgttcctg 
               
               
                   
               
               
                 tccagaccaaagtaccttgccttcacttaagtaatcaattcctaggttatattctgatgtcaaaggaagtcaaaagatgtgaaaaacaa 
               
               
                   
               
               
                 tttctgacccacaactcatgctttgtagatgactagatcaaaaaatttcagccatatcttaacagtgagtgaacaggaaatctcctctt 
               
               
                   
               
               
                 ttccctacatctgagatcccagcttctaagaccttcaattctcactcttgatgcaacagaccttggaagcatacaggagagctgaactt 
               
               
                   
               
               
                 ggtcaacaaaggagaaaagtttgttggcctccaaaggcacagctcaaacttttcaagccttctctaatcttaaaggtaaacaagggtct 
               
               
                   
               
               
                 catttctttgagaacttcagggaaaatagacaaggacttgcctggtgcttttggtaggggagcttgcactttccccctttctggaggaa 
               
               
                   
               
               
                 atatttatccccaggtagttccctttttgcaccagtggttctttgaagagacttccacctgggaacagttaaacagcaactacagggcc 
               
               
                   
               
               
                 ttgaactgcacactttcagtccggtcctcacagttgaaaagacctaagcttgtgcctgatttaagcctttttggtcataaaacattgaa 
               
               
                   
               
               
                 ttctaatctccctctcaaccctacagtcacccatttggtatattaaagatgtgttgtctactgtctagtatccctcaagtagtgtcagg 
               
               
                   
               
               
                 aattagtcatttaaatagtctgcaagccaggagtggtggctcatgtctgtaattccagcacttgagaggtagaagtgggaggactgctt 
               
               
                   
               
               
                 gagctcaagagtttgatattatcctggacaacatagcaagacctcgtctctacttaaaaaaaaaaaaaaaattagccaggcatgtgatg 
               
               
                   
               
               
                 tacacctgtagtcccagctactcaggaggccgaaatgggaggatcccttgagctcaggaggtcaaggctgcagtgagacatgatcttgc 
               
               
                   
               
               
                 cactgcactccagcctggacagcagagtgaaaccttgcctcacgaaacagaatacaaaaacaaacaaacaaaaaactgctccgcaatgc 
               
               
                   
               
               
                 gcttccttgatgctctaccacataggtctgggtactttgtacacattatctcattgctgttcataattgttagattaattttgtaatat 
               
               
                   
               
               
                 tgatattattcctagaaagctgaggcctcaagatgataacttttattttctggacttgtaatagctttctcttgtattcaccatgttgt 
               
               
                   
               
               
                 aactttcttagagtagtaacaatataaagttattgtgagtttttgcaaacacagcaaacacaacgacccatatagacattgatgtgaaa 
               
               
                   
               
               
                 ttgtctattgtcaatttatgggaaaacaagtatgtactttttctactaagccattgaaacaggaataacagaacaagattgaaagaata 
               
               
                   
               
               
                 cattttccgaaattacttgagtattatacaaagacaagcacgtggacctgggaggagggttattgtccatgactggtgtgtggagacaa 
               
               
                   
               
               
                 atgcaggtttataatagatgggatggcatctagcgcaatgactttgccatcacttttagagagctcttggggaccccagtacacaagag 
               
               
                   
               
               
                 gggacgcagggtatatgtagacatctcattctttttcttagtgtgagaataagaatagccatgacctgagtttatagacaatgagccct 
               
               
                   
               
               
                 tttctctctcccactcagcagctatgagatggcttgccctgcctctctactaggctgactcactccaaggcccagcaatgggcagggct 
               
               
                   
               
               
                 ctgtcagggctttgatagcactatctgcagagccagggccgagaaggggtggactccagagactctccctcccattcccgagcagggtt 
               
               
                   
               
               
                 tgcttatttatgcatttaaatgatatatttattttaaaagaaataacaggagactgcccagccctggctgtgacatggaaactatgtag 
               
               
                   
               
               
                 aatattttgggttccatttttttttccttctttcagttagaggaaaaggggctcactgcacatacactagacagaaagtcaggagcttt 
               
               
                   
               
               
                 gaatccaagcctgatcatttccatgtcatactgagaaagtccccacccttctctgagcctcagtttctctttttataagtaggagtctg 
               
               
                   
               
               
                 gagtaaatgatttccaatggctctcatttcaatacaaaatttccgtttattaaatgcatgagcttctgttactccaagactgagaagga 
               
               
                   
               
               
                 aattgaacctgagactcattgactggcaagatgtccccagaggctctcattcagcaataaaattctcaccttcacccaggcccactgag 
               
               
                   
               
               
                 tgtcagatttgcatgcactagttcacgtgtgtaaaaaggaggatgcttctttcctttgtattctcacatacctttaggaaagaacttag 
               
               
                   
               
               
                 cacccttcccacacagccatcccaataactcatttcagtgactcaacccttgactttataaaagtcttgggcagtatagagcagagatt 
               
               
                   
               
               
                 aagagtacagatgctggagccagaccacctgagtgattagtgactcagtttctcttagtagttgtatgactcagtttcttcatctgtaa 
               
               
                   
               
               
                 aatggagggttttttaattagtttgtttttgagaaagggtctcactctgtcacccaaatgggagtgtagtggcaaaatctcggctcact 
               
               
                   
               
               
                 gcaacttgcacttcccaggctcaagcggtcctcccacctcaacatcctgagtagctggaaccacaggtacacaccaccatacctcgcta 
               
               
                   
               
               
                 attttttgtatttttggtagagatggggtttcacatgttacacaggatggtctcagactccggagctcaagcaatctgcccacctcagc 
               
               
                   
               
               
                 cttccaaagtgctgggattataagcatgattacaggagttttaacaggctcataagattgttctgcagcccgagtgagttaatacatgc 
               
               
                   
               
               
                 aaagagtttaaagcagtgacttataaatgctaactactctagaaatgtttgctagtattttttgtttaactgcaatcattcttgctgca 
               
               
                   
               
               
                 ggtgaaaactagtgttctgtactttatgcccattcatctttaactgtaataataaaaataactgacatttattgaaggctatcagagac 
               
               
                   
               
               
                 tgtaattagtgctttgcataattaatcatatttaatactcttggattctttcaggtagatactattattatccccattttactacagtt 
               
               
                   
               
               
                 aaaaaaactacctctcaacttgctcaagcatacactctcacacacacaaacataaactactagcaaatagtagaattgagatttggtcc 
               
               
                   
               
               
                 taattatgtctttgctcactatccaataaatatttattgacatgtacttcttggcagtctgtatgctggatgctggggatacaaagatg 
               
               
                   
               
               
                 tttaaatttaagctccagtctctgcttccaaaggcctcccaggccaagttatccattcagaaagcattttttactctttgcattccact 
               
               
                   
               
               
                 gtttttcctaagtgactaaaaaattacactttattcgtctgtgtcctgctctgggatgatagtctgactttcctaacctgagcctaaca 
               
               
                   
               
               
                 tccctgacatcaggaaagactacaccatgtggagaaggggtggtggttttgattgctgctgtcttcagttagatggttaactttgtgaa 
               
               
                   
               
               
                 gttgaaaactgtggctctctggttgactgttagagttctggcacttgtcactatgcctattatttaacaaatgcatgaatgcttcagaa 
               
               
                   
               
               
                 tatgggaatattatcttctggaatagggaatcaagttatattatgtaacccaggattagaagattcttctgtgtgtaagaatttcataa 
               
               
                   
               
               
                 acattaagctgtctagcaaaagcaagggcttggaaaatctgtgagctcctcaccatatagaaagcttttaacccatcattgaataaatc 
               
               
                   
               
               
                 cctataggggatttctaccctgagcaaaaggctggtcttgattaattcccaaactcatatagctctgagaaagtctatgctgttaacgt 
               
               
                   
               
               
                 tttcttgtctgctaccccatcatatgcacaacaataaatgcaggcctaggcatgactgaaggctctctcataattcttggttgcatgaa 
               
               
                   
               
               
                 tcagattatcaacagaaatgttgagacaaactatggggaagcagggtatgaaagagctctgaatgaaatggaaaccgcaatgcttcctg 
               
               
                   
               
               
                 cccattcagggctccagcatgtagaaatctggggctttgtgaagactggcttaaaatcagaagccccattggataagagtagggaagaa 
               
               
                   
               
               
                 cctagagcctacgctgagcaggtttccttcatgtgacagggagcctcctgccccgaacttccagggatcctctcttaagtgtttcctgc 
               
               
                   
               
               
                 tggaatctcctcacttctatctggaaatggtttctccacagtccagcccctggctagttgaaagagttacccatgcagaggccctccta 
               
               
                   
               
               
                 gcatccagagactagtgcttagattcctactttcagcgttggacaacctggatccacttgcccagtgttcttccttagttcctaccttc 
               
               
                   
               
               
                 gaccttgatcctcctttatcttcctgaaccctgctgagatgatctatgtggggagaatggcttctttgagaaacatcttcttcgttagt 
               
               
                   
               
               
                 ggcctgcccctcattcccactttaatatccagaatcactataagaagaatataataagaggaataactcttattataggtaagggaaaa 
               
               
                   
               
               
                 ttaagaggcatacgtgatgggatgagtaagagaggagagggaaggattaatggacgataaaatctactactatttgttgagacctttta 
               
               
                   
               
               
                 tagtctaatcaattttgctattgttttccatcctcacgctaactccataaaaaaacactattattatctttattttgccatgacaagac 
               
               
                   
               
               
                 tgagctcagaagagtcaagcatttgcctaaggtcggacatgtcagaggcagtgccagacctatgtgagactctgcagctactgctcatg 
               
               
                   
               
               
                 ggccctgtgctgcactgatgaggaggatcagatggatggggcaatgaagcaaaggaatcattctgtggataaaggagacagccatgaag 
               
               
                   
               
               
                 aagtctatgactgtaaatttgggagcaggagtctctaaggacttggatttcaaggaattttgactcagcaaacacaagaccctcacggt 
               
               
                   
               
               
                 gactttgcgagctggtgtgccagatgtgtctatcagaggttccagggagggtggggtggggtcagggctggccaccagctatcagggcc 
               
               
                   
               
               
                 cagatgggttataggctggcaggctcagataggtggttaggtcaggttggtggtgctgggtggagtccatgactcccaggagccaggag 
               
               
                   
               
               
                 agatagaccatgagtagagggcagacatgggaaaggtgggggaggcacagcatagcagcatttttcattctactactacatgggactgc 
               
               
                   
               
               
                 tcccctatacccccagctaggggcaagtgccttgactcctatgttttcaggatcatcatctataaagtaagagtaataattgtgtctat 
               
               
                   
               
               
                 ctcatagggttattatgaggatcaaaggagatgcacactctctggaccagtggcctaacagttcaggacagagctatgggcttcctatg 
               
               
                   
               
               
                 tatgggtcagtggtctcaatgtagcaggcaagttccagaagatagcatcaaccactgttagagatatactgccagtctcagagcctgat 
               
               
                   
               
               
                 gttaatttagcaatgggctgggaccctcctccagtagaaccttctaaccagctgctgcagtcaaagtcgaatgcagctggttagacttt 
               
               
                   
               
               
                 ttttaatgaaagcttagctttcattaaagattaagctcctaagcagggcacagatgaaattgtctaacagcaactttgccatctaaaaa 
               
               
                   
               
               
                 aatctgacttcactggaaacatggaagcccaaggttctgaacatgagaaatttttaggaatctgcacaggagttgagagggaaacaaga 
               
               
                   
               
               
                 tggtgaagggactagaaaccacatgagagacacgaggaaatagtgtagatttaggctggaggtaaatgaaagagaagtgggaattaata 
               
               
                   
               
               
                 cttactgaaatctttctatatgtcaggtgccattttatgatatttaataatctcattacatatggtaattctgtgagatatgtattatt 
               
               
                   
               
               
                 gaacatactataattaatactaatgataagtaacacctcttgagtacttagtatatgctagaatcaaatttaagtttatcatatgaggc 
               
               
                   
               
               
                 cgggcacggtggctcatatatgggattacatgcctgtaatcccagcactttgggaggccaaggcaattggatcacctgaggtcaggagt 
               
               
                   
               
               
                 tccagaccagcctggccaacatggtgaaaccccttctctactaaaaaatacaaaaaatcagccaggtgtggtggcacgcgtctataatc 
               
               
                   
               
               
                 ccagctactcaggaggctgaggcaggagaatcacttgaacccaggaggtggaggttgcagtgagctaagattgcaccactgcactccag 
               
               
                   
               
               
                 cctaggcgacagagtgagactccatctcaaaaaaaaaaaaagaagtttattatatgaattaacttagttttactcacaccaatactcag 
               
               
                   
               
               
                 aagtagattattacctcatttattgatgaggagcccaatgtacttgtagtgtagatcaacttattgaaagcacaagctaataagtagac 
               
               
                   
               
               
                 aattagtaattagaagtcagatggtctgagctctcctactgtctacattacatgagctcttattaactggggactcgaaaatcaaagac 
               
               
                   
               
               
                 atgaaataatttgtccaagcttacagaaccaccaagtagtaaggctaggatgtagacccagttctgctacctctgaagacagtgttttt 
               
               
                   
               
               
                 tccacagcaaaacacaaactcagatattgtggatgcgagaaattagaagtagatattcctgccctgtggcccttgcttcttacttttac 
               
               
                   
               
               
                 ttcttgtcgattggaagttgtggtccaagccacagttgcagaccatacttcctcaaccataattgcatttcttcaggaaagtttgaggg 
               
               
                   
               
               
                 agaaaaaggtaaagaaaaatttagaaacaacttcagaataaagagattttctcttgggttacagagattgtcatatgacaaattataag 
               
               
                   
               
               
                 cagacacttgagaaaactgaaggcccatgcctgcccaaattaccctttgaccccttggtcaagctgcaactttggttaaagggagtgtt 
               
               
                   
               
               
                 tatgtgttatagtgttcatttactcttctggtctaacccattggctccgtcttcatcctgcagtgacctcagtgcctcagaaacataca 
               
               
                   
               
               
                 tatgtttgtctagtttaagtttgtgtgaaattctaactagcgtcaagaactgagggccctaaactatgctaggaatagtgctgtggtgc 
               
               
                   
               
               
                 tgtgataggtacacaagaaatgagaagaaactgcagattctctgcatctccctttgccgggtctgacaacaaagtttccccaaatttta 
               
               
                   
               
               
                 ccaatgcaagccatttctccatatgctaactactttaaaatcatttggggcttcacattgtctttctcatctgtaaaaagaatggaaga 
               
               
                   
               
               
                 actcattcctacagaactccctatgtcttccctgatgggctagagttcctctttctcaaaaattagccattattgtatttccttctaag 
               
               
                   
               
               
                 ccaaagctcagaggtcttgtattgcccagtgacatgcacactggtcaaaagtaggctaagtagaagggtactttcacaggaacagagag 
               
               
                   
               
               
                 caaaagaggtgggtgaatgagagggtaagtgagaaaagacaaatgagaagttacaacatgatggcttgttgtctaaatatctcctaggg 
               
               
                   
               
               
                 aattattgtgagaggtctgaatagtgttgtaaaataagctgaatctgctgccaacattaacagtcaagaaatacctccgaataactgta 
               
               
                   
               
               
                 cctccaattattctttaaggtagcatgcaactgtaatagttgcatgtatatatttatcataatactgtaacagaaaacacttactgaat 
               
               
                   
               
               
                 atatactgtgtccctagttctttacacaataaactaatctcatcctcataattctattagctaatacatattatcatcctatatttcag 
               
               
                   
               
               
                 agacttcaagaagttaagcaacttgctcaagatcatctaagaagtaggtggtatttctgggctcatttggcccctcctaatctctcatg 
               
               
                   
               
               
                 gcaacatggctgcctaaagtgttgattgccttaattcatcagggatgggctcatactcactgcagaccttaactggcatcctcttttct 
               
               
                   
               
               
                 tatgtgatctgcctgaccctagtagacttatgaaatttctgatgagaaaggagagaggagaaaggcagagctgactgtgatgagtgatg 
               
               
                   
               
               
                 aaggtgccttctcatctgggtaccagtggggcctctaagactaagtcactctgtctcactgtgtcttagccagttccttacagcttgcc 
               
               
                   
               
               
                 ctgatgggagatagagaatgggtatcctccaacaaaaaaataaattttcatttctcaaggtccaacttatgttttcttaatttttaaaa 
               
               
                   
               
               
                 aaatcttgaccattctccactctctaaaataatccacagtgagagaaacattcttttcccccatcccataaatacctctattaaatatg 
               
               
                   
               
               
                 gaaaatctgggcatggtgtctcacacctgtaatcccagcactttgggaggctgaggtgggtggactgcttggagctcaggagttcaaga 
               
               
                   
               
               
                 ccatcttggacaacatggtgataccctgcctctacaaaaagtacaaaaattagcctggcatggtggtgtgcacctgtaatcccagctat 
               
               
                   
               
               
                 tagggtggctgaggcaggagaattgcttgaacccgggaggcggaggttgcagtgagctgagatcgtgccactgcactccagcctggggg 
               
               
                   
               
               
                 acagagcacattataattaactgttattttttacttggactcttgtggggaataagatacatgttttattcttatttatgattcaagca 
               
               
                   
               
               
                 ctgaaaatagtgtttagcatccagcaggtgcttcaaaaccatttgctgaatgattactatactttttacaagctcagctccctctatcc 
               
               
                   
               
               
                 cttccagcatcctcatctctgattaaataagcttcagtttttccttagttcctgttacatttctgtgtgtctccattagtgacctccca 
               
               
                   
               
               
                 tagtccaagcatgagcagttctggccaggcccctgtcggggtcagtgccccacccccgccttctggttctgtgtaaccttctaagcaaa 
               
               
                   
               
               
                 ccttctggctcaagcacagcaatgctgagtcatgatgagtcatgctgaggcttagggtgtgtgcccagatgttctcagcctagagtgat 
               
               
                   
               
               
                 gactcctatctgggtccccagcaggatgcttacagggcagatggcaaaaaaaaggagaagctgaccacctgactaaaactccacctcaa 
               
               
                   
               
               
                 acggcatcataaagaaaatggatgcctgagacagaatgtgacatattctagaatatattatttcctgaatatatatatatatatacaca 
               
               
                   
               
               
                 tatacgtatatatatatatatatatatatttgttgttatcaattgccatagaatgattagttattgtgaatcaaatatttatcttgcag 
               
               
                   
               
               
                 gtggcctctatacctagaagcggcagaatcaggctttattaatacatgtgtatagatttttaggatctatacacatgtattaatatgaa 
               
               
                   
               
               
                 acaaggatatggaagaggaaggcatgaaaacaggaaaagaaaacaaaccttgtttgccattttaaggcacccctggacagctaggtggc 
               
               
                   
               
               
                 aaaaggcctgtgctgttagaggacacatgctcacatacggggtcagatctgacttggggtgctactgggaagctctcatcttaaggata 
               
               
                   
               
               
                 catctcaggccagtcttggtgcattaggaagatgtaggcaactctgatcctgagaggaaagaaacattcctccaggagagctaaaaggg 
               
               
                   
               
               
                 ttcacctgtgtgggtaactgtgaaggactacaagaggatgaaaaacaatgacagacagacataatgcttgtgggagaaaaaacaggagg 
               
               
                   
               
               
                 tcaaggggatagagaaggcttccagaagaatggctttgaagctggcttctgtaggagttcacagtggcaaagatgtttcagaaatgtga 
               
               
                   
               
               
                 catgacttaaggaactatacaaaaaggaacaaatttaaggagaggcagataaattagttcaacagacatgcaaggaattttcagatgaa 
               
               
                   
               
               
                 tgttatgtctccactgagcttcttgaggttagcagctgtgagggttttgcaggcccaggacccattacaggacctcacgtatacttgac 
               
               
                   
               
               
                 actgttttttgtattcatttgtgaatgaatgacctcttgtcagtctactcggtttcgctgtgaatgaatgatgtcttgtcagcctactt 
               
               
                   
               
               
                 ggtttcgctaagagcacagagagaagatttagtgatgctatgtaaaaacttcctttttggttcaagtgtatgtttgtgatagaaatgaa 
               
               
                   
               
               
                 gacaggctacatgatgcatatctaacataaacacaaacattaagaaaggaaatcaacctgaagagtatttatacagataacaaaataca 
               
               
                   
               
               
                 gagagtgagttaaatgtgtaataactgtggcacaggctggaatatgagccatttaaatcacaaattaattagaaaaaaaacagtgggga 
               
               
                   
               
               
                 aaaaattccatggatgggtctagaaagactagcattgttttaggttgagtggcagtgtttaaagggtgatatcagactaaacttgaaat 
               
               
                   
               
               
                 atgtggctaaataactagaatactctttattttttcgtatcatgaatagcagatatagcttgatggccccatgcttggtttaacatcct 
               
               
                   
               
               
                 tgctgttcctgacatgaaatccttaatttttgacaaaggggctattcattttcattttatattgggcctagaaattatgtagatggtcc 
               
               
                   
               
               
                 tgaggaaaagtttatagcttgtctatttctctctctaacatagttgtcagcacaatgcctaggctataggaagtactcaaagcttgtta 
               
               
                   
               
               
                 aattgaattctatccttcttattcaattctacacatggaggaaaaactcatcagggatggaggcacgcctctaaggaaggcaggtgtgg 
               
               
                   
               
               
                 ctctgcagtgtgattgggtacttgcaggacgaagggtggggtgggagtggctaaccttccattcctagtgcagaggtcacagcctaaac 
               
               
                   
               
               
                 atcaaattccttgaggtgcggtggctcactcctgtaatcacagcagtttgggacgccaaggtgggcagatcacttgaggtcaggagttg 
               
               
                   
               
               
                 gacaccagcccagccaacatagtgaaacctggtctctgcttaaaaatataaaaattagctggacgtggtgacgggagcctgtaatccaa 
               
               
                   
               
               
                 ctacttgggaggctgaggcaggagaatcgcttgaaccggggaggtggagtttgcactgagcagagatcatgccattgcactccagcctc 
               
               
                   
               
               
                 cagagcgagactctgtctaaagaaaaacgaaaacaaacaaacaaacaaacaaacaaaacccatcaaattccctgaccgaacagaattct 
               
               
                   
               
               
                 gtctgattgttctctgacttatctaccattttccctccttaaagaaactgtgaacttccttcagctagaggggcctggctcagaagcct 
               
               
                   
               
               
                 ctggtcagcatccaagaaatacttgatgtcactttggctaaaggtatgatgtgtagacaagctccagagatggtttctcatttccatat 
               
               
                   
               
               
                 ccacccacccagctttccaattttaaagccaattctgaggtagagactgtgatgaacaaacaccttgacaaaattcaacccaaagactc 
               
               
                   
               
               
                 actttgcctagcttcaaaatccttactctgacatatactcacagccagaaattagcatgcactagagtgtgcatgagtgcaacacacac 
               
               
                   
               
               
                 acacaccaattccatattctctgtcagaaaatcctgttggtttttcgtgaaaggatgttttcagaggctgaccccttgccttcacctcc 
               
               
                   
               
               
                 aatgctaccactctggtctaagtcactgtcaccaccacctaaattatagctgttgactcataacaatcttcctgcttctaccactgccc 
               
               
                   
               
               
                 cactacaatttcttcccaatatactatccaaattagtcttttcaaaatgtaagtcatatatggtcacctctttgttcaaagtcttctga 
               
               
                   
               
               
                 tagtttcctatatcatttataataaaaccaaatccttacaattctctacaatagttgttcatgcatatattatgtttattacagataca 
               
               
                   
               
               
                 tatatatagctctcatataaataaatatatatatttatgtgtatgtgtgtagagtgttttttcttacaactctatgatgtaggtattat 
               
               
                   
               
               
                 tagtgtcccaaattttataatttaggacttctatgatctcatcttttattctccccttcaccgaatctcatcctacattggccttattg 
               
               
                   
               
               
                 atattccttgaaaattctaagcatcttacatctttagggtatttacatttgccattccctatgccctaaatatttaatcatagtttcat 
               
               
                   
               
               
                 ataaatgggttcctcatcatctatgggtactctctcaggtgttaactttatagtgaggactttcctgccatactacttaaagtagcgat 
               
               
                   
               
               
                 accctttcaccctgtcctaatcacactctggccttcatttcagttttttttttttctccatagcacctaatctcattggtatataacat 
               
               
                   
               
               
                 gtttcatttgcttatttaatgtcaagctctttccactatcaagtccatgaaaacaggaactttattcctctattctgtttttgtgctgt 
               
               
                   
               
               
                 attcttagcaattttacaattttgaatgaatgaatgagcagtcaaacacatatacaactataattaaaaggatgtatgctgacacatcc 
               
               
                   
               
               
                 actgctatgcacacacaaagaaatcagtggagtagagctggaagtgctaagcctgcatagagctagttagccctccgcaggcagagcct 
               
               
                   
               
               
                 tgatgggattactgagttctagaattggactcatttgttttgtaggctgagatttgctcttgaaaacttgttctgaccaaaataaaagg 
               
               
                   
               
               
                 ctcaaaagatgaatatcgaaaccagggtgttttttacactggaatttataactagagcactcatgtttatgtaagcaattaattgtttc 
               
               
                   
               
               
                 atcagtcaggtaaaagtaaagaaaaactgtgccaaggcaggtagcctaatgcaatatgccactaaagtaaacattatttcataggtgtc 
               
               
                   
               
               
                 agatatggcttattcatccatcttcatgggaaggatggccttggcctggacatcagtgttatgtgaggttcaaaacacctctaggctat 
               
               
                   
               
               
                 aaggcaacagagctccttttttttttttctgtgctttcctggctgtccaaatctctaatgataagcatacttctattcaatgagaatat 
               
               
                   
               
               
                 tctgtaagattatagttaagaattgtgggagccattccgtctcttatagttaaatttgagcttcttttatgatcactgtttttttaata 
               
               
                   
               
               
                 tgctttaagttctggggtacatgtgccatggtggtttgctgcacccatcaacccgtcatctacattaggtatttctcctaatgctatcc 
               
               
                   
               
               
                 ttcccctagccccccacccccaacaggccccagtgtgtgatgttcccctccctgtgtccatggatcactggtttttttttgtttttttt 
               
               
                   
               
               
                 ttttttttaaagtctcagttaaatttttggaatgtaatttattttcctggtatcctaggacttgcaagttatctggtcactttagccct 
               
               
                   
               
               
                 cacgttttgatgataatcacatatttgtaaacacaacacacacacacacacacacacacatatatatatatataaaacatatatataca 
               
               
                   
               
               
                 taaacacacataacatatttatcgggcatttctgagcaactaatcatgcaggactctcaaacactaacctatagccttttctatgtatc 
               
               
                   
               
               
                 tacttgtgtagaaaccaagcgtggggactgagaaggcaatagcaggagcattctgactctcactgcctttagctaggcccctccctcat 
               
               
                   
               
               
                 cacagctcagcatagtcctgagctcttatctatatccacacacagtttctgacgctgcccagctatcaccatcccaagtctaaagaaaa 
               
               
                   
               
               
                 aaataatgggtttgcccatctctgttgattagaaaacaaaacaaaataaaataagcccctaagctcccagaaaacatgactaaaccagc 
               
               
                   
               
               
                 aagaagaagaaaatacaataggtatatgaggagactggtgacactagtgtctgaatgaggcttgagtacagaaaagaggctctagcagc 
               
               
                   
               
               
                 atagtggtttagaggagatgtttctttccttcacagatgccttagcctcaataagcttgcggttgtggaagtttactttcagaacaaac 
               
               
                   
               
               
                 tcctgtggggctagaattattgatggctaaaagaagcccgggggagggaaaaatcattcagcatcctcacccttagtgacacaaaacag 
               
               
                   
               
               
                 agggggcctggttttccatatttcctcatgatggatgatctcgttaatgaaggtggtctgacgagatcattgcttcttccatttaagcc 
               
               
                   
               
               
                 ttgctcacttgccaatcctcagttttaaccttctccagagaaatacacattttttattcaggaaacatactatgttatagtttcaatac 
               
               
                   
               
               
                 taaataatcaaagtactgaagatagcatgcataggcaagaaaaagtccttagctttatgttgctgttgtttcagaatttaaaaaagatc 
               
               
                   
               
               
                 accaagtcaaggacttctcagttctagcactagaggtggaatcttagcatataatcagaggtttttcaaaatttctagacataagattc 
               
               
                   
               
               
                 aaagccctgcacttaaaatagtctcatttgaattaactctttatataaattgaaagcacattctgaactacttcagagtattgttttat 
               
               
                   
               
               
                 ttctatgttcttagttcataaatacattaggcaatgcaatttaattaaaaaaacccaagaatttcttagaattttaatcatgaaaataa 
               
               
                   
               
               
                 atgaaggcatctttacttactcaaggtcccaaaaggtcaaagaaaccaggaaagtaaagctatatttcagcggaaaatgggatatttat 
               
               
                   
               
               
                 gagttttctaagttgacagactcaagttttaaccttcagtgcccatcatgtaggaaagtgtggcataactggctgattctggctttcta 
               
               
                   
               
               
                 ctcctttttcccattaaagatccctcctgcttaattaacattcacaagtaactctggttgtactttaggcacagtggctcccgaggtca 
               
               
                   
               
               
                 gtcacacaataggatgtctgtgctccaagttgccagagagagagattactcttgagaatgagcctcagccctggctcaaactcacctgc 
               
               
                   
               
               
                 aaacttcgtgagagatgaggcagaggtacactacgaaagcaacagttagaagctaaatgatgagaacacatggactcatagagggaaac 
               
               
                   
               
               
                 aacgcatactggggcctatcagagggtggagggtgagagaaggagaggatcaggaaaaatcactaatggatgctaagcgtaatacctga 
               
               
                   
               
               
                 gtgatgagatcatctatacaacaaacccccttgacattcatttatctatgtaacaaacctgcacatcctgtacatgtacccctgaactt 
               
               
                   
               
               
                 aaaataaaagttgaaaacaagaaagcaacagtttgaacacttgttatggtctattctctcattctttacaattacactagaaaatagcc 
               
               
                   
               
               
                 acaggcttcctgcaaggcagccacagaatttatgacttgtgatatccaagtcattcctggataatgcaaaatctaacacaaaatctagt 
               
               
                   
               
               
                 agaatcatttgcttacatctatttttgttctgagaatatagatttagatacataatggaagcagaataatttaaaatctggctaattta 
               
               
                   
               
               
                 gaatcctaagcagctcttttcctatcagtggtttacaagccttgtttatatttttcctattttaaaaataaaaataaagtaagttattt 
               
               
                   
               
               
                 gtggtaaagaatattcattaaagtatttatttcttagataataccatgaaaaacattcagtgaagtgaagggcctactttacttaacaa 
               
               
                   
               
               
                 gaatctaatttatataatttttcatactaatagcatctaagaacagtacaatatttgactcttcaggttaaacatatgtcataaattag 
               
               
                   
               
               
                 ccagaaagatttaagaaaatattggatgtttccttgtttaaattaggcatcttacagtttttagaatcctgcatagaacttaagaaatt 
               
               
                   
               
               
                 acaaatgctaaagcaaacccaaacaggcaggaattaatcttcatcgaatttgggtgtttctttctaaaagtcctttatacttaaatgtc 
               
               
                   
               
               
                 ttaagacatacatagattttattttactaattttaattatatagacaataaatgaatattcttactgattactttttctgactgtctaa 
               
               
                   
               
               
                 tctttctgatctatcctggatggccataacacttatctctctgaactttgggcttttaatataggaaagaaaagcaataatccattttt 
               
               
                   
               
               
                 catggtatctcatatgataaacaaataaaatgcttaaaaatgagcaggtgaagcaatttatcttgaaccaacaagcatcgaagcaataa 
               
               
                   
               
               
                 tgagactgcccgcagcctacctgacttctgagtcaggatttataagccttgttactgagacacaaacctgggcctttcaatgctataac 
               
               
                   
               
               
                 ctttcttgaagctcctccctaccacctttagccataaggaaacatggaatgggtcagatccctggatgcaagccaggtctggaaccata 
               
               
                   
               
               
                 ggcagtaaggagagaagaaaatgtgggctctgcaactggctccgagggagcaggagaggatcaaccccatactctgaatctaagagaag 
               
               
                   
               
               
                 actggtgtccatactctgaatgggaagaatgatgggattacccatagggcttgttttagggagaaacctgttctccaaactcttggcct 
               
               
                   
               
               
                 tgagatacctggtccttattccttggactttggcaatgtctgaccctcacattcaagttctgaggaagggccactgccttcatactgtg 
               
               
                   
               
               
                 gatctgtagcaaattccccctgaaaacccagagctgtatcttaattggttaaaaaaaattatattatctcaacgactgttcttctctga 
               
               
                   
               
               
                 gtagccaagctcagcttggttcaagctacaagcagctgagctgctttttgtctagtcattgttcttttatttcagtggatcaaatacgt 
               
               
                   
               
               
                 tctttccaaacctaggatcttgtcttcctaggctatatattttgtcccaggaagtcttaatctggggtccacagaacactagggggctg 
               
               
                   
               
               
                 gtgaagtttatagaaaaaaaatctgtatttttacttacatgtaactgaaatttagcattttcttctactttgaatgcaaaggacaaact 
               
               
                   
               
               
                 agaatgacatcatcagtacctattgcatagttataaagagaaaccacagatattttcatactacaccataggtattgcagatctttttg 
               
               
                   
               
               
                 tttttgtttttgtttgagatggagtttcgctcttattgcccaggctggagtgcagtggcatgatttcggctcactgcaacctccccttc 
               
               
                   
               
               
                 ctgcattcaagcaattctcctgccttggcctcctgagtagctggggattacaggcacctgccaccatgccagtctaatttttgtatttt 
               
               
                   
               
               
                 tagtagagatggggtttcgccatgttggccaggctggtcttgaactcctgacctcagatgatctgcccgccttggcctcctgaagtgct 
               
               
                   
               
               
                 gggattataggtgtgagccaccacgcctggcccattgcagatatttttaattcacatttatctgcatcactacttggatcttaaggtag 
               
               
                   
               
               
                 ctgtagacccaatcctagatctaatgctttcataaagaagcaaatataataaatactataccacaaatgtaatgtttgatgtctgataa 
               
               
                   
               
               
                 tgatatttcagtgtaattaaacttagcactcctatgtatattatttgatgcaataaaaacatatttttttagcacttacagtctgccaa 
               
               
                   
               
               
                 actggcctgtgacacaaaaaaagtttaggaattcctggttttgtctgtgttagccaatggttagaatatatgctcagaaagataccatt 
               
               
                   
               
               
                 ggttaatagctaaaagaaaatggagtagaaattcagtggcctggaataataacaatttgggcagtcattaagtcaggtgaagacttctg 
               
               
                   
               
               
                 gaatcatgggagaaaagcaagggagacattcttacttgccacaagtgtttttttttttttttttttttatcacaaacataagaaaatat 
               
               
                   
               
               
                 aataaataacaaagtcaggttatagaagagagaaacgctcttagtaaacttggaatatggaatccccaaaggcacttgacttgggagac 
               
               
                   
               
               
                 aggagccatactgctaagtgaaaaagacgaagaacctctagggcctgaacatacaggaaattgtaggaacagaaattcctagatctggt 
               
               
                   
               
               
                 ggggcaaggggagccataggagaaagaaatggtagaaatggatggagacggaggcagaggtgggcagatcatgaggtcaagagatcgag 
               
               
                   
               
               
                 accatcctggcaaacatggtgaaatcccgtctctactaaaaataaaaaaattagctgggcatggtggcatgcgcctgtagtcccagctg 
               
               
                   
               
               
                 ctcgggaggctgaggcaggagaatcgtttgaacccaggaggcgaaggttgcagtgagctgagatagtgccattgcactccagtctggca 
               
               
                   
               
               
                 acagagtgagactccgtctcaaaaaaaaaaaaaaaagaaagaaagaaaagaaaaagaaaaaagaaaaaataaatggatgtagaacaagc 
               
               
                   
               
               
                 cagaaggaggaactgggctggggcaatgagattatggtgatgtaagggacttttatagaattaacaatgctggaatttgtggaactctg 
               
               
                   
               
               
                 cttctattattcccccaatcattacttctgtcacattgatagttaaataatttctgtgaatttattccttgattctaaaatatgaggat 
               
               
                   
               
               
                 aatgacaatggtattataagggcagattaagtgatatagcatgagcaatattcttcaggcacatggatcgaattgaatacactgtaaat 
               
               
                   
               
               
                 cccaacttccagtttcagctctaccaagtaaagagctagcaagtcatcaaaatggggacatacagaaaaaaaaaaggacactagaggaa 
               
               
                   
               
               
                 taatataccctgactcctagcctgattaatatatcgattcacttttttctctgtttgatgacaaattctggctttaaataattttagga 
               
               
                   
               
               
                 ttttaggcttctcagctcccttcccagtgagaagtataagcaggacagacaggcaagcaagaagagagccccaggcaatactcacaaag 
               
               
                   
               
               
                 tagccaatgtcccctgtggtcatagagaaatgaaaagagagaggattctctggaagcactggatgtaatcttttctgtctgtcctctct 
               
               
                   
               
               
                 agggaatcaccccaaggtactgtactttgggattaaggctttagtcccactgtggactacttgctattctgttcagtttctagaaggaa 
               
               
                   
               
               
                 ctatgtacggtttttgtctccctagagaaactaaggtacagaagttttgtttacaatgcactccttaagagagctagaactgggtgaga 
               
               
                   
               
               
                 ttctgttttaacagctttattttcttttccttggccctgtttttgtcactgtcaccacctttaaggcaaatgttaaatgcgctttggct 
               
               
                   
               
               
                 gaaactttttttcctattttgagatttgctcctttatatgaggctttcttggaaaaggagaatgggagagatggatatcattttggaag 
               
               
                   
               
               
                 atgatgaagagggtaaaaaaggggacaaatggaaatttgtgttgcagatagatgaggagccaacaaaaaagagcctcaggatccagcac 
               
               
                   
               
               
                 acattatcacaaacttagtgtccatccatcactgctgaccctctccggacctgactccacccctgagggacacaggtcagccttgacca 
               
               
                   
               
               
                 atgacttttaagtaccatggagaacagggggccagaacttcggcagtaaagaataaaaggccagacagagaggcagcagcacatatctg 
               
               
                   
               
               
                 cttccgacacagctgcaatcactagcaagctctcaggcctggcatcatggtgcattttactgctgaggagaaggctgccgtcactagcc 
               
               
                   
               
               
                 tgtggagcaagatgaatgtggaagaggctggaggtgaagccttgggcaggtaagcattggttctcaatgcatgggaatgaagggtgaat 
               
               
                   
               
               
                 attaccctagcaagttgattgggaaagtcctcaagattttttgcatctctaattttgtatctgatatggtgtcatttcatagactcctc 
               
               
                   
               
               
                 gttgtttacccctggacccagagattttttgacagctttggaaacctgtcgtctccctctgccatcctgggcaaccccaaggtcaaggc 
               
               
                   
               
               
                 ccatggcaagaaggtgctgacttcctttggagatgctattaaaaacatggacaacctcaagcccgcctttgctaagctgagtgagctgc 
               
               
                   
               
               
                 actgtgacaagctgcatgtggatcctgagaacttcaaggtgagttcaggtgctggtgatgtgattttttggctttatattttgacatta 
               
               
                   
               
               
                 attgaagctcataatcttattggaaagaccaacaaagatctcagaaatcatgggtcgagcttgatgttagaacagcagacttctagtga 
               
               
                   
               
               
                 gcataaccaaaacttacatgattcagaactagtgacagtaaaggactactaacagcctgaattggcttaacttttcaggaaatcttgcc 
               
               
                   
               
               
                 agaacttgatgtgtttatcccagagaattgtattatagaattgtagacttgtgaaagaagaatgaaatttggcttttggtagatgaaag 
               
               
                   
               
               
                 tccatttcaaggaaatagaaatgccttattttatgtgggtcatgataattgaggtttagaaagagatttttgcaaaaaaaataaaagat 
               
               
                   
               
               
                 ttgctcaaagaaaaataagacacattttctaaaatatgttaaatttcccatcagtattgtgaccaagtgaaggcttgtttccgaatttg 
               
               
                   
               
               
                 ttggggattttaaactcccgctgagaactcttgcagcactcacattctacatttacaaaaattagacaattgcttaaagaaaaacaggg 
               
               
                   
               
               
                 agagagggaacccaataatactggtaaaatggggaagggggtgagggtgtaggtaggtagaatgttgaatgtagggctcatagaataaa 
               
               
                   
               
               
                 attgaacctaagctcatctgaattttttgggtgggcacaaaccttggaacagtttgaggtcagggttgtctaggaatgtaggtataaag 
               
               
                   
               
               
                 ccgtttttgtttgtttgtttgttttttcatcaagttgttttcggaaacttctactcaacatgcctgtgtgttattttgtcttttgccta 
               
               
                   
               
               
                 acagctcctgggtaacgtgatggtgattattctggctactcactttggcaaggagttcacccctgaagtgcaggctgcctggcagaagc 
               
               
                   
               
               
                 tggtgtctgctgtcgccattgccctggcccataagtaccactgagttctcttccagtttgcaggtgttcctgtgaccctgacaccctcc 
               
               
                   
               
               
                 ttctgcacatggggactgggcttggccttgagagaaagccttctgtttaataaagtacattttcttcagtaatcaaaaattgcaatttt 
               
               
                   
               
               
                 atcttctccatcttttactcttgtgttaaaaggaaaaagtgttcatgggctgagggatggagagaaacataggaagaaccaagagcttc 
               
               
                   
               
               
                 cttaagaaatgtatgggggcttgtaaaattaatgtggatgttatgggagaattccaggattccaaggaggatgatatgatggagaaaaa 
               
               
                   
               
               
                 tctttatcggggtgggaaaatggttaattaagtggacagagactcctaggcagtttttactgcaccggggaaagaaggagctgttagtg 
               
               
                   
               
               
                 gtacctgagaaagcagatttgtggtacatgtcacttttcattaaaaacaaaaacaaaacaaaacaaaacttcatagatatccaagatat 
               
               
                   
               
               
                 aggctagaattactattttaatttactcttatttacattttgaagtagctagcttgtcacatgttttatgaaattgatttggagataag 
               
               
                   
               
               
                 atgagtgtgtatcaacaatagcctgctctttccatgaaggattccattatttcatgggttagctgaagctaagacacatgatatcattg 
               
               
                   
               
               
                 tgcattatcttctgatagaatgtaacatgcactaaaataaagttagagttaggacctgagtgggaaagtttttggagagtgtgatgaag 
               
               
                   
               
               
                 actttccgtgggagatagaatactaataaaggcttaaattctaaaaccagcaagctagggcttcgtgacttgcatgaaactggctctct 
               
               
                   
               
               
                 ggaagtagaagggagagtaagacatacgtagaggactaggaaagaccagatagtacagggcctggctacaaaaatacaagcttttacta 
               
               
                   
               
               
                 tgctattgcaatactaaacgataagcattaggatgttaagtgactcaggaaataagattttgggaaaaagtaatctgcttatgtgcaca 
               
               
                   
               
               
                 aaatggattcaagtttgcagataaaataaaatatggatgatgattcaaggggacagatacaatggttcaaacccaagaggagcagtgag 
               
               
                   
               
               
                 tctgtggaatttgaaggatggacaaaggtggggtgagaaagacatagtattcgactgactgtgggagatgagaaggaagaaggaggtga 
               
               
                   
               
               
                 taaatgactgaaagctcccagactggtgaagataacaggaggaaaccatgcactgacctggtgactctcatgtgtgaagggtagaggga 
               
               
                   
               
               
                 tattaacagatttactttttaggaagtgctagattggtcagggagttttgaccttcaggtcttgtgtctttcatatcaaggaacctttg 
               
               
                   
               
               
                 cattttccaagttagagtgccatattttggcaaatataactttattagtaattttatagtgctctcacattgatcagactttttcctgt 
               
               
                   
               
               
                 gaattacttttgaatttggctgtatatatccagaatatgggagagagacaaataattattgtagttgcaggctatcaacaatactggtc 
               
               
                   
               
               
                 tctctgagccttataacctttcaatatgcccataaacagagtaaacagggattattcatggcactaaatattttcacctagtcagtcaa 
               
               
                   
               
               
                 caaatgggagcaatgtgcattttttgatacatatttttatatatttatggggtacatgtgatacttacatgcctagaacatgtgatgat 
               
               
                   
               
               
                 taagtctagatatttaggatatccattgctttgagcatttatcatttctatgtattgagaaaatttcaaatcctcatttctagccattt 
               
               
                   
               
               
                 tgaaatatataataaatagtaattaactatagtcaccctactcaaatatcaaacattatggcttaatccttctatccaactgtgtttgt 
               
               
                   
               
               
                 acctattaaccaacatctcttaaatcccctcccatacacactcacactttttccagcctctgataactatcattctactctctaccacc 
               
               
                   
               
               
                 atgagacccacttttttagctcccacagatgaataaaaacatgtgatatttgactttctgtatctggcttattttattatctatctctt 
               
               
                   
               
               
                 tggcataccaagagtttgtttttgttctgcttcagggctttcaattaacataatgacctctggttccatccatgttgctacaaatgaca 
               
               
                   
               
               
                 agatttcattctttttcatggcaaaatagtactgtgcaaaaatacaattttttaatccgttcatctgttgatagacacttaggttgatc 
               
               
                   
               
               
                 ccaaaccttaactattgtgaatagtgcttcaataaacatgagtgtaatgtgtccattggatatactgatttcctttcttttggataaat 
               
               
                   
               
               
                 aaccactagtgagattgctggattgtatgatagttctgtttttagtttactgagaaatcttcatactgttttccataatggttgtacta 
               
               
                   
               
               
                 ttttacattcccaccaacagtgtgtaagaaagagttcccttttctccatatcctcacaaggatctgttattttttgtcttttttgttaa 
               
               
                   
               
               
                 tagccgttttaactagagtaagtagatatctcattgtagttttgatttgcatttccctgatcattagtgatgttgagaattttttcata 
               
               
                   
               
               
                 tgtttgttggtcatttgtatatctttttctgagaattgtctgttcatgtccttagcctactttttattgggattgtttgttattttctt 
               
               
                   
               
               
                 gataatctatttgtgttcattttagagcctggatattattcttttgtcagatgtatagattgtgaagattttctcccactctgtgggtt 
               
               
                   
               
               
                 gtctgtttattctgcagactcttccttttgccatgcaaaagctctttagtttaatttagtcccagatattttctttgtttttatgtatt 
               
               
                   
               
               
                 tgcatttgtgttcttggtcatgaaatcctttcctaagccaatgtgtagaagggtttttccgatgttattttctagaattgttacagttt 
               
               
                   
               
               
                 cagggcttagatttaagtccttgatccatcttgagttgatttttgtataaggtgagagatgaagatccagtttcattctcctacatgta 
               
               
                   
               
               
                 gcttgccagctatccccgcaccatttgttgaatagggtgccctttccccactttatgtttttgtttgctttgtcaaagatcagttggat 
               
               
                   
               
               
                 gtaagtatttgagtttatttctgggttctctattctgttccattggtcgatgtgcctatttgtacaccagcatcatgctgttttggtga 
               
               
                   
               
               
                 ctatggccttattgtatagtttgaaatgaggtaatgtaatgccttcagatttgttcttttttttagacttgcttgtttattgggctctt 
               
               
                   
               
               
                 ttttggttccataagaattttaggattgttttttctagttctgtgaagactaatggtggtattttgatgggaattgcaatgaatttgta 
               
               
                   
               
               
                 ggttgcttctggcattatggccattttcacaatattgattctacccatctatgagaatggcatgtgtttccatttgtttgtgtcttata 
               
               
                   
               
               
                 tgattactttcagccgtgttttgtagttttccttgtagatgtctttcacctccttggttaggtatatattcctaagtttttgttttgtt 
               
               
                   
               
               
                 ttgttttgttttttgcagctattgtaaaaggggttgagttcttgattttattctcagcttggtcattgctggtatgtaagaaagcaact 
               
               
                   
               
               
                 cattggtgtacgttaattttgtatccagaaactttgctgaattattttatcagttctagggggttttggaggagtctttagagttttct 
               
               
                   
               
               
                 acatacacaatcatatcatcagcaaacagtgacagtttgactttctctttaacaatttggatgtgctttacttgtttctcttgtctgat 
               
               
                   
               
               
                 tgctcttgctaggacttccagtaatatgttaaagagaagtggtgagagtgggtatccttgtctcattccagttttcagacagaatgctt 
               
               
                   
               
               
                 ttaactttttcccattcaatataatgttggctgtgtgtttaccatagctggcttttattacattgaggtatgtcctttgtaaaccgatt 
               
               
                   
               
               
                 ttgctgagttttagtcataaagtgatgttgaattttgttgaatgcagtttctgtggctattgagataatcacatgatttttgtttccaa 
               
               
                   
               
               
                 ttctctttatgttgtgtatcacacttattgacttgcgtatgttaaaccatccgtgcatccctcgcatgaaacccacttgatcatgggtt 
               
               
                   
               
               
                 ttgatatgctgtcggatgctattagctagtattttgtcaaggatgttggcatctatgttcatcagggatattgatctgtagtgtttttt 
               
               
                   
               
               
                 ttttttggttatgttctttcccagttttggtattaaggtgatactggcttcatagaatgatttagggaggattctctctttctctatct 
               
               
                   
               
               
                 tgtagaatactgtcaataggattggtatcaattcttctttgaatgtctggtagaattcagctgtgaatctatctggtcctggacttttt 
               
               
                   
               
               
                 tgttgttggtaaatttttattatcatttcagtcttgctgcttattactggtctgttcagggtatctaattcttcctgacttaagctaga 
               
               
                   
               
               
                 gccctgtatctttccaggaattcgaacgtctcctttaggttttctagtttatgcatgtaaaggtgttcatagtagccttgaataatctt 
               
               
                   
               
               
                 ttgtatttctgtggtatcagtaatagtatctcctgttttgtttctaattgagtttatttgcacttctctcctcttttcttggttaatct 
               
               
                   
               
               
                 tgctaatggtctatcagttttatttatcttttcaaagaaccagctttttatttcatttagcttttgtatttttttgcagttgttttaat 
               
               
                   
               
               
                 ttcatttagttctcctcttatcttagttattccctttcttttgctgggttttggttctgtttgtttttgtttctctagtttcttgtggt 
               
               
                   
               
               
                 gtgaccttatattgtctgtctgtcctctttcagactctttgacatcgacatttagggctgtgaactttccttttagcaccatctttgct 
               
               
                   
               
               
                 gtatcctagaggttttgataggttgtgtcactattgtcggtcagttcaagtaattttgttgttcttattatactttaagttctgggata 
               
               
                   
               
               
                 catgtgcagaatgtgcaggtttgttacataggtatagatgtgccatggtggtttgctgcacccatcaacctgtcatctacattaggtat 
               
               
                   
               
               
                 ttcttttaatgttatccctctcctaaccccctcaccccccgacaggccctggtgtgtgatgttcccctccctgtgtccatgtgttctca 
               
               
                   
               
               
                 ttgttcaactcccacttatgagtgagaacgtgtggtgtttggtttctctgttcctgtgttagtttgctcagaatgatggtttccacctt 
               
               
                   
               
               
                 catccatgtccctgcaaagacatgaactcatcatttttatggctgcatagtattccatggtgtatatgtgccacattttctttatccat 
               
               
                   
               
               
                 tatatcgctgatggccatttgggttggttccaagtctttgctattgtgaatagtgccacaataaacatacgtgtgcacgtgtctttata 
               
               
                   
               
               
                 gtagaatgatttctaattctttgggtatatacccagtaatgggattgctgggtcaaacagtatttctggttctagatccttgaggaatc 
               
               
                   
               
               
                 gccacactgtcttccacaatggttgaactaatttacacacccatcaacagtgtaaaatttttcctattcttccacatcctctccagcac 
               
               
                   
               
               
                 cttttgtttcctgactttttaataattgccattctaactggcatgagatggtatctcattgtggttttgatttgcatttctctaatgac 
               
               
                   
               
               
                 cagtgatgatgagcttcttttcatgtgtttcttggccacataaatgacttctttagagaagcatctgttcatatcctttgtccactttt 
               
               
                   
               
               
                 tgatggggtcgttaggttttttcttgtaaatttgttgaagttctttgtagattttggatgttagccctttgtcagatggatagattgca 
               
               
                   
               
               
                 aaaattttctcccattctgtaggttgcctgttcactctgatgatagtcttttgctgtgcagaagctctttagtttaattagatcccata 
               
               
                   
               
               
                 tgtcaattttggcctttgttgtcattgcttttgatgttttagtcgtgaatttttgcccatgcctatgtcctgaatggtattgcctaggt 
               
               
                   
               
               
                 tatcttctaggatttttatggttttaggttgcacatttaagtctttaatccaccttgagttaatttttgtataaggtgtaaggaagggg 
               
               
                   
               
               
                 tacagtttcagttttatgcatattgctagccagtttttccagcaccatttattaaatagggaattctttctccattgcttttgtgatgt 
               
               
                   
               
               
                 ttgtcaaagatcagatggtcgtagatgtgtggcattatttctgaggcttctgttctgttccactggtctatatatctgttttggtacca 
               
               
                   
               
               
                 gtaccatgctgtttttgttactgtagccttgtagtatagtttgaagtcaggtagcatcatgcctccagctttgttctttttgtttagga 
               
               
                   
               
               
                 ttgtcttggctatatgggctcttttttgattccatatgacatttaaagtagttttttctaattctttgaaaaaagtcagtggtagcttg 
               
               
                   
               
               
                 atggggatagcattgaatctataaattactttgggcagtatggccattttaaagatattgattctttctatctatgagcatggaatgtt 
               
               
                   
               
               
                 tttccatttgtttgtgtcctctcttatttccttgagcagtgagtggtttgtagctctccttgaagaggttcttcacatcccttagaagt 
               
               
                   
               
               
                 tgtatttctaggtattttattttattctctttgcagcaattgtgaatgggagttcacccatgatttggctctctgcttgtctattattg 
               
               
                   
               
               
                 gtgtataggaacgcttgtgatttctgcacactgattttgtatcttgagactttgctgaagctgtttatcagcttaagattttgggctga 
               
               
                   
               
               
                 gatgacagggtcttctaaatatacaatcatgtcatctgcaaacagagacaatttgacttcctctcttcctatttgaatatgctttattt 
               
               
                   
               
               
                 ctttctcttgcctgattgtcctggcgagaacttccaatactatgttgagtaagagtggcgagagggcatccttgtcttgtgccggtttt 
               
               
                   
               
               
                 caaagcaaatgatttttaaatttccatcttgatttcattgttgacccaatgatcattcaggagcaggttatttaatttccctgtatttg 
               
               
                   
               
               
                 catggttttgaaggttccttttgtagttgatttccaattttattctactgtggtctgagagagtgcttgatataatttcaatttttaaa 
               
               
                   
               
               
                 aatttattgaggcttgttttgtggcatatcatatggcctatcttggagaaagttccatgtgctgatgaatagaatgtgtattctgcagt 
               
               
                   
               
               
                 tgttgggtagaatgtcctgtaaatatctgttaagtccatttgttctttaaatccattgtttctttgtagactgtcttgatgacctgcct 
               
               
                   
               
               
                 agtgcagtcagtggagtattgaagtcccccactattattatgttgctgtctagtctagtagtaattgttttataaatttgggatctcca 
               
               
                   
               
               
                 gtattagatgcatatatattaagaattgtaatattctcccattggacaagggcttttatcattatatgatgtccctctttgtctttttt 
               
               
                   
               
               
                 aactgctgtttctttaaagtttgttttgtctgacataagaatagctgctttggctcgcttttggtgtccatttgtgtggaatgtcattt 
               
               
                   
               
               
                 tccacccctttaccttaagtttatgtgagtccttatgtgttaggtgagtctcctgaaggcggcagataactggttggtgaattcttatt 
               
               
                   
               
               
                 cattctgcaattctgtatcttttaagtggagcatttagtccatttacattcaacatcagtattgaggtgtgaggtactattccattctt 
               
               
                   
               
               
                 cgtggtatttgttgcctgtgtatctttttatctgtatttttgttgtatatgtcctatgggatttatgctttaaagaggttctgttttga 
               
               
                   
               
               
                 tgtgcttccagggtttatttcaagatttagagctccttttatcagttcttgtagtgttggcttggtagtgccgaattctctcagcattt 
               
               
                   
               
               
                 gtttttctgaaaaacactgtgtattttcttcatttgtgaagcttagtttcactggatataaaattcttggctgataattgttttgttta 
               
               
                   
               
               
                 agaaggctgaagatagggccatattcacttctagcttttacggtttctgctgagaaatctgctgttaatctgataggttttctttcata 
               
               
                   
               
               
                 ggttacctggtagtttcacctcacagctcttaagattctctttgtctttagataactttggatactctgatgacaatgtacctaggcaa 
               
               
                   
               
               
                 tgatatttttgcaatgaatttcccaggtgtttattgagcttcttgtatttggatatctaggtctctagcaaggtgggggaagttttcct 
               
               
                   
               
               
                 tgattatttccctggataagttttccaaacttttagatttctcttctttctcaggaatgctgattattcttaggtttgattgtttaaca 
               
               
                   
               
               
                 taatcccagatttcttggaggctttgttcatattttcttattcttttttctttgtctttgttggattgggttaattcaaaaactttgtc 
               
               
                   
               
               
                 ttcaagctctgaatttcttctgcttggattctattgctgagactttctagagcattttgcatttctataagtgcatccattcatccatt 
               
               
                   
               
               
                 gtttcctgaagttttgaatgttttttatttatgctatctctttaactgaagatttctcccctcatttcttgtatcatatttttggtttt 
               
               
                   
               
               
                 tttaaaattggacttcaccttcctcggatgcctccttgattagcttaataactgaccttctgaattatttttcaggtaaatcagggatt 
               
               
                   
               
               
                 tcttcttggtttggatgcattgctggtgagctagtatgattttttggggggtgttaaagaaccttgtttttcatattaccagagttagt 
               
               
                   
               
               
                 tttctggttccttctcacttgggtaggctctgtcagagggaaagtctaggcctcaaggctgagacttttgtcccatgaggtgttccctt 
               
               
                   
               
               
                 gatgtagcacagtcccccttttcctaggcgtggggcttcctgagagccgaactgtagtgattgttatctctcttctggatctagccacc 
               
               
                   
               
               
                 catcaggtctaccagactccaggctggtactggggtttgtctgcacagagtcttgtgacgtgaaccatctgtgggtctctcagccatag 
               
               
                   
               
               
                 atacaaccacctgctccaatggaggtggcagaggatgaaatggactctgtgagggtccttacttttggttgttcaatgcactatttttg 
               
               
                   
               
               
                 tgctggttggcctcctgccaggaggtggcactttctagaaagcatcagcagaggcagtcaggtggtggtggctgggggggctggggcac 
               
               
                   
               
               
                 cctagaactcccaagaatatatgccctttgtcttcagctaccagggtgagtaaggaaggaccatcaggtgggggcaggactagtcgtgt 
               
               
                   
               
               
                 ctgagctcagagtctccttgggcaggtctttctgtggctactgtgggaggatgggggtgtagtttccaggtcaatggatttatgttcct 
               
               
                   
               
               
                 aggacaattatggctgcctctgctgtgtcatgcaggtcatcaggaaagtgggggaaagcaagcagtcacgtgacttgcccagctcccat 
               
               
                   
               
               
                 gcaactcaaaaggttggtctcacttccagcgtgcaccctcccccgcaacagcaccgaatctgtttccatgcagtcagtgagcaaggctg 
               
               
                   
               
               
                 agaacttgccccaggctaccagctgcgaaaccaagtagggctgtcctacttccctgccagtggagtctgcacaccaaattcatgtcccc 
               
               
                   
               
               
                 ccaccaacccccccactgcccagcccctagatctggccaggtggagattttctttttcctgtcatcttttcccagttcctctggcagcc 
               
               
                   
               
               
                 ctcccaaatgacccctgtgaggcaaggcagaaatggcttcctaggggacccagagagcccacagggcttttcccgctgcttcctctacc 
               
               
                   
               
               
                 cctgtattttgcttggccctctaaattgactcagctccaggtaaggtcagaatcttctcctgtggtctagatcttcaggttccccagtg 
               
               
                   
               
               
                 aggatgtgtgtttgggggtagacggtcccccttttccacttccacagtttgggcactcacaatatttggggtgtttcccgggtcctgca 
               
               
                   
               
               
                 ggagcaatctgcttctttcagagggtgtgtgcgttctctcagctttcttgatttatttctgcaggtggttctgcaaaaaaaattcctga 
               
               
                   
               
               
                 tgggagacttcacatgctgctctgtgcatccgagtgggagctgcaatgtacttctgctgcctcccatctgccatcaccctctaatttgt 
               
               
                   
               
               
                 cggtaatatgcatttttaatcaatctttttttctctctctctctttttcttctcccccaaaactatactgccctttgatatcaaggaat 
               
               
                   
               
               
                 caaggacgtgatgttgaggggtgggcagtggatacactctttaccccttagggagctatatctagatttagatattgccaattcaagat 
               
               
                   
               
               
                 aacttaattgaaagcaaattcataatgaataCaCaCaCaCaCaCacacatctgcatgacaagatttttaatagttgaaagaataactaa 
               
               
                   
               
               
                 taattgtccacaggcaataagggctttttaagcaaaacagttgtgataaacaggtcattcttagaatagtaatccagccaatagtacag 
               
               
                   
               
               
                 gttgcttagagattatgtcattaccagagttaaaattctataatggcttctcactccctaccactgaggacaagtttatgtccttaggt 
               
               
                   
               
               
                 ttatgcttccctgaaacaataccacctgctattctccactttacatatcaacggcactggttctttatctaactctctggcacagcagg 
               
               
                   
               
               
                 agtttgttttcttctgcttcagagctttgaatttactatttcagcttctaaactttatttggcaatgccttcccatggcagattccttc 
               
               
                   
               
               
                 tgtcattttgcctctgttcgaatactttctccttaatttcattcttagttaataatatctgaaattattttgttgtttaacttaattat 
               
               
                   
               
               
                 taattttatgtatgttctacctagattataatcttcagaggaaagttttattctctgacttatttaacttaaatgcccactactttaaa 
               
               
                   
               
               
                 aattatgacatttatttaacagatatttgctgaacaaatgtttgaaaatacatgggaaagaatgcttgaaaacacttgaaattgcttgt 
               
               
                   
               
               
                 gtaaagaaacagttttatcagttaggatttaatcaatgtcagaagcaatgatataggaaaaatcgaggaataagacagttatggataag 
               
               
                   
               
               
                 gagaaatcaacaaactcttaaaagatattgcctcaaaagcataagaggaaataagggtttatacatgacttttagaacactgccttggt 
               
               
                   
               
               
                 ttttggataaatggggaagttgtttgaaaacaggagggatcctagatattccttagtctgaggaggagcaattaagattcacttgttta 
               
               
                   
               
               
                 gaggctgggagtggtggctcacgcctgtaatcccagaattttgggaggccaaggcaggcagatcacctgaggtcaagagttcaagacca 
               
               
                   
               
               
                 acctggccaacatggtgaaatcccatctctacaaaaatacaaaaattagacaggcatgatggcaagtgcctgtaatcccagctacttgg 
               
               
                   
               
               
                 gaggctgaggaaggagaattgcttgaacctggaaggcaggagttgcagtgagccgagatcataccactgcactccagcctgggtgacag 
               
               
                   
               
               
                 aacaagactctgtctcaaaaaaaaaaaagagagattcaaaagattcacttgtttaggccttagcgggcttagacaccagtctctgacac 
               
               
                   
               
               
                 attcttaaaggtcaggctctacaaatggaacccaaccagactctcagatatggccaaagatctatacacacccatctcacagatcccct 
               
               
                   
               
               
                 atcttaaagagaccctaatttgggttcacctcagtctctataatctgtaccagcataccaataaaaatctttctcacccatccttagat 
               
               
                   
               
               
                 tgagagaagtcacttattattatgtgagtaactggaagatactgataagttgacaaatctttttctttcctttcttattcaacttttat 
               
               
                   
               
               
                 tttaacttccaaagaacaagtgcaatatgtgcagctttgttgcgcaggtcaacatgtatctttctggtcttttagccgcctaacacttt 
               
               
                   
               
               
                 gagcagatataagccttacacaggattatgaagtctgaaaggattccaccaatattattataattcctatcaacctgataggttagggg 
               
               
                   
               
               
                 aaggtagagctctcctccaataagccagatttccagagtttctgacgtcataatctaccaaggtcatggatcgagttcagagaaaaaac 
               
               
                   
               
               
                 aaaagcaaaaccaaacctaccaaaaaataaaaatcccaaagaaaaaataaagaaaaaaacagcatgaatacttcctgccatgttaagtg 
               
               
                   
               
               
                 gccaatatgtcagaaacagcactgagttacagataaagatgtctaaactacagtgacatcccagctgtcacagtgtgtggactattagt 
               
               
                   
               
               
                 caataaaacagtccctgcctcttaagagttgttttccatgcaaatacatgtcttatgtcttagaataagattccctaagaagtgaacct 
               
               
                   
               
               
                 agcatttatacaagataattaattctaatccatagtatctggtaaagagcattctaccatcatctttaccgagcatagaagagctacac 
               
               
                   
               
               
                 caaaaccctgggtcatcagccagcacatacacttatccagtgataaatacacatcatcgggtgcctacatacatacctgaatataaaaa 
               
               
                   
               
               
                 aaatacttttgctgagatgaaacaggcgtgatttatttcaaataggtacggataagtagatattgaagtaaggattcagtcttatatta 
               
               
                   
               
               
                 tattacataacattaatctattcctgcactgaaactgttgctttataggatttttcactacactaatgagaacttaagagataatggcc 
               
               
                   
               
               
                 taaaaccacagagagtatattcaaagataagtatagcacttcttatttggaaaccaatgcttactaaatgagactaagacgtgtcccat 
               
               
                   
               
               
                 caaaaatcctggacctatgcctaaaacacatttcacaatccctgaacttttcaaaaattggtacatgctttaactttaaactacaggcc 
               
               
                   
               
               
                 tcactggagctacagacaagaaggtgaaaaacggctgacaaaagaagtcctggtatcttctatggtgggagaagaaaactagctaaagg 
               
               
                   
               
               
                 gaagaataaattagagaaaaattggaatgactgaatcggaacaaggcaaaggctataaaaaaaattaagcagcagtatcctcttggggg 
               
               
                   
               
               
                 ccccttccccacactatctcaatgcaaatatctgtctgaaacggtccctggctaaactccacccatgggttggccagccttgccttgac 
               
               
                   
               
               
                 caatagccttgacaaggcaaacttgaccaatagtcttagagtatccagtgaggccaggggccggcggctggctagggatgaagaataaa 
               
               
                   
               
               
                 aggaagcacccttcagcagttccacacactcgcttctggaacgtctgaggttatcaataagctcctagtccagacgccatgggtcattt 
               
               
                   
               
               
                 cacagaggaggacaaggctactatcacaagcctgtggggcaaggtgaatgtggaagatgctggaggagaaaccctgggaaggtaggctc 
               
               
                   
               
               
                 tggtgaccaggacaagggagggaaggaaggaccctgtgcctggcaaaagtccaggtcgcttctcaggatttgtggcaccttctgactgt 
               
               
                   
               
               
                 caaactgttcttgtcaatctcacaggctcctggttgtctacccatggacccagaggttctttgacagctttggcaacctgtcctctgcc 
               
               
                   
               
               
                 tctgccatcatgggcaaccccaaagtcaaggcacatggcaagaaggtgctgacttccttgggagatgccataaagcacctggatgatct 
               
               
                   
               
               
                 caagggcacctttgcccagctgagtgaactgcactgtgacaagctgcatgtggatcctgagaacttcaaggtgagtccaggagatgttt 
               
               
                   
               
               
                 cagcactgttgcctttagtctcgaggcaacttagacaactgagtattgatctgagcacagcagggtgtgagctgtttgaagatactggg 
               
               
                   
               
               
                 gttgggagtgaagaaactgcagaggactaactgggctgagacccagtggcaatgttttagggcctaaggagtgcctctgaaaatctaga 
               
               
                   
               
               
                 tggacaactttgactttgagaaaagagaggtggaaatgaggaaaatgacttttctttattagatttcggtagaaagaactttcaccttt 
               
               
                   
               
               
                 cccctatttttgttattcgttttaaaacatctatctggaggcaggacaagtatggtcattaaaaagatgcaggcagaaggcatatattg 
               
               
                   
               
               
                 gctcagtcaaagtggggaactttggtggccaaacatacattgctaaggctattcctatatcagctggacacatataaaatgctgctaat 
               
               
                   
               
               
                 gcttcattacaaacttatatcctttaattccagatgggggcaaagtatgtccaggggtgaggaacaattgaaacatttgggctggagta 
               
               
                   
               
               
                 gattttgaaagtcagctctgtgtgtgtgtgtgtgtgtgtgcgcgcgtgtgtttgtgtgtgtgtgagagcgtgtgtttcttttaacgttt 
               
               
                   
               
               
                 tcagcctacagcatacagggttcatggtggcaagaagataacaagatttaaattatggccagtgactagtgctgcaagaagaacaacta 
               
               
                   
               
               
                 cctgcatttaatgggaaagcaaaatctcaggctttgagggaagttaacataggcttgattctgggtggaagcttggtgtgtagttatct 
               
               
                   
               
               
                 ggaggccaggctggagctctcagctcactatgggttcatctttattgtctcctttcatctcaacagctcctgggaaatgtgctggtgac 
               
               
                   
               
               
                 cgttttggcaatccatttcggcaaagaattcacccctgaggtgcaggcttcctggcagaagatggtgactggagtggccagtgccctgt 
               
               
                   
               
               
                 cctccagataccactgagctcactgcccatgatgcagagctttcaaggataggctttattctgcaagcaatcaaataataaatctattc 
               
               
                   
               
               
                 tgctaagagatcacacatggttgtcttcagttctttttttatgtctttttaaatatatgagccacaaagggttttatgttgagggatgt 
               
               
                   
               
               
                 gtttatgtgtatttatacatggctatgtgtgtttgtgtcatgtgcacactccacacttttttgtttacgttagatgtgggttttgatga 
               
               
                   
               
               
                 gcaaataaaagaactaggcaataaagaaacttgtacatgggagttctgcaagtgggagtaaaaggtgcaggagaaatctggttggaaga 
               
               
                   
               
               
                 aagacctctataggacaggactcctcagaaacagatgttttggaagagatggggaaaggttcagtgaagggggctgaacccccttccct 
               
               
                   
               
               
                 ggattgcagcacagcagcgaggaaggggctcaacgaagaaaaagtgttccaagctttaggaagtcaaggtttaggcagggatagccatt 
               
               
                   
               
               
                 ctattttattaggggcaatactatttccaacggcatctggcttttctcagcccttgtgaggctctacagggaggttgaggtgttagaga 
               
               
                   
               
               
                 tcagagcaggaaacaggtttttctttccacggtaactacaatgaagtgatccttactttactaaggaacttttcattttaagtgttgac 
               
               
                   
               
               
                 gcatgcctaaagaggtgaaattaatcccatacccttaagtctacagactggtcacagcatttcaaggaggagacctcattgtaagcttc 
               
               
                   
               
               
                 tagggaggtggggacttaggtgaaggaaatgagccagcagaagctcacaagtcagcatcagcgtgtcatgtctcagcagcagaacagca 
               
               
                   
               
               
                 cggtcagatgaaaatatagtgtgaagaatttgtataacattaattgagaaggcagattcactggagttcttatataattgaaagttaat 
               
               
                   
               
               
                 gcacgttaataagcaagagtttagtttaatgtgatggtgttatgaacttaacgcttgtgtctccagaaaattcacatgctgaatcccca 
               
               
                   
               
               
                 actcccaattggctccatttgtgggggaggctttggaaaagtaatcaggtttagaggagctcatgagagcagatccccatcatagaatt 
               
               
                   
               
               
                 attttcctcatcagaagcagagagattagccatttctcttccttctggtgaggacacagtgggaagtcagccacctgcaacccaggaag 
               
               
                   
               
               
                 agagccctgaccaggaaccagcagaaaagtgagaaaaaatcctgttgttgaagtcacccagtctatgctattttgttatagcaccttgc 
               
               
                   
               
               
                 actaagtaaggcagatgaagaaagagaaaaaaataagcttcggtgttcagtggattagaaaccatgtttatctcaggtttacaaatctc 
               
               
                   
               
               
                 cacttgtcctctgtgtttcagaataaaataccaactctactactctcatctgtaagatgcaaatagtaagcctgagcccttctgtctaa 
               
               
                   
               
               
                 ctttgaattctattttttcttcaacgtactttaggcttgtaatgtgtttatatacagtgaaatgtcaagttctttctttatatttcttt 
               
               
                   
               
               
                 ctttcttttttttcctcagcctcagagttttccacatgcccttcctactttcaggaacttctttctccaaacgtcttctgcctggctcc 
               
               
                   
               
               
                 atcaaatcataaaggacccacttcaaatgccatcactcactaccatttcacaattcgcactttctttctttgtcctttttttttttagt 
               
               
                   
               
               
                 aaaacaagtttataaaaaattgaaggaataaatgaatggctacttcataggcagagtagacgcaagggctactggttgccgatttttat 
               
               
                   
               
               
                 tgttatttttcaatagtatgctaaacaaggggtagattatttatgctgcccatttttagaccataaaagataacttcctgatgttgcca 
               
               
                   
               
               
                 tggcatttttttccttttaattttatttcatttcattttaatttcgaaggtacatgtgcaggatgtgcaggcttgttacatgggtaaat 
               
               
                   
               
               
                 gtgtgtctttctggccttttagccatctgtatcaatgagcagatataagctttacacaggatcatgaaggatgaaagaatttcaccaat 
               
               
                   
               
               
                 attataataatttcaatcaacctgatagcttaggggataaactaatttgaagatacagcttgcctccgataagccagaattccagagct 
               
               
                   
               
               
                 tctggcattataatctagcaaggttagagatcatggatcactttcagagaaaaacaaaaacaaactaaccaaaagcaaaacagaaccaa 
               
               
                   
               
               
                 aaaaccaccataaatacttcctaccctgttaatggtccaatatgtcagaaacagcactgtgttagaaataaagctgtctaaagtacact 
               
               
                   
               
               
                 aatattcgagttataatagtgtgtggactattagtcaataaaaacaacccttgcctctttagagttgttttccatgtacacgcacatct 
               
               
                   
               
               
                 tatgtcttagagtaagattccctgagaagtgaacctagcatttatacaagataattaattctaatccacagtacctgccaaagaacatt 
               
               
                   
               
               
                 ctaccatcatctttactgagcatagaagagctacgccaaaaccctgggtcatcagccagcacacacacttatccagtggtaaatacaca 
               
               
                   
               
               
                 tcatctggtgtatacatacatacctgaatatggaatcaaatatttttctaagatgaaacagtcatgatttatttcaaataggtacggat 
               
               
                   
               
               
                 aagtagatattgaggtaagcattaggtcttatattatgtaacactaatctattactgcgctgaaactgtggctttatagaaattgtttt 
               
               
                   
               
               
                 cactgcactattgagaaattaagagataatggcaaaagtcacaaagagtatattcaaaaagaagtatagcactttttccttagaaacca 
               
               
                   
               
               
                 ctgctaactgaaagagactaagatttgtcccgtcaaaaatcctggacctatgcctaaaacacatttcacaatccctgaacttttcaaaa 
               
               
                   
               
               
                 attggtacatgctttagctttaaactacaggcctcactggagctagagacaagaaggtaaaaaacggctgacaaaagaagtcctggtat 
               
               
                   
               
               
                 cctctatgatgggagaaggaaactagctaaagggaagaataaattagagaaaaactggaatgactgaatcggaacaaggcaaaggctat 
               
               
                   
               
               
                 aaaaaaaattagcagtatcctcttgggggccccttccccacactatctcaatgcaaatatctgtctgaaacggtccctggctaaactcc 
               
               
                   
               
               
                 acccatgggttggccagccttgccttgaccaatagccttgacaaggcaaacttgaccaatagtcttagagtatccagtgaggccagggg 
               
               
                   
               
               
                 ccggcggctggctagggatgaagaataaaaggaagcacccttcagcagttccacacactcgcttctggaacgtctgaggttatcaataa 
               
               
                   
               
               
                 gctcctagtccagacgccatgggtcatttcacagaggaggacaaggctactatcacaagcctgtggggcaaggtgaatgtggaagatgc 
               
               
                   
               
               
                 tggaggagaaaccctgggaaggtaggctctggtgaccaggacaagggagggaaggaaggaccctgtgcctggcaaaagtccaggtcgct 
               
               
                   
               
               
                 tctcaggatttgtggcaccttctgactgtcaaactgttcttgtcaatctcacaggctcctggttgtctacccatggacccagaggttct 
               
               
                   
               
               
                 ttgacagctttggcaacctgtcctctgcctctgccatcatgggcaaccccaaagtcaaggcacatggcaagaaggtgctgacttccttg 
               
               
                   
               
               
                 ggagatgccacaaagcacctggatgatctcaagggcacctttgcccagctgagtgaactgcactgtgacaagctgcatgtggatcctga 
               
               
                   
               
               
                 gaacttcaaggtgagtccaggagatgtttcagccctgttgcctttagtctcgaggcaacttagacaacggagtattgatctgagcacag 
               
               
                   
               
               
                 cagggtgtgagctgtttgaagatactggggttgggggtgaagaaactgcagaggactaactgggctgagacccagtggtaatgttttag 
               
               
                   
               
               
                 ggcctaaggagtgcctctaaaaatctagatggacaattttgactttgagaaaagagaggtggaaatgaggaaaatgacttttctttatt 
               
               
                   
               
               
                 agattccagtagaaagaactttcatctttccctcatttttgttgttttaaaacatctatctggaggcaggacaagtatggtcgttaaaa 
               
               
                   
               
               
                 agatgcaggcagaaggcatatattggctcagtcaaagtggggaactttggtggccaaacatacattgctaaggctattcctatatcagc 
               
               
                   
               
               
                 tggacacatataaaatgctgctaatgcttcattacaaacttatatcctttaattccagatgggggcaaagtatgtccaggggtgaggaa 
               
               
                   
               
               
                 caattgaaacatttgggctggagtagattttgaaagtcagctctgtgtgtgtgtgtgtgtgtgcgcgcgcgcgtgtgtgtgtgtgtgtc 
               
               
                   
               
               
                 agcgtgtgtttcttttaacgtcttcagcctacaacatacagggttcatggtggcaagaagatagcaagatttaaattatggccagtgac 
               
               
                   
               
               
                 tagtgcttgaaggggaacaactacctgcatttaatgggaaggcaaaatctcaggctttgagggaagttaacataggcttgattctgggt 
               
               
                   
               
               
                 ggaagcttggtgtgtagttatctggaggccaggctggagctctcagctcactatgggttcatctttattgtctcctttcatctcaacag 
               
               
                   
               
               
                 ctcctgggaaatgtgctggtgaccgttttggcaatccatttcggcaaagaattcacccctgaggtgcaggcttcctggcagaagatggt 
               
               
                   
               
               
                 gactgcagtggccagtgccctgtcctccagataccactgagctcactgcccatgattcagagctttcaaggataggctttattctgcaa 
               
               
                   
               
               
                 gcaatacaaataataaatctattctgctgagagatcacacatgattttcttcagctcttttttttacatctttttaaatatatgagcca 
               
               
                   
               
               
                 caaagggtttatattgagggaagtgtgtatgtgtatttctgcatgcctgtttgtgtttgtggtgtgtgcatgctcctcatttattttta 
               
               
                   
               
               
                 tatgagatgtgcattttgatgagcaaataaaagcagtaaagacacttgtacacgggagttctgcaagtgggagtaaatggtgtaggaga 
               
               
                   
               
               
                 aatccggtgggaagaaagacctctataggacaggacttctcagaaacagatgttttggaagagatgggaaaaggttcagtgaagacctg 
               
               
                   
               
               
                 ggggctggattgattgcagctgagtagcaaggatggttcttaaggaagggaaagtgttccaagctttaggaattcaaggtttagtcagg 
               
               
                   
               
               
                 tgtagcaattctattttattaggaggaatactatttctaatggcacttagcttttcacagcccttgtggatgcctaagaaagtgaaatt 
               
               
                   
               
               
                 aatcccatgccctcaagtgtgcagattggtcacagcatttcaagggagagacctcattgtaagactctgggggaggtggggacttaggt 
               
               
                   
               
               
                 gtaagaaatgaatcagcagaggctcacaagtcagcatgagcatgttatgtctgagaaacagaccagcactgtgagatcaaaatgtagtg 
               
               
                   
               
               
                 ggaagaatttgtacaacattaattggaaggcttacttaatggaatttttgtatagttggatgttagtgcatctctataagtaagagttt 
               
               
                   
               
               
                 aatatgatggtgttacggacctaatgtttgtgtctcctcaaaattcacatgctgaatccccaactcccaactgaccttatctgtggggg 
               
               
                   
               
               
                 aggcttttgaaaagtaattaggtttagatgagctcataagagcagatccccatcataaaattattttccttatcagaagcagagagaca 
               
               
                   
               
               
                 agccatttctctttcctcccggtgaggacacagtgagaagtccgccatctgcaatccaggaagagaaccctgaccacgagtcagccttc 
               
               
                   
               
               
                 agaaatgtgagaaaaaactctgttgttgaagccacccagtcttttgtattttgttatagcaccttgcactgagtaaggcagatgaagaa 
               
               
                   
               
               
                 ggagaaaaaaataagcttgggttttgagtggactacagaccatgtttatctcaggtttgcaaagctcccctcgtcccctatgtttcagt 
               
               
                   
               
               
                 ataaaatacctactctactactctcatctataagacccaaataataagcctgcgcccttctctctaactttgatttctcctatttttac 
               
               
                   
               
               
                 ttcaacatgctttactctagccttgtaatgtctttacatacagtgaaatgtaaagttctttattctttttttctttctttcttttttct 
               
               
                   
               
               
                 cctcagcctcagaatttggcacatgcccttccttctttcaggaacttctccaacatctctgcctggctccatcatatcataaaggtccc 
               
               
                   
               
               
                 acttcaaatgcagtcactaccgtttcagaatatgcactttctttcttttttgttttttgttttttttaagtcaaagcaaatttcttgag 
               
               
                   
               
               
                 agagtaaagaaataaacgaatgactactgcataggcagagcagccccgagggccgctggttgttccttttatggttatttcttgatgat 
               
               
                   
               
               
                 atgttaaacaagttttggattatttatgccttctctttttaggccatatagggtaactttctgacattgccatggcatttttcttttaa 
               
               
                   
               
               
                 tttaatttactgttaccttaaattcaggggtacacgtacaggatatgcaggtttgttttataggtaaaagtgtgccatggttttaatgg 
               
               
                   
               
               
                 gttttttttttcttgtaaagttgtttaagtttcttgtttactctggatattaggcctttgtcagaagaatagattggaaaatctttttc 
               
               
                   
               
               
                 ccattctgtagattgtctttcgctctgatggtagtttcttttgctgagcaggagctctttagtttaattagattccattggtcaatttt 
               
               
                   
               
               
                 tgcttttgctgcaattgcttttcacgctttcatcatgaaatctgtgcccgtgtttatatcatgaatagtattgccttgatttttttcta 
               
               
                   
               
               
                 ggctttttatagtttggggtttttcatttaagtctctaatccatctggagttaattttggataaggtataaggaaggagtccagtttca 
               
               
                   
               
               
                 tttttcagcatatggctagccagttctcccccatcatttattaaattgaaaatcctttccccattgcttgcttttgtcaggtttctaaa 
               
               
                   
               
               
                 agaccagatggttgtaggtacaatatgcagtttcttcaagtcatataataccatctgaaatctcttattaattcatttcttttagtatg 
               
               
                   
               
               
                 tatgctggtctcctctgctcactatagtgagggcaccattagccagagaatctgtctgtctagttcatgtaagattctcagaattaaga 
               
               
                   
               
               
                 aaaatggatggcatatgaatgaaacttcatggatgacatatggaatctaatatgtatttgttgaattaatgcataagatgcaacagaga 
               
               
                   
               
               
                 gaagttgacaactgcaatgataacctggtattgatgatataagagtctatagatcacagtagaagcaataatcatggaaaacaattgga 
               
               
                   
               
               
                 aatggggaacagccacaaacaagaaagaatcaatacttccaggaaagtgactgcaggtcacttttcctggagcgggtgagagaaaagtg 
               
               
                   
               
               
                 gaagttagcagtaactgctgaattcctggttggctgatggaaagatggggcagctgttcactggtacgcagggttttagatgtatgtac 
               
               
                   
               
               
                 ctaaggatatgaggtatggcaatgaacagaaattcttttgggaatgagttttagggccattaaaggacatgacctgaagtttcctctga 
               
               
                   
               
               
                 ggccagtccccacaactcaatataaatgtgtttcctgcatatagtcaaagttgccacttctttttcttcatatcatcgatctctgctct 
               
               
                   
               
               
                 taaagataatcttggttttgcctcaaactgtttgtcactacaaactttccccatgttcctaagtaaaacaggtaactgcctctcaacta 
               
               
                   
               
               
                 tatcaagtagactaaaatattgtgtctctaatatcagaaattcagctttaatatattgggtttaactctttgaaatttagagtctcctt 
               
               
                   
               
               
                 gaaatacacatgggggtgatttcctaaactttatttcttgtaaggatttatctcaggggtaacacacaaaccagcatcctgaacctcta 
               
               
                   
               
               
                 agtatgaggacagtaagccttaagaatataaaataaactgttcttctctctgccggtggaagtgtgccctgtctattcctgaaattgct 
               
               
                   
               
               
                 tgtttgagacgcatgagacgtgcagcacatgagacacgtgcagcagcctgtggaatattgtcagtgaagaatgtctttgcctgattaga 
               
               
                   
               
               
                 tataaagacaagttaaacacagcattagactatagatcaagcctgtgccagacacaaatgacctaatgcccagcacgggccacggaatc 
               
               
                   
               
               
                 tcctatcctcttgcttgaacagagcagcacacttctcccccaacactattagatgttctggcataattttgtagatatgtaggatttga 
               
               
                   
               
               
                 catggactattgttcaatgattcagaggaaatctcctttgttcagataagtacactgactactaaatggattaaaaaacacagtaataa 
               
               
                   
               
               
                 aacccagttttccccttacttccctagtttgtttcttattctgctttcttccaagttgatgctggatagaggtgtttatttctattcta 
               
               
                   
               
               
                 aaaagtgatgaaattggccgggcgcggtggctcacacctgtaatcccagcactttgggaggctgaggtgggcggatcacgaggtcagga 
               
               
                   
               
               
                 gatcaagaccatcctggctaacatggtgaaaccccatctctactaaaaatacaaaaaattagccagagacagtggcgggtgcctgtagt 
               
               
                   
               
               
                 cccagctactcgggaggctgaggcaggagaatggcgtgaacctgggaggcagagcttgcggtgagcagagatcgcgccactgcacactc 
               
               
                   
               
               
                 cagcctgggtgacaaagcgagactccatctcaaaaaaaaaaaaaaaaaaaagaaaaagaaagaaagaaagaaaaaaaaactgatgaaat 
               
               
                   
               
               
                 tgtgtattcaatgtagtctcaagagaattgaaaaccaagaaaggctgtggcttcttccacataaagcctggatgaataacaggataaca 
               
               
                   
               
               
                 cgttgttacattgtcacaactcctgatccaggaattgatggctaagatattcgtaattcttatccttttcagttgtaacttattcctat 
               
               
                   
               
               
                 ttgtcagcattcaggttattagcggctgctggcgaagtccttgagaaataaactgcacactggatggtgggggtagtgtaggaaaatgg 
               
               
                   
               
               
                 aggggaaggaagtaaagtttcaaattaagcctgaacagcaaagttcccctgagaaggccacctggattctatcagaaactcgaatgtcc 
               
               
                   
               
               
                 atcttgcaaaacttccttgcccaaaccccacccctggagtcacaacccacccttgaccaatagattcattttactgagggaggcaaagg 
               
               
                   
               
               
                 gctggtcaatagattcatttcactgggagaggcaaagggctgggggccagagaggagaagtaaaaagccacacatgaagcagcaatgca 
               
               
                   
               
               
                 ggcatgcttctggctcatctgtgatcaccaggaaactcccagatctgacactgtagtgcatttcactgctgacaagaaggctgctgcca 
               
               
                   
               
               
                 ccagcctgtgaagcaaggttaaggtgagaaggctggaggtgagattctgggcaggtaggtactggaagccgggacaaggtgcagaaagg 
               
               
                   
               
               
                 cagaaagtgtttctgaaagagggattagcccgttgtcttacatagtctgactttgcacctgctctgtgattatgactatcccacagtct 
               
               
                   
               
               
                 cctggttgtctacccatggacctagaggtactttgaaagttttggatatctgggctctgactgtgcaataatgggcaaccccaaagtca 
               
               
                   
               
               
                 aggcacatggcaagaaggtgctgatctccttcggaaaagctgttatgctcacggatgacctcaaaggcacctttgctacactgagtgac 
               
               
                   
               
               
                 ctgcactgtaacaagctgcacgtggaccctgagaacttcctggtgagtagtaagtacactcacgctttcttctttacccttagatattt 
               
               
                   
               
               
                 gcactatgggtacttttgaaagcagaggtggctttctcttgtgttatgagtcagctatgggatatgatatttcagcagtgggattttga 
               
               
                   
               
               
                 gagttatgttgctgtaaataacataactaaaatttggtagagcaaggactatgaataatggaaggccacttaccatttgatagctctga 
               
               
                   
               
               
                 aaaacacatcttataaaaaattctggccaaaatcaaactgagtgtttttggatgagggaacagaagttgagatagagaaaataacatct 
               
               
                   
               
               
                 ttcctttggtcagcgaaattttctataaaaattaatagtcacttttctgcatagtcctggaggttagaaaaagatcaactgaacaaagt 
               
               
                   
               
               
                 agtgggaagctgttaaaaagaggattgtttccctccgaatgatgatggtatacttttgtacgcatggtacaggattctttgttatgagt 
               
               
                   
               
               
                 gtttgggaaaattgtatgtatgtatgtatgtatgtatgtgatgactggggacttatcctatccattactgttccttgaagtactattat 
               
               
                   
               
               
                 cctactttttaaaaggacgaagtctctaaaaaaaaaatgaaacaatcacaatatgttggggtagtgagttggcatagcaagtaagagaa 
               
               
                   
               
               
                 ggataggacacaatgggaggtgcagggctgccagtcatattgaagctgatatctagcccataatggtgagagttgctcaaactctggtg 
               
               
                   
               
               
                 aaaaaggatgtaagtgttatatctatttactgcaagtccagcttgaggccttctattcactatgtaccattttcttttttatcttcact 
               
               
                   
               
               
                 ccctccccagctcttaggcaacgtgatattgattgttttggcaacccacttcagcgaggattttaccctacagatacaggcttcttggc 
               
               
                   
               
               
                 agtaactaacaaatgctgtggttaatgctgtagcccacaagaccactgagttccctgtccactatgtttgtacctatggtccactatgt 
               
               
                   
               
               
                 ttgtacctatgtcccaaaatctcatctcctttagatgggggaggttggggagaagagcagtatcctgcctgctgattcagttcctgcat 
               
               
                   
               
               
                 gataaaaatagaataaagaaatatgctctctaagaaatatcattgtactctttttctgtctttatattttaccctgattcagccaaaag 
               
               
                   
               
               
                 gacgcactatttctgatggaaatgagaatgttggagaatgggagtttaaggacagagaagatactttcttgcaatcctgcaagaaaaga 
               
               
                   
               
               
                 gagaactcgtgggtggatttagtggggtagttactcctaggaaggggaaatcgtctctagaataagacaatgtttttacagaaagggag 
               
               
                   
               
               
                 gtcaatggaggtactctttggaggtgtaagaggattgttggtagtgtgtagaggtatgttaggactcaaattagaagttctgtataggc 
               
               
                   
               
               
                 tattatttgtatgaaactcaggatatagctcatttggtgactgcagttcacttctacttattttaaacaacatattttttattatttat 
               
               
                   
               
               
                 aatgaagtggggatggggcttcctagagaccaatcaagggccaaaccttgaactttctcttaacgtcttcaatggtattaatagagaat 
               
               
                   
               
               
                 tatctctaaggcatgtgaactggctgtcttggttttcatctgtacttcatctgctacctctgtgacctgaaacatatttataattccat 
               
               
                   
               
               
                 taagctgtgcatatgatagatttatcatatgtattttccttaaaggatttttgtaagaactaattgaattgatacctgtaaagtcttta 
               
               
                   
               
               
                 tcacactacccaataaataataaatctctttgttcagctctctgtttctataaatatgtacaagttttattgtttttagtggtagtgat 
               
               
                   
               
               
                 tttattctctttctatatatatacacacacatgtgtgcattcataaatatatacaatttttatgaataaaaaattattagcaatcaata 
               
               
                   
               
               
                 ttgaaaaccactgatttttgtttatgtgagcaaacagcagattaaaaggctgagatttaggaaacagcacgttaagtcaagttgataga 
               
               
                   
               
               
                 ggagaatatggacatttaaaagaggcaggatgatataaaattagggaaactggatgcagagaccagatgaagtaagaaaaatagctatc 
               
               
                   
               
               
                 gttttgagcaaaaatcactgaagtttcttgcatatgagagtgacataataaatagggaaacgtagaaaattgattcacatgtatatata 
               
               
                   
               
               
                 tatatagaactgattagacaaagtctaacttgggtatagtcagaggagcttgctgtaattatattgaggtgatggataaagaactgaag 
               
               
                   
               
               
                 ttgatggaaacaatgaagttaagaaaaaaaatcgagtaagagaccattgtggcagtgattgcacagaactggaaaacattgtgaaacag 
               
               
                   
               
               
                 agagtcagagatgacagctaaaatccctgtctgtgaatgaaaagaaggaaatttattgacagaacagcaaatgcctacaagccccctgt 
               
               
                   
               
               
                 ttggatctggcaatgaacgtagccattctgtggcaatcacttcaaactcctgtacccaagacccttaggaagtatgtagcaccctcaaa 
               
               
                   
               
               
                 cctaaaacctcaaagaaagaggttttagaagatataataccctttcttctccagtttcattaatcccaaaacctctttctcaaagtatt 
               
               
                   
               
               
                 tcctctatgtgtccaccccaaagagctcacctcaccatatctcttgagtgggagcacatagataggcggtgctaccatctaacagcttc 
               
               
                   
               
               
                 tgaaattcctttgtcatatttttgagtccccactaataacccacaaagcagaataaataccagttgctcatgtacaataatcactcaac 
               
               
                   
               
               
                 tgctgtcttgtagcatacattaattaagcacattctttgaataattactgtgtccaaacaatcacactttaaaatctcacacttgtgct 
               
               
                   
               
               
                 atcccttgcccttctgaatgtcactctgtattttaaatgaagagatgagggttgaatttcctgtgttacttattgttcatttctcgatg 
               
               
                   
               
               
                 aggagttttcacattcacctttagtggaaaacacataagtacacatcttacaggaaaaatataccaaactgacatgtagcatgaatgct 
               
               
                   
               
               
                 tgtgcatgtagtcatataaaatcttgtagcaatgtaaacattctctgatatacacatacagatgtgtctatatgtctacacaatttctt 
               
               
                   
               
               
                 atgctccatgaacaaacattccatgcacacataagaacacacactgttacagatgcatacttgagtgcattgacaaaattaccccagtc 
               
               
                   
               
               
                 aatctagagaatttggatttctgcatttgactctgttagctttgtacatgctgttcatttactctgggtgatgtctttccctcattttg 
               
               
                   
               
               
                 ccttgtctatcttgtactcatactttaagtcctaacttatatgttatctcaactaagaagctatttttttttaattttaactgggctta 
               
               
                   
               
               
                 aagccctgtctataaactctgctacaattatgggctctttcttataatatttagtgtttttcctactaatgtacttaatctgctcattg 
               
               
                   
               
               
                 tatattcctaccactaaattttaacctcttttatggtagagacattgtcttgtaaactcttatttccctagtatttggagatgaaaaaa 
               
               
                   
               
               
                 aagattaaattatccaaaattagatctctcttttctacattatgagtattacactatccatagagaagtttgtttgagacctaaactga 
               
               
                   
               
               
                 ggaacctttggttctaaaatgactatgtgatatcttagtatttataggtcatgaggttccttcctctgcctctgctatagtttgattag 
               
               
                   
               
               
                 tcaacaagcatgtgtcatgcatttattcacatcagaatttcatacactaataagacatagtatcagaagtcagtttattagttatatca 
               
               
                   
               
               
                 gttagggtccatcaaggaaaggacaaaccattatcagttactcaacctagaattaaatacagctcttaatagttaattatccttgtatt 
               
               
                   
               
               
                 ggaagagctaaaatatcaaataaaggacagtgcagaaatctagatgttagtaacatcagaaaacctcttccgccattaggcctagaagg 
               
               
                   
               
               
                 gcagaaggagaaaatgtttataccaccagagtccagaaccagagcccataaccagaggtccactggattcagtgagctagtgggtgctc 
               
               
                   
               
               
                 cttggagagagccagaactgtctaatgggggcatcaaagtatcagccataaaaaaccataaaaaagactgtctgctgtaggagatccgt 
               
               
                   
               
               
                 tcagagagagagagagaccagaaataatcttgcttatgctttccctcagccagtgtttaccattgcagaatgtacatgcgactgaaagg 
               
               
                   
               
               
                 gtgaggaaacctgggaaatgtcagttcctcaaatacagagaacactgagggaaggatgagaaataaatgtgaaagcagacatgaatggt 
               
               
                   
               
               
                 aattgacagaaggaaactaggatgtgtccagtaaatgaataattacagtgtgcagtgattattgcaatgattaatgtattgataagata 
               
               
                   
               
               
                 atatgaaaacacagaattcaaacagcagtgaactgagattagaattgtggagagcactggcatttaagaatgtcacacttagaatgtgt 
               
               
                   
               
               
                 ctctaggcattgttctgtgcatatatcatctcaatattcattatctgaaaattatgaattaggtacaaagctcaaataatttatttttt 
               
               
                   
               
               
                 caggttagcaagaactttttttttttttttctgagatagagcattgctatggttgcccaggctggagtgcaatggcatgatccaggctc 
               
               
                   
               
               
                 actgcaacatctgcctcccaggttcaagcgattctcctgcctcagcctcccaagtagctggcactacaggcatgtgccaccaccatgcc 
               
               
                   
               
               
                 tggctaattttctatttttagtagatagggggtttcaccatgttggtcaggctgatctcgaactcctaacatcaggtgatccaccctcc 
               
               
                   
               
               
                 tcggcctctgaaagtgctgggatcacaggcgtgagccaccacacccagccaagaatgtgaattttgtagaaggatataacccatatttc 
               
               
                   
               
               
                 tctgaccctagagtccttagtatacctcccataccatgtggctcatcctccttacatacatttcccatctttcaccctaccttttcctt 
               
               
                   
               
               
                 tttgtttcagcttttcactgtgtcaaaatctagaaccttatctcctacctgctctgaaaccaacagcaagttgacttccattctaaccc 
               
               
                   
               
               
                 acattggcattacactaattaaaatcgatactgagttctaaaatcatcggggattttggggactatgtcttacttcatacttccttgag 
               
               
                   
               
               
                 atttcacattaaatgttggtgttcattaaaggtccttcatttaactttgtattcatcacactcttggattcacagttatatctaaactc 
               
               
                   
               
               
                 ttaaatacagcctgtataatcccaattcccaactctgatttctaacctctgacctccaacctcagtgccaaacccatatatcaaacaat 
               
               
                   
               
               
                 gtactgggcttatttatatagatgtcctataggcacctcagactcagcatgggtatttcacttgttatactaaaactgtttctcttcca 
               
               
                   
               
               
                 gtgttttccattttagtcattagatagctacttgcccattcaccaaggtcacagattaaaatcatttccctacctctaatcaacagttc 
               
               
                   
               
               
                 gattctgcttcaatttgtccctatctattaatcaccactcttactgcccagtcaggtcctcattgtttcctgaacaagagtagatgcta 
               
               
                   
               
               
                 ttctttccacttttagaccttatcctggctggatgcggtggctcaggcttgtaaacccagcactttgggaggccaaggcaggcagatca 
               
               
                   
               
               
                 cttgaggtcaggagttcaagaccagcctgaccaacatggtgaaaccccatctctactaaaaatacaaaatcagccgggcgtgtggtgca 
               
               
                   
               
               
                 tgcctgcagtcccagctattcaggtggctgaggcaggagaattgcttgaacccaggaggcagaggttgcggtgagcctagattgcacca 
               
               
                   
               
               
                 ttgcactctagcttgggcaatagggatgaaactccatctcagaagagaaaagaaaaaaagaccttattctgttatacaaatcctctcaa 
               
               
                   
               
               
                 tgcaatccatatagaataaacatgtaaccagatctcccaatgtgtaaaatcatttcaggtagaacagaattaaagtgaaaagccaagtc 
               
               
                   
               
               
                 tttggaattaacagacaaagatcaaataacagtcctcatggccttaagaatttacctaacattttttttagaatcaattttcttatata 
               
               
                   
               
               
                 tgaattggaaacataattcctccctcacaaacacattctaagattttaaggagatattgatgaagtacatcatctgtcatttttaacag 
               
               
                   
               
               
                 gtagtggtagtgattcacacagcacattatgatctgttcttgtatgttctgttccattctgtattcttgacctggttgtattctttctg 
               
               
                   
               
               
                 agctccagatccacatatctaagtacatctttttgcattttacaagagtgcatacaatacaatgtatccaagactgtatttctgatttt 
               
               
                   
               
               
                 atcgtaccactaaactcacaaatgtggccctattcttgtgttcacgactgacatcaccgtcatggtccaagtctgataatagaaatggc 
               
               
                   
               
               
                 attgtcactttcttccctactgcaacagaagcccagctatttgtctcccattttctctacttctaaaatacatttcttcactaagtgag 
               
               
                   
               
               
                 aataatcttttaaagacacaaatcaaaccatgccaccacctttcttgaattattcaatatctttcgttggcttccaggttacagaaaaa 
               
               
                   
               
               
                 taacttgtaacaaagtttaaaggtcattcatggctcctctctaccctattttataacatttccccttgtgatcagaatctcaggcacat 
               
               
                   
               
               
                 catccatctttctatatacaaataaagtcatatagtttgaactcacctctggttacttttaatcaaccaaatgctgtaaaatgcatttg 
               
               
                   
               
               
                 tatcgctacgtgttaagcagtagttgattcttttcatttctgtgtaatattctattctttgactataccgtaatttatcaattctactg 
               
               
                   
               
               
                 ttggtaagcatttaagtggctaccggtttgaggtttttatgattattgctgtcataagcatttctatacatgtctttggatacacacat 
               
               
                   
               
               
                 gcatgtgtttctgaatatctaaaaatgtaattgctaggtaatagacttatcaagcatccagcatttgtggatactattaaaggttttcc 
               
               
                   
               
               
                 aaaggggttatactattgtacagtgtcaccaacagagtttgagtttctattgatccatatcaccaccaaaatttgaactgtcagtctta 
               
               
                   
               
               
                 tctcttctcttgtctcttttttcctcttttttttccttcccttcccctctcttcgtttcttttctctcctcttctcttctttcctctct 
               
               
                   
               
               
                 tcccttccctttctctttctcttccctatcccttctcctctcctctcccctccttttttctcctctcctctccattatttatttttcct 
               
               
                   
               
               
                 tcttctcctccatcccttccatcctctctcttcccctcttccttccttcctttctccatttcttcctcctctttccttcaatccttcct 
               
               
                   
               
               
                 tttggatatgctcatgggtgtgtatttgtctgccattgtggcattatttgaattcagaaaagagtgaaaaactactgggatcttcattc 
               
               
                   
               
               
                 ctgggtctaattccacatttttttttaagaacacatctgtaaaaatgttctgtactagcatattcccaggaacttcgttaaatttaatc 
               
               
                   
               
               
                 tggctgaatatggtaaatctacttttcactttgcattctttctttagtcataccataattttaaacattcaaaatatttgtatataata 
               
               
                   
               
               
                 tttgattttatctgtcattaaaatgttaaccttaaaattcatgtttccagaacctatttcaataactggtaaataaacactattcattt 
               
               
                   
               
               
                 tttaaatattcttttaatggatatttatttcaatataataaaaaattagagttttattataggaagaatttaccaaaagaaggaggaag 
               
               
                   
               
               
                 caagcaagtttaaactgcagcaatagatttgtccattccaacctctcaaaattcccttggagacaaaaatctctagaggcaaagaagaa 
               
               
                   
               
               
                 ctttatattgagtcaacttgttaaaacatctgcttttagataagttttcttagtataaagtgacagaaacaaataagttaaactctaag 
               
               
                   
               
               
                 atacattccactatattagcctaaaacacttctgcaaaaatgaaactaggaggatatttttagaaacaactgctgaaagagatgcggtg 
               
               
                   
               
               
                 gggagatatgtagaggagaacagggtttctgagtcaagacacacatgacagaacagccaatctcagggcaagttaagggaatagtggaa 
               
               
                   
               
               
                 tgaaggttcatttttcattctcacaaactaatgaaaccctgcttatcttaaaccaacctgctcactggagcagggaggacaggaccagc 
               
               
                   
               
               
                 ataaaaggcagggcagagtcgactgttgcttacactttcttctgacataacagtgttcactagcaacctcaaacagacaccatggtgca 
               
               
                   
               
               
                 tctgactcctgaggagaagactgctgtcaatgccctgtggggcaaagtgaacgtggatgcagttggtggtgaggccctgggcaggttgg 
               
               
                   
               
               
                 tatcaaggttataagagaggctcaaggaggcaaatggaaactgggcatgtgtagacagagaagactcttgggtttctgataggcactga 
               
               
                   
               
               
                 ctctctgtcccttgggctgttttcctaccctcagattactggtggtctacccttggacccagaggttctttgagtcctttggggatctg 
               
               
                   
               
               
                 tcctctcctgatgctgttatgggcaaccctaaggtgaaggctcatggcaagaaggtgctaggtgcctttagtgatggcctggctcacct 
               
               
                   
               
               
                 ggacaacctcaagggcactttttctcagctgagtgagctgcactgtgacaagctgcacgtggatcctgagaacttcagggtgagtccag 
               
               
                   
               
               
                 gagatgcttcacttttctctttttactttctaatcttacattttggttcttttacctacctgctcttctcccacatttttgtcatttta 
               
               
                   
               
               
                 ctatattttatcatttaatgcttctaaaattttgttaattttttatttaaatattctgcattttttccttcctcacaatcttgctattt 
               
               
                   
               
               
                 taaattatttaatatcctgtctttctctcccaaccccctcccttcatttttccttctctaacaacaactcaaattatgcataccagctc 
               
               
                   
               
               
                 tcacctgctaattctgcacttagaataatccttttgtctctccacatgggtatgggagaggctccaactcaaagatgagaggcatagaa 
               
               
                   
               
               
                 tactgttttagaggctataaatcattttacaataaggaataattggaattttataaattctgtagtaaatggaatggaaaggaaagtga 
               
               
                   
               
               
                 atatttgattatgaaagactaggcagttacactggaggtggggcagaagtcgttgctaggagacagcccatcatcacactgattaatca 
               
               
                   
               
               
                 attaatttgtatctattaatctgtttatagtaattaatttgtatatgctatatacacatacaaaattaaaactaatttggaattaattt 
               
               
                   
               
               
                 gtatatagtattatacagcatatatagcatatatgtacatatatagactacatgctagttaagtacatagaggatgtgtgtgtatagat 
               
               
                   
               
               
                 atatgttatatgtatgcattcatatatgtacttatttatgctgatgggaataacctggggatcagttttgtctaagatttgggcagaaa 
               
               
                   
               
               
                 aaaatgggtgttggctcagtttctcagaagccagtctttatttctctgttaaccatatgcatgtatctgcctacctcttctccgcagct 
               
               
                   
               
               
                 cttgggcaatgtgctggtgtgtgtgctggcccgcaactttggcaaggaattcaccccacaaatgcaggctgcctatcagaaggtggtgg 
               
               
                   
               
               
                 ctggtgtggctaatgccctggctcacaagtaccattgagatcctggactgtttcctgataaccataagaagaccctatttccctagatt 
               
               
                   
               
               
                 ctattttctgaacttgggaacacaatgcctacttcaagggtatggcttctgcctaataaagaatgttcagctcaacttcctgattaatt 
               
               
                   
               
               
                 tcacttatttcatttttttgtccaggtgtgtaagaaggttcctgaggctctacagatagggagcacttgtttattttacaaagagtaca 
               
               
                   
               
               
                 tgggaaaagagaaaagcaagggaaccgtacaaggcattaatgggtgacacttctacctccaaagagcagaaattatcaagaactcttga 
               
               
                   
               
               
                 tacaaagataatactggcactgcagaggttctagggaagacctcaaccctaagacatagcctcaagggtaatgctacgattaaactcca 
               
               
                   
               
               
                 acaattactgagaaaataatgtgctcaattaaaggcataatgattactcaagacaatgttatgttgtctttcttcctccttcctttgcc 
               
               
                   
               
               
                 tgcacattgtagcccataatactataccccatcaagtgttcctgctccaagaaatagcttcctcctcttacttgccccagaacatctct 
               
               
                   
               
               
                 gtaaagaatttcctcttatcttcccatatttcagtcaagattcattgctcacgtattacttgtgacctctcttgaccccagccacaata 
               
               
                   
               
               
                 aacttctctatactacccaaaaaatctttccaaaccctcccccacaccattttttatatttttatatttttcttatttatttcatgcac 
               
               
                   
               
               
                 acacacacactccgtgctttataagcaattctgcctattctctaccttcttacatgcctactgtgcctcatattaaattcatcaatggg 
               
               
                   
               
               
                 cagaaagaaaatatttattcaagaaaacagtgaatgaatgaacgaatgagtaaatgagtaaatgaaggaatgattattccttgctttag 
               
               
                   
               
               
                 aacttctggaattagaggacaatattaataataccatcgcacagtgtttctttgttgttaatgctacaacatacaaagaggaagcatgc 
               
               
                   
               
               
                 agtaaacaaccgaacagttatttcctttctgatcataggagtaatatttttttccttgagcaccatttttgccataggtaaaattagaa 
               
               
                   
               
               
                 ggatttttagaactttctcagttgtatacatttttaaaaatctgtattatatgcatgttgattaattttaaacttacttgaatacctaa 
               
               
                   
               
               
                 acagaatctgttgtttccttgtgtttgaaagtgctttcacagtaactctgtctgtactgccagaatatactgacaatgtgttatagtta 
               
               
                   
               
               
                 actgttttgatcacaacattttgaattgactggcagcagaagctcttttatatccatgtgttttccttaagtcattatacatagtaggc 
               
               
                   
               
               
                 actgagaactctttatatctgaataagatatttaggaaccactggtttacatatcagaagcagagctactcagggcattttggggaaga 
               
               
                   
               
               
                 tcactttcacattcctgagcatagggaagttctcataagagtaagatattaaaaggagatacttgtgtggtattcgaaagacagtaaga 
               
               
                   
               
               
                 gagattgtagaccttatgatcttgatagggaaaacaaactacattcctttctccaaaagtcaaaaaaaaagagcaaatatagcttacta 
               
               
                   
               
               
                 taccttctattcctacaccattagaagtagtcagtgagtctaggcaagatgttggccctaaaaatccaaataccagagaattcatgaga 
               
               
                   
               
               
                 acatcacctggatgggacatgtgccgagcacacacaattactatatgctaggcattgctatcttcatattgaagatgaggaggtcaaga 
               
               
                   
               
               
                 gatgaaaaaagacttggcaccttgttgttatattaaaattatttgttagagtagagcttttgtaagagtctaggagtgtgggagctaaa 
               
               
                   
               
               
                 tgatgatacacatggacacaaaaaatagatcaacagacacccaggcctacttgagggttgagggtgggaagagggagacgatgaaaaag 
               
               
                   
               
               
                 aacctattgggtattaagttcatcactgagtgatgaaataatctgtacatcaagacccagtgatatgcaatttacctatataacttgta 
               
               
                   
               
               
                 catgtacccccaaatttaaaatgaaagttaaaacaaagtataggaatggaattaattcctcaagatttggctttaattttatttgataa 
               
               
                   
               
               
                 tttatcaaatggttgtttttcttttctcactatggcgttgctttataaactatgttcagtatgtctgaatgaaagggtgtgtgtgtgtg 
               
               
                   
               
               
                 tgaaagagagggagagaggaagggaagagaggacgtaataatgtgaatttgagttcatgaaaatttttcaataaaataatttaatgtca 
               
               
                   
               
               
                 ggagaattaagcctaatagtctcctaaatcatccatctcttgagcttcagagcagtcctctgaattaatgcctacatgtttgtaaaggg 
               
               
                   
               
               
                 tgttcagactgaagccaagattctacctctaaagagatgcaatctcaaatttatctgaagactgtacctctgctctccataaattgaca 
               
               
                   
               
               
                 ccatggcccacttaatgaggttaaaaaaaagctaattctgaatgaaaatctgagcccagtggaggaaatattaatgaacaaggtgcaga 
               
               
                   
               
               
                 ctgaaatataaatttttctgtaataattatgcatatactttagcaaagttctgtctatgttgactttattgctttttggtaagaaatac 
               
               
                   
               
               
                 aactttttaaagtgaactaaactatcctatttccaaactattttgtgtgtgtgcggtttgtttctatgggttctggttttcttggagca 
               
               
                   
               
               
                 tttttatttcattttaattaattaattctgagagctgctgagttgtgtttactgagagattgtgtatctgcgagagaagtctgtagcaa 
               
               
                   
               
               
                 gtagctagactgtgcttgacctaggaacatatacagtagattgctaaaatgtctcacttggggaattttagactaaacagtagagcatg 
               
               
                   
               
               
                 tataaaaatactctagtcaagtgctgcttttgaaacaaatgataaaaccacactcccatagatgagtgtcatgattttcatggaggaag 
               
               
                   
               
               
                 ttaatattcatcctctaagtatacccagactagggccattctgatataaaacattaggacttaagaaagattaatagactggagtaaag 
               
               
                   
               
               
                 gaaatggacctctgtctctctcgctgtctcttttttgaggacttgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgtgttgtggtcagtg 
               
               
                   
               
               
                 gggctggaataaaagtagaatagacctgcacctgctgtggcatccattcacagagtagaagcaagctcacaatagtgaagatgtcagta 
               
               
                   
               
               
                 agcttgaatagtttttcaggaactttgaatgctgatttagatttgaaactgaggctctgaccataaccaaatttgcactatttattgct 
               
               
                   
               
               
                 tcttgaaacttatttgcctggtatgcctgggcttttgatggtcttagtatagcttgcagccttgtccctgcagggtattatgggtaata 
               
               
                   
               
               
                 gaaagaaaagtctgcgttacactctagtcacactaagtaactaccattggaaaagcaacccctgccttgaagccaggatgatggtatct 
               
               
                   
               
               
                 gcagcagttgccaacacaagagaaggatccatagttcatcatttaaaaaagaaaacaaaatagaaaaaggaaaactatttctgagcata 
               
               
                   
               
               
                 agaagttgtagggtaagtctttaagaaggtgacaatttctgccaatcaggatttcaaagctcttgctttgacaattttggtctttcaga 
               
               
                   
               
               
                 atactataaatataacctatattataatttcataaagtctgtgcattttctttgacccaggatatttgcaaaagacatattcaaacttc 
               
               
                   
               
               
                 cgcagaacactttatttcacatatacatgcctcttatatcagggatgtgaaacagggtcttgaaaactgtctaaatctaaaacaatgct 
               
               
                   
               
               
                 aatgcaggtttaaatttaataaaataaaatccaaaatctaacagccaagtcaaatctgcatgttttaacatttaaaatattttaaagac 
               
               
                   
               
               
                 gtcttttcccaggattcaacatgtgaaatcttttctcagggatacacgtgtgcctagatcctcattgctttagttttttacagaggaat 
               
               
                   
               
               
                 gaatataaaaagaaaatacttaaattttatccctcttacctctataatcatacataggcataattttttaacctaggctccagatagcc 
               
               
                   
               
               
                 atagaagaaccaaacactttctgcgtgtgtgagaataatcagagtgagattttttcacaagtacctgatgagggttgagacaggtagaa 
               
               
                   
               
               
                 aaagtgagagatctctatttatttagcaataatagagaaagcatttaagagaataaagcaatggaaataagaaatttgtaaatttcctt 
               
               
                   
               
               
                 ctgataactagaaatagaggatccagtttcttttggttaacctaaattttatttcattttattgttttattttattttattttatttta 
               
               
                   
               
               
                 ttttgtgtaatcgtagtttcagagtgttagagctgaaaggaagaagtaggagaaacatgcaaagtaaaagtataacactttccttacta 
               
               
                   
               
               
                 aaccgacatgggtttccaggtaggggcaggattcaggatgactgacagggcccttagggaacactgagaccctacgctgacctcataaa 
               
               
                   
               
               
                 tgcttgctacctttgctgttttaattacatcttttaatagcaggaagcagaactctgcacttcaaaagtttttcctcacctgaggagtt 
               
               
                   
               
               
                 aatttagtacaaggggaaaaagtacagggggatgggagaaaggcgatcacgttgggaagctatagagaaagaagagtaaattttagtaa 
               
               
                   
               
               
                 aggaggtttaaacaaacaaaatataaagagaaataggaacttgaatcaaggaaatgattttaaaacgcagtattcttagtggactagag 
               
               
                   
               
               
                 gaaaaaaataatctgagccaagtagaagaccttttcccctcctacccctactttctaagtcacagaggctttttgttcccccagacact 
               
               
                   
               
               
                 cttgcagattagtccaggcagaaacagttagatgtccccagttaacctcctatttgacaccactgattaccccattgatagtcacactt 
               
               
                   
               
               
                 tgggttgtaagtgactttttatttatttgtatttttgactgcattaagaggtctctagttttttatctcttgtttcccaaaacctaata 
               
               
                   
               
               
                 agtaactaatgcacagagcacattgatttgtatttattctatttttagacataatttattagcatgcatgagcaaattaagaaaaacaa 
               
               
                   
               
               
                 caacaaatgaatgcatatatatgtatatgtatgtgtgtatatatacacacatatatatatatattttttcttttcttaccagaaggttt 
               
               
                   
               
               
                 taatccaaataaggagaagatatgcttagaaccgaggtagagttttcatccattctgtcctgtaagtattttgcatattctggagacgc 
               
               
                   
               
               
                 aggaagagatccatctacatatcccaaagctgaattatggtagacaaaactcttccacttttagtgcatcaacttcttatttgtgtaat 
               
               
                   
               
               
                 aagaaaattgggaaaacgatcttcaatatgcttaccaagctgtgattccaaatattacgtaaatacacttgcaaaggaggatgttttta 
               
               
                   
               
               
                 gtagcaatttgtactgatggtatggggccaagagatatatcttagagggagggctgagggtttgaagtccaactcctaagccagtgcca 
               
               
                   
               
               
                 gaagagccaaggacaggtacggctgtcatcacttagacctcaccctgtggagccacaccctagggttggccaatctactcccaggagca 
               
               
                   
               
               
                 gggagggcaggagccagggctgggcataaaagtcagggcagagccatctattgcttacatttgcttctgacacaactgtgttcactagc 
               
               
                   
               
               
                 aacctcaaacagacaccatggtgcatctgactcctgaggagaagtctgccgttactgccctgtggggcaaggtgaacgtggatgaagtt 
               
               
                   
               
               
                 ggtggtgaggccctgggcaggttggtatcaaggttacaagacaggtttaaggagaccaatagaaactgggcatgtggagacagagaaga 
               
               
                   
               
               
                 ctcttgggtttctgataggcactgactctctctgcctattggtctattttcccacccttaggctgctggtggtctacccttggacccag 
               
               
                   
               
               
                 aggttctttgagtcctttggggatctgtccactcctgatgctgttatgggcaaccctaaggtgaaggctcatggcaagaaagtgctcgg 
               
               
                   
               
               
                 tgcctttagtgatggcctggctcacctggacaacctcaagggcacctttgccacactgagtgagctgcactgtgacaagctgcacgtgg 
               
               
                   
               
               
                 atcctgagaacttcagggtgagtctatgggacgcttgatgttttctttccccttcttttctatggttaagttcatgtcataggaagggg 
               
               
                   
               
               
                 ataagtaacagggtacagtttagaatgggaaacagacgaatgattgcatcagtgtggaagtctcaggatcgttttagtttcttttattt 
               
               
                   
               
               
                 gctgttcataacaattgttttcttttgtttaattcttgctttctttttttttcttctccgcaatttttactattatacttaatgcctta 
               
               
                   
               
               
                 acattgtgtataacaaaaggaaatatctctgagatacattaagtaacttaaaaaaaaactttacacagtctgcctagtacattactatt 
               
               
                   
               
               
                 tggaatatatgtgtgcttatttgcatattcataatctccctactttattttcttttatttttaattgatacataatcattatacatatt 
               
               
                   
               
               
                 tatgggttaaagtgtaatgttttaatatgtgtacacatattgaccaaatcagggtaattttgcatttgtaattttaaaaaatgctttct 
               
               
                   
               
               
                 tcttttaatatacttttttgtttatcttatttctaatactttccctaatctctttctttcagggcaataatgatacaatgtatcatgcc 
               
               
                   
               
               
                 tctttgcaccattctaaagaataacagtgataatttctgggttaaggcaatagcaatatctctgcatataaatatttctgcatataaat 
               
               
                   
               
               
                 tgtaactgatgtaagaggtttcatattgctaatagcagctacaatccagctaccattctgcttttattttatggttgggataaggctgg 
               
               
                   
               
               
                 attattctgagtccaagctaggcccttttgctaatcatgttcatacctcttatcttcctcccacagctcctgggcaacgtgctggtctg 
               
               
                   
               
               
                 tgtgctggcccatcactttggcaaagaattcaccccaccagtgcaggctgcctatcagaaagtggtggctggtgtggctaatgccctgg 
               
               
                   
               
               
                 cccacaagtatcactaagctcgctttcttgctgtccaatttctattaaaggttcctttgttccctaagtccaactactaaactggggga 
               
               
                   
               
               
                 tattatgaagggccttgagcatctggattctgcctaataaaaaacatttattttcattgcaatgatgtatttaaattatttctgaatat 
               
               
                   
               
               
                 tttactaaaaagggaatgtgggaggtcagtgcatttaaaacataaagaaatgaagagctagttcaaaccttgggaaaatacactatatc 
               
               
                   
               
               
                 ttaaactccatgaaagaaggtgaggctgcaaacagctaatgcacattggcaacagcccctgatgcatatgccttattcatccctcagaa 
               
               
                   
               
               
                 aaggattcaagtagaggcttgatttggaggttaaagttttgctatgctgtattttacattacttattgttttagctgtcctcatgaatg 
               
               
                   
               
               
                 tcttttcactacccatttgcttatcctgcatctctcagccttgactccactcagttctcttgcttagagataccacctttcccctgaag 
               
               
                   
               
               
                 tgttccttccatgttttacggcgagatggtttctcctcgcctggccactcagccttagttgtctctgttgtcttatagaggtctacttg 
               
               
                   
               
               
                 aagaaggaaaaacaggggtcatggtttgactgtcctgtgagcccttcttccctgcctcccccactcacagtgacccggaatctgcagtg 
               
               
                   
               
               
                 ctagtctcccggaactatcactctttcacagtctgctttggaaggactgggcttagtatgaaaagttaggactgagaagaatttgaaag 
               
               
                   
               
               
                 gcggctttttgtagcttgatattcactactgtcttattaccctgtcataggcccaccccaaatggaagtcccattcttcctcaggatgt 
               
               
                   
               
               
                 ttaagattagcattcaggaagagatcagaggtctgctggctcccttatcatgtcccttatggtgcttctggctctgcagttattagcat 
               
               
                   
               
               
                 agtgttaccatcaaccaccttaacttcatttttcttattcaatacctaggtaggtagatgctagattctggaaataaaatatgagtctc 
               
               
                   
               
               
                 aagtggtccttgtcctctctcccagtcaaattctgaatctagttggcaagattctgaaatcaaggcatataatcagtaataagtgatga 
               
               
                   
               
               
                 tagaagggtatatagaagaattttattatatgagagggtgaaaccctcaaaatgaaatgaaatcagacccttgtcttacaccataaaca 
               
               
                   
               
               
                 aaaataaatttgaatgggttaaagaattaaactaagacctaaaaccataaaaatttttaaagaaatcaaaagaagaaaattctaatatt 
               
               
                   
               
               
                 cacgttgcagccgttttttgaatttgatatgagaagcaaaggcaacaaaaggaaaaataaagaagtgaggctacatcaaactaaaaaat 
               
               
                   
               
               
                 ttccacacaaaaaacaaaacaatgaacaaatgaaaggtgaaccatgaaatggcatatttgcaaaccaaatatttcttaaatattttggt 
               
               
                   
               
               
                 taatatccaaaatatataagaaacacagatgattcaataacaaacaaaaaattaaaaataggaaaataaaaaaattaaaaagaagaaaa 
               
               
                   
               
               
                 tcctgccatttatggcagaattgatgaacctggaggatgtaaaactaagaaaaataagcctgacacaaaaagacaaatactacacaacc 
               
               
                   
               
               
                 ttgctcatatgtgaaacataaaaaagtcactctcatggaaacagacagtagaggtatggtttccaggggttgggggtgggagaatcagg 
               
               
                   
               
               
                 aaactattactcaaagggtataaaatttcagttatgtgggatgaataaattctagatatctaatgtacagcatcgtgactgtagttaat 
               
               
                   
               
               
                 tgtactgtaagtatatttaaaatttgcaaagagagtagatttttttttttttttagatggagttttgctcttgttgtccaggctggagt 
               
               
                   
               
               
                 gcaatggcaagatcttggctcactgcaacctccgcctcctgggttcaagcaaatctcctgcctcagcctcccgagtagctgggattaca 
               
               
                   
               
               
                 ggcatgcgacaccatgcccagctaattttgtatttttagtagagacggggtttctccatgttggtcaggctgatccgcctgcctcggcc 
               
               
                   
               
               
                 acccaaagggctgggattacaggcgtgagccaccgggcctggccgagagtagatcttaaaagcatttaccacaagaaaaaggtaactat 
               
               
                   
               
               
                 gtgagataatgggtatgttaattagcttgattgtggtaatcatttcacaaggtatacatatattaaaacatcatgttgtacaccttaaa 
               
               
                   
               
               
                 tatatacaatttttatttgtgaatgatacctcaataaagttgaagaataataaaaaagaatagacatcacatgaattaaaaaactaaaa 
               
               
                   
               
               
                 aataaaaaaatgcatcttgatgattagaattgcattcttgatttttcagatacaaatatccatttgactgtttactcttttccaaaaca 
               
               
                   
               
               
                 atacaataaattttagcactttatcttcattttccccttcccaatctataattatatatatatatattttagatattttgtatagtttt 
               
               
                   
               
               
                 actccctagattttctagtgttattattaaatagtgaagaaatgtttacacttatgtacaaaatgttttgcatgcttttcttcatttct 
               
               
                   
               
               
                 aacattctctctaagtttattctatttttttctgattatccttaatattatctctttctgctggaaatacattgttacttttggtttat 
               
               
                   
               
               
                 ctaaaaatggcttcattttcttcattctaaaatcatgttaaattaataccactcatgtgtaagtaagatagtggaataaatagaaatcc 
               
               
                   
               
               
                 aaaaactaaatctcactaaaatataataatgtgatatataaaaatatagcttttaaatttagcttggaaataaaaaacaaacagtaatt 
               
               
                   
               
               
                 gaacaactatactttttgaaaagagtaaagtgaaatgcttaactgcatataccacaatcgattacacaattaggtgtgaaggtaaaatt 
               
               
                   
               
               
                 cagtcacgaaaaaactagaataaaaatatgggaagacatgtatataatcttagagataacactgttatttaattatcaacccaaagtag 
               
               
                   
               
               
                 aaactatcaagggagaaataaattcagtcaacaataaaagcatttaagaagttattctaggctgggagcggtggctcacacctgcaatt 
               
               
                   
               
               
                 gcagcactttgggaggcctagacaggcggatcacgacgtcaggagttcaagatcagcctggccaacatagtgaaacctcatcgctacta 
               
               
                   
               
               
                 aaaatataaaaacttagcctggcgtggtggcaggcatgtgtaatcccagcaatttgggaggctgaggcaggagaatcgcttgatcctgg 
               
               
                   
               
               
                 gaggcagaggttgcagtgagccaagattgtgccactgcattccagcccaggtgacagcatgagactccgtcacaaaaaaaaaagaaaaa 
               
               
                   
               
               
                 aaaaaggggggggggagcggtggagccaagatgaccgaataggaacagctccagtctatagctcccatcgtgagtgacgcagaagacgg 
               
               
                   
               
               
                 gtgatttctgcatttccaactgaggtaccaggttcatctcacagggaagtgccaggcagtgggtgcaggacagtaggtgcagtgcactg 
               
               
                   
               
               
                 tgcatgagccaaagcagggcgaggcatcacctcacccgggaagcacaaggggtcagggaattccctttcctagtcaaagaaaagggtga 
               
               
                   
               
               
                 cagatggcacctggaaaatcgggtcactcccgccctaatactgcgctcttccaacaagcttaacaaatggcacaccaggagattatatc 
               
               
                   
               
               
                 ccatgcctggctcagagggtcctacgcccatggagcctcgctcattgctagcacagcagtctgaggtcaaactgcaaggtggcagtgag 
               
               
                   
               
               
                 gctgggggaggggtgcccaccattgtccaggcttgagcaggtaaacaaagccgcctggaagctcgaactgggtggagcccaccacagct 
               
               
                   
               
               
                 caaggaggcctgcctgcctctgtaggctccacctctaggggcagggcacagacaaacaaaagacaacaagaacctctgcagacttaaat 
               
               
                   
               
               
                 gtccctgtctgacagctttgaagagagtagtggttctcccagcacatagcttcagatctgagaacaggcagactgcctcctcaagtggg 
               
               
                   
               
               
                 tccctgacccccgagtagcctaactgggaggcatcccccagtaggggcagactgacacctcacatggctggtactcctctaagacaaaa 
               
               
                   
               
               
                 cttccagaggaatgatcaggcagcagcatttgcggttcaccaatatccactgttctgcagccaccgctgttgatacccaggaaaacagc 
               
               
                   
               
               
                 ttctggagtggacctccagtaaactccaacagacctgcagctgagggtcctgactgttagaaggaaaactaacaaacagaaaggacatc 
               
               
                   
               
               
                 cacaccaaaaacccatctgtacatcgccatcatcaaagaccaaaggtagataaaaccataaagatggggaaaaagcagagcagaaaaac 
               
               
                   
               
               
                 tggacactctaaaaatgagagtgcctctcctcctccaaagtaacgcagctcctcaccagcaatggaacaaagctgggcagagaatgact 
               
               
                   
               
               
                 ttgacgagttgagagaggaaggcttcagaagatcaaactactccaagctaaaggaggaagttcgaacaaacggcaaagaagtaaaaaac 
               
               
                   
               
               
                 tttgaaaaaaaattagatgaatggataactagaataaccaatgcacagaagtccttaaaggacctgatggagctgaaaaccaaggcagg 
               
               
                   
               
               
                 agaactacgtgacaaatacacaagcctcagtaaccgatgagatcaactggaagaaagggtatcaatgacgaaagatgaaatgaatgaaa 
               
               
                   
               
               
                 tgaagcatgaagagaagtttagagaaaaaagaataaaaagaaacgaacaaagcctccaagaaatatgggactatgtgaaaagaccaaat 
               
               
                   
               
               
                 ctacatctaattggtgtagctgaaagtgatggggagaatggaaccaagttggaaaacactctgcaggatattatccaggagaacttccc 
               
               
                   
               
               
                 caatctagcaaggcaagcccaaattcacattcaggaaatacagagaacgccacaaagatactcctagagaaaagcaactccaagacaca 
               
               
                   
               
               
                 taactgtcagattcaccaaagttgaaatgaaggaaaaaatgttaagggcagccagagagaaaggtcgggttacccacaaagggaagccc 
               
               
                   
               
               
                 atcagactaacagctgatctatcggcagaaactctacaagccagaagaaagtgggggccaatattcaacattgttaaagaaaagaattt 
               
               
                   
               
               
                 tcaacccagaatttcatatccagccaaactaagcttcataagtgaaggagaaataaaatcctttacagacaagcaaatgctgagagatt 
               
               
                   
               
               
                 ttgtcaccaccaggcctgccctacaagagctcctgaaggaagcactaaacatggaaaggaacaactagtatcagccactgcaaaaacat 
               
               
                   
               
               
                 gccaaattgtaaagaccatcaaggctaggaagaaactgcatcaacgagcaaaataaccagctaacatcataatgacaggatcaaattca 
               
               
                   
               
               
                 tacataacaatactcaccttaaatgtaaataggctaaatgctccaattaaaagacacagactggcaaattggataaggagtcaagaccc 
               
               
                   
               
               
                 atctgtgttctgtattcaggaaacccatctcacgtgcagagacacacataggctcgaaataaaaggatggaggaatatctaccaagcaa 
               
               
                   
               
               
                 atggaaaacaaaaaaaggcaggggttgcaatcctagtctctgataaaacagattttaaaccaacaaagatcaaaagagacaaagaaggc 
               
               
                   
               
               
                 cattacataatggcaaagggatctattcaagaagaagaactaactatactaaatatatatgcacccaatacaggagcacccagattcat 
               
               
                   
               
               
                 aaaacaagtcctgagtgacctacaaagagacttagatgcccacacaataataatgggagactttaacaccccactgtcaacattagaca 
               
               
                   
               
               
                 gatcaacgagacagaaagttaacaaggatatccaggaattggactcagctctgcaccaagcagacctaatagacatctacagaactctc 
               
               
                   
               
               
                 caccccaaatcaacagaatatacattcttttcagcaccacaccacacctattccaaaactgaccacatagttggaagtaaagctctcct 
               
               
                   
               
               
                 cagcaaatgtaaaagaacagaaactataacaaactgtctctcagaccacagtgcaatcaaactagaactcaggattaagaaactcactc 
               
               
                   
               
               
                 aaaaccactcagctacatggaaactgaacagcctgctcctgaatgactactgggtacataacaaaatgaaggcagaaataaagatgttc 
               
               
                   
               
               
                 tttgaaaccaacgagaacaaagacacaacacaccagaatctctgagacacattcaaagcagtgtgtagagggaaatttatagcactaaa 
               
               
                   
               
               
                 tgcccacaagggaaagcaggaaagatctaaaattgacaccctaacatcacaattaaaaaactagagaagcaggagcaaacacattcaaa 
               
               
                   
               
               
                 agctaacagaagacaagaaataactaagatcagagcagaagtgaaggacatagagacacaaaaaaacccttcaaaaaaatcaatgaatc 
               
               
                   
               
               
                 cagaagctgtttttttgaaaagatcaacaaaattgatagactgctagcaagactaataaagaagaaaagagagaagaatcaaatagacg 
               
               
                   
               
               
                 caataaaaaatgacacggggtatcaccactgatcccacagaaatacaaactaccgtcagagaatactataaacacctctacgcaaataa 
               
               
                   
               
               
                 actagaaaatctagaagaaatggataaattcctcgacacatacactctgccaagactaaaccaggaagaagttgtatctctgaatagac 
               
               
                   
               
               
                 caataacaggctctgaaattgaggcaataattaatagcttatcaaccaaaaaaagtccgggaccagtaggattcatagccgaattctac 
               
               
                   
               
               
                 cagaggtacaaggaggagctggtaccattccttctgaaactattccaatcaatagaaaaagagggaatcctccctaactcattttatga 
               
               
                   
               
               
                 ggccagcatcatcctgataccaaagcctgacagagacacaacaaaaaaagagaatgttacaccaatatccttgatgaacattgatgcaa 
               
               
                   
               
               
                 aaatcctcaataaaatactggcaaactgatccaccatgatcaagtgggcttcatccctgccatgcaaggctggttcaacatacgaaaat 
               
               
                   
               
               
                 caataaacataatccagcatataaacagaaccaaagacacaaaccatatgattatctcaatagatgcagaaaaggcctttgacaaaatt 
               
               
                   
               
               
                 caacaacgcttcatgctaaaaactctcaataaattaggtattgatgggacatatctcaaaataataagagctatctatgacaaacccac 
               
               
                   
               
               
                 agccaatatcatactgagtggacaaaaactggaagcattccctttgaaaactggcacaaggcagggatgccctctctcaccactcctat 
               
               
                   
               
               
                 tcaacatagtgttgtaagttctggccagggcaatcaggcaggagaaggaaataaagggcattcaattaggaaaagaggaagtgaaattg 
               
               
                   
               
               
                 tccctgtttgcagatgacatgattgtatatctagaaaaccccattgtctcagcccaaaatctccttaagctgataagcaacttcagcaa 
               
               
                   
               
               
                 agtctcaggatataaaatcagtgtgcaaaaatcacaagtattcctatgcaccaataacagacaaacagagagccaaatcatgagtgaac 
               
               
                   
               
               
                 tcccattcacaattgcttcaaagagaataaaatacctaggaatccaacttacaagggatgtgaaggacctcttcaaggagaactacaaa 
               
               
                   
               
               
                 ccactgctcaatgaaataaaagaggatacaaacaaatggaagaacattccatgctcatgggtaggaagaatcaatatcgtgaaaatggt 
               
               
                   
               
               
                 catactgcccaaggtaatttatagattcaatgccatccccatcaagctaccaatgactttcttcacagaactggaaaaaactactttaa 
               
               
                   
               
               
                 agttcatatggaaccaaaaaagagcccacatcaccaaggcaatcctaagccaaaagaacaaagctggaggcatcacgctacctgacttc 
               
               
                   
               
               
                 aaactatactacaatgctacggtaaccaaaacagcatggtactggtaccaaaacagagatctagaccaatggaacagaacagagccctc 
               
               
                   
               
               
                 agaaataatgccgcatatctacaactatctgatctttgacaaacctgagagaaacaagcaatggggaaaggattccctatttaataaat 
               
               
                   
               
               
                 ggtgctgggaaaactggctagccatatgtagaaagctgaaactggatcccttccttacaccttatacaaaaattaattcaagatggatt 
               
               
                   
               
               
                 aaagacttacatgttagacctaaaaccataaaaaccctagaaaaaaacctaggcaataccattcaggacataggcatgggcaaggactt 
               
               
                   
               
               
                 catgtctaaaacaccaaaagcaatggcaacaaaagacaaaatggacaaacgggatctaattaaactaaagagcttctgcacagctaaag 
               
               
                   
               
               
                 aaactaccatcagagtgaacaggcaacctacaaaatgggagaaaatttttgcaatctactcatctgacaaagggctaatatccagaatc 
               
               
                   
               
               
                 tacaatgaactcaaacaaatttacaagaaaaaacaaacaaccccatcaaaaagtgggcaaaggatatgaacagacacttcgcaaaagaa 
               
               
                   
               
               
                 gacatttatgtaatcaaaaaacacatgaaaaaatgctcatcatcactagccatcagagaaatgcaaatcaaaaccacaatgagatacca 
               
               
                   
               
               
                 tctcacaccagttagaatggcgatcattaaaaagtcaggaaacaacaggtgctggagaggatgtggagaaacaggaacaacttttacac 
               
               
                   
               
               
                 tgttggtgggactgtaaactagttcaaccattgcggaagtcagtgtggcaattcctcaggaatctagaactagaaataccatttgaccc 
               
               
                   
               
               
                 agccatcccattactgggtacatacccaaaggattataaatcatgctgctataaagacacatgcacacgtatgtttattgcagcactat 
               
               
                   
               
               
                 tcacaatagcaaagacttggaaccaacccaaatgtccaacaacgatagactggattaagaaaatgtggcacatatacaccatggaatac 
               
               
                   
               
               
                 tatgcagccataaaaaatgatgagttcatgtcctttgtagggacatggatgaagctggaaactatcattctcagcaaactatcacaagg 
               
               
                   
               
               
                 agaataaaccaaacaccgcatgttctcactcataggtgggaattgaacaatgagaacacatggacacatgaagaggaacatcacactct 
               
               
                   
               
               
                 ggggactgttatggggtggggggcaggggcagggatagcactaggagatatacctaatgctaaatgacgagttaatgggtgcagcacac 
               
               
                   
               
               
                 caacatggcacatgtatacatatataacaaacctgcatgttgtgcacatgtaccctaaaacttgaagtataataataaaaaaaagttat 
               
               
                   
               
               
                 cctattaaaactgatctcacacatccgtagagccattatcaagtctttctctttgaaatagacagaaatttagtgttttctcagtcagt 
               
               
                   
               
               
                 taac 
               
            
           
         
       
     
     Five 5′ hypersensitive site (HS) sites (HS1-HS5) and one 3′ HS site have been identified in the human β-globin LCR (Stamatoyannopoulos et al., (2001)). The 5′ HSs 1-4 are Dnase I hypersensitive sites. The HS2 and HS3 elements are the most powerful single elements within the LCR (Ellis et al., EMBO J. (1996), 15:562-568; Collis et al.,  EMBO J . (1990) 9:233-240), as corroborated by many groups. Deleting HS2 in the context of PYAC in transgenic mice severely affects HS site formation as well as expression of all of the human β-globin genes at every developmental stage (Bungert et al.,  Mol. Cell Biol . (1999); 19:3062-3072). It was reported that deletion of HS2 minimally reduced the expression of the embryonic εy and βhi globin genes in yolk sac-derived erythrocytes (Ley et al., Ann. N.Y. Acad. Sci. (1998); 850:45-53; Hug et al.,  Mol. Cell Biol.  (1996); 26:2906-2912). HS2 functions primarily as an enhancer. 
     In certain embodiments, the β-globin LCR comprises a HS2 region. In non-limiting example, the β-globin LCR comprises a HS2 region, a HS3 region, and a HS4 region. In certain embodiments, the HS2 region, HS3 region and HS4 region within the β-globin LCR are contiguous. 
     In certain embodiments, the β-globin LCR consists essentially of a HS2 region, a HS3 region and a HS4 region. In another embodiment, the β-globin LCR comprises two introduced GATA-1 binding sites at the junction between the HS3 region and the HS4 region. The HS3 region can lie between the HS2 region and the HS4 region. The length and the sequence of the HS2 region can vary. The HS2 region can have a length of from about 400 bp to about 1000 bp, e.g., from about 400 bp to about 500 bp, from about 500 bp to about 600 bp, from about 600 bp to about 700 bp, from about 700 bp to about 800 bp, from about 800 bp to about 850 bp, from about 800 bp to about 820 bp, from about 820 bp to about 830 bp, from about 830 bp to about 840 bp, from about 840 bp to about 850 bp, from about 850 bp to about 900 bp, from about 800 bp to about 900 bp, or from about 900 bp to about 1000 bp. 
     In certain embodiments, the HS2 region has a length of from about 850 bp to about 900 bp. In certain non-limiting embodiments, the HS2 region has a length of about 860 bp. In certain non-limiting embodiments, the HS2 region comprises or has the nucleotide sequence set forth in SEQ ID NO:9, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 9] 
               
               
                 GTATATGTGTATATATATATATATATATTCAGGAAATAATATATTCTAGA 
               
               
                   
               
               
                 ATATGTCACATTCTGTCTCAGGCATCCATTTTCTTTATGATGCCGTTTGA 
               
               
                   
               
               
                 GGTGGAGTTTTAGTCAGGTGGTCAGCTTCTCCttttttttGCCATCTGCC 
               
               
                   
               
               
                 CTGTAAGCATCCTGCTGGGGACCCAGATAGGAGTCATCACTCTAGGCTGA 
               
               
                   
               
               
                 GAACATCTGGGCACACACCCTAAGCCTCAGCATGACTCATCATGACTCAG 
               
               
                   
               
               
                 CATTGCTGTGCTTGAGCCAGAAGGTTTGCTTAGAAGGTTACACAGAACCA 
               
               
                   
               
               
                 GAAGGCGGGGGTGGGGCACTGACCCCGACAGGGGCCTGGCCAGAACTGCT 
               
               
                   
               
               
                 CATGCTTGGACTATGGGAGGTCACTAATGGAGACACACAGAAATGTAACA 
               
               
                   
               
               
                 GGAACTAAGGAAAAACTGAAGCTTATTTAATCAGAGATGAGATGCTGGAA 
               
               
                   
               
               
                 GGGATAGAGGGAGCTGAGCTTGTAAAAAGTATAGTAATCATTCAGCAAAT 
               
               
                   
               
               
                 GGTTTTGAAGCACCTGCTGGATGCTAAACACTATTTTCAGTGCTTGAATC 
               
               
                   
               
               
                 ATAAATAAGAATAAAACATGTATCTTATTCCCCACAAGAGTCCAAGTAAA 
               
               
                   
               
               
                 AAATAACAGTTAATTATAATGTGCTCTGTCCCCCAGGCTGGAGTGCAGTG 
               
               
                   
               
               
                 GCACGATCTCAGCTCACTGCAACCTCCGCCTCCCGGGTTCAAGCAATTCT 
               
               
                   
               
               
                 CCTGCCTCAGCCACCCTAATAGCTGGGATTACAGGTGCACACCACCATGC 
               
               
                   
               
               
                 CAGGCTAATTTTTGTACTTTTTGTAGAGGCAGGGTATCACCATGTTGTCC 
               
               
                   
               
               
                 AAGATGGTCTTGAACTCCTGAGCTCCAAGCAGTCCACCCACCTCAGCCTC 
               
               
                   
               
               
                 CCAAAGTGCT 
               
            
           
         
       
     
     In certain embodiments, the HS2 region has a length of from about 800 bp to about 850 bp. In certain embodiments, the HS2 region has a length of about 840 bp. In certain embodiments, the HS2 region has a length of about 820 bp. In certain embodiments, the HS2 region has a length of about 816 bp. 
     In certain non-limiting embodiments, the HS2 region comprises or has the nucleotide sequence that is a consecutive portion of SEQ ID NO:9. In certain embodiments, the consecutive portion of SEQ ID NO: 9 is at least about 600 bp, or at least about 700 bp, or at least about 800 bp, or at least about 820 bp, and up to about 860 bp in length. Alternatively or additionally, in non-limiting various embodiments, the HS2 region comprises or has nucleotides 1 to 860, 20 to 860, 30 to 860, 40 to 860, 45 to 860, 50 to 860, 100 to 860, or 200 to 860 of SEQ ID NO: 9. In certain embodiments, the HS2 region comprises or has nucleotides 45 to 860 of SEQ ID NO: 9. 
     In certain embodiments, the HS2 region comprises or has a nucleotide sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 99%, or at least about 100% homologous to the nucleotide sequence set forth in SEQ ID NO:9 or a consecutive portion thereof. In certain embodiments, the HS2 region comprises or has a nucleotide sequence that is at least about 99% homologous to the nucleotide sequence set forth in SEQ ID NO:9. In certain embodiments, the HS2 region comprises or has a nucleotide sequence that is about 100% homologous to nucleotides 45 to 860 of SEQ ID NO: 9, e.g., the HS2 region has a nucleotide sequence that includes up to 5, up to 4, up to 3, up to 2, or up to 1 mutations of nucleotides 45 to 860 of SEQ ID NO: 9. In certain embodiments, the HS2 region comprises or has the nucleotide sequence set forth in SEQ ID NO: 33, which is provided below. 
     
       
         
           
               
            
               
                 (SEQ ID NO: 33) 
               
               
                 TCTAGAATATGTCACATTCTGTCTCAGGCATCCATTTTCTTTATGATGCC 
               
               
                   
               
               
                 GTTTGAGGTGGAGTTTTAGTCAGGTGGTCAGCTTCTCCttttttttGCCA 
               
               
                   
               
               
                 TCTGCCCTGTAAGCATCCTGCTGGGGACCCAGATAGGAGTCATCACTCTA 
               
               
                   
               
               
                 GGCTGAGAACATCTGGGCACACACCCTAAGCCTCAGCATGACTCATCATG 
               
               
                   
               
               
                 ACTCAGCATTGCTGTGCTTGAGCCAGAAGGTTTGCTTAGAAGGTTACACA 
               
               
                   
               
               
                 GAACCAGAAGGCGGGGGTGGGGCACTGACCCCGACAGGGGCCTGGCCAGA 
               
               
                   
               
               
                 ACTGCTCATGCTTGGACTATGGGAGGTCACTAATGGAGACACACAGAAAT 
               
               
                   
               
               
                 GTAACAGGAACTAAGGAAAAACTGAAGCTTATTTAATCAGAGATGAGATG 
               
               
                   
               
               
                 CTGGAAGGGATAGAGGGAGCTGAGCTTGTAAAAAGTATAGTAATCATTCA 
               
               
                   
               
               
                 GCAAATGGTTTTGAAGCACCTGCTGGATGCTAAACACTATTTTCAGTGCT 
               
               
                   
               
               
                 TGAATCATAAATAAGAACAAAACATGTATCTTATTCCCCACAAGAGTCCA 
               
               
                   
               
               
                 AGTAAAAAATAACAGTTAATTATAATGTGCTCTGTCCCCCAGGCTGGAGT 
               
               
                   
               
               
                 GCAGTGGCACGATCTCAGCTCACTGCAACCTCCGCCTCCCGGGTTCAAGC 
               
               
                   
               
               
                 AATTCTCCTGCCTCAGCCACCCTAATAGCTGGGATTACAGGTGCACACCA 
               
               
                   
               
               
                 CCATGCCAGGCTAATTTTTGTACTTTTTGTAGAGGCAGGGTATCACCATG 
               
               
                   
               
               
                 TTGTCCAAGATGGTCTTGAACTCCTGAGCTCCAAGCAGTCCACCCACCTC 
               
               
                   
               
               
                 AGCCTCCCAAAGTGCT. 
               
            
           
         
       
     
     In certain embodiments, the HS2 region comprises or has a nucleotide sequence that is a modification of the nucleotide sequence set forth in SEQ ID NO: 9 or a consecutive portion thereof. Non-limiting modifications include deletion, mutations, additions, or combinations thereof. In certain embodiments, the modification comprises one or more mutation, e.g., up to 5, up to 4, up to 3, up to 2, or up to 1 mutations. In certain embodiments, the modification comprises one mutation. In certain embodiments, the one or more mutation is located within a polyadenylation site (or polyadenylation signal motif), e.g., a polyadenylation site on SEQ ID NO: 9. In certain embodiments, the polyadenylation site has the nucleotide sequence AATAAA. In certain embodiments, the one or more mutation comprises a mismatch mutation T&gt;C at a polyadenylation site having the nucleotide sequence AATAAA, e.g., the mutated sequence is AACAAA. In certain embodiments, the polyadenylation site has the nucleotide sequence ATTAAA. In certain embodiments, the one or more mutation comprises a mismatch mutation T&gt;C at a polyadenylation site having the nucleotide sequence ATTAAA, e.g., the mutated sequence is ATCAAA or ACTAAA. In certain embodiments, the one or more mutation comprises two mismatch mutations T&gt;C at a polyadenylation site having the nucleotide sequence ATTAAA, e.g., the mutated sequence is ACCAAA. The inventors discovered that a modification (e.g., one or more mismatch mutation) at a polyadenylation site can preempt premature termination of RNA transcription, thereby increasing the titer of the expression cassette (e.g., expression vector) without compromising the expression of the globin gene. 
     In certain embodiments, the HS2 region has a length of about 650 bp (e.g., 646 bp). In certain embodiments, the HS2 region has a length of about 420 bp (e.g., 423 bp). 
     The length and the sequence of the HS3 region can vary. The HS3 region can have a length of from about 200 bp to about 1400 bp, e.g., from about 200 bp to about 300 bp, from about 300 bp to about 400 bp, from about 400 bp to about 500 bp, from about 500 bp to about 600 bp, from about 600 bp to about 700 bp, from about 700 bp to about 800 bp, from about 800 bp to about 900 bp, from about 900 bp to about 1000 bp, from about 1000 bp to about 1100 bp, from about 1100 bp to about 1200 bp, from about 1200 bp to about 1300 bp, or from about 1300 bp to about 1400 bp. In certain embodiments, the HS3 region has a length of about 1300 bp. In certain non-limiting embodiments, the HS3 region has a length of 1308 bp. In certain non-limiting embodiments, the HS3 region has a length of 1301 bp. In certain non-limiting embodiments, the HS3 region comprises or has the nucleotide sequence set forth in SEQ ID NO:5, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 5] 
               
               
                 AAGCTTTCATTAAAAAAAGTCTAACCAGCTGCATTCGACTTTGACTGCAG 
               
               
                   
               
               
                 CAGCTGGTTAGAAGGTTCTACTGGAGGAGGGTCCCAGCCCATTGCTAAAT 
               
               
                   
               
               
                 TAACATCAGGCTCTGAGACTGGCAGTATATCTCTAACAGTGGTTGATGCT 
               
               
                   
               
               
                 ATCTTCTGGAACTTGCCTGCTACATTGAGACCACTGACCCATACATAGGA 
               
               
                   
               
               
                 AGCCCATAGCTCTGTCCTGAACTGTTAGGCCACTGGTCCAGAGAGTGTGC 
               
               
                   
               
               
                 ATCTCCTTTGATCCTCATAATAACCCTATGAGATAGACACAATTATTACT 
               
               
                   
               
               
                 CTTACTTTATAGATGATGATCCTGAAAACATAGGAGTCAAGGCACTTGCC 
               
               
                   
               
               
                 CCTAGCTGGGGGTATAGGGGAGCAGTCCCATGTAGTAGTAGAATGAAAAA 
               
               
                   
               
               
                 TGCTGCTATGCTGTGCCTCCCCCACCTTTCCCATGTCTGCCCTCTACTCA 
               
               
                   
               
               
                 TGGTCTATCTCTCCTGGCTCCTGGGAGTCATGGACTCCACCCAGCACCAC 
               
               
                   
               
               
                 CAACCTGACCTAACCACCTATCTGAGCCTGCCAGCCTATAACCCATCTGG 
               
               
                   
               
               
                 GCCCTGATAGCTGGTGGCCAGCCCTGACCCCACCCCACCCTCCCTGGAAC 
               
               
                   
               
               
                 CTCTGATAGACACATCTGGCACACCAGCTCGCAAAGTCACCGTGAGGGTC 
               
               
                   
               
               
                 TTGTGTTTGCTGAGTCAAAATTCCTTGAAATCCAAGTCCTTAGAGACTCC 
               
               
                   
               
               
                 TGCTCCCAAATTTACAGTCATAGACTTCTTCATGGCTGTCTCCTTTATCC 
               
               
                   
               
               
                 ACAGAATGATTCCTTTGCTTCATTGCCCCATCCATCTGATCCTCCTCATC 
               
               
                   
               
               
                 AGTGCAGCACAGGGCCCATGAGCAGTAGCTGCAGAGTCTCACATAGGTCT 
               
               
                   
               
               
                 GGCACTGCCTCTGACATGTCCGACCTTAGGCAAATGCTTGACTCTTCTGA 
               
               
                   
               
               
                 GCTCAGTCTTGTCATGGCAAAATAAAGATAATAATAGTGTTTTTTTATGG 
               
               
                   
               
               
                 AGTTAGCGTGAGGATGGAAAACAATAGCAAAATTGATTAGACTATAAAAG 
               
               
                   
               
               
                 GTCTCAACAAATAGTAGTAGATTTTATCATCCATTAATCCTTCCCTCTCC 
               
               
                   
               
               
                 TCTCTTACTCATCCCATCACGTATGCCTCTTAATTTTCCCTTACCTATAA 
               
               
                   
               
               
                 TAAGAGTTATTCCTCTTATTATATTCTTCTTATAGTGATTCTGGATATTA 
               
               
                   
               
               
                 AAGTGGGAATGAGGGGCAGGCCACTAACGAAGAAGATGTTTCTCAAAGAA 
               
               
                   
               
               
                 GCCATTCTCCCCACATAGATCATCTCAGCAGGGTTCAGGAAGATAAAGGA 
               
               
                   
               
               
                 GGATCAAGGTCGAAGGTAGGAACTAAGGAAGAACACTGGGCAAGTGGATC 
               
               
                   
               
               
                 C 
               
            
           
         
       
     
     In certain embodiments, the HS2 region comprises or has the nucleotide sequence that is a consecutive portion of SEQ ID NO: 5. In certain embodiments, the consecutive portion of SEQ ID NO: 5 is at least about 600 bp, or at least about 700 bp, or at least about 800 bp, at least about 900 bp, at least about 1000 bp, at least about 1100 bp, or at least about 1200 bp, and up to about 1300 bp in length. 
     In certain embodiments, the HS3 region comprises or has a nucleotide sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 99%, or at least about 100% homologous to the nucleotide sequence set forth in SEQ ID NO: 5 or a consecutive portion thereof. In certain embodiments, the HS3 region comprises or has a nucleotide sequence that is at least about 99% homologous to the nucleotide sequence set forth in SEQ ID NO: 5. In certain embodiments, the HS3 region comprises or has a nucleotide sequence that is about 100% homologous to the nucleotide sequence set forth in SEQ ID NO: 5, e.g., the HS3 region has a nucleotide sequence that includes up to 5, up to 4, up to 3, up to 2, or up to 1 mutations of SEQ ID NO: 5. In certain embodiments, the HS3 region comprises or has the nucleotide sequence set forth in SEQ ID NO: 34, which is provided below. 
     
       
         
           
               
            
               
                 (SEQ ID NO: 34) 
               
               
                 AAGCTTTCATCaaaaaaaGTCTAACCAGCTGCATTCGACTTTGACTGCAG 
               
               
                   
               
               
                 CAGCTGGTTAGAAGGTTCTACTGGAGGAGGGTCCCAGCCCATTGCTAAAT 
               
               
                   
               
               
                 TAACATCAGGCTCTGAGACTGGCAGTATATCTCTAACAGTGGTTGATGCT 
               
               
                   
               
               
                 ATCTTCTGGAACTTGCCTGCTACATTGAGACCACTGACCCATACATAGGA 
               
               
                   
               
               
                 AGCCCATAGCTCTGTCCTGAACTGTTAGGCCACTGGTCCAGAGAGTGTGC 
               
               
                   
               
               
                 ATCTCCTTTGATCCTCATAATAACCCTATGAGATAGACACAATTATTACT 
               
               
                   
               
               
                 CTTACTTTATAGATGATGATCCTGAAAACATAGGAGTCAAGGCACTTGCC 
               
               
                   
               
               
                 CCTAGCTGGGGGTATAGGGGAGCAGTCCCATGTAGTAGTAGAATGAAAAA 
               
               
                   
               
               
                 TGCTGCTATGCTGTGCCTCCCCCACCTTTCCCATGTCTGCCCTCTACTCA 
               
               
                   
               
               
                 TGGTCTATCTCTCCTGGCTCCTGGGAGTCATGGACTCCACCCAGCACCAC 
               
               
                   
               
               
                 CAACCTGACCTAACCACCTATCTGAGCCTGCCAGCCTATAACCCATCTGG 
               
               
                   
               
               
                 GCCCTGATAGCTGGTGGCCAGCCCTGACCCCACCCCACCCTCCCTGGAAC 
               
               
                   
               
               
                 CTCTGATAGACACATCTGGCACACCAGCTCGCAAAGTCACCGTGAGGGTC 
               
               
                   
               
               
                 TTGTGTTTGCTGAGTCAAAATTCCTTGAAATCCAAGTCCTTAGAGACTCC 
               
               
                   
               
               
                 TGCTCCCAAATTTACAGTCATAGACTTCTTCATGGCTGTCTCCTTTATCC 
               
               
                   
               
               
                 ACAGAATGATTCCTTTGCTTCATTGCCCCATCCATCTGATCCTCCTCATC 
               
               
                   
               
               
                 AGTGCAGCACAGGGCCCATGAGCAGTAGCTGCAGAGTCTCACATAGGTCT 
               
               
                   
               
               
                 GGCACTGCCTCTGACATGTCCGACCTTAGGCAAATGCTTGACTCTTCTGA 
               
               
                   
               
               
                 GCTCAGTCTTGTCATGGCAAAACAAAGATAATAATAGTGtttttttATGG 
               
               
                   
               
               
                 AGTTAGCGTGAGGATGGAAAACAATAGCAAAATTGATTAGACTATAAAAG 
               
               
                   
               
               
                 GTCTCAACAAATAGTAGTAGATTTTATCATCCATTAATCCTTCCCTCTCC 
               
               
                   
               
               
                 TCTCTTACTCATCCCATCACGTATGCCTCTTAATTTTCCCTTACCTATAA 
               
               
                   
               
               
                 TAAGAGTTATTCCTCTTATTATATTCTTCTTATAGTGATTCTGGATATCA 
               
               
                   
               
               
                 AAGTGGGAATGAGGGGCAGGCCACTAACGAAGAAGATGTTTCTCAAAGAA 
               
               
                   
               
               
                 GCCATTCTCCCCACATAGATCATCTCAGCAGGGTTCAGGAAGATAAAGGA 
               
               
                   
               
               
                 GGATCAAGGTCGAAGGTAGGAACTAAGGAAGAACACTGGGCAAGTGGATC 
               
               
                   
               
               
                 C. 
               
            
           
         
       
     
     In certain embodiments, the HS3 region comprises or has a nucleotide sequence that is a modification of the nucleotide sequence set forth in SEQ ID NO: 5 or a consecutive portion thereof. Non-limiting modifications include deletion, mutations, additions, or combinations thereof. In certain embodiments, the modification comprises one or more mutation, e.g., up to 5, up to 4, up to 3, up to 2, or up to 1 mutations. In certain embodiments, the modification comprises three mutations. In certain embodiments, the one or more mutation is located within a polyadenylation site (or polyadenylation signal motif), e.g., a polyadenylation site on SEQ ID NO: 5. In certain embodiments, the polyadenylation site has the nucleotide sequence AATAAA. In certain embodiments, the one or more mutation comprises a mismatch mutation T&gt;C at a polyadenylation site having the nucleotide sequence AATAAA, e.g., the mutated sequence is AACAAA. In certain embodiments, the polyadenylation site has the nucleotide sequence ATTAAA. In certain embodiments, the one or more mutation comprises a mismatch mutation T&gt;C at a polyadenylation site having the nucleotide sequence ATTAAA, e.g., the mutated sequence is ATCAAA or ACTAAA. In certain embodiments, the one or more mutation comprises two mismatch mutations T&gt;C at a polyadenylation site having the nucleotide sequence ATTAAA, e.g., the mutated sequence is ACCAAA. 
     A modification (e.g., one or more mismatch mutation) at a polyadenylation site can preempt premature termination of RNA transcription, thereby increasing the titer of the expression cassette (e.g., expression vector) without compromising the expression of the globin gene. 
     In certain embodiments, the HS3 region has a length of about 850 bp (e.g., 845 bp). In certain embodiments, the HS3 region has a length of from about 280 bp to about 290 bp (e.g., 280 bp and 287 bp). 
     Similarly, the length and the sequence of the HS4 region can vary. The HS4 region can have a length of from about 200 bp to about 1200 bp, e.g., from about 200 bp to about 300 bp, from about 300 bp to about 400 bp, from about 400 bp to about 500 bp, from about 500 bp to about 600 bp, from about 600 bp to about 700 bp, from about 700 bp to about 800 bp, from about 800 bp to about 900 bp, from about 900 bp to about 1000 bp, from about 1000 bp to about 1100 bp, or from about 1100 bp to about 1200 bp. 
     In certain embodiments, the HS4 region has a length of about 1.0 kb or more. In certain embodiments, the HS4 region has a length of about 1.1 kb. In certain embodiments, the HS4 region has a length of about 1150 bp (e.g., 1153 bp). In certain non-limiting embodiments, the HS4 region has a length of 1100 bp. In certain non-limiting embodiments, the HS4 region has a length of 1065 bp. In certain non-limiting embodiments, the HS4 region has the nucleotide sequence set forth in SEQ ID NO:6, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 6] 
               
               
                 TGAGCCCCTTTTCCTCTAACTGAAAGAAGGAAAAAAAAAATGGAACCCAA 
               
               
                   
               
               
                 AATATTCTACATAGTTTCCATGTCACAGCCAGGGCTGGGCAGTCTCCTGT 
               
               
                   
               
               
                 TATTTCTTTTAAAATAAATATATCATTTAAATGCATAAATAAGCAAACCC 
               
               
                   
               
               
                 TGCTCGGGAATGGGAGGGAGAGTCTCTGGAGTCCACCCCTTCTCGGCCCT 
               
               
                   
               
               
                 GGCTCTGCAGATAGTGCTATCAAAGCCCTGACAGAGCCCTGCCCATTGCT 
               
               
                   
               
               
                 GGGCCTTGGAGTGAGTCAGCCTAGTAGAGAGGCAGGGCAAGCCATCTCAT 
               
               
                   
               
               
                 AGCTGCTGAGTGGGAGAGAGAAAAGGGCTCATTGTCTATAAACTCAGGTC 
               
               
                   
               
               
                 ATGGCTATTCTTATTCTCACACTAAGAAAAAGAATGAGATGTCTACATAT 
               
               
                   
               
               
                 ACCCTGCGTCCCCTCTTGTGTACTGGGGCCCCCAAGAGCTCTCTAAAAGT 
               
               
                   
               
               
                 GATGGCAAAGTCATTGCGCTAGATGCCATCCCATCTATTATAAACCTGCA 
               
               
                   
               
               
                 TTTGTCTCCACACACCAGTCATGGACAATAACCCTCCTCCCAGGTCCACG 
               
               
                   
               
               
                 TGCTTGTCTTTGTATAATACTCAAGTAATTTCGGAAAATGTATTCTTTCA 
               
               
                   
               
               
                 ATCTTGTTCTGTTATTCCTGTTTCAATGGCTTAGTAGAAAAAGTACATAC 
               
               
                   
               
               
                 TTGTTTTCCCATAAATTGACAATAGACAATTTCACATCAATGTCTATATG 
               
               
                   
               
               
                 GGTCGTTGTGTTTGCTGTGTTTGCAAAAACTCACAATAACTTTATATTGT 
               
               
                   
               
               
                 TACTACTCTAAGAAAGTTACAACATGGTGAATACAAGAGAAAGCTATTAC 
               
               
                   
               
               
                 AAGTCCAGAAAATAAAAGTTATCATCTTGAGGCCTCAGCTTTCTAGGAAT 
               
               
                   
               
               
                 AATATCAATATTACAAAATTTAATCTAACAATTATGAACAGCAATGAGAT 
               
               
                   
               
               
                 AATATGTACAAAGTACCCAGACCTATGTGGTAGAGCATCAAGGAAGCGCA 
               
               
                   
               
               
                 TTGCGGAGCAGTTTTTTGTTTGTTTGTTTTTGTATTCTGTTTCGTGAGGC 
               
               
                   
               
               
                 AAGGTTTCACTCTGCTGTCCAGGCTGGAGTGCAGTGGCAAGATCATGTCT 
               
               
                   
               
               
                 CACTGCAGCCTTGAC 
               
            
           
         
       
     
     In certain non-limiting embodiments, the HS4 region has the nucleotide sequence set forth in SEQ ID NO:7, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 7] 
               
               
                 TGAGCCCCTTTTCCTCTAACTGAAAGAAGGAAAAAAAAAATGGAACCCAA 
               
               
                   
               
               
                 AATATTCTACATAGTTTCCATGTCACAGCCAGGGCTGGGCAGTCTCCTGT 
               
               
                   
               
               
                 TATTTCTTTTAAAATAAATATATCATTAAATGCATAAATAAGCAAACCCT 
               
               
                   
               
               
                 GCTCGGGAATGGGAGGGAGAGTCTCTGGAGTCCACCCCTTCTCGGCCCTG 
               
               
                   
               
               
                 GCTCTGCAGATAGTGCTATCAAAGCCCTGACAGAGCCCTGCCCATTGCTG 
               
               
                   
               
               
                 GGCCTTGGAGTGAGTCAGCCTAGTAGAGAGGCAGGGCAAGCCATCTCATA 
               
               
                   
               
               
                 GCTGCTGAGTGGGAGAGAGAAAAGGGCTCATTGTCTATAAACTCAGGTCA 
               
               
                   
               
               
                 TGGCTATTCTTATTCTCACACTAAGAAAAAGAATGAGATGTCTACATATA 
               
               
                   
               
               
                 CCCTGCGTCCCCTCTTGTGTACTGGGGCCCCCAAGAGCTCTCTAAAAGTG 
               
               
                   
               
               
                 ATGGCAAAGTCATTGCGCTAGATGCCATCCCATCTATTATAAACCTGCAT 
               
               
                   
               
               
                 TTGTCTCCACACACCAGTCATGGACAATAACCCTCCTCCCAGGTCCACGT 
               
               
                   
               
               
                 GCTTGTCTTTGTATAATACTCAAGTAATTTCGGAAAATGTATTCTTTCAA 
               
               
                   
               
               
                 TCTTGTTCTGTTATTCCTGTTTCAATGGCTTAGTAGAAAAAGTACATACT 
               
               
                   
               
               
                 TGTTTTCCCATAAATTGACAATAGACAATTTCACATCAATGTCTATATGG 
               
               
                   
               
               
                 GTCGTTGTGTTTGCTGTGTTTGCAAAAACTCACAATAACTTTATATTGTT 
               
               
                   
               
               
                 ACTACTCTAAGAAAGTTACAACATGGTGAATACAAGAGAAAGCTATTACA 
               
               
                   
               
               
                 AGTCCAGAAAATAAAAGTTATCATCTTGAGGCCTCAGCTTTCTAGGaATA 
               
               
                   
               
               
                 ATATCAATATTACAAAATTAATCTAACAATTATGAACAGCAATGAGATAA 
               
               
                   
               
               
                 TATGTACAAAGTACCCAGACCTATGTGGTAGAGCATCAAGGAAGCGCATT 
               
               
                   
               
               
                 GCGGAGCAGTTTTTTGTTTGTTTGTTTTTGTATTCTGTTTCGTGAGGCAA 
               
               
                   
               
               
                 GGTTTCACTCTGCTGTCCAGGCTGGAGTGCAGTGGCAAGATCATGTCTCA 
               
               
                   
               
               
                 CTGCAGCCTTGACAC 
               
            
           
         
       
     
     In certain embodiments, the HS4 region has a length of less than about 1.0 kb, less than about 900 bp, or less than about 800 bp. In certain embodiments, the HS4 region has a length of from about 700 bp to about 800 bp, e.g., from about 700 bp to about 750 bp, or from about 750 bp to about 800 bp. In certain embodiments, the HS4 region has a length of about 750 bp. In certain embodiments, the HS4 region has a length of about 754 bp. 
     In certain embodiments, the HS4 region comprises or has the nucleotide sequence that is a consecutive portion of SEQ ID NO: 6 or 7. In certain embodiments, the consecutive portion of SEQ ID NO: 6 or 7 is at least about 600 bp or at least about 700 bp and up to about 800 bp or up to about 900 bp, or up to about 1.0 kb in length. Alternatively or additionally, in non-limiting various embodiments, the HS4 region comprises or has nucleotides 1 to 900, 100 to 900, 100 to 1000, 110 to 900, 115 to 900, 115 to 890, 115 to 880, 115 to 870, 115 to 868, 115 to 860, 115 to 1000 of SEQ ID NO: 6 or 7. In certain embodiments, the HS4 region comprises or has nucleotides 115 to 868 of SEQ ID NO: 6. 
     In certain embodiments, the HS4 region comprises or consists essentially of a core sequence of the HS4 region. In certain embodiments, the HS4 region or consists essentially of the full length of a 280 bp core sequence of a human HS4 region (e.g., the 280 bp core sequence of human HS4 disclosed in Pruzina et al.,  Nucleic Acids Research  (1991); 19:7:1413-1419, which is incorporated by reference in its entirety). 
     In certain embodiments, the HS4 region does not comprise the nucleotide sequence set forth in SEQ ID NO: 50, which is provided below. 
     
       
         
           
               
            
               
                 [SEQ ID NO: 50] 
               
               
                 TGAGCCCCTTTTCCTCTAACTGAAAGAAGGAAAAAAAAAATGGAACC 
               
               
                   
               
               
                 CAAAATATTCTACATAGTTTCCATGTCACAGCCAGGGCTGGGCAGTC 
               
               
                   
               
               
                 TCCTGTTATTTCTTTTAAAA 
               
            
           
         
       
     
     In certain embodiments, the HS4 region does not comprise the nucleotide sequence set forth in SEQ ID NO: 51, which is provided below. 
     
       
         
           
               
            
               
                 [SEQ ID NO: 51] 
               
               
                 TTTAATCTAACAATTATGAACAGCAATGAGATAATATGTACAAAGTA 
               
               
                   
               
               
                 CCCAGACCTATGTGGTAGAGCATCAAGGAAGCGCATTGCGGAGCAGT 
               
               
                   
               
               
                 TTTTTGTTTGTTTGTTTTTGTATTCTGTTTCGTGAGGCAAGGTTTCA 
               
               
                   
               
               
                 CTCTGCTGTCCAGGCTGGAGTGCAGTGGCAAGATCATGTCTCACTGC 
               
               
                   
               
               
                 AGCCTTGAC 
               
            
           
         
       
     
     In certain embodiments, the HS4 region does not comprise the nucleotide sequence set forth in SEQ ID NO: 50 or the nucleotide sequence set forth in SEQ ID NO: 51. 
     In certain embodiments, the HS4 region comprises or has a nucleotide sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 99%, or at least about 100% homologous to the nucleotide sequence set forth in SEQ ID NO: 6 or a consecutive portion thereof. In certain embodiments, the HS4 region comprises or has a nucleotide sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 99%, or at least about 100% homologous to the nucleotide sequence set forth in SEQ ID NO: 7 or a consecutive portion thereof. In certain embodiments, the HS4 region comprises or has a nucleotide sequence that is at least about 99% homologous to the nucleotide sequence set forth in SEQ ID NO: 6 or 7. In certain embodiments, the HS4 region comprises or has a nucleotide sequence that is at least about 99% (e.g., about 100%) homologous to nucleotides 115 to 868 of SEQ ID NO: 6, e.g., the HS4 region has a nucleotide sequence that includes up to 5, up to 4, up to 3, up to 2, or up to 1 mutations of nucleotides 115 to 868 of SEQ ID NO: 6. In certain embodiments, the HS4 region comprises or has the nucleotide sequence set forth in SEQ ID NO: 35, which is provided below. 
     
       
         
           
               
            
               
                 [SEQ ID NO: 35] 
               
               
                 TAAATATATCATTTAAATGCATAAATAAGCAAACCCTGCTCGGGAAT 
               
               
                   
               
               
                 GGGAGGGAGAGTCTCTGGAGTCCACCCCTTCTCGGCCCTGGCTCTGC 
               
               
                   
               
               
                 AGATAGTGCTATCAAAGCCCTGACAGAGCCCTGCCCATTGCTGGGCC 
               
               
                   
               
               
                 TTGGAGTGAGTCAGCCTAGTAGAGAGGCAGGGCAAGCCATCTCATAG 
               
               
                   
               
               
                 CTGCTGAGTGGGAGAGAGAAAAGGGCTCATTGTCTATAAACTCAGGT 
               
               
                   
               
               
                 CATGGCTATTCTTATTCTCACACTAAGAAAAAGAATGAGATGTCTAC 
               
               
                   
               
               
                 ATATACCCTGCGTCCCCTCTTGTGTACTGGGGCCCCCAAGAGCTCTC 
               
               
                   
               
               
                 TAAAAGTGATGGCAAAGTCATTGCGCTAGATGCCATCCCATCTATTA 
               
               
                   
               
               
                 TAAACCTGCATTTGTCTCCACACACCAGTCATGGACAATAACCCTCC 
               
               
                   
               
               
                 TCCCAGGTCCACGTGCTTGTCTTTGTATAATACTCAAGTAATTTCGG 
               
               
                   
               
               
                 AAAATGTATTCTTTCAATCTTGTTCTGTTATTCCTGTTTCAATGGCT 
               
               
                   
               
               
                 TAGTAGAAAAAGTACATACTTGTTTTCCCATAAATTGACAATAGACA 
               
               
                   
               
               
                 ATTTCACATCAATGTCTATATGGGTCGTTGTGTTTGCTGTGTTTGCA 
               
               
                   
               
               
                 AAAACTCACAATAACTTTATATTGTTACTACTCTAAGAAAGTTACAA 
               
               
                   
               
               
                 CATGGTGAATACAAGAGAAAGCTATTACAAGTCCAGAAAACAAAAGT 
               
               
                   
               
               
                 TATCATCTTGAGGCCTCAGCTTTCTAGGAATAATATCAATATTACAA 
               
               
                   
               
               
                 AA. 
               
            
           
         
       
     
     In certain embodiments, the HS4 region comprises or has a nucleotide sequence that is a modification of the nucleotide sequence set forth in SEQ ID NO: 6 or a consecutive portion thereof. In certain embodiments, the HS4 region comprises or has a nucleotide sequence that is a modification of the nucleotide sequence set forth in SEQ ID NO: 7 or a consecutive portion thereof. Non-limiting modifications include deletion, mutations, additions, or combinations thereof. In certain embodiments, the modification comprises one or more mutation, e.g., up to 5, up to 4, up to 3, up to 2, or up to 1 mutations. In certain embodiments, the modification comprises one mutation. In certain embodiments, the one or more mutation is located within a polyadenylation site (or polyadenylation signal motif), e.g., a polyadenylation site on SEQ ID NO: 6 or a polyadenylation site on SEQ ID NO: 7. In certain embodiments, the polyadenylation site has the nucleotide sequence AATAAA. In certain embodiments, the one or more mutation comprises a mismatch mutation T&gt;C at a polyadenylation site having the nucleotide sequence AATAAA, e.g., the mutated sequence is AACAAA. In certain embodiments, the polyadenylation site has the nucleotide sequence ATTAAA. In certain embodiments, the one or more mutation comprises a mismatch mutation T&gt;C at a polyadenylation site having the nucleotide sequence ATTAAA, e.g., the mutated sequence is ATCAAA or ACTAAA. In certain embodiments, the one or more mutation comprises two mismatch mutations T&gt;C at a polyadenylation site having the nucleotide sequence ATTAAA, e.g., the mutated sequence is ACCAAA. A modification (e.g., one or more mismatch mutation) at a polyadenylation site can preempt premature termination of RNA transcription, thereby increasing the titer of the expression cassette (e.g., expression vector) without compromising the expression of the globin gene. 
     In certain embodiments, the HS4 region has a length of less than about 500 bp. In certain embodiments, the HS4 region has a length of about 450 bp. In certain non-limiting embodiments, the HS4 region has a length of about 446 bp. In certain non-limiting embodiments, the HS4 region comprises or has the nucleotide sequence set forth in SEQ ID NO:8, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 8] 
               
               
                 TGGAACCCAAAATATTCTACATAGTTTCCATGTCACAGCCAGGGCTG 
               
               
                   
               
               
                 GGCAGTCTCCTGTTATTTCTTTTAAAATAAATATATCATTTAAATGC 
               
               
                   
               
               
                 ATAAATAAGCAAACCCTGCTCGGGAATGGGAGGGAGAGTCTCTGGAG 
               
               
                   
               
               
                 TCCACCCCTTCTCGGCCCTGGCTCTGCAGATAGTGCTATCAAAGCCC 
               
               
                   
               
               
                 TGACAGAGCCCTGCCCATTGCTGGGCCTTGGAGTGAGTCAGCCTAGT 
               
               
                   
               
               
                 AGAGAGGCAGGGCAAGCCATCTCATAGCTGCTGAGTGGGAGAGAGAA 
               
               
                   
               
               
                 AAGGGCTCATTGTCTATAAACTCAGGTCATGGCTATTCTTATTCTCA 
               
               
                   
               
               
                 CACTAAGAAAAAGAATGAGATGTCTACATATACCCTGCGTCCCCTCT 
               
               
                   
               
               
                 TGTGTACTGGGGTCCCCAAGAGCTCTCTAAAAGTGATGGCAAAGTCA 
               
               
                   
               
               
                 TTGCGCTAGATGCCATCCCATCT 
               
            
           
         
       
     
     In certain embodiments, the HS4 region has a length of about 280 bp (e.g., 283 bp). In certain embodiments, the HS4 region has a length of about 240 bp (e.g., 243 bp). 
     In certain non-limiting embodiments, the β-globin LCR comprises or consists essentially of a HS2 region having the nucleotide sequence set forth in SEQ ID NO:9 or a consecutive portion thereof, a modification of SEQ ID NO: 9, a modification of a consecutive portion of SEQ ID NO: 9 (e.g., SEQ ID NO: 33), SEQ ID NO:20, SEQ ID NO:21; a HS3 region having the nucleotide sequence set forth in SEQ ID NO:5 or a modification of SEQ ID NO: 5 (e.g., SEQ ID NO: 34); and a HS4 region having the nucleotide sequence set forth in SEQ ID NO:6 or a consecutive portion thereof, SEQ ID NO:7 or a consecutive portion thereof, a modification of SEQ ID NO: 6, a modification of a consecutive portion of SEQ ID NO: 6 (e.g., SEQ ID NO: 35), a modification of SEQ ID NO: 7, a modification of a consecutive portion of SEQ ID NO: 7, or SEQ ID NO:8, and the β-globin LCR does not comprise a HS1 region. 
     In certain non-limiting embodiments, the β-globin LCR comprises or consists essentially of a HS2 region having the nucleotide sequence set forth in SEQ ID NO:9, a HS3 region having the nucleotide sequence set forth in SEQ ID NO:5, and a HS4 region having the nucleotide sequence set forth in SEQ ID NO:7, as shown in  FIG. 1 . 
     In certain non-limiting embodiments, the β-globin LCR comprises or consists essentially of a HS2 region having nucleotides 45 to 860 of SEQ ID NO: 9; a HS3 region having the nucleotide sequence set forth in SEQ ID NO: 5; and a HS4 region having nucleotides 115 to 868 of SEQ ID NO: 6, and the β-globin LCR does not comprise a HS1 region, e.g., the β-globin LCR of SNS23.B87.A1 shown in  FIG. 15 . 
     In certain non-limiting embodiments, the β-globin LCR comprises or consists essentially of a HS2 region having the nucleotide sequence set forth in SEQ ID NO: 33; a HS3 region having the nucleotide sequence set forth in SEQ ID NO: 34; and a HS4 region having the nucleotide sequence set forth in SEQ ID NO: 35, and the β-globin LCR does not comprise a HS1 region, e.g., the β-globin LCR of SNS23.2.B87.A1 shown in  FIGS. 13 and 14 . 
     In certain non-limiting embodiments, the β-globin LCR further comprises a HS1 region, i.e., a β-globin LCR comprising a HS1 region, a HS2 region, a HS3 region, and a HS4 region. In certain embodiments, the HS1 region, HS2 region, HS3 region and HS4 region within the β-globin LCR are contiguous. In certain non-limiting embodiments, the β-globin LCR consisting essentially of a HS1 region, a HS2 region, a HS3 region and a HS4 region. In certain embodiments, the β-globin LCR comprises two introduced GATA-1 binding sites at the junction between the HS3 region and the HS4 region. 
     The length and the sequence of the HS1 region can vary. In certain embodiments, the HS1 region is from about 300 bp to about 1500 bp in length, e.g., from about 300 bp to about 1100 bp in length. In certain embodiments, the HS1 region has a length of about 1.0 kb or more, e.g., about 1.1 kb, about 1.2 kb, about 1.3 kb, about 1.4 kb, or about 1.5 kb. In certain embodiments, the HS1 region has a length of about 1.1 kb. In certain non-limiting embodiments, the HS1 region has a length of 1074 bp. In certain non-limiting embodiments, the HS1 region comprises or has the nucleotide sequence set forth in SEQ ID NO:2, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 2] 
               
               
                 AAGTAAACTTCCACAACCGCAAGCTTATTGAGGCTAAGGCATCTGTG 
               
               
                   
               
               
                 AAGGAAAGAAACATCTCCTCTAAACCACTATGCTGCTAGAGCCTCTT 
               
               
                   
               
               
                 TTCTGTACTCAAGCCTCATTCAGACACTAGTGTCACCAGTCTCCTCA 
               
               
                   
               
               
                 TATACCTATTGTATTTTCTTCTTCTTGCTGGTTTAGTCATGTTTTCT 
               
               
                   
               
               
                 GGGAGCTTAGGGGCTTATTTTATTTTGTTTTGTTTTCTAATCAACAG 
               
               
                   
               
               
                 AGATGGGCAAACCCATTATTTTTTTCTTTAGACTTGGGATGGTGATA 
               
               
                   
               
               
                 GCTGGGCAGCGTCAGAAACTGTGTGTGGATATAGATAAGAGCTCGGA 
               
               
                   
               
               
                 CTATGCTGAGCTGTGATGAGGGAGGGACCTAGCCAAAGGCAGTGAGA 
               
               
                   
               
               
                 GTCAGAATGCTCCTGCTATTGCCTTCTCAGTCCCCACGCTTGGTTTC 
               
               
                   
               
               
                 TACACAAGTAGATACATAGAAAAGGCTATAGGTTAGTGTTTGAGAGT 
               
               
                   
               
               
                 CCTGCATGAGTTAGTTGCTCAGAAATGCCCGATAAATATGTTATGTG 
               
               
                   
               
               
                 TGTTTATGTATATATATGTTTTATATATATATATGTGTGTGTGTGTG 
               
               
                   
               
               
                 TGTGTGTGTGTTGTGTTTACAAATATGTGATTATCATCAAAACGTGA 
               
               
                   
               
               
                 GGGCTAAAGTGACCAGATAACTTGCAGGTCCTAGGATACCAGGAAAA 
               
               
                   
               
               
                 TAAATTACATTCCAAAAATTTAACTGAGACTTTAAAAAAAAAAAAAA 
               
               
                   
               
               
                 AAAAAAAAAAAAAACCAGTGATCCATGGACACAGGGAGGGGAACATC 
               
               
                   
               
               
                 ACACACTGGGGCCTGTTGGGGGTGGGGGGCTAGGGGAAGGATAGCAT 
               
               
                   
               
               
                 TAGGAGAAATACCTAATGTAGATGACGGGTTGATGGGTGCAGCAAAC 
               
               
                   
               
               
                 CACCATGGCACATGTACCCCAGAACTTAAAGCATATTAAAAAAACAG 
               
               
                   
               
               
                 TGATCATAAAAGAAGCTCAAATTTAACTATAAGAGACGGAATGGCTC 
               
               
                   
               
               
                 CCACAATTCTTAACTATAATCTTACAGAATATTCTCATTGAATAGAA 
               
               
                   
               
               
                 GTATGCTTATCATTAGAGATTTGGACAGCCAGGAAAGCACAGAAAAA 
               
               
                   
               
               
                 AAAAAAAGGAGCTCTGTTGCCTTATAGCCTAGAGGTGTTT 
               
            
           
         
       
     
     In certain embodiments, the HS1 region has a length of less than about 1.0 kb, e.g., from about 400 bp to about 700 bp, from about 400 bp to about 500 bp, from about 500 bp to about 600 bp, from about 600 bp to about 700 bp, from about 700 bp to about 800 bp, from about 800 bp to about 900 bp, or from about 900 bp to about 1.0 kb. In certain embodiments, the HS1 region has a length of less than about 700 bp. In certain embodiments, the HS1 region has a length of about 600 bp. In certain non-limiting embodiments, the HS1 region has a length of 602 bp. In certain non-limiting embodiments, the HS1 region comprises or has the nucleotide sequence set forth in SEQ ID NO:3, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 3] 
               
               
                 GGCATCTGTGAAGGAAAGAAACATCTCCTCTAAACCACTATGCTGCT 
               
               
                   
               
               
                 AGAGCCTCTTTTCTGTACTCAAGCCTCATTCAGACACTAGTGTCACC 
               
               
                   
               
               
                 AGTCTCCTCATATACCTATTGTATTTTCTTCTTCTTGCTGGTTTAGT 
               
               
                   
               
               
                 CATGTTTTCTGGGAGCTTAGGGGCTTATTTTATTTTGTTTTGTTTTC 
               
               
                   
               
               
                 TAATCAACAGAGATGGGCAAACCCATTATTTTTTTCTTTAGACTTGG 
               
               
                   
               
               
                 GATGGTGATAGCTGGGCAGCGTCAGAAACTGTGTGTGGATATAGATA 
               
               
                   
               
               
                 AGAGCTCGGACTATGCTGAGCTGTGATGAGGGAGGGACCTAGCCAAA 
               
               
                   
               
               
                 GGCAGTGAGAGTCAGAATGCTCCTGCTATTGCCTTCTCAGTCCCCAC 
               
               
                   
               
               
                 GCTTGGTTTCTACACAAGTAGATACATAGAAAAGGCTATAGGTTAGT 
               
               
                   
               
               
                 GTTTGAGAGTCCTGCATGAGTTAGTTGCTCAGAAATGCCCGATAAAT 
               
               
                   
               
               
                 ATGTTATGTGTGTTTATGTATATATATGTTTTATATATATATATGTG 
               
               
                   
               
               
                 TGTGTGTGTGTGTGTGTGTGTTGTGTTTACAAATATGTGATTATCAT 
               
               
                   
               
               
                 CAAAACGTGAGGGCTAAAGTGACCAGATAACTTGCAGG 
               
            
           
         
       
     
     In certain embodiments, the HS1 region has a length of less than about 500 bp. In certain embodiments, the HS1 region has a length of about 490 bp. In certain non-limiting embodiments, the HS1 region has a length of 489 bp. In certain non-limiting embodiments, the HS1 region comprises or has the nucleotide sequence set forth in SEQ ID NO:4, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 4] 
               
               
                 GGCATCTGTGAAGGAAAGAAACATCTCCTCTAAACCACTATGCTGCT 
               
               
                   
               
               
                 AGAGCCTCTTTTCTGTACTCAAGCCTCATTCAGACACTAGTGTCACC 
               
               
                   
               
               
                 AGTCTCCTCATATACCTATTGTATTTTCTTCTTCTTGCTGGTTTAGT 
               
               
                   
               
               
                 CATGTTTTCTGGGAGCTTAGGGGCTTATTTTATTTTGTTTTGTTTTC 
               
               
                   
               
               
                 TAATCAACAGAGATGGGCAAACCCATTATTTTTTTCTTTAGACTTGG 
               
               
                   
               
               
                 GATGGTGATAGCTGGGCAGCGTCAGAAACTGTGTGTGGATATAGATA 
               
               
                   
               
               
                 AGAGCTCGGACTATGCTGAGCTGTGATGAGGGAGGGACCTAGCCAAA 
               
               
                   
               
               
                 GGCAGTGAGAGTCAGAATGCTCCTGCTATTGCCTTCTCAGTCCCCAC 
               
               
                   
               
               
                 GCTTGGTTTCTACACAAGTAGATACATAGAAAAGGCTATAGGTTAGT 
               
               
                   
               
               
                 GTTTGAGAGTCCTGCATGAGTTAGTTGCTCAGAAATGCCCGATAAAT 
               
               
                   
               
               
                 ATGTTATGTGTGTTTATGT 
               
            
           
         
       
     
     In certain non-limiting embodiments, the β-globin LCR comprises or consists essentially of a HS1 region disclosed herein, a HS2 region disclosed herein, a HS3 region disclosed herein, and a HS4 region disclosed herein. 
     In certain non-limiting embodiments, the β-globin LCR comprises or consists essentially of a HS1 region having the nucleotide sequence set forth in SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 22 or SEQ ID NO: 23; a HS2 region having the nucleotide sequence set forth in SEQ ID NO: 9 or a consecutive portion thereof, a modification of SEQ ID NO: 9, a modification of a consecutive portion of SEQ ID NO: 9 (e.g., SEQ ID NO: 33), SEQ ID NO:20, SEQ ID NO:21; a HS3 region having the nucleotide sequence set forth in SEQ ID NO: 5 or a modification thereof (e.g., SEQ ID NO: 34); and a HS4 region having the nucleotide sequence set forth in SEQ ID NO: 6 or a consecutive portion thereof, SEQ ID NO: 7 or a consecutive portion thereof, a modification of SEQ ID NO: 6, a modification of a consecutive portion of SEQ ID NO: 6 (e.g., SEQ ID NO: 35), a modification of SEQ ID NO: 7, or a modification of a consecutive portion of SEQ ID NO: 7, or SEQ ID NO:8. 
     In certain non-limiting embodiments, the β-globin LCR comprises or consists essentially of a HS1 region having the nucleotide sequence set forth in SEQ ID NO: 3; a HS2 region having nucleotides 45 to 860 of SEQ ID NO: 9; a HS3 region having the nucleotide sequence set forth in SEQ ID NO: 5; and a HS4 region having nucleotides 115 to 868 of SEQ ID NO: 6 e.g., the β-globin LCR of SNS22.B87.A1 shown in  FIG. 15 . 
     In certain non-limiting embodiments, the β-globin LCR comprises or consists essentially of a HS1 region having the nucleotide sequence set forth in SEQ ID NO: 3; a HS2 region having nucleotide sequence set forth in SEQ ID NO: 33; a HS3 region having the nucleotide sequence set forth in SEQ ID NO: 34; and a HS4 region having the nucleotide sequence set forth in SEQ ID NO: 35, e.g., as the β-globin LCR of SNS22.2.B87.A1 shown in  FIG. 13 . 
     Recent studies have shown that HS2 is not erythroid-specific, but is expressed in other cell lines and lineages (See Example 3 and  FIG. 7 ) and is also present in undifferentiated human embryonic stem cells (Chang et al., Stem cell reviews (2013); 9:397-407). Due to the non-erythroid activity of HS2, HS2-containing globin vectors may pose a risk for their safe use in clinical treatment, e.g., for treating thalassemia and sickle cell patients. In certain embodiments, the β-globin LCR does not comprise a HS2 region. In certain embodiments, the β-globin LCR does not comprise a core sequence of HS2. A core sequence of HS2 provides position independent, high level expression. In addition, a core sequence of HS2 sustains the enhancer activity of HS2. For example, the core sequence of HS2 enhances the transcription of a globin gene (e.g., human β-globin gene). Additionally, a core sequence of HS2 comprises one or more binding sites or binding motifs for ubiquitous as well as tissue-specific (e.g., erythroid-specific) proteins (e.g., transcription factors), including, but not limited to, members of AP1 family of proteins (e.g., NF-E2), GATA-1 (also known as “NF-E1” or “NFE1”), Krüppel-like Zn finger proteins (e.g., ubiquitous proteins Sp1 and YY1, and erythroid-restricted factor erythroid Krüppel-like factor (EKLF)), and basic helix-loop-helix (bHLH) proteins (E boxes) (e.g., USF and TALI). AP1 binding sites are required for enhancement and induction (Moi and Kan (1990); Ney et al., (1990); Talbot and Grosveld (1991)). Furthermore, binding of NF-E2 can cause disruption of in vitro reconstituted chromatin at HS2 (Armstrong and Emerson (1996)). Mutations in the GATA-1 binding sites can cause a reduction in enhancer activity of HS2 in transgenic mice (Caterina et al., (1994)). Although both AP1 (e.g., AP1/NF-E2) and GATA1 binding sites are important for core function, mice lacking these factors do not show impaired globin gene expression (Weiss et al., 1994). 
     In certain embodiments, the β-globin LCR does not comprise the full length of a core sequence of HS2. In certain embodiments, the core sequence of a HS2 region is a core sequence of human HS2. In certain non-limiting embodiments, the core sequence of human HS2 comprises a tandem pair of binding sites for members of AP1 family of proteins (e.g., NF-E2) (referred to as “AP1/NF-E2” binding sites) (e.g., GCTGAGTCA, and GATGAGTCA), one binding site for Kruppel-like Zn finger proteins (e.g., AGGGTGTGT), one GATA-1 binding site (e.g., CTATCT), and three E boxes (CANNTG, e.g., CAGATG, and CACCTG). In certain non-limiting embodiments, the β-globin LCR does not comprise the full length of a 388 bp core sequence of human HS2, which has the nucleotide sequence set forth in SEQ ID NO:20 provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 20] 
               
               
                 TAAGCTTCAGTTTTTCCTTAGTTCCTGTTACATTTCTGTGTGTCTCC 
               
               
                   
               
               
                 ATTAGTGACCTCCCATAGTCCAAGCATGAGCAGTTCTGGCCAGGCCC 
               
               
                   
               
               
                 CTGTCGGGGTCAGTGCCCCACCCCCGCCTTCTGGTTCTGTGTAACCT 
               
               
                   
               
               
                 TCTAAGCAAACCTTCTGGCTCAAGCACAGCAATGCTGAGTCATGATG 
               
               
                   
               
               
                 AGTCATGCTGAGGCTTAGGGTGTGTGCCCAGATGTTCTCAGCCTAGA 
               
               
                   
               
               
                 GTGATGACTCCTATCTGGGTCCCCAGCAGGATGCTTACAGGGCAGAT 
               
               
                   
               
               
                 GGCAAAAAAAAGGAGAAGCTGACCACCTGACTAAAACTCCACCTCAA 
               
               
                   
               
               
                 ACGGCATCATAAAGAAAATGGATGCCTGAGACAGAATGTGACATATT 
               
               
                   
               
               
                 CTAGAATATATT 
               
            
           
         
       
     
     The nucleotide sequence set forth in SEQ ID NO:20 corresponds to nucleotides position 16671 to position 17058 of SEQ ID NO:19 (GenBank Access No.: NG_000007.3). In SEQ ID NO:20, one AP1/NF-E2 binding site having the nucleotide sequence of GCTGAGTCA is located at position 175 to position 183, one AP1/NF-E2 binding site having the nucleotide sequence of GATGAGTCA is located at position 185 to position 193, one binding site for Krüppel-like Zn finger proteins having the nucleotide sequence of AGGGTGTGT is located as position 205 to position 213, two E boxes, each of which have the nucleotide sequence of CAGATG, is located at position 217 to position 222, and position 278 to position 283, one GATA-1 binding site having the nucleotide sequence of CTATCT is located at position 246 to position 251, one E box having the nucleotide sequence of CACCTG is located at position 306 to position 311. 
     In certain non-limiting embodiments, the β-globin LCR does not comprise the full length of a 387 bp core sequence of human HS2, which has the nucleotide sequence set forth in SEQ ID NO:21 provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 21] 
               
               
                 TAAGCTTCAGTTTTTCCTTAGTTCCTGTTACATTTCTGTGTGTCTCC 
               
               
                   
               
               
                 ATTAGTGACCTCCCATAGTCCAAGCATGAGCAGTTCTGGCCAGGCCC 
               
               
                   
               
               
                 CTGTCGGGGTCAGTGCCCCACCCCCGCCTTCTGGTTCTGTGTAACCT 
               
               
                   
               
               
                 TCTAAGCAAACCTTCTGGCTCAAGCACAGCAATGCTGAGTCATGATG 
               
               
                   
               
               
                 AGTCATGCTGAGGCTAGGGTGTGTGCCCAGATGTTCTCAGCCTAGAG 
               
               
                   
               
               
                 TGATGACTCCTATCTGGGTCCCCAGCAGGATGCTTACAGGGCAGATG 
               
               
                   
               
               
                 GCAAAAAAAAGGAGAAGCTGACCACCTGACTAAAACTCCACCTCAAA 
               
               
                   
               
               
                 CGGCATCATAAAGAAAATGGATGCCTGAGACAGAATGTGACATATTC 
               
               
                   
               
               
                 TAGAATATATT 
               
            
           
         
       
     
     In SEQ ID NO:21, one AP1/NF-E2 binding site having the nucleotide sequence of GCTGAGTCA is located at position 175 to position 183, one AP1/NF-E2 binding site having the nucleotide sequence of GATGAGTCA is located at position 185 to position 193, one binding site for Krüppel-like Zn finger proteins having the nucleotide sequence of AGGGTGTGT is located as position 204 to position 212, two E boxes, each of which have the nucleotide sequence of CAGATG, is located at position 216 to position 221, and position 277 to position 282, one GATA-1 binding site having the nucleotide sequence of CTATCT is located at position 245 to position 250, one E box having the nucleotide sequence of CACCTG is located at position 305 to position 310. 
     In certain embodiments, the β-globin LCR does not comprise a HS2 region that comprises a core sequence of HS2. A HS2 region that comprises a core sequence of HS2 can vary in length and sequence. In non-limiting examples, a HS2 region that comprises a core sequence of HS2 is from about 400 bp to about 1000 bp, e.g., from about 400 bp to about 500 bp, from about 500 bp to about 600 bp, from about 600 bp to about 700 bp, from about 700 bp to about 800 bp, from about 800 bp to about 900 bp, or from about 900 bp to about 1000 bp, in length. In certain non-limiting embodiments, the β-globin LCR does not comprise a 840 bp HS2 region (e.g., the HS2 region comprised in the globin vector TNS9 disclosed in U.S. Pat. No. 7,541,179). In certain non-limiting embodiments, the β-globin LCR does not comprise a 860 bp HS2 region. In certain non-limiting embodiments, the β-globin LCR does not comprise an about 650 bp HS2 region. In certain non-limiting embodiments, the β-globin LCR does not comprise a 646 bp HS2 region (e.g., the HS2 region comprised in the globin vector LentiGlobin™, also known as “β 87 ”). In certain non-limiting embodiments, the β-globin LCR does not comprise an about 420 bp HS2 region. In certain non-limiting embodiments, the β-globin LCR does not comprise a 423 bp HS2 region (e.g., the HS2 region comprised in the globin vector disclosed in Sadelain et al.,  Proc. Nat&#39;l Acad. Sci . ( USA ) (1995); 92:6728-6732). 
     In certain embodiments, the β-globin LCR does not comprise a HS2 region that sustains the enhancer activity of HS2. In certain embodiments, the β-globin LCR does not comprise a HS2 region that is capable of enhancing the transcription of a globin gene (e.g., human β-globin gene). In non-limiting examples, the β-globin LCR does not comprise a HS2 region whose ability to enhance the transcription of a globin gene (e.g., human β-globin gene) is no less than about 60%, no less than about 70%, no less than about 80%, no less than about 90%, or no less than about 95% in comparison to a native HS2 region. 
     In certain embodiments, the β-globin LCR does not comprise a HS2 region that comprises one, two, three, four, five, six or seven of the following binding sites: two (a tandem pair of) AP1/NF-E2 binding sites (e.g., GCTGAGTCA, and GATGAGTCA), one binding site for Kruppel-like Zn finger proteins (e.g., AGGGTGTGT), one GATA-1 binding site (e.g., CTATCT), and three E boxes (CANNTG, e.g., CAGATG, and CACCTG). In certain embodiments, the β-globin LCR does not comprise a HS2 region that comprises six of the above-described binding sites. For example, in certain embodiments, the β-globin LCR does not comprise a HS2 region that comprises two AP1/NF-E2 binding sites, one binding site for Kruppel-like Zn finger proteins, one GATA-1 binding site, and two not three E boxes. In certain embodiments, the β-globin LCR does not comprise a HS2 region that comprises one not two AP1/NF-E2 binding site, one binding site for Kruppel-like Zn finger proteins, one GATA-1 binding site, and three E boxes. In certain embodiments, the β-globin LCR does not comprise a HS2 region that comprises two AP1/NF-E2 binding sites, one GATA-1 binding site, and three E boxes and does not comprise one binding site for Kruppel-like Zn finger proteins. In certain embodiments, the β-globin LCR does not comprise a HS2 region that comprises two AP1/NF-E2 binding sites, one binding site for Kruppel-like Zn finger proteins, and three E boxes, and does not comprise one GATA-1 binding site. 
     In certain embodiments, the β-globin LCR comprises a HS1 region, a HS3 region, and a HS4 region, and does not comprise a HS2 region. In certain embodiments, the HS1 region, HS3 region and HS4 region within the β-globin LCR are contiguous. In certain non-limiting embodiments, the β-globin LCR consists essentially of a HS1 region, a HS3 region and a HS4 region. In certain embodiments, the β-globin LCR comprises two introduced GATA-1 binding sites at the junction between the HS3 region and the HS4 region. The HS3 region can lie between the HS1 region and the HS4 region. The HS1, HS3, and HS4 regions can be any HS1, HS3, and HS4 regions disclosed herein. The HS2 region can also be any HS2 region disclosed herein. 
     In certain non-limiting embodiments, the β-globin LCR comprises or consists essentially of a HS1 region disclosed herein, a HS3 region disclosed herein, and a HS4 region disclosed herein, and does not comprise a HS2 region. 
     In certain non-limiting embodiments, the β-globin LCR comprises or consists essentially of a HS1 region having the nucleotide sequence set forth in SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:22 or SEQ ID NO:23; a HS3 region having the nucleotide sequence set forth in SEQ ID NO:5 or a modification thereof (e.g., SEQ ID NO: 34); and a HS4 region having the nucleotide sequence set forth in SEQ ID NO:6 or a consecutive portion thereof, SEQ ID NO:7 or a consecutive portion thereof, a modification of SEQ ID NO: 6, a modification of a consecutive portion of SEQ ID NO: 6 (e.g., SEQ ID NO: 35), a modification of SEQ ID NO: 7, or a modification of a consecutive portion of SEQ ID NO: 7, or SEQ ID NO:8, and the β-globin LCR does not comprise a HS2 region. 
     In certain non-limiting embodiments, the β-globin LCR comprises or consists essentially of a HS1 region having the nucleotide sequence set forth in SEQ ID NO:2, a HS3 region having the nucleotide sequence set forth in SEQ ID NO:5, and a HS4 region having the nucleotide sequence set forth in SEQ ID NO:6, and the β-globin LCR does not comprise a HS2 region, as shown in  FIG. 2 . 
     In certain non-limiting embodiments, the β-globin LCR comprises or consists essentially of a HS1 region having the nucleotide sequence set forth in SEQ ID NO:3, a HS3 region having the nucleotide sequence set forth in SEQ ID NO:5, and a HS4 region having the nucleotide sequence set forth in SEQ ID NO:8, and the β-globin LCR does not comprise a HS2 region, as shown in  FIG. 3 . 
     In certain non-limiting embodiments, the β-globin LCR comprises or consists essentially of a HS1 region having the nucleotide sequence set forth in SEQ ID NO:4, a HS3 region having the nucleotide sequence set forth in SEQ ID NO:5, and a HS4 region having the nucleotide sequence set forth in SEQ ID NO:8, and the β-globin LCR does not comprise a HS2 region. 
     In certain non-limiting embodiments, the β-globin LCR comprises or consists essentially of a HS1 region having the nucleotide sequence set forth in SEQ ID NO: 3; a HS3 region having the nucleotide sequence set forth in SEQ ID NO: 5; and a HS4 region having nucleotides 115 to 868 of SEQ ID NO: 6 e.g., the β-globin LCR of SNS24.B87.A1 shown in  FIG. 15 . 
     In certain non-limiting embodiments, the β-globin LCR comprises or consists essentially of a HS1 region having the nucleotide sequence set forth in SEQ ID NO: 3; a HS3 region having the nucleotide sequence set forth in SEQ ID NO: 34; and a HS4 region having the nucleotide sequence set forth in SEQ ID NO: 35, e.g., as the β-globin LCR of SNS24.2.B87.A1 shown in  FIG. 13 . 
     In certain embodiments, the β-globin LCR does not comprise a HS1 region or a HS2 region. In certain embodiments, the β-globin LCR does not comprise a core sequence of HS1. A core sequence of HS1 sustains the activity of HS1, e.g., enhancer activity, or functioning as a facilitator or regulatory element to tether the enhancer activity of other HS regions, e.g., HS2-4. In addition, a core sequence of HS1 comprises one or more binding sites or binding motifs for ubiquitous as well as tissue-specific (e.g., erythroid-specific) proteins (e.g., transcription factors), including, but not limited to, GATA-1, and Krüppel-like Zn finger proteins (e.g., erythroid-restricted factor EKLF). 
     In certain embodiments, the β-globin LCR does not comprise the full length of a core sequence of HS1. In certain embodiments, the core sequence of a HS1 region is a core sequence of human HS1. In certain non-limiting embodiments, the core sequence of human HS1 comprises two GATA-1 binding sites (e.g., TTATCT, and CTATCA), and one binding site for EKLF (e.g., CCACACACA). In certain embodiments, the β-globin LCR does not comprise the full length of a 286 bp core sequence of human HS1. In certain non-limiting embodiments, the 286 bp core sequence of human HS1 has the nucleotide sequence set forth in SEQ ID NO:22 provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 22] 
               
               
                 CTGAGCAACTAACTCATGCAGGACTCTCAAACACTAACCTATAGCCT 
               
               
                   
               
               
                 TTTCTATGTATCTACTTGTGTAGAAACCAAGCGTGGGGACTGAGAAG 
               
               
                   
               
               
                 GCAATAGCAGGAGCATTCTGACTCTCACTGCCTTTGGCTAGGTCCCT 
               
               
                   
               
               
                 CCCTCATCACAGCTCAGCATAGTCCGAGCTCTTATCTATATCCACAC 
               
               
                   
               
               
                 ACAGTTTCTGACGCTGCCCAGCTATCACCATCCCAAGTCTAAAGAAA 
               
               
                   
               
               
                 AAAATAATGGGTTTGCCCATCTCTGTTGATTAGAAAACAAAACAAAA 
               
               
                   
               
               
                 TAAA 
               
            
           
         
       
     
     In SEQ ID NO:22, one GATA-1 binding site having the nucleotide sequence of TTATCT is located at position 173 to position 178, one GATA-1 binding site having the nucleotide sequence of CTATCA located at position 210 to position 215, and one binding site for EKLF having the nucleotide sequence of CCACACACA is located at position 183 to position 191. 
     In certain non-limiting embodiments, the 286 bp core sequence of human HS1 has the nucleotide sequence set forth in SEQ ID NO:23 provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 23] 
               
               
                 CTGAGCAACTAATCATGCAGGACTCTCAAACACTAACCTATAGCCTT 
               
               
                   
               
               
                 TTCTATGTATCTACTTGTGTAGAAACCAAGCGTGGGGACTGAGAAGG 
               
               
                   
               
               
                 CAATAGCAGGAGCATTCTGACTCTCACTGCCTTTAGCTAGGCCCCTC 
               
               
                   
               
               
                 CCTCATCACAGCTCAGCATAGTCCTGAGCTCTTATCTATATCCACAC 
               
               
                   
               
               
                 ACAGTTTCTGACGCTGCCCAGCTATCACCATCCCAAGTCTAAAGAAA 
               
               
                   
               
               
                 AAAATAATGGGTTTGCCCATCTCTGTTGATTAGAAAACAAAACAAAA 
               
               
                   
               
               
                 TAAA 
               
            
           
         
       
     
     The nucleotide sequence set forth in SEQ ID NO:23 corresponds to nucleotides position 21481 to position 21766 of SEQ ID NO:19 (GenBank Access No.: NG_000007.3). In SEQ ID NO:23, one GATA-1 binding site having the nucleotide sequence of TTATCT is located at position 173 to position 178, one GATA-1 binding site having the nucleotide sequence of CTATCA located at position 210 to position 215, and one binding site for EKLF having the nucleotide sequence of CCACACACA is located at position 183 to position 191. 
     In certain embodiments, the β-globin LCR does not comprise a HS1 region that comprises a core sequence of HS1. A HS1 region that comprises a core sequence of HS1 can vary in length and sequence. In non-limiting examples, a HS1 region that comprises a core sequence of HS1 is from about 300 bp to about 1200 bp, e.g., from about 300 bp to about 400 bp, from about 400 bp to about 500 bp, from about 500 bp to about 600 bp, from about 600 bp to about 700 bp, from about 700 bp to about 800 bp, from about 800 bp to about 900 bp, from about 900 bp to about 1000 bp, from about 1000 bp to about 1100 bp, or from about 1100 bp to about 1200 bp, in length. In certain non-limiting embodiments, the β-globin LCR does not comprise an about 1.0 kb bp HS1 region. In certain non-limiting embodiments, the β-globin LCR does not comprise an about 1.1 kb HS1 region. 
     In certain embodiments, the β-globin LCR does not comprise a HS1 region that sustains the activity of HS1, e.g., enhancer activity, or functioning as a facilitator or regulatory element to tether the enhancer activity of other HS regions, e.g., HS2-4. In certain embodiments, the β-globin LCR does not comprise a HS1 region that is capable of enhancing the transcription of a globin gene (e.g., human β-globin gene). In non-limiting examples, the β-globin LCR does not comprise a HS1 region whose ability to enhance the transcription of a globin gene (e.g., human β-globin gene) is no less than about 60%, no less than about 70%, no less than about 80%, no less than about 90%, or no less than about 95% in comparison to a native HS1 region. In non-limiting examples, the β-globin LCR does not comprise a HS1 region whose ability to tether the enhancer activity of one or more of HS2-HS4 is no less than about 60%, no less than about 70%, no less than about 80%, no less than about 90%, or no less than about 95% in comparison to a native HS1 region. 
     In certain embodiments, the β-globin LCR does not comprise a HS1 region that comprises one, two, or three of the following binding sites: two GATA-1 binding sites (e.g., TTATCT, and CTATCA), and one binding site for EKLF (e.g., CCACACACA). In certain embodiments, the β-globin LCR does not comprise a HS1 region that comprises two of the above-described binding sites. For example, in certain embodiments, the β-globin LCR does not comprise a HS1 region that comprises two GATA-1 binding sites and does not comprise one binding site for EKLF. In certain embodiments, the β-globin LCR does not comprise a HS1 region that comprises one not two AP1/NF-E2 binding site and one binding site for EKLF. 
     In certain embodiments, the β-globin LCR comprises a HS3 region and a HS4 region, and the β-globin LCR does not comprise a HS1 region or a HS2 region. In certain embodiments, the HS3 region and HS4 region within the β-globin LCR are contiguous. In certain non-limiting embodiments, the β-globin LCR consisting essentially of a HS3 region and a HS4 region. In another embodiment, the β-globin LCR comprises two introduced GATA-1 binding sites at the junction between the HS3 region and the HS4 region. The HS3 region can lie between the globin gene or functional portion thereof and the HS4 region. 
     In certain embodiments, the β-globin LCR comprises a HS3 region having the nucleotide sequence set forth in SEQ ID NO:5 and a HS4 region having the nucleotide sequence set forth in SEQ ID NO:6, SEQ ID NO:7 or SEQ ID NO:8, and the β-globin LCR does not comprise a HS1 region or a HS2 region. 
     In certain non-limiting embodiments, the β-globin LCR comprises a HS3 region having the nucleotide sequence set forth in SEQ ID NO:5 and a HS4 region having the nucleotide sequence set forth in SEQ ID NO:6, and the β-globin LCR does not comprise a HS1 region or a HS2 region, as shown in  FIG. 4 . 
     In certain embodiments, the β-globin LCR does not comprise a HS1 region, a HS2 region, or a HS4 region. In certain embodiments, the β-globin LCR does not comprise the full length of a core sequence of HS4. In certain embodiments, the core sequence of a HS4 region is a core sequence of human HS4. In certain embodiments, the β-globin LCR does not comprise the full length of a 280 bp core sequence of human HS4 (e.g., the 280 bp core sequence of human HS4 disclosed in Pruzina et al.,  Nucleic Acids Research  (1991); 19:7:1413-1419). 
     In certain embodiments, the β-globin LCR does not comprise a HS4 region that comprises a core sequence of HS4. A HS4 region that comprises a core sequence of HS4 can vary in length and sequence. In non-limiting examples, a HS4 region that comprises a core sequence of HS4 is from about 300 bp to about 1200 bp, e.g., from about 300 bp to about 400 bp, from about 400 bp to about 500 bp, from about 500 bp to about 600 bp, from about 600 bp to about 700 bp, from about 700 bp to about 760 bp, from about 700 bp to about 800 bp, from about 800 bp to about 900 bp, from about 900 bp to about 1000 bp, from about 1000 bp to about 1100 bp, or from about 1100 bp to about 1200 bp, in length. In certain non-limiting embodiments, the β-globin LCR does not comprise an about 1.0 kb HS4 region. In certain non-limiting embodiments, the β-globin LCR does not comprise an about 1.1 kb HS4 region. In certain non-limiting embodiments, the β-globin LCR does not comprise an about 760 bp HS4 region. In certain non-limiting embodiments, the β-globin LCR does not comprise an about 754 bp HS4 region. 
     In certain embodiments, the β-globin LCR does not comprise a HS4 region that sustains the activity of HS4, e.g., enhancer activity. In certain embodiments, the β-globin LCR does not comprise a HS4 region that is capable of enhancing the transcription of a globin gene (e.g., human β-globin gene), conferring position independent expression, and/or increasing β-globin transgene expression. In certain embodiments, the expression cassette comprises at least one erythroid-specific enhancer, which can compensate the activities of HS1, HS2, and/or HS4 regions. 
     In certain embodiments, the β-globin LCR comprises a HS3 region, and the β-globin LCR does not comprise a HS1 region, a HS2 region or a HS4 region. In certain embodiments, the β-globin LCR consisting essentially of a HS3 region. In certain embodiments, the β-globin LCR comprises a HS3 region having the nucleotide sequence set forth in SEQ ID NO: 5 and a HS4 region having the nucleotide sequence set forth in SEQ ID NO:6, SEQ ID NO:7 or SEQ ID NO:8, and the β-globin LCR does not comprise a HS1 region or a HS2 region. In certain embodiments, the expression cassette comprises at least one erythroid-specific enhancer. 
     In certain non-limiting embodiments, the β-globin LCR comprises a HS3 region having the nucleotide sequence set forth in SEQ ID NO:5, and the β-globin LCR does not comprise a HS1 region, a HS2 region or a HS4 region. In certain embodiments, the expression cassette comprises at least one erythroid-specific enhancer. 
     In certain non-limiting embodiments, the β-globin LCR comprises a HS3 region having the nucleotide sequence set forth in SEQ ID NO: 34, and the β-globin LCR does not comprise a HS1 region, a HS2 region or a HS4 region, e.g., the β-globin LCR of SNS27.2.B87.A1 shown in  FIG. 13 . In certain embodiments, the expression cassette comprises at least one erythroid-specific enhancer, e.g., SNS27.2.B87.A1 shown in  FIG. 13 . 
     3.2. Globin Gene 
     In accordance with the presently disclosed subject matter, the expression cassette comprises a globin gene or a functional portion thereof. The globin gene can be a β-globin gene, a γ-globin gene, or a δ-globin gene. In certain embodiments, the expression cassette comprises a human β-globin gene. In accordance with the presently disclosed subject matter, the human β-globin gene can be a wild-type human β-globin gene, a deleted human β-globin gene comprising one or more deletions of intron sequences, or a mutated human β-globin gene encoding at least one anti-sickling amino acid residue. In certain non-limiting embodiments, a presently disclosed expression cassette comprises a wild-type human β-globin gene. A wild-type human β-globin gene comprises three exons (exon 1, exon 2, and exon 3). In certain embodiments, a presently disclosed expression cassette comprises a non-wild-type (mutated or modified) human β A -globin gene. In certain embodiments, a presently disclosed expression cassette comprises a human β-globin gene with a deletion in intron 2 (IVS2). In certain embodiments, the deletion in IVS2 is about 370 bp. The deletion in IVS2 can eliminate AT-rich (ATR) sequences that comprise a cryptic polyadenylation site responsible of premature termination of the transcription. In certain embodiments, a presently disclosed expression cassette comprises a human β A -globin gene encoding a threonine to glutamine mutation at codon 87 (β A-T87Q ). The glutamine residue at position 87 in the gamma-globin chain augments the anti-sickling activity of the gamma chain relative to the beta chain, while preserving adult oxygen-binding characteristics of the beta chain (Nagel et al.,  Proc. Natl. Acad. Sci. U.S.A . (1979); 76:670-672). In certain embodiments, a functional portion of a globin gene has at least about 80%, at least about 90%, at least about 95%, at least about 99% or at least about 100% identity to a corresponding wild-type reference polynucleotide sequence. 
     In certain embodiments, the human β A -globin gene is a human β A -globin gene encoding a threonine to glutamine mutation at codon 87 (β A-T87Q ). In certain embodiments, the human β A -globin gene encoding a threonine to glutamine mutation at codon 87 (β A-T87Q ) further comprises a deletion in intron 2 (e.g., an about 370 bp deletion). In certain embodiments, the human β A -globin gene encoding a threonine to glutamine mutation at codon 87 (β A-T87Q ) comprises the nucleotide sequence set forth in SEQ ID NO: 36, which is provided below. 
     
       
         
           
               
            
               
                 [SEQ ID NO: 36] 
               
               
                 gc aatgaaaata aatgtttttt attaggcaga atccagatgc 
               
               
                   
               
               
                 tcaaggccct tcataatatc ccccagttta gtagttggac 
               
               
                   
               
               
                 ttagggaaca aaggaacctt taatagaaat tggacagcaa 
               
               
                   
               
               
                 gaaagcgagc ttagtgatac ttgtgggcca gggcattagc 
               
               
                   
               
               
                 cacaccagcc accactttct gataggcagc ctgcactggt 
               
               
                   
               
               
                 ggggtgAATT CTTTGCCAAA GTGATGGGCC AGCACACAGA 
               
               
                   
               
               
                 CCAGCACGTT GCCCAGGAGC TGTGGGAGGA AGATAAGAGG 
               
               
                   
               
               
                 TATGAACATG ATTAGCAAAA GGGCCTAGCT TGGACTCAGA 
               
               
                   
               
               
                 ATAATCCAGC CTTATCCCAA CCATAAAATA AAAGCAGAAT 
               
               
                   
               
               
                 GGTAGCTGGA TTGTAGCTGC TATTAGCAAT ATGAAACCTC 
               
               
                   
               
               
                 TTACATCAGT TACAATTTAT ATGCAGAAAT ATTTATATGC 
               
               
                   
               
               
                 AGAAATATTG CTATTGCCTT AACCCAGAAA TTATCACTGT 
               
               
                   
               
               
                 TATTCTTTAG AATGGTGCAA AGAGGCATGA TACATTGTAT 
               
               
                   
               
               
                 CATTATTGCC CTGAAAGAAA GAGATTAGGG AAAGTATTAG 
               
               
                   
               
               
                 AAATAAGATA AACAAAAAAG TATATTAAAA GAAGAAAGCA 
               
               
                   
               
               
                 TTTTTTAAAA TTACAAATGC AAAATTACCC TGATTTGGTC 
               
               
                   
               
               
                 AATATGTGTA CCCTGTTACT TCTCCCCTTC CTATGACATG 
               
               
                   
               
               
                 AACTTAACCA TAGAAAAGAA GGGGAAAGAA AACATCAAGG 
               
               
                   
               
               
                 GTCCCATAGA CTCACCCTGA AGTTCTCAGG ATCCACGTGC 
               
               
                   
               
               
                 AGCTTGTCAC AGTGCAGCTC ACTCAGctgG GCAAAGGTGC 
               
               
                   
               
               
                 CCTTGAGGTT GTCCAGGTGA GCCAGGCCAT CACTAAAGGC 
               
               
                   
               
               
                 ACCGAGCACT TTCTTGCCAT GAGCCTTCAC CTTAGGGTTG 
               
               
                   
               
               
                 CCCATAACAG CATCAGGAGT GGACAGATCC CCAAAGGACT 
               
               
                   
               
               
                 CAAAGAACCT CTGGGTCCAA GGGTAGACCA CCAGCAGCCT 
               
               
                   
               
               
                 AAGGGTGGGA AAATAGACCA ATAGGCAGAG AGAGTCAGTG 
               
               
                   
               
               
                 CCTATCAGAA ACCCAAGAGT CTTCTCTGTC TCCACATGCC 
               
               
                   
               
               
                 CAGTTTCTAT TGGTCTCCTT AAACCTGTCT TGTAACCTTG 
               
               
                   
               
               
                 ATACCAACCT GCCCAGGGCC TCACCACCAA CTTCATCCAC 
               
               
                   
               
               
                 GTTCACCTTG CCCCACAGGG CAGTAACGGC AGACTTCTCC 
               
               
                   
               
               
                 TCAGGAGTCA GGTGCACCAT GGTGTCTGTT TGAGGTTGCT 
               
               
                   
               
               
                 AGTGAACACA GTTGTGTCAG AAGCAAATGT 
               
            
           
         
       
     
     In certain embodiments, the human β A -globin gene encoding a threonine to glutamine mutation at codon 87 (β A-T87Q ) comprises the nucleotide sequence set forth in SEQ ID NO: 53, which is provided below. 
     
       
         
           
               
            
               
                 [SEQ ID NO: 53] 
               
               
                 gc aatgaaaata aatgtttttt attaggcaga atccagatgc 
               
               
                   
               
               
                 tcaaggccct tcataatatc ccccagttta gtagttggac 
               
               
                   
               
               
                 ttagggaaca aaggaacctt taatagaaat tggacagcaa 
               
               
                   
               
               
                 gaaagcgagc ttagtgatac ttgtgggcca gggcattagc 
               
               
                   
               
               
                 cacaccagcc accactttct gataggcagc ctgcactggt 
               
               
                   
               
               
                 ggggtgAATT CTTTGCCAAA GTGATGGGCC AGCACACAGA 
               
               
                   
               
               
                 CCAGCACGTT GCCCAGGAGC TGTGGGAGGA AGATAAGAGG 
               
               
                   
               
               
                 TATGAACATG ATTAGCAAAA GGGCCTAGCT TGGACTCAGA 
               
               
                   
               
               
                 ATAATCCAGC CTTATCCCAA CCATAAAATA AAAGCAGAAT 
               
               
                   
               
               
                 GGTAGCTGGA TTGTAGCTGC TATTAGCAAT ATGAAACCTC 
               
               
                   
               
               
                 TTACATCAGT TACAATTTAT ATGCAGAAAT ATTTATATGC 
               
               
                   
               
               
                 AGAAATATTG CTATTGCCTT AACCCAGAAA TTATCACTGT 
               
               
                   
               
               
                 TATTCTTTAG AATGGTGCAA AGAGGCATGA TACATTGTAT 
               
               
                   
               
               
                 CATTATTGCC CTGAAAGAAA GAGATTAGGG AAAGTATTAG 
               
               
                   
               
               
                 AAATAAGATA AACAAAAAAG TATATTAAAA GAAGAAAGCA 
               
               
                   
               
               
                 TTTTTTAAAA TTACAAATGC AAAATTACCC TGATTTGGTC 
               
               
                   
               
               
                 AATATGTGTA CCCTGTTACT TCTCCCCTTC CTATGACATG 
               
               
                   
               
               
                 AACTTAACCA TAGAAAAGAA GGGGAAAGAA AACATCAAGG 
               
               
                   
               
               
                 GTCCCATAGA CTCACCCTGA AGTTCTCAGG ATCCACGTGC 
               
               
                   
               
               
                 AGCTTGTCAC AGTGCAGCTC ACTCAGTTGG GCAAAGGTGC 
               
               
                   
               
               
                 CCTTGAGGTT GTCCAGGTGA GCCAGGCCAT CACTAAAGGC 
               
               
                   
               
               
                 ACCGAGCACT TTCTTGCCAT GAGCCTTCAC CTTAGGGTTG 
               
               
                   
               
               
                 CCCATAACAG CATCAGGAGT GGACAGATCC CCAAAGGACT 
               
               
                   
               
               
                 CAAAGAACCT CTGGGTCCAA GGGTAGACCA CCAGCAGCCT 
               
               
                   
               
               
                 AAGGGTGGGA AAATAGACCA ATAGGCAGAG AGAGTCAGTG 
               
               
                   
               
               
                 CCTATCAGAA ACCCAAGAGT CTTCTCTGTC TCCACATGCC 
               
               
                   
               
               
                 CAGTTTCTAT TGGTCTCCTT AAACCTGTCT TGTAACCTTG 
               
               
                   
               
               
                 ATACCAACCT GCCCAGGGCC TCACCACCAA CTTCATCCAC 
               
               
                   
               
               
                 GTTCACCTTG CCCCACAGGG CAGTAACGGC AGACTTCTCC 
               
               
                   
               
               
                 TCAGGAGTCA GGTGCACCAT GGTGTCTGTT TGAGGTTGCT 
               
               
                   
               
               
                 AGTGAACACA GTTGTGTCAG AAGCAAATGT 
               
            
           
         
       
     
     In certain non-limiting embodiments, the human β-globin gene is a human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 (β A-E22A ). In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 (β A-E22A ) further comprises a deletion in intron 2 (e.g., an about 370 bp deletion). In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 (β A-E22A ) comprises the nucleotide sequence set forth in SEQ ID NO: 54, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 54] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaag 
               
               
                   
               
               
                 gcccttcataatatcccccagtttagtagttggacttagggaacaaa 
               
               
                   
               
               
                 ggaacctttaatagaaattggacagcaagaaagcgagcttagtgata 
               
               
                   
               
               
                 cttgtgggccagggcattagccacaccagccaccactttctgatagg 
               
               
                   
               
               
                 cagcctgcactggtggggtgAATTCTTTGCCAAAGTGATGGGCCAGC 
               
               
                   
               
               
                 ACACAGACCAGCACGTTGCCCAGGAGCTGTGGGAGGAAGATAAGAGG 
               
               
                   
               
               
                 TATGAACATGATTAGCAAAAGGGCCTAGCTTGGACTCAGAATAATCC 
               
               
                   
               
               
                 AGCCTTATCCCAACCATAAAATAAAAGCAGAATGGTAGCTGGATTGT 
               
               
                   
               
               
                 AGCTGCTATTAGCAATATGAAACCTCTTACATCAGTTACAATTTATA 
               
               
                   
               
               
                 TGCAGAAATATTTATATGCAGAAATATTGCTATTGCCTTAACCCAGA 
               
               
                   
               
               
                 AATTATCACTGTTATTCTTTAGAATGGTGCAAAGAGGCATGATACAT 
               
               
                   
               
               
                 TGTATCATTATTGCCCTGAAAGAAAGAGATTAGGGAAAGTATTAGAA 
               
               
                   
               
               
                 ATAAGATAAACAAAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAA 
               
               
                   
               
               
                 ATTACAAATGCAAAATTACCCTGATTTGGTCAATATGTGTACCCTGT 
               
               
                   
               
               
                 TACTTCTCCCCTTCCTATGACATGAACTTAACCATAGAAAAGAAGGG 
               
               
                   
               
               
                 GAAAGAAAACATCAAGGGTCCCATAGACTCACCCTGAAGTTCTCAGG 
               
               
                   
               
               
                 ATCCACGTGCAGCTTGTCACAGTGCAGCTCACTCAGtgtGGCAAAGG 
               
               
                   
               
               
                 TGCCCTTGAGgttGTCCAGGTGAGCCAGGCCATCACTAAAGGCACCG 
               
               
                   
               
               
                 AGCACTTTCTTGCCATGAGCCTTCACCTTAGGGTTGCCCATAACAGC 
               
               
                   
               
               
                 ATCAGGAGTGGACAGATCCCCAAAGGACTCAAAGAACCTCTGGGTCC 
               
               
                   
               
               
                 AAGGGTAGACCACCAGCAGCCTAAGGGTGGGAAAATAGACCAATAGG 
               
               
                   
               
               
                 CAGAGAGAGTCAGTGCCTATCAGAAACCCAAGAGTCTTCTCTGTCTC 
               
               
                   
               
               
                 CACATGCCCAGTTTCTATTGGTCTCCTTAAACCTGTCTTGTAACCTT 
               
               
                   
               
               
                 GATACCAACCTGCCCAGGGCCTCACCACCAACggcATCCACGTTCAC 
               
               
                   
               
               
                 CTTGCCCCACAGGGCAGTAACGGCAGACTTCTCCTCAGGAGTCAGGT 
               
               
                   
               
               
                 GCACCATGGTGTCTGTTTGAGGTTGCTAGTGAACACAGTTGTGTCAG 
               
               
                   
               
               
                 AAGCAAATGT 
               
            
           
         
       
     
     In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 (β A-E22A ) comprises the nucleotide sequence set forth in SEQ ID NO: 55, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 55] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaag 
               
               
                   
               
               
                 gcccttcataatatcccccagtttagtagttggacttagggaacaaa 
               
               
                   
               
               
                 ggaacctttaatagaaattggacagcaagaaagcgagcttagtgata 
               
               
                   
               
               
                 cttgtgggccagggcattagccacaccagccaccactttctgatagg 
               
               
                   
               
               
                 cagcctgcactggtggggtgAATTCTTTGCCAAAGTGATGGGCCAGC 
               
               
                   
               
               
                 ACACAGACCAGCACGTTGCCCAGGAGCTGTGGGAGGAAGATAAGAGG 
               
               
                   
               
               
                 TATGAACATGATTAGCAAAAGGGCCTAGCTTGGACTCAGAATAATCC 
               
               
                   
               
               
                 AGCCTTATCCCAACCATAAAATAAAAGCAGAATGGTAGCTGGATTGT 
               
               
                   
               
               
                 AGCTGCTATTAGCAATATGAAACCTCTTACATCAGTTACAATTTATA 
               
               
                   
               
               
                 TGCAGAAATATTTATATGCAGAAATATTGCTATTGCCTTAACCCAGA 
               
               
                   
               
               
                 AATTATCACTGTTATTCTTTAGAATGGTGCAAAGAGGCATGATACAT 
               
               
                   
               
               
                 TGTATCATTATTGCCCTGAAAGAAAGAGATTAGGGAAAGTATTAGAA 
               
               
                   
               
               
                 ATAAGATAAACAAAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAA 
               
               
                   
               
               
                 ATTACAAATGCAAAATTACCCTGATTTGGTCAATATGTGTACCCTGT 
               
               
                   
               
               
                 TACTTCTCCCCTTCCTATGACATGAACTTAACCATAGAAAAGAAGGG 
               
               
                   
               
               
                 GAAAGAAAACATCAAGGGTCCCATAGACTCACCCTGAAGTTCTCAGG 
               
               
                   
               
               
                 ATCCACGTGCAGCTTGTCACAGTGCAGCTCACTCAGtgtGGCAAAGG 
               
               
                   
               
               
                 TGCCCTTGAGgttGTCCAGGTGAGCCAGGCCATCACTAAAGGCACCG 
               
               
                   
               
               
                 AGCACTTTCTTGCCATGAGCCTTCACCTTAGGGTTGCCCATAACAGC 
               
               
                   
               
               
                 ATCAGGAGTGGACAGATCCCCAAAGGACTCAAAGAACCTCTGGGTCC 
               
               
                   
               
               
                 AAGGGTAGACCACCAGCAGCCTAAGGGTGGGAAAATAGACCAATAGG 
               
               
                   
               
               
                 CAGAGAGAGTCAGTGCCTATCAGAAACCCAAGAGTCTTCTCTGTCTC 
               
               
                   
               
               
                 CACATGCCCAGTTTCTATTGGTCTCCTTAAACCTGTCTTGTAACCTT 
               
               
                   
               
               
                 GATACCAACCTGCCCAGGGCCTCACCACCAACTGCATCCACGTTCAC 
               
               
                   
               
               
                 CTTGCCCCACAGGGCAGTAACGGCAGACTTCTCCTCAGGAGTCAGGT 
               
               
                   
               
               
                 GCACCATGGTGTCTGTTTGAGGTTGCTAGTGAACACAGTTGTGTCAG 
               
               
                   
               
               
                 AAGCAAATGT 
               
            
           
         
       
     
     In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 (β A-E22A ) comprises the nucleotide sequence set forth in SEQ ID NO: 56, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 56] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaaggcc 
               
               
                   
               
               
                 cttcataatatcccccagtttagtagttggacttagggaacaaaggaacc 
               
               
                   
               
               
                 tttaatagaaattggacagcaagaaagcgagcttagtgatacttgtgggc 
               
               
                   
               
               
                 cagggcattagccacaccagccaccactttctgataggcagcctgcactg 
               
               
                   
               
               
                 AgtggggtgAATTCTTTGCCAAAGTGATGGGCCAGCACACGACCAGCACG 
               
               
                   
               
               
                 TTGCCCAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAA 
               
               
                   
               
               
                 AAGGGCCTAGCTTGGACTCAGAATAATCCAGCCTTATCCCAACCATAAAA 
               
               
                   
               
               
                 TAAAAGCAGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACC 
               
               
                   
               
               
                 TCTTACATCAGTTACAATTTATATGCAGAAATATTTATATGCAGAAATAT 
               
               
                   
               
               
                 TGCTATTGCCTTAACCCAGAAATTATCACTGTTATTCTTTAGAATGGTGC 
               
               
                   
               
               
                 AAAGAGGCATGATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAG 
               
               
                   
               
               
                 GGAAAGTATTAGAAATAAGATAAACAAAAAAGTATATTAAAAGAAGAAAG 
               
               
                   
               
               
                 CATTTTTTAAAATTACAAATGCAAAATTACCCTGATTTGGTCAATATGTG 
               
               
                   
               
               
                 TACCCTGTTACTTCTCCCCTTCCTATGACATGAACTTAACCATAGAAAAG 
               
               
                   
               
               
                 AAGGGGAAAGAAAACATCAAGGGTCCCATAGACTCACCCTGAAGTTCTCA 
               
               
                   
               
               
                 GGATCCACGTGCAGCTTGTCACAGTGCAGCTCACTCAGtgtGGCAAAGGT 
               
               
                   
               
               
                 GCCCTTGAGgttGTCCAGGTGAGCCAGGCCATCACTAAAGGCACCGAGCA 
               
               
                   
               
               
                 CTTTCTTGCCATGAGCCTTCACCTTAGGGTTGCCCATAACAGCATCAGGA 
               
               
                   
               
               
                 GTGGACAGATCCCCAAAGGACTCAAAGAACCTCTGGGTCCAAGGGTAGAC 
               
               
                   
               
               
                 CACCAGCAGCCTAAGGGTGGGAAAATAGACCAATAGGCAGAGAGAGTCAG 
               
               
                   
               
               
                 TGCCTATCAGAAACCCAAGAGTCTTCTCTGTCTCCACATGCCCAGTTTCT 
               
               
                   
               
               
                 ATTGGTCTCCTTAAACCTGTCTTGTAACCTTGATACCAACCTGCCCAGGG 
               
               
                   
               
               
                 CCTCACCACCAACAGCATCCACGTTCACCTTGCCCCACAGGGCAGTAACG 
               
               
                   
               
               
                 GCAGACTTCTCCTCAGGAGTCAGGTGCACCATGGTGTCTGTTTGAGGTTG 
               
               
                   
               
               
                 CTAGTGAACACAGTTGTGTCAGAAGCAAATGT 
               
            
           
         
       
     
     In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 (β A-E22A ) comprises the nucleotide sequence set forth in SEQ ID NO: 57, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 57] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaaggccc 
               
               
                   
               
               
                 ttcataatatcccccagtttagtagttggacttagggaacaaaggaacctt 
               
               
                   
               
               
                 taatagaaattggacagcaagaaagcgagcttagtgatacttgtgggccag 
               
               
                   
               
               
                 ggcattagccacaccagccaccactttctgataggcagcctgcactggtgg 
               
               
                   
               
               
                 ggtgAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACGTTGCC 
               
               
                   
               
               
                 CAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAAAAGGGC 
               
               
                   
               
               
                 CTAGCTTGGACTCAGAATAATCCAGCCTTATCCCAACCATAAAATAAAAGC 
               
               
                   
               
               
                 AGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACCTCTTACAT 
               
               
                   
               
               
                 CAGTTACAATTTATATGCAGAAATATTTATATGCAGAAATATTGCTATTGC 
               
               
                   
               
               
                 CTTAACCCAGAAATTATCACTGTTATTCTTTAGAATGGTGCAAAGAGGCAT 
               
               
                   
               
               
                 GATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAGGGAAAGTATTA 
               
               
                   
               
               
                 GAAATAAGATAAACAAAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAAA 
               
               
                   
               
               
                 TTACAAATGCAAAATTACCCTGATTTGGTCAATATGTGTACCCTGTTACTT 
               
               
                   
               
               
                 CTCCCCTTCCTATGACATGAACTTAACCATAGAAAAGAAGGGGAAAGAAAA 
               
               
                   
               
               
                 CATCAAGGGTCCCATAGACTCACCCTGAAGTTCTCAGGATCCACGTGCAGC 
               
               
                   
               
               
                 TTGTCACAGTGCAGCTCACTCAGtgtGGCAAAGGTGCCCTTGAGgttGTCC 
               
               
                   
               
               
                 AGGTGAGCCAGGCCATCACTAAAGGCACCGAGCACTTTCTTGCCATGAGCC 
               
               
                   
               
               
                 TTCACCTTAGGGTTGCCCATAACAGCATCAGGAGTGGACAGATCCCCAAAG 
               
               
                   
               
               
                 GACTCAAAGAACCTCTGGGTCCAAGGGTAGACCACCAGCAGCCTAAGGGTG 
               
               
                   
               
               
                 GGAAAATAGACCAATAGGCAGAGAGAGTCAGTGCCTATCAGAAACCCAAGA 
               
               
                   
               
               
                 GTCTTCTCTGTCTCCACATGCCCAGTTTCTATTGGTCTCCTTAAACCTGTC 
               
               
                   
               
               
                 TTGTAACCTTGATACCAACCTGCCCAGGGCCTCACCACCAACCGCATCCAC 
               
               
                   
               
               
                 GTTCACCTTGCCCCACAGGGCAGTAACGGCAGACTTCTCCTCAGGAGTCAG 
               
               
                   
               
               
                 GTGCACCATGGTGTCTGTTTGAGGTTGCTAGTGAACACAGTTGTGTCAGAA 
               
               
                   
               
               
                 GCAAATGT 
               
            
           
         
       
     
     In certain non-limiting embodiments, the human β-globin gene is a human β A -globin gene encoding an asparagine to lysine mutation at codon 80 (β −N80K ). In certain embodiments, the human β A -globin gene encoding an asparagine to lysine mutation at codon 80 (β −N80K ) further comprises a deletion in intron 2 (e.g., an about 370 bp deletion). In certain embodiments, the human β A -globin gene encoding an asparagine to lysine mutation at codon 80 (β −N80K ) comprises the nucleotide sequence set forth in SEQ ID NO: 58, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 58] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaaggcc 
               
               
                   
               
               
                 cttcataatatcccccagtttagtagttggacttagggaacaaaggaacc 
               
               
                   
               
               
                 tttaatagaaattggacagcaagaaagcgagcttagtgatacttgtgggc 
               
               
                   
               
               
                 cagggcattagccacaccagccaccactttctgataggcagcctgcactg 
               
               
                   
               
               
                 gtggggtgAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACG 
               
               
                   
               
               
                 TTGCCCAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAA 
               
               
                   
               
               
                 AAGGGCCTAGCTTGGACTCAGAATAATCCAGCCTTATCCCAACCATAAAA 
               
               
                   
               
               
                 TAAAAGCAGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACC 
               
               
                   
               
               
                 TCTTACATCAGTTACAATTTATATGCAGAAATATTTATATGCAGAAATAT 
               
               
                   
               
               
                 TGCTATTGCCTTAACCCAGAAATTATCACTGTTATTCTTTAGAATGGTGC 
               
               
                   
               
               
                 AAAGAGGCATGATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAG 
               
               
                   
               
               
                 GGAAAGTATTAGAAATAAGATAAACAAAAAAGTATATTAAAAGAAGAAAG 
               
               
                   
               
               
                 CATTTTTTAAAATTACAAATGCAAAATTACCCTGATTTGGTCAATATGTG 
               
               
                   
               
               
                 TACCCTGTTACTTCTCCCCTTCCTATGACATGAACTTAACCATAGAAAAG 
               
               
                   
               
               
                 AAGGGGAAAGAAAACATCAAGGGTCCCATAGACTCACCCTGAAGTTCTCA 
               
               
                   
               
               
                 GGATCCACGTGCAGCTTGTCACAGTGCAGCTCACTCAGtgtGGCAAAGGT 
               
               
                   
               
               
                 GCCCTTGAGcttGTCCAGGTGAGCCAGGCCATCACTAAAGGCACCGAGCA 
               
               
                   
               
               
                 CTTTCTTGCCATGAGCCTTCACCTTAGGGTTGCCCATAACAGCATCAGGA 
               
               
                   
               
               
                 GTGGACAGATCCCCAAAGGACTCAAAGAACCTCTGGGTCCAAGGGTAGAC 
               
               
                   
               
               
                 CACCAGCAGCCTAAGGGTGGGAAAATAGACCAATAGGCAGAGAGAGTCAG 
               
               
                   
               
               
                 TGCCTATCAGAAACCCAAGAGTCTTCTCTGTCTCCACATGCCCAGTTTCT 
               
               
                   
               
               
                 ATTGGTCTCCTTAAACCTGTCTTGTAACCTTGATACCAACCTGCCCAGGG 
               
               
                   
               
               
                 CCTCACCACCAACttcATCCACGTTCACCTTGCCCCACAGGGCAGTAACG 
               
               
                   
               
               
                 GCAGACTTCTCCTCAGGAGTCAGGTGCACCATGGTGTCTGTTTGAGGTTG 
               
               
                   
               
               
                 CTAGTGAACACAGTTGTGTCAGAAGCAAATGT  
               
            
           
         
       
     
     In certain embodiments, the human β A -globin gene encoding an asparagine to lysine mutation at codon 80 (β −N80K ) comprises the nucleotide sequence set forth in SEQ ID NO: 59, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 59] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaaggcc 
               
               
                   
               
               
                 cttcataatatcccccagtttagtagttggacttagggaacaaaggaacc 
               
               
                   
               
               
                 tttaatagaaattggacagcaagaaagcgagcttagtgatacttgtgggc 
               
               
                   
               
               
                 cagggcattagccacaccagccaccactttctgataggcagcctgcactg 
               
               
                   
               
               
                 gtggggtgAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACG 
               
               
                   
               
               
                 TTGCCCAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAA 
               
               
                   
               
               
                 AAGGGCCTAGCTTGGACTCAGAATAATCCAGCCTTATCCCAACCATAAAA 
               
               
                   
               
               
                 TAAAAGCAGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACC 
               
               
                   
               
               
                 TCTTACATCAGTTACAATTTATATGCAGAAATATTTATATGCAGAAATAT 
               
               
                   
               
               
                 TGCTATTGCCTTAACCCAGAAATTATCACTGTTATTCTTTAGAATGGTGC 
               
               
                   
               
               
                 AAAGAGGCATGATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAG 
               
               
                   
               
               
                 GGAAAGTATTAGAAATAAGATAAACAAAAAAGTATATTAAAAGAAGAAAG 
               
               
                   
               
               
                 CATTTTTTAAAATTACAAATGCAAAATTACCCTGATTTGGTCAATATGTG 
               
               
                   
               
               
                 TACCCTGTTACTTCTCCCCTTCCTATGACATGAACTTAACCATAGAAAAG 
               
               
                   
               
               
                 AAGGGGAAAGAAAACATCAAGGGTCCCATAGACTCACCCTGAAGTTCTCA 
               
               
                   
               
               
                 GGATCCACGTGCAGCTTGTCACAGTGCAGCTCACTCAGtgtGGCAAAGGT 
               
               
                   
               
               
                 GCCCTTGAGTTTGTCCAGGTGAGCCAGGCCATCACTAAAGGCACCGAGCA 
               
               
                   
               
               
                 CTTTCTTGCCATGAGCCTTCACCTTAGGGTTGCCCATAACAGCATCAGGA 
               
               
                   
               
               
                 GTGGACAGATCCCCAAAGGACTCAAAGAACCTCTGGGTCCAAGGGTAGAC 
               
               
                   
               
               
                 CACCAGCAGCCTAAGGGTGGGAAAATAGACCAATAGGCAGAGAGAGTCAG 
               
               
                   
               
               
                 TGCCTATCAGAAACCCAAGAGTCTTCTCTGTCTCCACATGCCCAGTTTCT 
               
               
                   
               
               
                 ATTGGTCTCCTTAAACCTGTCTTGTAACCTTGATACCAACCTGCCCAGGG 
               
               
                   
               
               
                 CCTCACCACCAACttcATCCACGTTCACCTTGCCCCACAGGGCAGTAACG 
               
               
                   
               
               
                 GCAGACTTCTCCTCAGGAGTCAGGTGCACCATGGTGTCTGTTTGAGGTTG  
               
               
                   
               
               
                 CTAGTGAACACAGTTGTGTCAGAAGCAAATGT 
               
            
           
         
       
     
     In certain non-limiting embodiments, the human β-globin gene is human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and an asparagine to lysine mutation at codon 80. In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and an asparagine to lysine mutation at codon 80 further comprises a deletion in intron 2 (e.g., an about 370 bp deletion). In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and an asparagine to lysine mutation at codon 80 comprises the nucleotide sequence set forth in SEQ ID NO: 60, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 60] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaaggcc 
               
               
                   
               
               
                 cttcataatatcccccagtttagtagttggacttagggaacaaaggaacc 
               
               
                   
               
               
                 tttaatagaaattggacagcaagaaagcgagcttagtgatacttgtgggc 
               
               
                   
               
               
                 cagggcattaGCCACACCAGCCACCACTTTCTGATAGGCAGCCTGCACTG 
               
               
                   
               
               
                 GTGGGGTGAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACG 
               
               
                   
               
               
                 TTGCCCAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAA 
               
               
                   
               
               
                 AAGGGCCTAGCTTGGACTCAGAATAATCCAGCCTTATCCCAACCATAAAA 
               
               
                   
               
               
                 TAAAAGCAGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACC 
               
               
                   
               
               
                 TCTTACATCAGTTACAATTTATATGCAGAAATATTTATATGCAGAAATAT 
               
               
                   
               
               
                 TGCTATTGCCTTAACCCAGAAATTATCACTGTTATTCTTTAGAATGGTGC 
               
               
                   
               
               
                 AAAGAGGCATGATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAG 
               
               
                   
               
               
                 GGAAAGTATTAGAAATAAGATAAACAAAAAAGTATATTAAAAGAAGAAAG 
               
               
                   
               
               
                 CATTTTTTAAAATTACAAATGCAAAATTACCCTGATTTGGTCAATATGTG 
               
               
                   
               
               
                 TACCCTGTTACTTCTCCCCTTCCTATGACATGAACTTAACCATAGAAAAG 
               
               
                   
               
               
                 AAGGGGAAAGAAAACATCAAGGGTCCCATAGACTCACCCTGAAGTTCTCA 
               
               
                   
               
               
                 GGATCCACGTGCAGCTTGTCACAGTGCAGCTCACTCAGTGTGGCAAAGGT 
               
               
                   
               
               
                 GCCCTTGAGCTTGTCCAGGTGAGCCAGGCCATCACTAAAGGCACCGAGCA 
               
               
                   
               
               
                 CTTTCTTGCCATGAGCCTTCACCTTAGGGTTGCCCATAACAGCATCAGGA 
               
               
                   
               
               
                 GTGGACAGATCCCCAAAGGACTCAAAGAACCTCTGGGTCCAAGGGTAGAC 
               
               
                   
               
               
                 CACCAGCAGCCTAAGGGTGGGAAAATAGACCAATAGGCAGAGAGAGTCAG 
               
               
                   
               
               
                 TGCCTATCAGAAACCCAAGAGTCTTCTCTGTCTCCACATGCCCAGTTTCT 
               
               
                   
               
               
                 ATTGGTCTCCTTAAACCTGTCTTGTAACCTTGATACCAACCTGCCCAGGG 
               
               
                   
               
               
                 CCTCACCACCAACGGCATCCACGTTCACCTTGCCCCACAGGGCAGTAACG 
               
               
                   
               
               
                 GCAGACTTCTCCTCAGGAGTCAGGTGCACCATGGTGTCTGTTTGAGGTTG 
               
               
                   
               
               
                 CTAGTGAACACAGTTGTGTCAGAAGCAAATGT 
               
            
           
         
       
     
     In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and an asparagine to lysine mutation at codon 80 comprises the nucleotide sequence set forth in SEQ ID NO: 61, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 61] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaaggccc 
               
               
                   
               
               
                 ttcataatatcccccagtttagtagttggacttagggaacaaaggaacctt 
               
               
                   
               
               
                 taatagaaattggacagcaagaaagcgagcttagtgatacttgtgggccag 
               
               
                   
               
               
                 ggcattagccacaccagccaccactttctgataggcagcctgcactggtgg 
               
               
                   
               
               
                 ggtgAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACGTTGCC 
               
               
                   
               
               
                 CAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAAAAGGGC 
               
               
                   
               
               
                 CTAGCTTGGACTCAGAATAATCCAGCCTTATCCCAACCATAAAATAAAAGC 
               
               
                   
               
               
                 AGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACCTCTTACAT 
               
               
                   
               
               
                 CAGTTACAATTTATATGCAGAAATATTTATATGCAGAAATATTGCTATTGC 
               
               
                   
               
               
                 CTTAACCCAGAAATTATCACTGTTATTCTTTAGAATGGTGCAAAGAGGCAT 
               
               
                   
               
               
                 GATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAGGGAAAGTATTA 
               
               
                   
               
               
                 GAAATAAGATAAACAAAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAAA 
               
               
                   
               
               
                 TTACAAATGCAAAATTACCCTGATTTGGTCAATATGTGTACCCTGTTACTT 
               
               
                   
               
               
                 CTCCCCTTCCTATGACATGAACTTAACCATAGAAAAGAAGGGGAAAGAAAA 
               
               
                   
               
               
                 CATCAAGGGTCCCATAGACTCACCCTGAAGTTCTCAGGATCCACGTGCAGC 
               
               
                   
               
               
                 TTGTCACAGTGCAGCTCACTCAGtgtGGCAAAGGTGCCCTTGAGTttGTCC 
               
               
                   
               
               
                 AGGTGAGCCAGGCCATCACTAAAGGCACCGAGCACTTTCTTGCCATGAGCC 
               
               
                   
               
               
                 TTCACCTTAGGGTTGCCCATAACAGCATCAGGAGTGGACAGATCCCCAAAG 
               
               
                   
               
               
                 GACTCAAAGAACCTCTGGGTCCAAGGGTAGACCACCAGCAGCCTAAGGGTG 
               
               
                   
               
               
                 GGAAAATAGACCAATAGGCAGAGAGAGTCAGTGCCTATCAGAAACCCAAGA 
               
               
                   
               
               
                 GTCTTCTCTGTCTCCACATGCCCAGTTTCTATTGGTCTCCTTAAACCTGTC 
               
               
                   
               
               
                 TTGTAACCTTGATACCAACCTGCCCAGGGCCTCACCACCAACggcATCCAC 
               
               
                   
               
               
                 GTTCACCTTGCCCCACAGGGCAGTAACGGCAGACTTCTCCTCAGGAGTCAG 
               
               
                   
               
               
                 GTGCACCATGGTGTCTGTTTGAGGTTGCTAGTGAACACAGTTGTGTCAGAA 
               
               
                   
               
               
                 GCAAATGT 
               
            
           
         
       
     
     In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and an asparagine to lysine mutation at codon 80 comprises the nucleotide sequence set forth in SEQ ID NO: 62, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 62] 
               
               
                 GCAATGAAAATAAATGTTTTTTATTAGGCAGAATCCAGATGCTCAAGGCC 
               
               
                   
               
               
                 CTTCATAATATCCCCCAGTTTAGTAGTTGGACTTAGGGAACAAAGGAACC 
               
               
                   
               
               
                 TTTAATAGAAATTGGACAGCAAGAAAGCGAGCTTAGTGATACTTGTGGGC 
               
               
                   
               
               
                 CAGGGCATTAGCCACACCAGCCACCACTTTCTGATAGGCAGCCTGCACTG 
               
               
                   
               
               
                 GTGGGGTGAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACG 
               
               
                   
               
               
                 TTGCCCAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAA 
               
               
                   
               
               
                 AAGGGCCTAGCTTGGACTCAGAATAATCCAGCCTTATCCCAACCATAAAA 
               
               
                   
               
               
                 TAAAAGCAGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACC 
               
               
                   
               
               
                 TCTTACATCAGTTACAATTTATATGCAGAAATATTTATATGCAGAAATAT 
               
               
                   
               
               
                 TGCTATTGCCTTAACCCAGAAATTATCACTGTTATTCTTTAGAATGGTGC 
               
               
                   
               
               
                 AAAGAGGCATGATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAG 
               
               
                   
               
               
                 GGAAAGTATTAGAAATAAGATAAACAAAAAAGTATATTAAAAGAAGAAAG 
               
               
                   
               
               
                 CATTTTTTAAAATTACAAATGCAAAATTACCCTGATTTGGTCAATATGTG 
               
               
                   
               
               
                 TACCCTGTTACTTCTCCCCTTCCTATGACATGAACTTAACCATAGAAAAG 
               
               
                   
               
               
                 AAGGGGAAAGAAAACATCAAGGGTCCCATAGACTCACCCTGAAGTTCTCA 
               
               
                   
               
               
                 GGATCCACGTGCAGCTTGTCACAGTGCAGCTCACTCAGTGTGGCAAAGGT 
               
               
                   
               
               
                 GCCCTTGAGCTTGTCCAGGTGAGCCAGGCCATCACTAAAGGCACCGAGCA 
               
               
                   
               
               
                 CTTTCTTGCCATGAGCCTTCACCTTAGGGTTGCCCATAACAGCATCAGGA 
               
               
                   
               
               
                 GTGGACAGATCCCCAAAGGACTCAAAGAACCTCTGGGTCCAAGGGTAGAC 
               
               
                   
               
               
                 CACCAGCAGCCTAAGGGTGGGAAAATAGACCAATAGGCAGAGAGAGTCAG 
               
               
                   
               
               
                 TGCCTATCAGAAACCCAAGAGTCTTCTCTGTCTCCACATGCCCAGTTTCT 
               
               
                   
               
               
                 ATTGGTCTCCTTAAACCTGTCTTGTAACCTTGATACCAACCTGCCCAGGG 
               
               
                   
               
               
                 CCTCACCACCAACTGCATCCACGTTCACCTTGCCCCACAGGGCAGTAACG 
               
               
                   
               
               
                 GCAGACTTCTCCTCAGGAGTCAGGTGCACCATGGTGTCTGTTTGAGGTTG 
               
               
                   
               
               
                 CTAGTGAACACAGTTGTGTCAGAAGCAAATGT 
               
            
           
         
       
     
     In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and an asparagine to lysine mutation at codon 80 comprises the nucleotide sequence set forth in SEQ ID NO: 63, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 63] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaaggccc 
               
               
                   
               
               
                 ttcataatatcccccagtttagtagttggacttagggaacaaaggaacctt 
               
               
                   
               
               
                 taatagaaattggacagcaagaaagcgagcttagtgatacttgtgggccag 
               
               
                   
               
               
                 ggcattagccacaccagccaccactttctgataggcagcctgcactggtgg 
               
               
                   
               
               
                 ggtgAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACGTTGCC 
               
               
                   
               
               
                 CAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAAAAGGGC 
               
               
                   
               
               
                 CTAGCTTGGACTCAGAATAATCCAGCCTTATCCCAACCATAAAATAAAAGC 
               
               
                   
               
               
                 AGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACCTCTTACAT 
               
               
                   
               
               
                 CAGTTACAATTTATATGCAGAAATATTTATATGCAGAAATATTGCTATTGC 
               
               
                   
               
               
                 CTTAACCCAGAAATTATCACTGTTATTCTTTAGAATGGTGCAAAGAGGCAT 
               
               
                   
               
               
                 GATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAGGGAAAGTATTA 
               
               
                   
               
               
                 GAAATAAGATAAACAAAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAAA 
               
               
                   
               
               
                 TTACAAATGCAAAATTACCCTGATTTGGTCAATATGTGTACCCTGTTACTT 
               
               
                   
               
               
                 CTCCCCTTCCTATGACATGAACTTAACCATAGAAAAGAAGGGGAAAGAAAA 
               
               
                   
               
               
                 CATCAAGGGTCCCATAGACTCACCCTGAAGTTCTCAGGATCCACGTGCAGC 
               
               
                   
               
               
                 TTGTCACAGTGCAGCTCACTCAGTGTGGCAAAGGTGCCCTTGAGCTTGTCC 
               
               
                   
               
               
                 AGGTGAGCCAGGCCATCACTAAAGGCACCGAGCACTTTCTTGCCATGAGCC 
               
               
                   
               
               
                 TTCACCTTAGGGTTGCCCATAACAGCATCAGGAGTGGACAGATCCCCAAAG 
               
               
                   
               
               
                 GACTCAAAGAACCTCTGGGTCCAAGGGTAGACCACCAGCAGCCTAAGGGTG 
               
               
                   
               
               
                 GGAAAATAGACCAATAGGCAGAGAGAGTCAGTGCCTATCAGAAACCCAAGA 
               
               
                   
               
               
                 GTCTTCTCTGTCTCCACATGCCCAGTTTCTATTGGTCTCCTTAAACCTGTC 
               
               
                   
               
               
                 TTGTAACCTTGATACCAACCTGCCCAGGGCCTCACCACCAACAGCATCCAC 
               
               
                   
               
               
                 GTTCACCTTGCCCCACAGGGCAGTAACGGCAGACTTCTCCTCAGGAGTCAG 
               
               
                   
               
               
                 GTGCACCATGGTGTCTGTTTGAGGTTGCTAGTGAACACAGTTGTGTCAGAA 
               
               
                   
               
               
                 GCAAATGT 
               
            
           
         
       
     
     In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and an asparagine to lysine mutation at codon 80 comprises the nucleotide sequence set forth in SEQ ID NO: 64, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 64] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaaggccc 
               
               
                   
               
               
                 ttcataatatcccccagtttagtagttggacttagggaacaaaggaacctt 
               
               
                   
               
               
                 taatagaaattggacagcaagaaagcgagcttagtgatacttgtgggccag 
               
               
                   
               
               
                 ggcattagccacaccagccaccactttctgataggcagcctgcactggtgg 
               
               
                   
               
               
                 ggtgAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACGTTGCC 
               
               
                   
               
               
                 CAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAAAAGGGC 
               
               
                   
               
               
                 CTAGCTTGGACTCAGAATAATCCAGCCTTATCCCAACCATAAAATAAAAGC 
               
               
                   
               
               
                 AGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACCTCTTACAT 
               
               
                   
               
               
                 CAGTTACAATTTATATGCAGAAATATTTATATGCAGAAATATTGCTATTGC 
               
               
                   
               
               
                 CTTAACCCAGAAATTATCACTGTTATTCTTTAGAATGGTGCAAAGAGGCAT 
               
               
                   
               
               
                 GATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAGGGAAAGTATTA 
               
               
                   
               
               
                 GAAATAAGATAAACAAAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAAA 
               
               
                   
               
               
                 TTACAAATGCAAAATTACCCTGATTTGGTCAATATGTGTACCCTGTTACTT 
               
               
                   
               
               
                 CTCCCCTTCCTATGACATGAACTTAACCATAGAAAAGAAGGGGAAAGAAAA 
               
               
                   
               
               
                 CATCAAGGGTCCCATAGACTCACCCTGAAGTTCTCAGGATCCACGTGCAGC 
               
               
                   
               
               
                 TTGTCACAGTGCAGCTCACTCAGTGTGGCAAAGGTGCCCTTGAGCTTGTCC 
               
               
                   
               
               
                 AGGTGAGCCAGGCCATCACTAAAGGCACCGAGCACTTTCTTGCCATGAGCC 
               
               
                   
               
               
                 TTCACCTTAGGGTTGCCCATAACAGCATCAGGAGTGGACAGATCCCCAAAG 
               
               
                   
               
               
                 GACTCAAAGAACCTCTGGGTCCAAGGGTAGACCACCAGCAGCCTAAGGGTG 
               
               
                   
               
               
                 GGAAAATAGACCAATAGGCAGAGAGAGTCAGTGCCTATCAGAAACCCAAGA 
               
               
                   
               
               
                 GTCTTCTCTGTCTCCACATGCCCAGTTTCTATTGGTCTCCTTAAACCTGTC 
               
               
                   
               
               
                 TTGTAACCTTGATACCAACCTGCCCAGGGCCTCACCACCAACCGCATCCAC 
               
               
                   
               
               
                 GTTCACCTTGCCCCACAGGGCAGTAACGGCAGACTTCTCCTCAGGAGTCAG 
               
               
                   
               
               
                 GTGCACCATGGTGTCTGTTTGAGGTTGCTAGTGAACACAGTTGTGTCAGAA  
               
               
                   
               
               
                 GCAAATGT 
               
            
           
         
       
     
     In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and an asparagine to lysine mutation at codon 80 comprises the nucleotide sequence set forth in SEQ ID NO: 65, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 65] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaaggcc 
               
               
                   
               
               
                 cttcataatatcccccagtttagtagttggacttagggaacaaaggaacc 
               
               
                   
               
               
                 tttaatagaaattggacagcaagaaagcgagcttagtgatacttgtgggc 
               
               
                   
               
               
                 cagggcattagccacaccagccaccactttctgataggcagcctgcactg 
               
               
                   
               
               
                 gtggggtgAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACG 
               
               
                   
               
               
                 TTGCCCAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAA 
               
               
                   
               
               
                 AAGGGCCTAGCTTGGACTCAGAATAATCCAGCCTTATCCCAACCATAAAA 
               
               
                   
               
               
                 TAAAAGCAGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACC 
               
               
                   
               
               
                 TCTTACATCAGTTACAATTTATATGCAGAAATATTTATATGCAGAAATAT 
               
               
                   
               
               
                 TGCTATTGCCTTAACCCAGAAATTATCACTGTTATTCTTTAGAATGGTGC 
               
               
                   
               
               
                 AAAGAGGCATGATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAG 
               
               
                   
               
               
                 GGAAAGTATTAGAAATAAGATAAACAAAAAAGTATATTAAAAGAAGAAAG 
               
               
                   
               
               
                 CATTTTTTAAAATTACAAATGCAAAATTACCCTGATTTGGTCAATATGTG 
               
               
                   
               
               
                 TACCCTGTTACTTCTCCCCTTCCTATGACATGAACTTAACCATAGAAAAG 
               
               
                   
               
               
                 AAGGGGAAAGAAAACATCAAGGGTCCCATAGACTCACCCTGAAGTTCTCA 
               
               
                   
               
               
                 GGATCCACGTGCAGCTTGTCACAGTGCAGCTCACTCAGTGTGGCAAAGGT 
               
               
                   
               
               
                 GCCCTTGAGTTTGTCCAGGTGAGCCAGGCCATCACTAAAGGCACCGAGCA 
               
               
                   
               
               
                 CTTTCTTGCCATGAGCCTTCACCTTAGGGTTGCCCATAACAGCATCAGGA 
               
               
                   
               
               
                 GTGGACAGATCCCCAAAGGACTCAAAGAACCTCTGGGTCCAAGGGTAGAC 
               
               
                   
               
               
                 CACCAGCAGCCTAAGGGTGGGAAAATAGACCAATAGGCAGAGAGAGTCAG 
               
               
                   
               
               
                 TGCCTATCAGAAACCCAAGAGTCTTCTCTGTCTCCACATGCCCAGTTTCT 
               
               
                   
               
               
                 ATTGGTCTCCTTAAACCTGTCTTGTAACCTTGATACCAACCTGCCCAGGG 
               
               
                   
               
               
                 CCTCACCACCAACTGCATCCACGTTCACCTTGCCCCACAGGGCAGTAACG 
               
               
                   
               
               
                 GCAGACTTCTCCTCAGGAGTCAGGTGCACCATGGTGTCTGTTTGAGGTTG 
               
               
                   
               
               
                 CTAGTGAACACAGTTGTGTCAGAAGCAAATGT 
               
            
           
         
       
     
     In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and an asparagine to lysine mutation at codon 80 comprises the nucleotide sequence set forth in SEQ ID NO: 66, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 66] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaaggccc 
               
               
                   
               
               
                 ttcataatatcccccagtttagtagttggACTTAGGGAACAAAGGAACCTT 
               
               
                   
               
               
                 TAATAGAAATTGGACAGCAAGAAAGCGAGCTTAGTGATACTTGTGGGCCAG 
               
               
                   
               
               
                 GGCATTAGCCACACCAGCCACCACTTTCTGATAGGCAGCCTGCACTGGTGG 
               
               
                   
               
               
                 GGTGAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACGTTGCC 
               
               
                   
               
               
                 CAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAAAAGGGC 
               
               
                   
               
               
                 CTAGCTTGGACTCAGAATAATCCAGCCTTATCCCAACCATAAAATAAAAGC 
               
               
                   
               
               
                 AGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACCTCTTACAT 
               
               
                   
               
               
                 CAGTTACAATTTATATGCAGAAATATTTATATGCAGAAATATTGCTATTGC 
               
               
                   
               
               
                 CTTAACCCAGAAATTATCACTGTTATTCTTTAGAATGGTGCAAAGAGGCAT 
               
               
                   
               
               
                 GATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAGGGAAAGTATTA 
               
               
                   
               
               
                 GAAATAAGATAAACAAAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAAA 
               
               
                   
               
               
                 TTACAAATGCAAAATTACCCTGATTTGGTCAATATGTGTACCCTGTTACTT 
               
               
                   
               
               
                 CTCCCCTTCCTATGACATGAACTTAACCATAGAAAAGAAGGGGAAAGAAAA 
               
               
                   
               
               
                 CATCAAGGGTCCCATAGACTCACCCTGAAGTTCTCAGGATCCACGTGCAGC 
               
               
                   
               
               
                 TTGTCACAGTGCAGCTCACTCAGTGTGGCAAAGGTGCCCTTGAGTTTGTCC 
               
               
                   
               
               
                 AGGTGAGCCAGGCCATCACTAAAGGCACCGAGCACTTTCTTGCCATGAGCC 
               
               
                   
               
               
                 TTCACCTTAGGGTTGCCCATAACAGCATCAGGAGTGGACAGATCCCCAAAG 
               
               
                   
               
               
                 GACTCAAAGAACCTCTGGGTCCAAGGGTAGACCACCAGCAGCCTAAGGGTG 
               
               
                   
               
               
                 GGAAAATAGACCAATAGGCAGAGAGAGTCAGTGCCTATCAGAAACCCAAGA 
               
               
                   
               
               
                 GTCTTCTCTGTCTCCACATGCCCAGTTTCTATTGGTCTCCTTAAACCTGTC 
               
               
                   
               
               
                 TTGTAACCTTGATACCAACCTGCCCAGGGCCTCACCACCAACAGCATCCAC 
               
               
                   
               
               
                 GTTCACCTTGCCCCACAGGGCAGTAACGGCAGACTTCTCCTCAGGAGTCAG 
               
               
                   
               
               
                 GTGCACCATGGTGTCTGTTTGAGGTTGCTAGTGAACACAGTTGTGTCAGAA 
               
               
                   
               
               
                 GCAAATGT 
               
            
           
         
       
     
     In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and an asparagine to lysine mutation at codon 80 comprises the nucleotide sequence set forth in SEQ ID NO: 67, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 67] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaaggcc 
               
               
                   
               
               
                 cttcataatatcccccagtttagtagttggacttagggaacaaaggaacc 
               
               
                   
               
               
                 tttaatagaaattggacagcaagaaagcgagcttagtgatacttgtgggc 
               
               
                   
               
               
                 cagggcattagccacaccagccaccactttctgataggcagcctgcactg 
               
               
                   
               
               
                 gtggggtgAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACG 
               
               
                   
               
               
                 TTGCCCAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAA 
               
               
                   
               
               
                 AAGGGCCTAGCTTGGACTCAGAATAATCCAGCCTTATCCCAACCATAAAA 
               
               
                   
               
               
                 TAAAAGCAGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACC 
               
               
                   
               
               
                 TCTTACATCAGTTACAATTTATATGCAGAAATATTTATATGCAGAAATAT 
               
               
                   
               
               
                 TGCTATTGCCTTAACCCAGAAATTATCACTGTTATTCTTTAGAATGGTGC 
               
               
                   
               
               
                 AAAGAGGCATGATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAG 
               
               
                   
               
               
                 GGAAAGTATTAGAAATAAGATAAACAAAAAAGTATATTAAAAGAAGAAAG 
               
               
                   
               
               
                 CATTTTTTAAAATTACAAATGCAAAATTACCCTGATTTGGTCAATATGTG 
               
               
                   
               
               
                 TACCCTGTTACTTCTCCCCTTCCTATGACATGAACTTAACCATAGAAAAG 
               
               
                   
               
               
                 AAGGGGAAAGAAAACATCAAGGGTCCCATAGACTCACCCTGAAGTTCTCA 
               
               
                   
               
               
                 GGATCCACGTGCAGCTTGTCACAGTGCAGCTCACTCAGTGTGGCAAAGGT 
               
               
                   
               
               
                 GCCCTTGAGTTTGTCCAGGTGAGCCAGGCCATCACTAAAGGCACCGAGCA 
               
               
                   
               
               
                 CTTTCTTGCCATGAGCCTTCACCTTAGGGTTGCCCATAACAGCATCAGGA 
               
               
                   
               
               
                 GTGGACAGATCCCCAAAGGACTCAAAGAACCTCTGGGTCCAAGGGTAGAC 
               
               
                   
               
               
                 CACCAGCAGCCTAAGGGTGGGAAAATAGACCAATAGGCAGAGAGAGTCAG 
               
               
                   
               
               
                 TGCCTATCAGAAACCCAAGAGTCTTCTCTGTCTCCACATGCCCAGTTTCT 
               
               
                   
               
               
                 ATTGGTCTCCTTAAACCTGTCTTGTAACCTTGATACCAACCTGCCCAGGG 
               
               
                   
               
               
                 CCTCACCACCAACCGCATCCACGTTCACCTTGCCCCACAGGGCAGTAACG 
               
               
                   
               
               
                 GCAGACTTCTCCTCAGGAGTCAGGTGCACCATGGTGTCTGTTTGAGGTTG  
               
               
                   
               
               
                 CTAGTGAACACAGTTGTGTCAGAAGCAAATGT 
               
            
           
         
       
     
     In certain non-limiting embodiments, the human β-globin gene is a human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and a threonine to glutamine mutation at codon 87. In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and a threonine to glutamine mutation at codon 87 further comprises a deletion in intron 2 (e.g., an about 370 bp deletion). In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and a threonine to glutamine mutation at codon 87 comprises the nucleotide sequence set forth in SEQ ID NO: 68, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 68] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaaggcc 
               
               
                   
               
               
                 cttcataatatcccccagtttagtagttggacttagggaacaaaggaacc 
               
               
                   
               
               
                 tttaatagaaattggacagcaagaaagcgagcttagtgatacttgtgggc 
               
               
                   
               
               
                 cagggcattagccacaccagccaccactttctgataggcagcctgcactg 
               
               
                   
               
               
                 gtggggtgAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACG 
               
               
                   
               
               
                 TTGCCCAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAA 
               
               
                   
               
               
                 AAGGGCCTAGCTTGGACTCAGAATAATCCAGCCTTATCCCAACCATAAAA 
               
               
                   
               
               
                 TAAAAGCAGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACC 
               
               
                   
               
               
                 TCTTACATCAGTTACAATTTATATGCAGAAATATTTATATGCAGAAATAT 
               
               
                   
               
               
                 TGCTATTGCCTTAACCCAGAAATTATCACTGTTATTCTTTAGAATGGTGC 
               
               
                   
               
               
                 AAAGAGGCATGATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAG 
               
               
                   
               
               
                 GGAAAGTATTAGAAATAAGATAAACAAAAAAGTATATTAAAAGAAGAAAG 
               
               
                   
               
               
                 CATTTTTTAAAATTACAAATGCAAAATTACCCTGATTTGGTCAATATGTG 
               
               
                   
               
               
                 TACCCTGTTACTTCTCCCCTTCCTATGACATGAACTTAACCATAGAAAAG 
               
               
                   
               
               
                 AAGGGGAAAGAAAACATCAAGGGTCCCATAGACTCACCCTGAAGTTCTCA 
               
               
                   
               
               
                 GGATCCACGTGCAGCTTGTCACAGTGCAGCTCACTCAGCTGGGCAAAGGT 
               
               
                   
               
               
                 GCCCTTGAGGTTGTCCAGGTGAGCCAGGCCATCACTAAAGGCACCGAGCA 
               
               
                   
               
               
                 CTTTCTTGCCATGAGCCTTCACCTTAGGGTTGCCCATAACAGCATCAGGA 
               
               
                   
               
               
                 GTGGACAGATCCCCAAAGGACTCAAAGAACCTCTGGGTCCAAGGGTAGAC 
               
               
                   
               
               
                 CACCAGCAGCCTAAGGGTGGGAAAATAGACCAATAGGCAGAGAGAGTCAG 
               
               
                   
               
               
                 TGCCTATCAGAAACCCAAGAGTCTTCTCTGTCTCCACATGCCCAGTTTCT 
               
               
                   
               
               
                 ATTGGTCTCCTTAAACCTGTCTTGTAACCTTGATACCAACCTGCCCAGGG 
               
               
                   
               
               
                 CCTCACCACCAACGGCATCCACGTTCACCTTGCCCCACAGGGCAGTAACG 
               
               
                   
               
               
                 GCAGACTTCTCCTCAGGAGTCAGGTGCACCATGGTGTCTGTTTGAGGTTG  
               
               
                   
               
               
                 CTAGTGAACACAGTTGTGTCAGAAGCAAATGT 
               
            
           
         
       
     
     In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and a threonine to glutamine mutation at codon 87 comprises the nucleotide sequence set forth in SEQ ID NO: 69, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 69] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaaggccc 
               
               
                   
               
               
                 ttcataatatcccccagtttagtagttggacttagggaacaaaggaacctt 
               
               
                   
               
               
                 taatagaaattggacagcaagaaagcgagcttagtgatacttgtgggccag 
               
               
                   
               
               
                 ggcattagccacaccagccaccactttctgataggcagcctgcactggtgg 
               
               
                   
               
               
                 ggtgAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACGTTGCC 
               
               
                   
               
               
                 CAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAAAAGGGC 
               
               
                   
               
               
                 CTAGCTTGGACTCAGAATAATCCAGCCTTATCCCAACCATAAAATAAAAGC 
               
               
                   
               
               
                 AGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACCTCTTACAT 
               
               
                   
               
               
                 CAGTTACAATTTATATGCAGAAATATTTATATGCAGAAATATTGCTATTGC 
               
               
                   
               
               
                 CTTAACCCAGAAATTATCACTGTTATTCTTTAGAATGGTGCAAAGAGGCAT 
               
               
                   
               
               
                 GATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAGGGAAAGTATTA 
               
               
                   
               
               
                 GAAATAAGATAAACAAAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAAA 
               
               
                   
               
               
                 TTACAAATGCAAAATTACCCTGATTTGGTCAATATGTGTACCCTGTTACTT 
               
               
                   
               
               
                 CTCCCCTTCCTATGACATGAACTTAACCATAGAAAAGAAGGGGAAAGAAAA 
               
               
                   
               
               
                 CATCAAGGGTCCCATAGACTCACCCTGAAGTTCTCAGGATCCACGTGCAGC 
               
               
                   
               
               
                 TTGTCACAGTGCAGCTCACTCAGTTGGGCAAAGGTGCCCTTGAGGTTGTCC 
               
               
                   
               
               
                 AGGTGAGCCAGGCCATCACTAAAGGCACCGAGCACTTTCTTGCCATGAGCC 
               
               
                   
               
               
                 TTCACCTTAGGGTTGCCCATAACAGCATCAGGAGTGGACAGATCCCCAAAG 
               
               
                   
               
               
                 GACTCAAAGAACCTCTGGGTCCAAGGGTAGACCACCAGCAGCCTAAGGGTG 
               
               
                   
               
               
                 GGAAAATAGACCAATAGGCAGAGAGAGTCAGTGCCTATCAGAAACCCAAGA 
               
               
                   
               
               
                 GTCTTCTCTGTCTCCACATGCCCAGTTTCTATTGGTCTCCTTAAACCTGTC 
               
               
                   
               
               
                 TTGTAACCTTGATACCAACCTGCCCAGGGCCTCACCACCAACGGCATCCAC 
               
               
                   
               
               
                 GTTCACCTTGCCCCACAGGGCAGTAACGGCAGACTTCTCCTCAGGAGTCAG 
               
               
                   
               
               
                 GTGCACCATGGTGTCTGTTTGAGGTTGCTAGTGAACACAGTTGTGTCAGAA 
               
               
                   
               
               
                 GCAAATGT 
               
            
           
         
       
     
     In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and a threonine to glutamine mutation at codon 87 comprises the nucleotide sequence set forth in SEQ ID NO: 70, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 70] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaaggccc 
               
               
                   
               
               
                 ttcataatatcccccagtttagtagttggacttagggaacaaaggaacctt 
               
               
                   
               
               
                 taatagaaattggacagcaagaaagcgagcttagtgatacttgtgggccag 
               
               
                   
               
               
                 ggcattagccacaccagccaccactttctgataggcagcctgcactggtgg 
               
               
                   
               
               
                 ggtgAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACGTTGCC 
               
               
                   
               
               
                 CAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAAAAGGGC 
               
               
                   
               
               
                 CTAGCTTGGACTCAGAATAATCCAGCCTTATCCCAACCATAAAATAAAAGC 
               
               
                   
               
               
                 AGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACCTCTTACAT 
               
               
                   
               
               
                 CAGTTACAATTTATATGCAGAAATATTTATATGCAGAAATATTGCTATTGC 
               
               
                   
               
               
                 CTTAACCCAGAAATTATCACTGTTATTCTTTAGAATGGTGCAAAGAGGCAT 
               
               
                   
               
               
                 GATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAGGGAAAGTATTA 
               
               
                   
               
               
                 GAAATAAGATAAACAAAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAAA 
               
               
                   
               
               
                 TTACAAATGCAAAATTACCCTGATTTGGTCAATATGTGTACCCTGTTACTT 
               
               
                   
               
               
                 CTCCCCTTCCTATGACATGAACTTAACCATAGAAAAGAAGGGGAAAGAAAA 
               
               
                   
               
               
                 CATCAAGGGTCCCATAGACTCACCCTGAAGTTCTCAGGATCCACGTGCAGC 
               
               
                   
               
               
                 TTGTCACAGTGCAGCTCACTCAGCTGGGCAAAGGTGCCCTTGAGGTTGTCC 
               
               
                   
               
               
                 AGGTGAGCCAGGCCATCACTAAAGGCACCGAGCACTTTCTTGCCATGAGCC 
               
               
                   
               
               
                 TTCACCTTAGGGTTGCCCATAACAGCATCAGGAGTGGACAGATCCCCAAAG 
               
               
                   
               
               
                 GACTCAAAGAACCTCTGGGTCCAAGGGTAGACCACCAGCAGCCTAAGGGTG 
               
               
                   
               
               
                 GGAAAATAGACCAATAGGCAGAGAGAGTCAGTGCCTATCAGAAACCCAAGA 
               
               
                   
               
               
                 GTCTTCTCTGTCTCCACATGCCCAGTTTCTATTGGTCTCCTTAAACCTGTC 
               
               
                   
               
               
                 TTGTAACCTTGATACCAACCTGCCCAGGGCCTCACCACCAACTGCATCCAC 
               
               
                   
               
               
                 GTTCACCTTGCCCCACAGGGCAGTAACGGCAGACTTCTCCTCAGGAGTCAG 
               
               
                   
               
               
                 GTGCACCATGGTGTCTGTTTGAGGTTGCTAGTGAACACAGTTGTGTCAGAA 
               
               
                   
               
               
                 GCAAATGT 
               
            
           
         
       
     
     In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and a threonine to glutamine mutation at codon 87 comprises the nucleotide sequence set forth in SEQ ID NO: 71, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 71] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaagg 
               
               
                   
               
               
                 cccttcataatatcccccagtttagtagttggacttagggaacaaagg 
               
               
                   
               
               
                 aacctttaatagaaattggacagcaagaaagcgagcttagtgatactt 
               
               
                   
               
               
                 gtgggccagggcattagccacaccagccaccactttctgataggcagc 
               
               
                   
               
               
                 ctgcactggtggggtgAATTCTTTGCCAAAGTGATGGGCCAGCACACA 
               
               
                   
               
               
                 GACCAGCACGTTGCCCAGGAGCTGTGGGAGGAAGATAAGAGGTATGAA 
               
               
                   
               
               
                 CATGATTAGCAAAAGGGCCTAGCTTGGACTCAGAATAATCCAGCCTTA 
               
               
                   
               
               
                 TCCCAACCATAAAATAAAAGCAGAATGGTAGCTGGATTGTAGCTGCTA 
               
               
                   
               
               
                 TTAGCAATATGAAACCTCTTACATCAGTTACAATTTATATGCAGAAAT 
               
               
                   
               
               
                 ATTTATATGCAGAAATATTGCTATTGCCTTAACCCAGAAATTATCACT 
               
               
                   
               
               
                 GTTATTCTTTAGAATGGTGCAAAGAGGCATGATACATTGTATCATTAT 
               
               
                   
               
               
                 TGCCCTGAAAGAAAGAGATTAGGGAAAGTATTAGAAATAAGATAAACA 
               
               
                   
               
               
                 AAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAAATTACAAATGCAA 
               
               
                   
               
               
                 AATTACCCTGATTTGGTCAATATGTGTACCCTGTTACTTCTCCCCTTC 
               
               
                   
               
               
                 CTATGACATGAACTTAACCATAGAAAAGAAGGGGAAAGAAAACATCAA 
               
               
                   
               
               
                 GGGTCCCATAGACTCACCCTGAAGTTCTCAGGATCCACGTGCAGCTTG 
               
               
                   
               
               
                 TCACAGTGCAGCTCACTCAGCTGGGCAAAGGTGCCCTTGAGGTTGTCC 
               
               
                   
               
               
                 AGGTGAGCCAGGCCATCACTAAAGGCACCGAGCACTTTCTTGCCATGA 
               
               
                   
               
               
                 GCCTTCACCTTAGGGTTGCCCATAACAGCATCAGGAGTGGACAGATCC 
               
               
                   
               
               
                 CCAAAGGACTCAAAGAACCTCTGGGTCCAAGGGTAGACCACCAGCAGC 
               
               
                   
               
               
                 CTAAGGGTGGGAAAATAGACCAATAGGCAGAGAGAGTCAGTGCCTATC 
               
               
                   
               
               
                 AGAAACCCAAGAGTCTTCTCTGTCTCCACATGCCCAGTTTCTATTGGT 
               
               
                   
               
               
                 CTCCTTAAACCTGTCTTGTAACCTTGATACCAACCTGCCCAGGGCCTC 
               
               
                   
               
               
                 ACCACCAACAGCATCCACGTTCACCTTGCCCCACAGGGCAGTAACGGC 
               
               
                   
               
               
                 AGACTTCTCCTCAGGAGTCAGGTGCACCATGGTGTCTGTTTGAGGTTG 
               
               
                   
               
               
                 CTAGTGAACACAGTTGTGTCAGAAGCAAATGT 
               
            
           
         
       
     
     In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and a threonine to glutamine mutation at codon 87 comprises the nucleotide sequence set forth in SEQ ID NO: 72, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 72] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaagg 
               
               
                   
               
               
                 cccttcataatatcccccagtttagtagttggacttagggaacaaagg 
               
               
                   
               
               
                 aacctttaatagaaattggacagcaagaaagcgagcttagtgatactt 
               
               
                   
               
               
                 gtgggccagggcattagccacaccagccaccactttctgataggcagc 
               
               
                   
               
               
                 ctgcactggtggggtgAATTCTTTGCCAAAGTGATGGGCCAGCACACA 
               
               
                   
               
               
                 GACCAGCACGTTGCCCAGGAGCTGTGGGAGGAAGATAAGAGGTATGAA 
               
               
                   
               
               
                 CATGATTAGCAAAAGGGCCTAGCTTGGACTCAGAATAATCCAGCCTTA 
               
               
                   
               
               
                 TCCCAACCATAAAATAAAAGCAGAATGGTAGCTGGATTGTAGCTGCTA 
               
               
                   
               
               
                 TTAGCAATATGAAACCTCTTACATCAGTTACAATTTATATGCAGAAAT 
               
               
                   
               
               
                 ATTTATATGCAGAAATATTGCTATTGCCTTAACCCAGAAATTATCACT 
               
               
                   
               
               
                 GTTATTCTTTAGAATGGTGCAAAGAGGCATGATACATTGTATCATTAT 
               
               
                   
               
               
                 TGCCCTGAAAGAAAGAGATTAGGGAAAGTATTAGAAATAAGATAAACA 
               
               
                   
               
               
                 AAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAAATTACAAATGCAA 
               
               
                   
               
               
                 AATTACCCTGATTTGGTCAATATGTGTACCCTGTTACTTCTCCCCTTC 
               
               
                   
               
               
                 CTATGACATGAACTTAACCATAGAAAAGAAGGGGAAAGAAAACATCAA 
               
               
                   
               
               
                 GGGTCCCATAGACTCACCCTGAAGTTCTCAGGATCCACGTGCAGCTTG 
               
               
                   
               
               
                 TCACAGTGCAGCTCACTCAGCTGGGCAAAGGTGCCCTTGAGGTTGTCC 
               
               
                   
               
               
                 AGGTGAGCCAGGCCATCACTAAAGGCACCGAGCACTTTCTTGCCATGA 
               
               
                   
               
               
                 GCCTTCACCTTAGGGTTGCCCATAACAGCATCAGGAGTGGACAGATCC 
               
               
                   
               
               
                 CCAAAGGACTCAAAGAACCTCTGGGTCCAAGGGTAGACCACCAGCAGC 
               
               
                   
               
               
                 CTAAGGGTGGGAAAATAGACCAATAGGCAGAGAGAGTCAGTGCCTATC 
               
               
                   
               
               
                 AGAAACCCAAGAGTCTTCTCTGTCTCCACATGCCCAGTTTCTATTGGT 
               
               
                   
               
               
                 CTCCTTAAACCTGTCTTGTAACCTTGATACCAACCTGCCCAGGGCCTC 
               
               
                   
               
               
                 ACCACCAACCGCATCCACGTTCACCTTGCCCCACAGGGCAGTAACGGC 
               
               
                   
               
               
                 AGACTTCTCCTCAGGAGTCAGGTGCACCATGGTGTCTGTTTGAGGTTG 
               
               
                   
               
               
                 CTAGTGAACACAGTTGTGTCAGAAGCAAATGT 
               
            
           
         
       
     
     In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and a threonine to glutamine mutation at codon 87 comprises the nucleotide sequence set forth in SEQ ID NO: 73, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 73] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaagg 
               
               
                   
               
               
                 cccttcataatatcccccagtttagtagttggacttagggaacaaagg 
               
               
                   
               
               
                 aacctttaatagaaattggacagcaagaaagcgagcttagtgatactt 
               
               
                   
               
               
                 gtgggccagggcattagccacaccagccaccactttctgataggcagc 
               
               
                   
               
               
                 ctgcactggtggggtgAATTCTTTGCCAAAGTGATGGGCCAGCACACA 
               
               
                   
               
               
                 GACCAGCACGTTGCCCAGGAGCTGTGGGAGGAAGATAAGAGGTATGAA 
               
               
                   
               
               
                 CATGATTAGCAAAAGGGCCTAGCTTGGACTCAGAATAATCCAGCCTTA 
               
               
                   
               
               
                 TCCCAACCATAAAATAAAAGCAGAATGGTAGCTGGATTGTAGCTGCTA 
               
               
                   
               
               
                 TTAGCAATATGAAACCTCTTACATCAGTTACAATTTATATGCAGAAAT 
               
               
                   
               
               
                 ATTTATATGCAGAAATATTGCTATTGCCTTAACCCAGAAATTATCACT 
               
               
                   
               
               
                 GTTATTCTTTAGAATGGTGCAAAGAGGCATGATACATTGTATCATTAT 
               
               
                   
               
               
                 TGCCCTGAAAGAAAGAGATTAGGGAAAGTATTAGAAATAAGATAAACA 
               
               
                   
               
               
                 AAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAAATTACAAATGCAA 
               
               
                   
               
               
                 AATTACCCTGATTTGGTCAATATGTGTACCCTGTTACTTCTCCCCTTC 
               
               
                   
               
               
                 CTATGACATGAACTTAACCATAGAAAAGAAGGGGAAAGAAAACATCAA 
               
               
                   
               
               
                 GGGTCCCATAGACTCACCCTGAAGTTCTCAGGATCCACGTGCAGCTTG 
               
               
                   
               
               
                 TCACAGTGCAGCTCACTCAGTTGGGCAAAGGTGCCCTTGAGGTTGTCC 
               
               
                   
               
               
                 AGGTGAGCCAGGCCATCACTAAAGGCACCGAGCACTTTCTTGCCATGA 
               
               
                   
               
               
                 GCCTTCACCTTAGGGTTGCCCATAACAGCATCAGGAGTGGACAGATCC 
               
               
                   
               
               
                 CCAAAGGACTCAAAGAACCTCTGGGTCCAAGGGTAGACCACCAGCAGC 
               
               
                   
               
               
                 CTAAGGGTGGGAAAATAGACCAATAGGCAGAGAGAGTCAGTGCCTATC 
               
               
                   
               
               
                 AGAAACCCAAGAGTCTTCTCTGTCTCCACATGCCCAGTTTCTATTGGT 
               
               
                   
               
               
                 CTCCTTAAACCTGTCTTGTAACCTTGATACCAACCTGCCCAGGGCCTC 
               
               
                   
               
               
                 ACCACCAACTGCATCCACGTTCACCTTGCCCCACAGGGCAGTAACGGC 
               
               
                   
               
               
                 AGACTTCTCCTCAGGAGTCAGGTGCACCATGGTGTCTGTTTGAGGTTG 
               
               
                   
               
               
                 CTAGTGAACACAGTTGTGTCAGAAGCAAATGT 
               
            
           
         
       
     
     In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and a threonine to glutamine mutation at codon 87 comprises the nucleotide sequence set forth in SEQ ID NO: 74, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 74] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaagg 
               
               
                   
               
               
                 cccttcataatatcccccagtttagtagttggacttagggaacaaagg 
               
               
                   
               
               
                 aacctttaatagaaattggacagcaagaaagcgagcttagtgatactt 
               
               
                   
               
               
                 gtgggccagggcattaGCCACACCAGCCACCACTTTCTGATAGGCAGC 
               
               
                   
               
               
                 CTGCACTGGTGGGGTGAATTCTTTGCCAAAGTGATGGGCCAGCACACA 
               
               
                   
               
               
                 GACCAGCACGTTGCCCAGGAGCTGTGGGAGGAAGATAAGAGGTATGAA 
               
               
                   
               
               
                 CATGATTAGCAAAAGGGCCTAGCTTGGACTCAGAATAATCCAGCCTTA 
               
               
                   
               
               
                 TCCCAACCATAAAATAAAAGCAGAATGGTAGCTGGATTGTAGCTGCTA 
               
               
                   
               
               
                 TTAGCAATATGAAACCTCTTACATCAGTTACAATTTATATGCAGAAAT 
               
               
                   
               
               
                 ATTTATATGCAGAAATATTGCTATTGCCTTAACCCAGAAATTATCACT 
               
               
                   
               
               
                 GTTATTCTTTAGAATGGTGCAAAGAGGCATGATACATTGTATCATTAT 
               
               
                   
               
               
                 TGCCCTGAAAGAAAGAGATTAGGGAAAGTATTAGAAATAAGATAAACA 
               
               
                   
               
               
                 AAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAAATTACAAATGCAA 
               
               
                   
               
               
                 AATTACCCTGATTTGGTCAATATGTGTACCCTGTTACTTCTCCCCTTC 
               
               
                   
               
               
                 CTATGACATGAACTTAACCATAGAAAAGAAGGGGAAAGAAAACATCAA 
               
               
                   
               
               
                 GGGTCCCATAGACTCACCCTGAAGTTCTCAGGATCCACGTGCAGCTTG 
               
               
                   
               
               
                 TCACAGTGCAGCTCACTCAGTTGGGCAAAGGTGCCCTTGAGGTTGTCC 
               
               
                   
               
               
                 AGGTGAGCCAGGCCATCACTAAAGGCACCGAGCACTTTCTTGCCATGA 
               
               
                   
               
               
                 GCCTTCACCTTAGGGTTGCCCATAACAGCATCAGGAGTGGACAGATCC 
               
               
                   
               
               
                 CCAAAGGACTCAAAGAACCTCTGGGTCCAAGGGTAGACCACCAGCAGC 
               
               
                   
               
               
                 CTAAGGGTGGGAAAATAGACCAATAGGCAGAGAGAGTCAGTGCCTATC 
               
               
                   
               
               
                 AGAAACCCAAGAGTCTTCTCTGTCTCCACATGCCCAGTTTCTATTGGT 
               
               
                   
               
               
                 CTCCTTAAACCTGTCTTGTAACCTTGATACCAACCTGCCCAGGGCCTC 
               
               
                   
               
               
                 ACCACCAACAGCATCCACGTTCACCTTGCCCCACAGGGCAGTAACGGC 
               
               
                   
               
               
                 AGACTTCTCCTCAGGAGTCAGGTGCACCATGGTGTCTGTTTGAGGTTG 
               
               
                   
               
               
                 CTAGTGAACACAGTTGTGTCAGAAGCAAATGT 
               
            
           
         
       
     
     In certain embodiments, the human β A -globin gene encoding a glutamic acid to alanine mutation at codon 22 and a threonine to glutamine mutation at codon 87 comprises the nucleotide sequence set forth in SEQ ID NO: 75, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 75] 
               
               
                 gcaatgaaaataaatgttttttattaggcagaatccagatgctcaagg 
               
               
                   
               
               
                 cccttcataatatcccccagtttagtagttggACTTAGGGAACAAAGG 
               
               
                   
               
               
                 AACCTTTAATAGAAATTGGACAGCAAGAAAGCGAGCTTAGTGATACTT 
               
               
                   
               
               
                 GTGGGCCAGGGCATTAGCCACACCAGCCACCACTTTCTGATAGGCAGC 
               
               
                   
               
               
                 CTGCACTGGTGGGGTGAATTCTTTGCCAAAGTGATGGGCCAGCACACA 
               
               
                   
               
               
                 GACCAGCACGTTGCCCAGGAGCTGTGGGAGGAAGATAAGAGGTATGAA 
               
               
                   
               
               
                 CATGATTAGCAAAAGGGCCTAGCTTGGACTCAGAATAATCCAGCCTTA 
               
               
                   
               
               
                 TCCCAACCATAAAATAAAAGCAGAATGGTAGCTGGATTGTAGCTGCTA 
               
               
                   
               
               
                 TTAGCAATATGAAACCTCTTACATCAGTTACAATTTATATGCAGAAAT 
               
               
                   
               
               
                 ATTTATATGCAGAAATATTGCTATTGCCTTAACCCAGAAATTATCACT 
               
               
                   
               
               
                 GTTATTCTTTAGAATGGTGCAAAGAGGCATGATACATTGTATCATTAT 
               
               
                   
               
               
                 TGCCCTGAAAGAAAGAGATTAGGGAAAGTATTAGAAATAAGATAAACA 
               
               
                   
               
               
                 AAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAAATTACAAATGCAA 
               
               
                   
               
               
                 AATTACCCTGATTTGGTCAATATGTGTACCCTGTTACTTCTCCCCTTC 
               
               
                   
               
               
                 CTATGACATGAACTTAACCATAGAAAAGAAGGGGAAAGAAAACATCAA 
               
               
                   
               
               
                 GGGTCCCATAGACTCACCCTGAAGTTCTCAGGATCCACGTGCAGCTTG 
               
               
                   
               
               
                 TCACAGTGCAGCTCACTCAGTTGGGCAAAGGTGCCCTTGAGGTTGTCC 
               
               
                   
               
               
                 AGGTGAGCCAGGCCATCACTAAAGGCACCGAGCACTTTCTTGCCATGA 
               
               
                   
               
               
                 GCCTTCACCTTAGGGTTGCCCATAACAGCATCAGGAGTGGACAGATCC 
               
               
                   
               
               
                 CCAAAGGACTCAAAGAACCTCTGGGTCCAAGGGTAGACCACCAGCAGC 
               
               
                   
               
               
                 CTAAGGGTGGGAAAATAGACCAATAGGCAGAGAGAGTCAGTGCCTATC 
               
               
                   
               
               
                 AGAAACCCAAGAGTCTTCTCTGTCTCCACATGCCCAGTTTCTATTGGT 
               
               
                   
               
               
                 CTCCTTAAACCTGTCTTGTAACCTTGATACCAACCTGCCCAGGGCCTC 
               
               
                   
               
               
                 ACCACCAACCGCATCCACGTTCACCTTGCCCCACAGGGCAGTAACGGC 
               
               
                   
               
               
                 AGACTTCTCCTCAGGAGTCAGGTGCACCATGGTGTCTGTTTGAGGTTG 
               
               
                   
               
               
                 CTAGTGAACACAGTTGTGTCAGAAGCAAATGT 
               
            
           
         
       
     
     In certain embodiments, the site of the human β-globin gene IVS2 deletion is located at nucleotides 650-652 of SEQ ID NO: 36. In certain embodiments, nucleotides 1 to 261 of SEQ ID NO: 36 is the nucleotide sequence of exon 3 of the human β-globin gene. In certain embodiments, nucleotides 738 to 960 of SEQ ID NO: 36 is the nucleotide sequence of exon 2 of the human β-globin gene. certain embodiments, nucleotides 1091 to 1232 of SEQ ID NO: 36 is the nucleotide sequence of exon 1 of the human β-globin gene. 
     3.3. Promoters 
     In accordance with the presently disclosed subject matter, the expression cassette can further comprise a β-globin promoter. In certain embodiments, the β-globin promoter is positioned between the globin gene or functional portion thereof and the β-globin LCR. The length and the sequence of the β-globin promoter can vary. In certain embodiments, the β-globin promoter is from about 100 bp to about 1600 bp in length, e.g., from about 200 bp to about 700 bp, from about 100 bp to about 200 bp, from about 200 bp to about 300 bp, from about 300 bp to about 400 bp, from about 400 bp to about 500 bp, from about 500 bp to about 600 bp, from about 600 bp to about 700 bp, from about 700 bp to about 800 bp, from about 800 bp to about 900 bp, from about 900 bp to about 1000 bp, from about 1000 bp to about 1100 bp, from about 1100 bp to about 1200 bp, from about 1200 bp to about 1300 bp, from about 1300 bp to about 1400 bp, from about 1400 bp to about 1500 bp, or from about 1500 bp to about 1600 bp in length. In certain embodiments, the β-globin promoter a human β-globin promoter that is about 130 bp, about 613 bp, about 265 bp, or about 1555 bp, in length. In certain embodiments, the β-globin promoter is a human β-globin promoter that is about 613 bp in length. In certain non-limiting embodiments, the human β-globin promoter has the nucleotide sequence set forth in SEQ ID NO:10, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 10] 
               
               
                 AAGCAATAGATGGCTCTGCCCTGACTTTTATGCCCAGCCCTGGCTCCT 
               
               
                   
               
               
                 GCCCTCCCTGCTCCTGGGAGTAGATTGGCCAACCCTAGGGTGTGGCTC 
               
               
                   
               
               
                 CACAGGGTGAGGTCTAAGTGATGACAGCCGTACCTGTCCTTGGCTCTT 
               
               
                   
               
               
                 CTGGCACTGGCTTAGGAGTTGGACTTCAAACCCTCAGCCCTCCCTCTA 
               
               
                   
               
               
                 AGATATATCTCTTGGCCCCATACCATCAGTACAAATTGCTACTAAAAA 
               
               
                   
               
               
                 CATCCTCCTTTGCAAGTGTATTTACGTAATATTTGGAATCACAGCTTG 
               
               
                   
               
               
                 GTAAGCATATTGAAGATCGTTTTCCCAATTTTCTTATTACACAAATAA 
               
               
                   
               
               
                 GAAATTGATGCACTAAAAGTGGAAGAGTTTTGTCTACCATAATTCAGC 
               
               
                   
               
               
                 TTTGGGATATGTAGATGGATCTCTTCCTGCGTCTCCAGAATATGCAAA 
               
               
                   
               
               
                 ATACTTACAGGACAGAATGGATGAAAACTCTACCTCAGTTCTAAGCAT 
               
               
                   
               
               
                 ATCTTCTCCTTATTTGGATTAAAACCTTCTGGTAAGAAAAGAAAAAAA 
               
               
                   
               
               
                 ATATATATATATATGTGTATATATACACACATACATATACATATATAT 
               
               
                   
               
               
                 GCATTCATTTGTTGTTGTTTTTCTTAATTTGCTCATG 
               
            
           
         
       
     
     In certain embodiments, the β-globin promoter is a human β-globin promoter that is about 265 bp in length. In certain non-limiting embodiments, the human β-globin promoter has the nucleotide sequence set forth in SEQ ID NO:11. 
     
       
         
           
               
            
               
                 [SEQ ID NO: 11] 
               
               
                 AAGCAATAGATGGCTCTGCCCTGACTTTTATGCCCAGCCCTGGCTCCT 
               
               
                   
               
               
                 GCCCTCCCTGCTCCTGGGAGTAGATTGGCCAACCCTAGGGTGTGGCTC 
               
               
                   
               
               
                 CACAGGGTGAGGTCTAAGTGATGACAGCCGTACCTGTCCTTGGCTCTT 
               
               
                   
               
               
                 CTGGCACTGGCTTAGGAGTTGGACTTCAAACCCTCAGCCCTCCCTCTA 
               
               
                   
               
               
                 AGATATATCTCTTGGCCCCATACCATCAGTACAAATTGCTACTAAAAA 
               
               
                   
               
               
                 CATCCTCCTTTGCAAGTGTATTTAC 
               
            
           
         
       
     
     3.4. Human β-globin 3′ Enhancers Additionally or alternatively, a presently disclosed expression cassette can further comprise a human β-globin 3′ enhancer. In certain embodiments, the human β-globin 3′ enhancer is positioned in the upstream of the globin gene or functional portion thereof. In certain embodiments, the β-globin 3′ enhancer is from about 500 bp to about 1000 bp in length, e.g., from about 500 bp to about 600 bp, from about 600 bp to about 700 bp, from about 700 bp to about 800 bp, or from about 800 bp to about 900 bp in length. In certain embodiments, the human β-globin 3′ enhancer is about 879 bp in length. In certain embodiments, the human β-globin 3′ enhancer has the nucleotide sequence set forth in SEQ ID NO:12, which is provided below. 
     
       
         
           
               
            
               
                 [SEQ ID NO: 12] 
               
               
                 TAGGTATTGAATAAGAAAAATGAAGTTAAGGTGGTTGATGGTAACACT 
               
               
                   
               
               
                 ATGCTAATAACTGCAGAGCCAGAAGCACCATAAGGGACATGATAAGGG 
               
               
                   
               
               
                 AGCCAGCAGACCTCTGATCTCTTCCTGAATGCTAATCTTAAACATCCT 
               
               
                   
               
               
                 GAGGAAGAATGGGACTTCCATTTGGGGTGGGCCTATGATAGGGTAATA 
               
               
                   
               
               
                 AGACAGTAGTGAATATCAAGCTACAAAAAGCCCCCTTTCAAATTCTTC 
               
               
                   
               
               
                 TCAGTCCTAACTTTTCATACTAAGCCCAGTCCTTCCAAAGCAGACTGT 
               
               
                   
               
               
                 GAAAGAGTGATAGTTCCGGGAGACTAGCACTGCAGATTCCGGGTCACT 
               
               
                   
               
               
                 GTGAGTGGGGGAGGCAGGGAAGAAGGGCTCACAGGACAGTCAAACCAT 
               
               
                   
               
               
                 GCCCCCTGTTTTTCCTTCTTCAAGTAGACCTCTATAAGACAACAGAGA 
               
               
                   
               
               
                 CAACTAAGGCTGAGTGGCCAGGCGAGGAGAAACCATCTCGCCGTAAAA 
               
               
                   
               
               
                 CATGGAAGGAACACTTCAGGGGAAAGGTGGTATCTCTAAGCAAGAGAA 
               
               
                   
               
               
                 CTGAGTGGAGTCAAGGCTGAGAGATGCAGGATAAGCAAATGGGTAGTG 
               
               
                   
               
               
                 AAAAGACATTCATGAGGACAGCTAAAACAATAAGTAATGTAAAATACA 
               
               
                   
               
               
                 GCATAGCAAAACTTTAACCTCCAAATCAAGCCTCTACTTGAATCCTTT 
               
               
                   
               
               
                 TCTGAGGGATGAATAAGGCATAGGCATCAGGGGCTGTTGCCAATGTGC 
               
               
                   
               
               
                 ATTAGCTGTTTGCAGCCTCACCTTCTTTCATGGAGTTTAAGATATAGT 
               
               
                   
               
               
                 GTATTTTCCCAAGGTTTGAACTAGCTCTTCATTTCTTTATGTTTTAAA 
               
               
                   
               
               
                 TGCACTGACCTCCCACATTCCCTTTTTAGTAAAATATTCAGAAATAAT 
               
               
                   
               
               
                 TTAAATACATCATTG 
               
            
           
         
       
     
     In certain embodiments, the human β-globin 3′ enhancer has the nucleotide sequence set forth in SEQ ID NO: 78, which is provided below. 
     
       
         
           
               
               
            
               
                   
                 [SEQ ID NO: 78] 
               
               
                   
                 CTAGGTATTGA ATAAGAAAAA TGAAGTTAAG GTGGTTGATG 
               
               
                   
                   
               
               
                   
                 GTAACACTAT GCTAATAACT GCAGAGCCAG AAGCACCATA 
               
               
                   
                   
               
               
                   
                 AGGGACATGA TAAGGGAGCC AGCAGACCTC TGATCTCTTC 
               
               
                   
                   
               
               
                   
                 CTGAATGCTA ATCTTAAACA TCCTGAGGAA GAATGGGACT 
               
               
                   
                   
               
               
                   
                 TCCATTTGGG GTGGGCCTAT GATAGGGTAA TAAGACAGTA 
               
               
                   
                   
               
               
                   
                 GTGAATATCA AGCTACAAAA AGCCCCCTTT CAAATTCTTC 
               
               
                   
                   
               
               
                   
                 TCAGTCCTAA CTTTTCATAC TAAGCCCAGT CCTTCCAAAG 
               
               
                   
                   
               
               
                   
                 CAGACTGTGA AAGAGTGATA GTTCCGGGAG ACTAGCACTG 
               
               
                   
                   
               
               
                   
                 CAGATTCCGG GTCACTGTGA GTGGGGGAGG CAGGGAAGAA 
               
               
                   
                   
               
               
                   
                 GGGCTCACAG GACAGTCAAA CCATGCCCCC TGTTTTTCCT 
               
               
                   
                   
               
               
                   
                 TCTTCAAGTA GACCTCTATA AGACAACAGA GACAACTAAG 
               
               
                   
                   
               
               
                   
                 GCTGAGTGGC CAGGCGAGGA GAAACCATCT CGCCGTAAAA 
               
               
                   
                   
               
               
                   
                 CATGGAAGGA ACACTTCAGG GGAAAGGTGG TATCTCTAAG 
               
               
                   
                   
               
               
                   
                 CAAGAGAACT GAGTGGAGTC AAGGCTGAGA GATGCAGGAT 
               
               
                   
                   
               
               
                   
                 AAGCAAATGG GTAGTGAAAA GACATTCATG AGGACAGCTA 
               
               
                   
                   
               
               
                   
                 AAACAATAAG TAATGTAAAA TACAGCATAG CAAAACTTTA 
               
               
                   
                   
               
               
                   
                 ACCTCCAAAT CAAGCCTCTA CTTGAATCCT TTTCTGAGGG 
               
               
                   
                   
               
               
                   
                 ATGAATAAGG CATAGGCATC AGGGGCTGTT GCCAATGTGC 
               
               
                   
                   
               
               
                   
                 ATTAGCTGTT TGCAGCCTCA CCTTCTTTCA TGGAGTTTAA 
               
               
                   
                   
               
               
                   
                 GATATAGTGT ATTTTCCCAA GGTTTGAACT AGCTCTTCAT 
               
               
                   
                   
               
               
                   
                 TTCTTTATGT TTTAAATGCA CTGACCTCCC ACATTCCCTT 
               
               
                   
                   
               
               
                   
                 TTTAGTAAAA TATTCAGAAA TAATTTAAAT ACATCATT 
               
            
           
         
       
     
     3.5. Erythroid-Specific Enhancers 
     Furthermore, a presently disclosed expression cassette can further comprise at least one erythroid-specific enhancer. The presently disclosed expression cassette allows for expression of a globin gene (e.g., human β-globin gene) in erythroid-specific fashion. The erythroid-specific enhancer can enhance the expression of the globin gene in erythroid-specific fashion. For example, the erythroid-specific enhancer lack enhancer activity in non-erythroid tissues. In particularly, for the β-globin LCR that lacks a HS2 region, which primarily functions as an expression enhancer, the addition of one or more erythroid-specific enhancers can compensate the enhancing activity of a HS2 region. Additionally, for an expression cassette that lacks a β-globin LCR (e.g., SNS26.B87.A1) or the β-globin LCR consists essentially of a HS3 region and does not comprise a HS1 region, a HS2 region, or a HS4 region (e.g., SN27.2B87.A1), the addition of one or more erythroid-specific enhancers are capable of driving the expression of the β-globin gene. See e.g., Table 1 and  FIG. 16 . Furthermore, the presently disclosed erythroid-specific enhancers do not decrease or reduce the titer of a vector comprising the expression cassette. The length of the erythroid-specific enhancer can vary, e.g., from about 100 bp to about 200 bp, from about 100 bp to about 120 bp, from about 120 bp to about 140 bp, from about 140 bp to about 200 (e.g., from about 140 bp to about 150 bp, from about 150 bp to about 160 bp, from about 160 bp to about 170 bp, from about 170 bp to about 180 bp, from about 180 bp to about 190 bp, or from about 190 bp to about 200 bp). In certain embodiments, the erythroid-specific enhancer has a length of from about 140 bp to about 200 bp. In certain non-limiting embodiments, the erythroid-specific enhancer has a length of 152 bp, which has the nucleotide sequence set forth in SEQ ID NO:13, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 13] 
               
               
                 TCTCCCACGCCCTGGTCTCAGCTTGGGGAGTGGTCAGACCCCAATGGC 
               
               
                   
               
               
                 GATAAACTCTGGCAACTTTATCTGTGcaCTGCAGGCTCAGCCCCAAca 
               
               
                   
               
               
                 GCTTTAGCTTTCACAAGCAGGCAGGGGAAGGGAAACACATATCTCCAG 
               
               
                   
               
               
                 ATATGAGG 
               
            
           
         
       
     
     In certain non-limiting embodiments, the erythroid-specific enhancer has a length of 157 bp, which has the nucleotide sequence set forth in SEQ ID NO:14, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 14] 
               
               
                 CTAAACCCCTCCCCCACCCTAGCCCCAAGCTTCATCTTAGCTCCACTC 
               
               
                   
               
               
                 CTGACCCTATCCAGCTAAAGGTCCCCACCCAGCTCCTGCCTATCTAGT 
               
               
                   
               
               
                 CATTGCATATGGCAAGACTTGAAAGTCCTATCTCAAAGCAGCAGAATT 
               
               
                   
               
               
                 ATCAGCTACGACT 
               
            
           
         
       
     
     In certain non-limiting embodiments, the erythroid-specific enhancer has a length of 141 bp, which has the nucleotide sequence set forth in SEQ ID NO:15, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 15] 
               
               
                 CCATCCCCCAGCACTCCCTGCCCCCACAGCCCAGACTTGACCAACTCC 
               
               
                   
               
               
                 CAGCTccGCCTGGGACTTCCAGATATGGGGCCCCACCCTTGCAGGCCT 
               
               
                   
               
               
                 TGGGGACGCTGAAGATATTGACTATCTGCGTGCCggAAAAGGGTG 
               
            
           
         
       
     
     In certain non-limiting embodiments, the erythroid-specific enhancer has a length of 171 bp, which has the nucleotide sequence set forth in SEQ ID NO:16, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 16] 
               
               
                 AAAGGCTGGGGGTGGGAGTAGCGGATTTGAAGCACTTGTTGGCCTACA 
               
               
                   
               
               
                 GAGGTGTGGCAAGCAGAGCACCTCAGAACTCAGGCGTACTGCCCGCCG 
               
               
                   
               
               
                 CCCGAGCCCTGCGAGGGCCGATAGCGAGGGTGTGGCCCTTATCTGCAC 
               
               
                   
               
               
                 CCAGCAGAGCGCCGGCGGGGTACGGTC 
               
            
           
         
       
     
     In certain non-limiting embodiments, the erythroid-specific enhancer has a length of 195 bp, which has the nucleotide sequence set forth in SEQ ID NO:17, which is provided below: 
     
       
         
           
               
            
               
                 [SEQ ID NO: 17] 
               
               
                 CAGTTGCCTCAGCTGAGTATGTCTTCTAAAGATAATGTCGATTGTGTA 
               
               
                   
               
               
                 TGGCTGATGGGATTCTAGGACCAAGCAAGAGGTTTTTTTTTTTCCCCC 
               
               
                   
               
               
                 ACATACTTAACGTTTCTATATTTCTATTTGAATTCGACTGGACAGTTC 
               
               
                   
               
               
                 CATTTGAATTATTTCTCTCTCTCTCTCTCTCTGACACATTTTATCTTG 
               
               
                   
               
               
                 CCA 
               
            
           
         
       
     
     The erythroid-specific enhancer can be located within the β-globin LCR, e.g., between any two of the HS1, HS2, HS3, and HS4 regions. In certain embodiments, the erythroid-specific enhancer is located between the HS1 and HS3 regions within the β-globin LCR. 
     Alternatively, the erythroid-specific enhancer can be located upstream or downstream of the insulator, e.g., where the expression cassette does not comprise a β-globin LCR. In certain embodiments, the erythroid-specific enhancer is located upstream of the erythroid-specific enhancer. 
     Furthermore, the erythroid-specific enhancer can be located upstream or downstream of any of the HS1, HS2, HS3, and HS4 regions. In certain embodiments, the erythroid-specific enhancer is located upstream of the HS3 region. 
     In certain embodiments, the β-globin LCR comprises or consists essentially of a HS1 region having the nucleotide sequence set forth in SEQ ID NO: 3; a HS3 region having the nucleotide sequence set forth in SEQ ID NO: 5; a HS4 region having nucleotides 115 to 868 of SEQ ID NO: 6; and an erythroid-specific enhancer having the nucleotide sequence set forth in SEQ ID NO: 15, wherein the erythroid-specific enhancer is positioned between the HS1 region and the HS3 region, e.g., the β-globin LCR of SNS24.B87.A1 shown in  FIG. 15 . 
     In certain non-limiting embodiments, the β-globin LCR comprises or consists essentially of a HS1 region having the nucleotide sequence set forth in SEQ ID NO: 3; a HS3 region having the nucleotide sequence set forth in SEQ ID NO: 34; and a HS4 region having the nucleotide sequence set forth in SEQ ID NO: 35; and an erythroid-specific enhancer having the nucleotide sequence set forth in SEQ ID NO: 15, wherein the erythroid-specific enhancer is positioned between the HS1 region and the HS3 region, e.g., as the β-globin LCR of SNS22.2.B87.A1 shown in  FIG. 13 . 
     Erythroid-specific enhancers can be identified and determined by any suitable methods known in the art. The erythroid-specific enhancers can be positioned at the 3′ UTR (downstream) or the 5′ UTR (downstream) of the β-globin LCR. In certain embodiments, the at least one erythroid-specific enhancer is positioned in the 5′ UTR of the β-globin LCR, e.g., the upstream of the HS3 region. The expression cassette can comprise one, two, three, four, or five erythroid-specific enhancers. In certain embodiments, the expression cassette comprises one erythroid-specific enhancer. In certain embodiments, the expression cassette comprises two erythroid-specific enhancers. In certain embodiments, the expression cassette comprises three erythroid-specific enhancers. In certain embodiments, the expression cassette comprises four erythroid-specific enhancers. In certain non-limiting embodiments, the expression cassette comprises five erythroid-specific enhancers. 
     3.6. Insulators 
     In accordance with the presently disclosed subject matter, the expression cassette comprises at least one of the above-described insulators. In certain embodiments, a presently disclosed expression cassette comprises at least one insulator comprising the CTCF binding site sequence set forth in SEQ ID NO:18, for example, but not limited to, an insulator comprising the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, or SEQ ID NO: 32 or a fragment thereof. In certain embodiments, a presently disclosed expression cassette comprises at least one insulator comprising or having the nucleotide sequence set forth in SEQ ID NO: 1. In various non-limiting embodiments, the insulator can be incorporated or inserted into one or both LTRs or elsewhere in the region of a presently disclosed expression cassette that integrates into the cellular genome. In certain embodiments, the insulator is positioned at the 3′ end of the expression cassette. In certain embodiments, the insulator is positioned at the 5′ end of the expression cassette. In certain embodiments, the expression cassette comprises two of the insulator disclosed herein, e.g., an insulator comprising the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, or SEQ ID NO: 32 or a fragment thereof, where one insulator is positioned at the 3′ end and the other insulator is positioned at the 5′ end of the expression cassette. 
     The presently disclosed insulators possess powerful enhancer blocking activity. In certain embodiments, the insulators possess barrier activity in addition to enhancer blocking activity. The presently disclosed insulators substantially decrease the risks of insertional mutagenesis and genotoxicity associated with viral vectors. Furthermore, when a presently disclosed insulator is incorporated into a vector, the insulator does not adversely effect vector titers of the vector. In certain embodiments, the insulators increase the in vivo expression of the globin gene or functional portion thereof. 
     3.7. Exemplary Expression Cassettes 
     For the purpose of illustration and not limitation,  FIGS. 1-4, 13, 14, and 15  show recombinant vectors comprising exemplary expression cassettes in accordance with certain embodiments of the presently disclosed subject matter. 
       FIG. 1  shows a recombinant vector comprising a presently disclosed expression cassette that comprises a human β A-T87Q  globin gene, which is operably linked to a β-globin LCR that comprises a 860 bp HS2 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:9), a 1301 bp HS3 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:5), and a 1065 bp HS4 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:7). 
       FIG. 2  shows one exemplary recombinant vector comprising an expression cassette in accordance with one embodiment of the presently disclosed subject matter.  FIG. 2  shows a recombinant vector comprising a presently disclosed expression cassette that comprises a human β A-T87Q  globin gene, which is operably linked to a β-globin LCR that comprises a 1.1 kb HS1 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:2), a 1301 bp HS3 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:5), and a 1065 bp HS4 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:6). 
       FIG. 3  shows one exemplary recombinant vector comprising an expression cassette in accordance with one embodiment of the presently disclosed subject matter.  FIG. 3  shows a recombinant vector comprising a presently disclosed expression cassette that comprises a human β A-T87Q  globin gene, which is operably linked to a β-globin LCR that comprises a 602 bp HS1 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:3), a 1301 bp HS3 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:5), and a 446 bp HS4 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:8). 
       FIG. 4  shows one exemplary recombinant vector comprising an expression cassette in accordance with one embodiment of the presently disclosed subject matter.  FIG. 4  shows a recombinant vector comprising a presently disclosed expression cassette that comprises a human β A-T87Q  globin gene, which is operably linked to a β-globin LCR that comprises a 1301 bp HS3 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:5), and a 1065 bp HS4 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:6). The expression cassette shown in  FIG. 4  also comprises the following five erythroid-specific enhancers (shown as “EE5” in  FIG. 4 ): one erythroid-specific enhancer having the nucleotide sequence set forth in SEQ ID NO:13, one erythroid-specific enhancer having the nucleotide sequence set forth in SEQ ID NO:14, one erythroid-specific enhancer having the nucleotide sequence set forth in SEQ ID NO:15, one erythroid-specific enhancer having the nucleotide sequence set forth in SEQ ID NO:16, and one erythroid-specific enhancer having the nucleotide sequence set forth in SEQ ID NO:17. 
       FIG. 13  shows the following five exemplary recombinant vectors: SNS22.2.B87A1, which comprises an expression cassette that comprises a human β A-T87Q  globin gene, which is operably linked to a β-globin LCR that comprises a 602 bp HS1 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:3), a 816 bp HS2 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:33), a 1301 bp HS3 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:34), and a 754 bp HS4 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:35); 
     SNS23.2.B87A1, which comprises an expression cassette that comprises a human β A-T87Q  globin gene, which is operably linked to a β-globin LCR that comprises a 816 bp HS2 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:33), a 1301 bp HS3 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:34), and a 754 bp HS4 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:35), wherein the β-globin LCR does not comprise a HS1 region; 
     SNS24.2.B87A1, which comprises an expression cassette that comprises a human β A-T87Q  globin gene, which is operably linked to a β-globin LCR that comprises a 602 bp HS1 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:3), a 1301 bp HS3 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:34), a 754 bp HS4 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:35); and an erythroid-specific enhancer having the nucleotide sequence set forth in SEQ ID NO:15, wherein the β-globin LCR does not comprise a HS2 region; 
     SNS26.B87A1, which comprises an expression cassette that comprises a human β A-T87Q  globin gene, and an erythroid-specific enhancer having the nucleotide sequence set forth in SEQ ID NO:15, wherein the expression cassette does not comprise a β-globin LCR; and 
     SNS27.2.B87.A1, which comprises an expression cassette that comprises an insulator comprising the nucleotide sequence set forth in SEQ ID NO: 1, and a human β A-T87Q  globin gene that is operably linked to a β-globin LCR comprising a 1301 bp HS3 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:34), and an erythroid-specific enhancer having the nucleotide sequence set forth in SEQ ID NO:15, wherein the β-globin LCR does not comprise a HS1 region, a HS2 region, or a HS4 region. 
       FIG. 15  shows five exemplary recombinant vectors including SNS22.B87A1, which comprises an expression cassette that comprises a human β A-T87Q  globin gene, which is operably linked to a β-globin LCR that comprises a 602 bp HS1 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:3), a 816 bp HS2 region (e.g., one having nucleotides 45 to 860 of SEQ ID NO: 9), a 1301 bp HS3 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:5), and a 754 bp HS4 region (e.g., one having nucleotides 115 to 868 of SEQ ID NO: 6); SNS23.B87A1, which comprises an expression cassette that comprises a human β A-T87Q  globin gene, which is operably linked to a β-globin LCR that comprises a 816 bp HS2 region (e.g., one having nucleotides 45 to 860 of SEQ ID NO: 9), a 1301 bp HS3 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:5), and a 754 bp HS4 region (e.g., one having nucleotides 115 to 868 of SEQ ID NO: 6); SNS24.B87A1, which comprises an expression cassette that comprises a human β A-T87Q  globin gene, which is operably linked to a β-globin LCR that comprises a 602 bp HS1 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:3), a 1301 bp HS3 region (e.g., one having the nucleotide sequence set forth in SEQ ID NO:5), a 754 bp HS4 region (e.g., one having nucleotides 115 to 868 of SEQ ID NO: 6); and an erythroid-specific enhancer having the nucleotide sequence set forth in SEQ ID NO:15; and 
     SNS26.B87.A1, which is also disclosed in  FIG. 13  as described above. 
     As shown in  FIGS. 1-4, 13, and 15 , each of the expression cassettes comprises an insulator, wherein the insulator has the nucleotide sequence set forth in SEQ ID NO:1 (i.e., insulator A1). In addition, as shown in  FIGS. 1-4, 13, 15 , each of the expression cassettes comprises a human β-globin 3′ enhancer that is positioned upstream of the human β-globin gene, wherein the human β-globin 3′ enhancer has a length of 879 bp and has the nucleotide sequence set forth in SEQ ID NO: 12. Additionally, as shown in  FIGS. 13 and 15 , each of SNS22.B87.A1, SNS23.B87.A1, SNS24.B87.A1, SNS22.2.B87.A1, SNS23.2.B87.A1, SNS24.2.B87.A1, SNS26.B87.A1, and SNS27.2B87.A1 comprises a 265 bp human β-globin promoter, wherein the human β-globin promoter has the nucleotide sequence set forth in SEQ ID NO: 11. TNS9.B87.A1 shown in  FIG. 15  comprises a 613 bp human β-globin promoter, wherein the human β-globin promoter has the nucleotide sequence set forth in SEQ ID NO: 10. 
     III. Vectors, Nucleases and CRISPR-Cas Systems 
     The presently disclosed subject matter provides vectors and delivery systems (e.g., a non-naturally occurring or engineered nucleases or a CRISPR-Cas system) comprising the above-described expression cassettes. The vectors and delivery systems are suitable delivery vehicles for the stable introduction of globin gene (e.g., human β-globin) into the genome of a broad range of target cells to increase expression of the globin protein (human β-globin protein) in the cell. 
     In certain embodiments, the vector is a retroviral vector (e.g., gamma retroviral vector or a lentiviral vector) that is employed for the introduction or transduction of the above-described expression cassette into the genome of a host cell (e.g., a hematopoietic stem cell, an embryonic stem cell, an induced pluripotent stem cell, or a hemogenic endothelium cell). In certain embodiments, the retroviral vector comprises an expression cassette that comprises one of the above-described insulators, e.g., insulator A1. The insulator can be positioned at the 3′ or the 5′ end of the expression cassette. In certain embodiments, the insulator is positioned at the 3′ end of the expression cassette. During reverse transcription and vector integration, the insulator positioned at the 3′ end is copied into the 5′ end of the expression cassette. The resulting topology places copies of the insulator between the genomic regions located at the 5′ LTR and the 3′ LTR of the integrated virus and enhancer activity from the 5′ LTR and internal package promoter, but does not contain the enhancer in the 3′ LTR. This topology can decrease genotoxicity, thereby resulting in decreased tumor formation and increased survival of the animals. 
     In certain embodiments, the vector is an Adeno-associated virus (AAV) vector. Non-limiting examples of AAV vectors include AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, and AAV11. In certain embodiments, the vector is an AAV6 vector. 
     In certain embodiments, the recombinant vector is a retroviral vector. In certain embodiments, the retroviral vector is a lentiviral vector. In certain embodiments, the recombinant vector is a self-inactivating (SIN) lentiviral vector in which the enhancer and promoter region of the 3′ long terminal repeat (“LTR”) is modified (e.g., a 133 bp deletion in the U3 region). SIN vectors can infect and integrate into the genome of non-dividing cells in vivo with an efficacy similar to that of wild-type vectors. SIN lentivirus vectors are disclosed in Miyoshi et al.,  J. Virol . (October 1998); 72:8150-8157, which is incorporated by reference in its entirety. SIN vectors can reduce the risk of creating undesired replication-competent virus because there is no complete U3 region in the 3′ LTR, thereby eliminating the ability of the virus to be passed (see Miyoshi 1998). 
     In certain embodiments, the retroviral vector is a gamma-retroviral vector. In certain embodiments, the retroviral vector is a SIN gamma-retroviral vector. 
     In certain embodiments, the recombinant vector further comprises one or more of a rev-responsive element (“RRE”), a central polypurine tract (“cPPT”), a 5′ LTR (in which the U3 region of the 5′ LTR comprises a heterologous promoter (e.g., replaced with a cytomegalovirus (CMV) HIV heterologous promoter) resulting in Tat-independent transcription with no decreases in virus titer), and a 3′ SIN LTR (e.g., a 3′ SIN LTR comprising a 389 bp deletion in the U3 region). 
     In certain embodiments, the recombinant vector further comprises a Woodchuck hepatitis post-regulatory element (WPRE) in the 3′ long terminal repeat (LTR) of the vector (e.g., 3′ to the R region in the 3′ LTR of the vector). In certain embodiments, the recombinant vector further comprises a bovine growth hormone polyadenylation signal in addition to the WPRE in the 3′ long terminal repeat (LTR) of the vector (e.g., 3′ to the R region in the 3′ LTR of the vector). The WPRE can increase the titer of the recombinant vector (e.g., can increase the vector titer by at least about 5 folds). Addition of a bovine growth hormone (“BGH”) polyadenylation (“polyA”) signal (“BGH/polyA”) to the WPRE can further increase the titer of the recombinant vector. In certain embodiments, the WPRE and BGH/polyA are eliminated after the vector is transfused to target cells, and thus, are not present in the proviruses. In certain embodiments, the WPRE and the BGH/polyA are not comprised within the expression cassette, and thus, not transferred to the cells transduced with the recombinant vector. 
     As shown in  FIGS. 1-4, 13, and 15 , the recombinant vector can further comprises a rev-responsive element (“RRE”), a central polypurine tract (“cPPT”), a 5′ LTR (in which the U3 region of the 5′ LTR comprises a heterologous promoter (e.g., replaced with a cytomegalovirus (CMV) HIV heterologous promoter) resulting in Tat-independent transcription with no decreases in virus titer), a 3′ SIN LTR comprising a 389 bp deletion in the U3 region, a WPRE, and a BGH/polyA in the 3′ long terminal repeat (LTR) of the vector (e.g., 3′ to the R region in the 3′ LTR). The heterologous promoter (e.g., a CMV/HIV heterologous promoter) can increase the RNA transcription and the production of the vector. The heterologous promoter (e.g., a CMV/HIV heterologous promoter) is eliminated after reverse transcription, and thus, is not present in the proviruses. 
     In certain non-limiting embodiments, a presently disclosed expression cassette can be cloned into a retroviral vector and expression can be driven from its endogenous promoter, from the retroviral long terminal repeat, or from an alternative internal promoter. Combinations of retroviral vector and an appropriate packaging line are also suitable, where the capsid proteins will be functional for infecting human cells. Various amphotropic virus-producing cell lines are known, including, but not limited to, PA12 (Miller, et al. (1985)  Mol. Cell. Biol.  5:431-437); PA317 (Miller, et al. (1986)  Mol. Cell. Biol.  6:2895-2902); and CRIP (Danos, et al. (1988)  Proc. Natl. Acad. Sci. USA  85:6460-6464). Non-amphotropic particles are suitable too, e.g., particles pseudotyped with VSVG, RD114 or GALV envelope and any other known in the art. 
     Suitable methods of transduction also include direct co-culture of the cells with producer cells, e.g., by the method of Bregni, et al. (1992)  Blood  80:1418-1422, or culturing with viral supernatant alone or concentrated vector stocks with or without appropriate growth factors and polycations, e.g., by the method of Xu, et al. (1994)  Exp. Hemat.  22:223-230; and Hughes, et al. (1992)  J. Clin. Invest.  89:1817. 
     Transducing viral vectors can be used to express a globin gene (e.g., a human β-globin gene) in a host cell (e.g., hematopoietic stem cells, an embryonic stem cell, or an induced pluripotent stem cell). In certain embodiments, the chosen vector exhibits high efficiency of infection and stable integration and expression (see, e.g., Cayouette et al., Human Gene Therapy (1997); 8:423-430; Kido et al., Current Eye Research (1996); 15:833-844; Bloomer et al., Journal of Virology (1997); 71:6641-6649; Naldini et al., Science (1996); 272:263 267; and Miyoshi et al., Proc. Natl. Acad. Sci. U.S.A. 94:10319, 1997). Other viral vectors that can be used include, for example, adenoviral, lentiviral, and adeno-associated viral vectors, vaccinia virus, a bovine papilloma virus, or a herpes virus, such as Epstein-Barr Virus (also see, for example, the vectors of Miller,  Human Gene Therapy  (1990); 15-14; Friedman,  Science  (1989); 244:1275-1281; Eglitis et al.,  BioTechniques  6:608-614, 1988; Tolstoshev et al.,  Current Opinion in Biotechnology  (1990); 1:55-61; Sharp,  The Lancet  (1991); 337: 1277-1278; Cornetta et al.,  Nucleic Acid Research and Molecular Biology  (1987) 36:311-322; Anderson,  Science  (1984); 226:401-409; Moen,  Blood Cells  (1991); 17:407-416; Miller et al.,  Biotechnology  (1989); 7:980-990; Le Gal La Salle et al.,  Science  (1993); 259:988-990; and Johnson,  Chest  (1995); 107:775-83S). Retroviral vectors are particularly well developed and have been used in clinical settings (Rosenberg et al.,  N. Engl. J. Med  (1990); 323:370; Anderson et al., U.S. Pat. No. 5,399,346). 
     The requirement for efficient delivery and integration make retroviral vectors suitable for transducing a presently disclosed expression cassette. Retroviral vectors can be derived from three genera of the retroviridae: the γ-retroviruses (also known as C-type murine retroviruses or oncoretroviruses), the lentiviruses, and the spumaviruses (also known as foamy viruses). Several reviews detailing molecular approaches for the generation of replication-defective retroviral particles are available (Cornetta et al. (2005); Cockrell &amp; Kafri (2007)). The vector itself, which encodes the therapeutic transgene or cDNA, retains the minimal viral sequences needed to enable packaging in viral particles in a packaging cell line, reverse transcription, and integration. The packaging cell expresses the necessary structural proteins and enzymes that are required to assemble an infectious recombinant particle that contains the vector sequence and the machinery needed for its reverse transcription and integration in the transduced cell. 
     While the manufacturing aspects of all retroviral vector types follow the same general principles, γ-retroviral, lentiviral and spumaviral vectors differ in some of their intrinsic biological properties. Gamma-retroviruses, including the prototypic murine leukaemia viruses (MLV), effectively infect many cell types but are unable to integrate in cells that do not proceed to S phase soon after infection. In contrast, lentiviruses and their vector derivatives can transduce nondividing cells (Follenzi &amp; Naldini, 2002; Salmon &amp; Trono, 2002) owing to their ability to translocate to the nucleus and integrate in the absence of cell division (Lewis &amp; Emerman, 1994; Goff, 2001). Another fundamental attribute of lentiviral vectors is their relative genomic stability, as established for globin lentiviral vectors (May et al., 2000), which contrasts with the genomic instability of MLV-based globin vectors (Leboulch et al., 1994; Sadelain et al., 1995). Lentiviral and foamy vectors further provide a greater packaging capacity (Kumar et al., 2001; Rethwilm, 2007). All three vector types have been used successfully for the transduction of cytokineactivated HSCs (Miyoshi et al., 1999; Josephson et al., 2002; Leurs et al., 2003). 
     These three vector systems differ in their integration patterns. The integration pattern of retroviruses is semi-random and biased towards genes and their vicinity in approximately two-thirds of all integration events (Schroder et al., 2002; Wu et al., 2003; Mitchell et al., 2004; De Palma et al., 2005; Trobridge et al., 2006). There are however subtle and possibly significant differences in their exact distribution. Gamma-retroviruses have a propensity to integrate upstream of transcribed genes, whereas lentiviruses and lentiviral vectors target the entire transcribed gene sequence. Foamy vectors appear to be less prone to intragenic integration (Trobridge et al., 2006). In certain embodiments, the vector comprising the expression cassette is a lentivirus vector. The vectors can be derived from human immunodeficiency-1 (HIV-1), human immunodeficiency-2 (HIV-2), simian immunodeficiency virus (SIV), feline immunodeficiency virus (Hy), bovine immunodeficiency virus (BIV), Jembrana Disease Virus (JDV), equine infectious anemia virus (EIAV), caprine arthritis encephalitis virus (CAEV) and the like. In one non-limiting embodiment, the lentiviral vector is an HIV vector. HIV-based constructs are the most efficient at transduction of human cells. 
     The semi-random pattern of vector integration exposes patients to the risk of insertional oncogenesis when the vector trans-activates a neighboring oncogene. This may result in clonal expansion (Ott et al, 2006; Cavazzana-Calvo et al, 2010), myelodysplasia (Stein et al, 2010) or leukaemia (Hacein-Bey-Abina et al, 2003, 2008; Howe et al, 2008). Targeted gene delivery strategies, utilizing a non-naturally occurring or engineered nuclease (including, but not limited to, Zinc-finger nuclease (ZNFs), meganuclease, transcription activator-like effector nuclease (TALEN)), or a CRISPR-Cas system, can reduce or even eliminate the concern of insertional oncogenesis that is inherent to the use of retroviral vectors. 
     Eukaryotic cells utilize two distinct DNA repair mechanisms in response to DNA double strand breaks (DSBs): Homologous recombination (HR) and non-homologous end-joining (NHEJ). The activation of the HR repair machinery depends on the cell cycle status, and it is restricted to the S and G2 phases; in contrast, the NHEJ pathway is active throughout the cell cycle. Mechanistically, HR is an error-free DNA repair mechanism, because it requires a homologous template to repair the damaged DNA strand. On the other hand, NHEJ is a template-independent repair mechanism that is imprecise, due to DNA end processing during repair that leads to insertions or deletions at the DNA break site (Moynahan &amp; Jasin, 2010). Because of its homology-based mechanism, HR has been used as a tool to site-specifically engineer the genome of different species. From a therapeutic perspective, HR has been successfully used to repair mutated genes, thus offering a promising approach to cell-mediated treatment of monogenic diseases (Porteus et al, 2006). 
     Gene targeting by HR requires the use of two homology arms that flank the transgene/target site of interest. Generally, standard plasmid DNAs have been used to deliver 5-10 kb homology arms along with transgenes for positive and negative selection. This method is commonly used to knock-out/knockin genes in mouse embryonic stem (mES) cells (Capecchi, 2005;  FIG. 2B ). In human cells, the use of this approach has allowed gene targeting with efficiencies in the order of 10 −6 , which are lower than in mES cells and are not therapeutically practical. HR efficiency can be increased by the introduction of DNA-doubled stranded breaks (DSBs) at the target site using specific rare-cutting endonucleases, resulting in over 1,000-fold increase in correct gene targeting (Jasin, 1996). The discovery of this phenomenon prompted the development of methods to create site-specific DSBs in the genome of different species. Various chimeric enzymes have been designed for this purpose over the last decade, namely zinc-finger nucleases (ZFNs), meganucleases, and transcription activator-like effector nucleases (TALENs). 
     ZFNs are modular chimeric proteins that contain a ZF-based DNA binding domain (DBD) and a FokI nuclease domain (Porteus &amp; Carroll, 2005). DBD is usually composed of three ZF domains, each with 3-base pair specificity; the FokI nuclease domain provides a DNA nicking activity, which is targeted by two flanking ZFNs. Owing to the modular nature of the DBD, any site in a genome could be targeted in principle. However, as a single ZFN can bind and nick DNA, there is potential for a high number of off-target effects, resulting in the activation of the NHEJ pathway that may either introduce insertions/deletions or integrate the targeting vector in a non-specific manner. Obligate FokI domains that can nick their respective DNA strand only when they form a heterodimer were recently reported (Doyon et al, 2011). The use of such obligate ZFNs can reduce the genotoxic effects of this approach. 
     Meganucleases (MNs)/homing endonucleases (HEs) are dsDNA nucleases that recognize and cleave large DNA sites (14-40 bp) with low cleavage frequencies in eukaryotic genomes (Paques &amp; Duchateau, 2007). Although this limits the potential target sites, MN-DNA structures have been used as a guide to specifically modify DNA-interacting residues in order to change the MN specificity (Marcaida et al, 2010). I-CreI has been successfully engineered to generate chimeric meganucleases that target the human XPC and RAG1 genes, and they have been shown to stimulate HR activity in mammalian cells with no evident genotoxicity (Redondo et al, 2008; Grizot et al, 2009). The genotoxicity of this approach will need to be compared to that of ZFNs and TALE nucleases. 
     TALENs are similar ZFN except that the DBD is derived from transcription activator-like effcetors (TALEs), which are virulent factors used by phytopathogenic bacteria (Herbers, 1992). The TALE DBD is modular, and it is composed of 34-residue repeats, and its DNA specificity is determined by the number and order of repeats (Herbers, 1992). Each repeat binds a single nucleotide in the target sequence through only two residues (Boch, 2011). The advantage over ZFN technology is the rapid construction of DBDs. 
     A number of studies have used these chimeric enzymes to stimulate HR for either gene addition or gene repair at their target site (Paques &amp; Duchateau, 2007; Urnov et al, 2010). Porteus designed a ZFN to a half site sequence from the human HBB that surrounds the sickle cell mutation nucleotide (Porteus, 2006). This ZFN targets the sequence and stimulates HR at a chimeric DNA target when combined with a ZFN targeting the Zif268 binding site. There have been recent advances in targeting genes in cord blood CD34 +  cells. Use of non-integrating lentiviruses to deliver ZFNs and the donor DNA in these cells to target the CCR5 gene was reported in Lombardo et al, 2007. Lombardo et al, 2007 showed gene addition at this locus with correct targeting in 80% of the positively selected cells. 
     The presently disclosed subject matter provides a non-naturally occurring or engineered nuclease comprising a presently disclosed expression cassette, as described above. Suitable nucleases include, but are not limited to, ZFNs, meganucleases, and TALENs. A presently disclosed nuclease comprises a DNA binding domain and a nuclease cleavage domain. The DNA binding domain of the nuclease can be engineered to bind to a sequence of choice, e.g., a predetermined site. An engineered DNA binding domain can have a distinct binding specificity, compared to a naturally occurring nuclease. Engineering methods include, but are not limited to, rational design and various types of selection. Any suitable cleavage domain can be operatively linked to a DNA-binding domain to form a nuclease. For example, Zinc-finger protein (ZFP) DNA-binding domains can be fused to nuclease cleavage domains to create ZFNs—a functional entity that is able to recognize its intended nucleic acid target through its engineered ZFP DNA binding domain and cause the DNA to be cut near the ZFP binding site via the nuclease activity. See, e.g., Kim et al.  Proc Nat&#39;l Acad Sci USA  (1996); 93(3):1156-1160. Likewise, TALE DNA-binding domains can be fused to nuclease cleavage domains to create TALENs. See, e.g., U.S. Publication No. 20110301073. 
     The cleavage domain can be heterologous to the DNA-binding domain, e.g., a meganuclease DNA-binding domain and cleavage domain from a different nuclease. Heterologous cleavage domains can be obtained from any endonuclease or exonuclease. Exemplary endonucleases from which a cleavage domain can be derived include, but are not limited to, restriction endonucleases and homing endonucleases. See, for example, 2002-2003 Catalog, New England Biolabs, Beverly, Mass.; and Belfort et al. (1997)  Nucleic Acids Res.  25:3379-3388. Additional enzymes which cleave DNA are known (e.g., S1 Nuclease; mung bean nuclease; pancreatic DNase I; micrococcal nuclease; yeast HO endonuclease; see also Linn et al. (eds.) Nucleases, Cold Spring Harbor Laboratory Press, 1993). One or more of these enzymes (or functional regions thereof) can be used as a source of cleavage domains and cleavage half-domains. 
     Similarly, a cleavage half-domain can be derived from the above-described nuclease that requires dimerization for cleavage activity. In general, two fusion proteins are required for cleavage if the fusion proteins comprise cleavage half-domains. Alternatively, a single protein comprising two cleavage half-domains can be used. The two cleavage half-domains can be derived from the same endonuclease (or functional portions thereof), or each cleavage half-domain can be derived from a different endonuclease (or functional portions thereof). 
     In certain embodiments, the nuclease comprises an expression cassette that comprises two of the insulators disclosed herein, e.g., two of the insulator having the nucleotide sequence set forth in SEQ ID NO: 1, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, or SEQ ID NO: 31. One of the two insulators is positioned at the 3′ end of the expression cassette, and the other insulator is positioned at the 5′ end of the expression cassette. 
     The presently disclosed subject matter also provides a non-naturally occurring or engineer CRISPR-Cas system comprising the above-described expression cassette. The CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-Cas (CRISPR Associated) system is an engineered nuclease system based on a bacterial system that can be used for genome engineering. It is based on part of the adaptive immune response of many bacteria and archea. When a virus or plasmid invades a bacterium, segments of the invader&#39;s DNA are converted into CRISPR RNAs (crRNA) by the “immune” response. The crRNA then associates, through a region of partial complementarity, with another type of RNA called tracrRNA to guide a CRISPR-Cas nuclease to a region homologous to the crRNA in the target DNA called a “proto spacer”. The CRISPR-Cas nuclease cleaves the DNA to generate blunt ends at the DSB at sites specified by a 20-nucleotide guide sequence contained within the crRNA transcript. The CRISPR-Cas nuclease requires both the crRNA and the tracrRNA for site specific DNA recognition and cleavage. This system has been engineered such that the crRNA and tracrRNA can be combined into one molecule (the “single guide RNA”); and the crRNA equivalent portion of the single guide RNA can be engineered to guide the CRISPR-Cas nuclease to target any desired sequence (see Jinek et al.,  Science  (2012); 337:816-821). Thus, the CRISPR-Cas system can be engineered to create a DSB at a desired target in a genome. In certain embodiments, the CRISPR-Cas system comprises a CRISPR-Cas nuclease and a single-guide RNA. Suitable examples of CRISPR-Cas nucleases include, but are not limited to, Cas1, Cas1B, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8, Cas9 (also known as Csn1 and Csx12), Cas10, Csy1, Csy2, Csy3, Cse1, Cse2, Csc1, Csc2, Csa5, Csn2, Csm2, Csm3, Csm4, Csm5, Csm6, Cmr1, Cmr3, Cmr4, Cmr5, Cmr6, Csb1, Csb2, Csb3, Csx17, Csx14, Csx10, Csx16, CsaX, Csx3, Csx1, Csx15, Csf1, Csf2, Csf3, Csf4, homologs thereof, or modified versions thereof. These CRISPR-Cas nucleases are known; for example, the amino acid sequence of  S. pyogenes  Cas9 protein may be found in the SwissProt database under accession number Q99ZW2. In some embodiments, the CRISPR-Cas nuclease has DNA cleavage activity, e.g., Cas9. In certain embodiments, the CRISPR-Cas nuclease is Cas9. The CRISPR-Cas nuclease can direct cleavage of one or both strands at the location of a target sequence (e.g., a genomic safe harbor site). Additionally, the CRISPR-Cas nuclease can direct cleavage of one or both strands within about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 50, 100, 200, 500, or more base pairs from the first or last nucleotide of a target sequence. 
     The presently disclosed nucleases and CRISPR-Cas system allow for targeted delivery of the expression cassette. In certain embodiments, a presently disclosed CRISPR-Cas system or the DNA binding domain of a presently disclosed nuclease binds to a genomic safe harbor site. A nuclease or the CRISPR-Cas system generates a double strand break at the genomic safe harbor site. Genomic safe harbor sites are intragenic or extragenic regions of the human genome that are able to accommodate the predictable expression of newly integrated DNA without adverse effects on the host cell or organism. A useful safe harbor must permit sufficient transgene expression to yield desired levels of the vector-encoded protein or non-coding RNA. A genomic safe harbor site also must not predispose cells to malignant transformation nor alter cellular functions. Methods for identifying genomic safe harbor sites are described in Sadelain et al., “Safe Harbours for the integration of new DNA in the human genome,” Nature Reviews (2012); 12:51-58; Papapetrou et al., “Genomic safe harbors permit high β-globin transgene expression in thalassemia induced pluripotent stem cells”  Nat Biotechnol . (2011) January; 29(1):73-8, which are incorporated by reference in their entireties. A presently disclosed genomic safe harbor site meets one or more (one, two, three, four, or five) of the following five criteria: (1) distance of at least 50 kb from the 5′ end of any gene (e.g., from the 5′ end of the gene), (ii) distance of at least 300 kb from any cancer-related gene, (iii) within an open/accessible chromatin structure (measured by DNA cleavage with natural or engineered nucleases), (iv) location outside a gene transcription unit and (v) location outside ultraconserved regions (UCRs), microRNA or long non-coding RNA of the human genome. As the most common insertional oncogenesis event is transactivation of neighboring tumor-promoting genes, the first two criteria exclude the portion of the human genome located near promoters of genes, in particular, cancer-related genes, which are genes functionally implicated in human cancers or the human homologs of genes implicated in cancer in model organisms. Proximity to miRNA genes is one exclusion criterion because miRNAs are implicated in the regulation of many cellular processes, including cell proliferation and differentiation. As vector integration within a transcription unit can disrupt gene function through the loss of function of a tumor suppressor gene or the generation of an aberrantly spliced gene product, the fourth (iv) criterion excludes all sites located inside transcribed genes. UCRs, which are regions that are highly conserved over multiple vertebrates and known to be enriched for enhancers and exons, and long non-coding RNAs, are also excluded. In certain embodiments, the genomic safe harbor site is an extragenic genomic safe harbor site. In certain embodiments, the genomic safe harbor site is located on chromosome 1. 
     The presently disclosed subject matter also provides polynucleotides encoding the above-described nucleases, vectors comprising the polynucleotides encoding the above-described nucleases, polynucleotides encoding the above-described CRISPR-Cas system, and vectors comprising the polynucleotides encoding the above-described CRISPR-Cas system. 
     The nucleases and polynucleotides encoding these nucleases, and the CRISPR-Cas system and polynucleotides encoding the CRISPR-Cas system can be delivered in vivo or ex vivo by any suitable means. For example, nucleases and CRISPR-Cas system as described herein can be delivered to a cell (e.g., a hematopoietic stem cell, an embryonic stem cell, an induced pluripotent stem cell, or an hemogenic endothelium cell) by a vector comprising polynucleotides encoding the nuclease or the CRISPR-Cas system. Any vectors can be used including, but not limited to, plasmid vectors, retroviral vectors (e.g., γ-retroviral vectors, lentiviral vectors and foamy viral vectors), adenovirus vectors, poxvirus vectors; herpes virus vectors and adena-associated virus vectors, etc. In certain embodiments, the vector comprising a polynucleotide encoding an above-described nuclease or an above-described CRISPR-Cas system is a lentiviral vector. In one particular embodiment, the lentiviral vector is a non-integrating lentiviral vector. Examples of non-integrating lentiviral vector are described in Ory et al. (1996)  Proc. Natl. Acad. Sci. USA  93:11382-11388; Dull et al., (1998)  J. Viral.  72:8463-8471; Zuffery et al. (1998)  J. Viral.  72:9873-9880; Follenzi et al., (2000)  Nature Genetics  25:217-222; U.S. Patent Publication No 2009/054985. 
     Additionally, non-viral approaches (e.g., single-stranded DNA) can also be employed for the expression of a globin gene in cells. For example, a nucleic acid molecule can be introduced into a cell by administering the nucleic acid in the presence of lipofection (Feigner et al.,  Proc. Natl. Acad. Sci. U.S.A.  84:7413, 1987; Ono et al.,  Neuroscience Letters  17:259, 1990; Brigham et al.,  Am. J. Med. Sci.  298:278, 1989; Staubinger et al.,  Methods in Enzymology  101:512, 1983), asialoorosomucoid-polylysine conjugation (Wu et al., Journal of Biological Chemistry 263:14621, 1988; Wu et al.,  Journal of Biological Chemistry  264:16985, 1989), or by micro-injection under surgical conditions (Wolff et al., Science 247:1465, 1990). Other non-viral means for gene transfer include transfection in vitro using calcium phosphate, DEAE dextran, electroporation, and protoplast fusion. Liposomes can also be potentially beneficial for delivery of DNA into a cell. Transplantation of normal genes into the affected tissues of a subject can also be accomplished by transferring a normal nucleic acid into a cultivatable cell type ex vivo (e.g., an autologous or heterologous primary cell or progeny thereof), after which the cell (or its descendants) are injected into a targeted tissue or are injected systemically. Recombinant receptors can also be derived or obtained using transposases. Transient expression may be obtained by RNA electroporation. 
     IV. Cells 
     Genetic modification of cells (e.g., hematopoietic stem cells, embryonic stem cells, induced pluripotent stem cells, and hemogenic endothelium cells) can be accomplished by transducing a substantially homogeneous cell composition with a recombinant DNA or RNA construct (e.g., a vector or a delivery system comprising the above-described expression cassette). The presently disclosed subject matter provides cells transduced with the above-described expression cassettes, cells transduced with the above-described vectors, and cells transduced with the above-described nucleases or with vectors comprising polynucleotides encoding the nucleases, and cell transduced with the above-described CARISPR-Cas system or with vectors comprising polynucleotides encoding the CARISPR-Cas system, which are collectively referred to as “transduced cells”. As described above, the vectors, nucleases and CRISPR-Cas system are employed for transduction of the expression cassette to the cells to express a globin gene (e.g., a human β-globin gene). In certain embodiments, the transduced cells are administered to a subject to treat and/or prevent a hematopoietic disease, disorder, or condition. The presently disclosed insulators can enhance the efficiency of the transduction of the expression cassette to cells. 
     Suitable transduced cells include, but are not limited to, stem cells, progenitor cells, and differentiated cells. As used herein, the term “progenitor” or “progenitor cells” refers to cells that have the capacity to self-renew and to differentiate into more mature cells. Progenitor cells have a reduced potency compared to pluripotent and multipotent stem cells. Many progenitor cells differentiate along a single lineage, but may also have quite extensive proliferative capacity. 
     In certain embodiments, the transduced cells are stem cells. Stem cells have the ability to differentiate into the appropriate cell types when administered to a particular biological niche, in vivo. A stem cell is an undifferentiated cell capable of (1) long term self-renewal, or the ability to generate at least one identical copy of the original cell, (2) differentiation at the single cell level into multiple, and in some instance only one, specialized cell type and (3) of in vivo functional regeneration of tissues. Stem cells are sub-classified according to their developmental potential as totipotent, pluripotent, multipotent and oligo/unipotent. As used herein, the term “pluripotent” means the ability of a cell to form all lineages of the body or soma (i.e., the embryo proper). For example, embryonic stem cells are a type of pluripotent stem cells that are able to form cells from each of the three germs layers, the ectoderm, the mesoderm, and the endoderm. As used herein, the term “multipotent” refers to the ability of an adult stem cell to form multiple cell types of one lineage. For example, hematopoietic stem cells are capable of forming all cells of the blood cell lineage, e.g., lymphoid and myeloid cells. 
     In certain embodiments, the transduced cells are embryonic stem cells, bone marrow stem cells, umbilical cord stem cells, placental stem cells, mesenchymal stem cells, neural stem cells, liver stem cells, pancreatic stem cells, cardiac stem cells, kidney stem cells, and/or hematopoietic stem cells. In certain embodiments, the transduced cells are hematopoietic stem cells (HSCs). HSCs give rise to committed hematopoietic progenitor cells (HPCs) that are capable of generating the entire repertoire of mature blood cells over the lifetime of an organism. The term “hematopoietic stem cell” or “HSC” refers to multipotent stem cells that give rise to all blood cell types of an organism, including myeloid (e.g., monocytes and macrophages, neutrophils, basophils, eosinophils, erythrocytes, megakaryocytes/platelets, dendritic cells), and lymphoid lineages (e.g., T-cells, B-cells, NK-cells). When transplanted into lethally irradiated animals or humans, hematopoietic stem and progenitor cells can repopulate the erythroid, neutrophil-macrophage, megakaryocyte and lymphoid hematopoietic cell pool. 
     HSCs can be isolated or collected from bone marrow, umbilical cord blood, or peripheral blood. HSCs can be identified according to certain phenotypic or genotypic markers. For example, HSCs can be identified by their small size, lack of lineage (lin) markers, low staining (side population) with vital dyes such as rhodamine 123 (rhodamineDULL, also called rholo) or Hoechst 33342, and presence of various antigenic markers on their surface, many of which belong to the cluster of differentiation series (e.g., CD34, CD38, CD90, CD133, CD105, CD45, Terl 19, and c-kit, the receptor for stem cell factor). In certain embodiments, the transduced cell is a CD34 +  HSC. 
     In certain embodiments, the transduced cell is an embryonic stem cell. In certain embodiments, the transduced cell is an induced pluripotent stem cell. In certain embodiments, the transduced cell is a hemogenic endothelium cell. 
     While HSCs are the natural vehicle for restoring long-term hematopoiesis, their use has some important limitations. The first is their relative scarcity, which can eventually preclude autologous HSC therapy when the harvested cellular product is too small. The second is the difficulty to perform biosafety testing such as integration site analysis and consequently to select cells with chosen integration sites, because adult HSCs cannot be replicated in vitro. The third limitation is that homologous recombination using current technologies is practically impossible thus compromising the advent of gene correction. All of these limitations are ultimately due to the fact that adult HSCs cannot be expanded in vitro without losing their stem cell potency. These limitations explain the importance of viral vectors such as gamma-retroviral and lentiviral vectors, which are remarkably quick and efficient in achieving stable gene transfer. 
     Use of ESs and induced pluripotent stem (iPS) cells for globin gene therapy is disclosed in Moi et al.,  Haematol  Mar. 1, 2008; 93(3):325-330. Embryonic stem (ES) cells are amenable to gene targeting and correction, which requires unlimited in vitro cell division without losing multipotency. Chang et al.,  Proc Natl Acad Sci USA  2006; 103:1036-40 provided proof of principle of the feasibility of the homologous recombination approach in mice with sickle cell anemia. Takahashi et al.  Cell  2006; 126:663-76 reported the successful reprogramming of fibroblasts to an embryonic stem-like state. Cells obtained by this reverse-differentiation process, called induced pluripotent stem (iPS) cells, were produced by exposing embryonic or young adult bulk fibroblast cultures to gamma-retroviral vectors encoding 4 transcription factors, which are physiologically active in the embryonic stem cells, but generally turned off when differentiation progresses. The cultured cells formed colonies similar to ES cell colonies. These findings have been confirmed and extended by others to both mouse and human fibroblasts (Meissner et al., Nat Biotechnol 2007; 25:1177-81; Nakagawa et al., Nat Biotechnol 2007; 26:101-6; Okita et al., Nature 2007; 448:313-7; Park et al., Nature 2007; 451:141-6; Takahashi et al., Nat Protoc 2007; 2:3081-9; Takahashi K et al., Cell 2007; 131:861-72; Wernig et al., Nature 2007; 448:318-24; Yu J et al., Science 2007; 318: 1917-20). Rudolf Jaenisch and co-workers achieved a successful gene therapy in a mouse model of sickle cell disease, using homologous recombination in ES-like iPS cells (Hanna et al., Science 2007; 318:1920-3). The process has so far been mostly applied to fibroblast harvested from a skin biopsy, which are then induced to become iPS by transduction with retroviral vectors that encode four stem cell transcription factors. iPS are amenable to the correction of the SC mutation by standard homologous recombination techniques and can then be differentiated in vitro into unlimited amounts of hematopoietic stem cells. The whole process ends with the autologous transplantation of the corrected HSC into the original mouse donor, which will now be cured of its SC disease. This technique is not only useful for homologous recombination, but can also enhance lentiviral-mediated globin gene transfer for the treatment of β-thalassemia by providing a means to perform detailed integration site analysis and adequate in vitro cell expansion before infusing cells into the recipient. 
     The cell of the presently disclosed subject matter can be autologous (“self”) or non-autologous (“non-self,” e.g., allogeneic, syngeneic or xenogeneic). As used herein, “autologous” refers to cells from the same subject. As used herein, “allogeneic” refers to cells of the same species that differ genetically to the cell in comparison. As used herein, “syngeneic” refers to cells of a different subject that are genetically identical to the cell in comparison. As used herein, “xenogeneic” refers to cells of a different species to the cell in comparison. In certain embodiments, the cell is autologous, e.g., a cell transduced with the presently disclosed expression cassette is administered to a subject from whom the cell is collected, e.g., the cell is collected from bone marrow, umbilical cord blood, peripheral blood, and/or adipose tissue of the subject. In certain embodiments, the cell is obtained or collected from bone marrow of a subject. 
     In certain embodiments, prior to transduction with the expression cassette, the cell is pre-stimulated, e.g., in the presence of one or more cytokines (e.g., IL-3, IL-1α, IL-6, Kit ligand (also known as “Stem Cell Factor (SCF)”), and Flt-3 ligand), and/or one or more glycoproteins (e.g., thrombopoietin and fibronectin). In certain embodiments, the cell is pre-stimulated in the presence of Flt-3 ligand, SCF, thrombopoietin, interleukin-3, and fibronectin. The cell can be pre-stimulated for about 24 hours or longer, e.g., about 48 hours, or about 36 hours. Subsequently, the cell is transduced with a presently disclosed expression cassette, or a vector or another delivery system comprising such expression cassette. Transduction can be performed on a fresh cell, or on a frozen cell. Genomic DNA of the cell is isolated to determine the vector copy number and analyze the integration site or integrated vector structure, e.g., by South blot analysis and/or by Quantitative PCR. For quantification of globin mRNA (e.g., human β-globin transgene analysis), total RNA is extracted from the cell. Quantitative primer extension assay can be used for quantification of globin mRNA. 
     V. Compositions and Formulations 
     The presently disclosed subject matter provides pharmaceutical compositions comprising a presently disclosed transduced cell as described above and a pharmaceutically acceptable carrier. As used herein, “pharmaceutically acceptable carrier” includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible, including pharmaceutically acceptable cell culture media. The pharmaceutically acceptable carrier can be suitable for parenteral (e.g., intravenous, intramuscular, subcutaneous, or intraperitoneal), spinal or epidermal administration (e.g., by injection, infusion or implantation). Depending on the route of administration, the active compound, e.g., the transduced cell, may be coated in a material to protect the compound from the action of acids and other natural conditions that may inactivate the compound. 
     Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the transduced cells, use thereof in the pharmaceutical compositions of the invention is contemplated. 
     The pharmaceutical compositions of the presently disclosed subject matter can further comprise one or more polypeptides, polynucleotides, vectors comprising the same, transduced cells, etc., as described herein, formulated in pharmaceutically-acceptable or physiologically-acceptable solutions for administration to a cell or an animal, either alone, or in combination with one or more other modalities of therapy. If desired, the pharmaceutical compositions of the presently disclosed subject matter can be administered in combination with other agents, including, but not limited to, cytokines, growth factors, hormones, small molecules or various pharmaceutically-active agents. Any additional agents that do not adversely affect the ability of the composition to deliver the intended gene therapy can be included in the compositions. 
     In the pharmaceutical compositions of the presently disclosed subject matter, formulation of pharmaceutically-acceptable excipients and carrier solutions is well known to those of ordinary skill in the art, as is the development of suitable dosing and treatment regimens for using the particular compositions described herein in a variety of treatment regimens, including, e.g., oral, parenteral, intravenous, intranasal, and intramuscular administration and formulation. 
     The pharmaceutical compositions of the presently disclosed subject matter can be delivered parenterally (e.g., intravenously, intramuscularly, or intraperitoneally) as described, for example, in U.S. Pat. Nos. 5,543,158; 5,641,515 and 5,399,363. Solutions of the active compounds as free base or pharmacologically acceptable salts may be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions may also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms. 
     Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. The use of such media and agents for pharmaceutically active substances is known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the pharmaceutical compositions of the invention is contemplated. Supplementary active compounds can also be incorporated into the compositions. 
     Therapeutic compositions typically must be sterile and stable under the conditions of manufacture and storage. The composition can be formulated as a solution, microemulsion, liposome, or other ordered structure suitable to high drug concentration. The pharmaceutically acceptable carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent that delays absorption, for example, monostearate salts and gelatin. 
     The pharmaceutical compositions of the presently disclosed subject matter can be conveniently provided as sterile liquid preparations, e.g., isotonic aqueous solutions, suspensions, emulsions, dispersions, or viscous compositions, which can be buffered to a selected pH. Liquid preparations are normally easier to prepare than gels, other viscous compositions, and solid compositions. Additionally, liquid compositions are somewhat more convenient to administer, especially by injection. Viscous compositions, on the other hand, can be formulated within the appropriate viscosity range to provide longer contact periods with specific tissues. Liquid or viscous compositions can comprise carriers, which can be a solvent or dispersing medium containing, for example, water, saline, phosphate buffered saline, polyol (for example, glycerol, propylene glycol, liquid polyethylene glycol, and the like) and suitable mixtures thereof. 
     Sterile injectable solutions can be prepared by incorporating the compositions of the presently disclosed subject matter in the required amount of the appropriate solvent with various amounts of the other ingredients, as desired. Such compositions may be in admixture with a suitable carrier, diluent, or excipient such as sterile water, physiological saline, glucose, dextrose, or the like. The compositions can also be lyophilized. The compositions can contain auxiliary substances such as wetting, dispersing, or emulsifying agents (e.g., methylcellulose), pH buffering agents, gelling or viscosity enhancing additives, preservatives, flavoring agents, colors, and the like, depending upon the route of administration and the preparation desired. Standard texts, such as “REMINGTON&#39;S PHARMACEUTICAL SCIENCE”, 17th edition, 1985, incorporated herein by reference, may be consulted to prepare suitable preparations, without undue experimentation. 
     Various additives which enhance the stability and sterility of the compositions, including antimicrobial preservatives, antioxidants, chelating agents, and buffers, can be added. Prevention of the action of microorganisms can be ensured by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, and the like. Prolonged absorption of the injectable pharmaceutical form can be brought about by the use of agents delaying absorption, for example, alum inurn monostearate and gelatin. 
     The compositions can be isotonic, i.e., they can have the same osmotic pressure as blood and lacrimal fluid. The desired isotonicity of the compositions of the presently disclosed subject matter can be accomplished using sodium chloride, or other pharmaceutically acceptable agents such as dextrose, boric acid, sodium tartrate, propylene glycol or other inorganic or organic solutes. Sodium chloride is preferred particularly for buffers containing sodium ions. 
     For parenteral administration in an aqueous solution, for example, the solution should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by sterilization microfiltration. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying (lyophilization) that yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof. 
     In certain embodiments, the compositions can be delivered by intranasal sprays, inhalation, and/or other aerosol delivery vehicles. Methods for delivering genes, polynucleotides, and peptide compositions directly to the lungs via nasal aerosol sprays are described, e.g., in U.S. Pat. Nos. 5,756,353 and 5,804,212. Methods of delivering drugs using lysophosphatidyl-glycerol compounds are described, e.g., in U.S. Pat. No. 5,725,871. Transmucosal drug delivery in the form of a polytetrafluoroetheylene support matrix is described, e.g., in U.S. Pat. No. 5,780,045. The compositions of the presently disclosed subject matter can be formulated for delivery either encapsulated in a lipid particle, a liposome, a vesicle, a nanosphere, a nanoparticle or the like. The formulation and use of such delivery vehicles can be carried out using known and conventional techniques. The formulations and compositions of the presently disclosed subject matter can comprise one or more repressors and/or activators comprising a combination of any number of polypeptides, polynucleotides, and small molecules, as described herein, formulated in pharmaceutically-acceptable or physiologically-acceptable solutions (e.g., culture medium) for administration to a cell or an animal, either alone, or in combination with one or more other modalities of therapy. 
     In certain aspects, the presently disclosed subject matter provides formulations or compositions suitable for the delivery of viral vector systems (i.e., viral-mediated transduction) including, but not limited to, retroviral (e.g., lentiviral) vectors. Exemplary formulations for ex vivo delivery can also include the use of various transfection agents known in the art, such as calcium phosphate, electoporation, heat shock and various liposome formulations (i.e., lipid-mediated transfection). Liposomes are lipid bilayers entrapping a fraction of aqueous fluid. DNA spontaneously associates to the external surface of cationic liposomes (by virtue of its charge) and these liposomes will interact with the cell membrane. 
     The skilled artisan can readily determine the amount of cells and optional additives, vehicles, and/or carrier in compositions and to be administered in methods of the presently disclosed subject matter. Typically, any additives (in addition to the transduced cell(s) and/or agent(s)) are present in an amount of from about 0.001% to about 50% by weight) solution in phosphate buffered saline, and the active ingredient is present in the order of micrograms to milligrams, such as from about 0.0001 wt % to about 5 wt %, from about 0.0001 wt % to about 1 wt %, from about 0.0001 wt % to about 0.05 wt %, from about 0.001 wt % to about 20 wt %, from about 0.01 wt % to about 10 wt %, or from about 0.05 wt % to about 5 wt %. For any composition to be administered to an animal or human, and for any particular method of administration, toxicity should be determined, such as by determining the lethal dose (LD) and LD50 in a suitable animal model e.g., rodent such as mouse; and, the dosage of the composition(s), concentration of components therein and timing of administering the composition(s), which elicit a suitable response. Such determinations do not require undue experimentation from the knowledge of the skilled artisan, this disclosure and the documents cited herein. And, the time for sequential administrations can be ascertained without undue experimentation. 
     VI. Uses and Methods 
     Vectors and other delivery systems (nucleases and CRISPR-Cas systems) comprising the presently disclosed expression cassette provide improved methods of gene therapy. As used herein, the term “gene therapy” refers to the introduction of a polynucleotide into a cell&#39;s genome that restores, corrects, or modifies the gene and/or expression of the gene. In various non-limiting embodiments, a presently disclosed vector or other delivery system (e.g., a nuclease or a CRISPR-Cas system) comprises an expression cassette comprising a globin gene or a functional portion thereof that encodes a globin protein (e.g., human β globin protein), which provides curative, preventative, or ameliorative benefits to a subject diagnosed with or that is suspected of having a disease, disorder, or condition of the hematopoietic system. The vector or other delivery systems (e.g., a nuclease and the CRISPR-Cas system) can infect and transduce the cell in vivo, ex vivo, or in vitro. In ex vivo and in vitro embodiments, the transduced cells can then be administered to a subject in need of therapy. The presently disclosed subject matter contemplates that the vectors and other delivery systems (e.g., nucleases or CRISPR-Cas systems), viral particles, and transduced cells of the presently disclosed subject matter are be used to treat, prevent, and/or ameliorate a disease, disorder, or condition of the hematopoietic system in a subject, e.g., a hemoglobinopathy. 
     As used herein, the term “hemoglobinopathy” or “hemoglobinopathic condition” includes any disorder involving the presence of an abnormal hemoglobin molecule in the blood. Examples of hemoglobinopathies included, but are not limited to, hemoglobin C disease, hemoglobin sickle cell disease (SCD), sickle cell anemia, and thalassemias. Also included are hemoglobinopathies in which a combination of abnormal hemoglobins are present in the blood (e.g., sickle cell/Hb-C disease). 
     As used herein, “thalassemia” refers to a hereditary disorder characterized by defective production of hemoglobin. Examples of thalassemias include α- and β-thalassemia. β-thalassemias are caused by a mutation in the beta globin chain, and can occur in a major or minor form. In the major form of β-thalassemia, children are normal at birth, but develop anemia during the first year of life. The mild form of β-thalassemia produces small red blood cells and the thalassemias are caused by deletion of a gene or genes from the globin chain. α-thalassemia typically results from deletions involving the HBA1 and HBA2 genes. Both of these genes encode α-globin, which is a component (subunit) of hemoglobin. There are two copies of the HBA1 gene and two copies of the HBA2 gene in each cellular genome. As a result, there are four alleles that produce α-globin. The different types of a thalassemia result from the loss of some or all of these alleles. Hb Bart syndrome, the most severe form of a thalassemia, results from the loss of all four α-globin alleles. HbH disease is caused by a loss of three of the four [alpha]-globin alleles. In these two conditions, a shortage of [alpha]-globin prevents cells from making normal hemoglobin. Instead, cells produce abnormal forms of hemoglobin called hemoglobin Bart (Hb Bart) or hemoglobin H (HbH). These abnormal hemoglobin molecules cannot effectively carry oxygen to the body&#39;s tissues. The substitution of Hb Bart or HbH for normal hemoglobin causes anemia and the other serious health problems associated with a thalassemia. 
     As used herein, the term “sickle cell disease” refers to a group of autosomal recessive genetic blood disorders, which results from mutations in a globin gene and which is characterized by red blood cells that assume an abnormal, rigid, sickle shape. They are defined by the presence of β S -gene coding for a β-globin chain variant in which glutamic acid is substituted by valine at amino acid position 6 of the peptide, and second β-gene that has a mutation that allows for the crystallization of HbS leading to a clinical phenotype. As used herein, the term “sickle cell anemia” refers to a specific form of sickle cell disease in patients who are homozygous for the mutation that causes HbS. Other common forms of sickle cell disease include HbS/β-thalassemia, HbS/HbC and HbS/HbD. 
     In certain embodiments, gene therapy methods of the presently disclosed subject mater are used to treat, prevent, or ameliorate a hemoglobinopathy that is selected from the group consisting of: hemoglobin C disease, hemoglobin sickle cell disease (SCD), sickle cell anemia, hereditary anemia, thalassemia, β-thalassemia, thalassemia major, thalassemia intermedia, α-thalassemia, and hemoglobin H disease. In certain embodiments, the hemoglobinopathy is β-thalassemia. In certain embodiments, the hemoglobinopathy is sickle cell anemia 
     In various non-limiting embodiments, vectors or other delivery systems (e.g., nucleases or CRISPR-Cas systems) comprising a presently disclosed expression cassette are administered by direct injection to a cell, tissue, or organ of a subject in need of gene therapy, in vivo. In certain embodiments, cells are transduced in vitro or ex vivo with vectors or other delivery systems (e.g., nucleases or CRISPR-Cas systems) of the presently disclosed subject matter, and optionally expanded ex vivo. The transduced cells are then administered to a subject in need of gene therapy, e.g., within a pharmaceutical formulation disclosed herein. 
     The presently disclosed subject matter provides a method of providing a transduced cell to a subject. In various non-limiting embodiments, the method comprises administering (e.g., parenterally) one or more cells (a population of cells) transduced with a presently disclosed expression cassette or a vector or another delivery system (e.g., a nuclease or CRISPR-Cas system) comprising such expression cassette to the subject. 
     The presently disclosed subject matter provides a method of treating a hemoglobinopathy in a subject. In various non-limiting embodiments, the method comprises administering an effective amount of a presently disclosed transduced cell or a population of the presently disclosed transduced cells (e.g., HSCs, embryonic stem cells, or iPSCs) to the subject. 
     For treatment, the amount administered is an amount effective in producing the desired effect. An effective amount can be provided in one or a series of administrations. An effective amount can be provided in a bolus or by continuous perfusion. An “effective amount” (or “therapeutically effective amount”) is an amount sufficient to affect a beneficial or desired clinical result upon treatment. An effective amount can be administered to a subject in one or more doses. In terms of treatment, an effective amount is an amount that is sufficient to palliate, ameliorate, stabilize, reverse or slow the progression of the disease, or otherwise reduce the pathological consequences of the disease. The effective amount is generally determined by the physician on a case-by-case basis and is within the skill of one in the art. Several factors are typically taken into account when determining an appropriate dosage to achieve an effective amount. These factors include age, sex and weight of the subject, the condition being treated, the severity of the condition and the form and effective concentration of the immunoresponsive cells administered. 
     In certain embodiments, following administration of one or more of the presently disclosed transduced cells, peripheral blood of the subject is collected and hemoglobin levels is measured. A therapeutically relevant level of hemoglobin is produced following administration of one or more of the presently disclosed transduced cells. Therapeutically relevant level of hemoglobin is a level of hemoglobin that is sufficient (1) to improve or correct anemia, (2) to restore the ability of the subject to produce red blood cells containing normal hemoglobin, (3) to correct ineffective erythropoiesis in the subject, (4) to correct extra-medullary hematopoiesis (e.g., splenic and hepatic extra-medullary hematopoiesis), and/or (5) to reduce iron accumulation, e.g., in peripheral tissues and organs. Therapeutically relevant level of hemoglobin can be at least about 7 g/dL Hb, at least about 7.5 g/dL Hb, at least about 8 g/dL Hb, at least about 8.5 g/dL Hb, at least about 9 g/dL Hb, at least about 9.5 g/dL Hb, at least about 10 g/dL Hb, at least about 10.5 g/dL Hb, at least about 11 g/dL Hb, at least about 11.5 g/dL Hb, at least about 12 g/dL Hb, at least about 12.5 g/dL Hb, at least about 13 g/dL Hb, at least about 13.5 g/dL Hb, at least about 14 g/dL Hb, at least about 14.5 g/dL Hb, or at least about 15 g/dL Hb. Additionally or alternatively, therapeutically relevant level of hemoglobin can be from about 7 g/dL Hb to about 7.5 g/dL Hb, from about 7.5 g/dL Hb to about 8 g/dL Hb, from about 8 g/dL Hb to about 8.5 g/dL Hb, from about 8.5 g/dL Hb to about 9 g/dL Hb, from about 9 g/dL Hb to about 9.5 g/dL Hb, from about 9.5 g/dL Hb to about 10 g/dL Hb, from about 10 g/dL Hb to about 10.5 g/dL Hb, from about 10.5 g/dL Hb to about 11 g/dL Hb, from about 11 g/dL Hb to about 11.5 g/dL Hb, from about 11.5 g/dL Hb to about 12 g/dL Hb, from about 12 g/dL Hb to about 12.5 g/dL Hb, from about 12.5 g/dL Hb to about 13 g/dL Hb, from about 13 g/dL Hb to about 13.5 g/dL Hb, from about 13.5 g/dL Hb to about 14 g/dL Hb, from about 14 g/dL Hb to about 14.5 g/dL Hb, from about 14.5 g/dL Hb to about 15 g/dL Hb, from about 7 g/dL Hb to about 8 g/dL Hb, from about 8 g/dL Hb to about 9 g/dL Hb, from about 9 g/dL Hb to about 10 g/dL Hb, from about 10 g/dL Hb to about 11 g/dL Hb, from about 11 g/dL Hb to about 12 g/dL Hb, from about 12 g/dL Hb to about 13 g/dL Hb, from about 13 g/dL Hb to about 14 g/dL Hb, from about 14 g/dL Hb to about 15 g/dL Hb, from about 7 g/dL Hb to about 9 g/dL Hb, from about 9 g/dL Hb to about 11 g/dL Hb, from about 11 g/dL Hb to about 13 g/dL Hb, or from about 13 g/dL Hb to about 15 g/dL Hb. In certain embodiments, the therapeutically relevant level of hemoglobin is maintained in the subject for at least about 6 months, for at least about 12 months (or 1 year), for at least about 24 months (or 2 years). In certain embodiments, the therapeutically relevant level of hemoglobin is maintained in the subject for up to about 6 months, for up to about 12 months (or 1 year), for up to about 24 months (or 2 years). In certain embodiments, the therapeutically relevant level of hemoglobin is maintained in the subject for about 6 months, for about 12 months (or 1 year), for about 24 months (or 2 years). In certain embodiments, the therapeutically relevant level of hemoglobin is maintained in the subject for from about 6 months to about 12 months (e.g., from about 6 months to about 8 months, from about 8 months to about 10 months, from about 10 months to about 12 months), from about 12 months to about 18 months (e.g., from about 12 months to about 14 months, from about 14 months to about 16 months, or from about 16 months to about 18 months), or from about 18 months to about 24 months (e.g., from about 18 months to about 20 months, from about 20 months to about 22 months, or from about 22 months to about 24 months). 
     In certain embodiments, the method comprises administering one or more cells transduced with a recombinant vector comprising a presently disclosed expression cassette as described above. The vector copy number of the recombinant vector in the cells that provide for the therapeutically relevant level of hemoglobin (e.g., 9-10 g/dL) in the subject is from about 0.5 to about 2, from about 0.5 to about 1, or from about 1 to about 2 vector copy number per cell. In certain embodiments, the vector copy number of the presently disclosed vector is about 0.5, about 0.6, about 0.7, about 0.8, about 0.9, about 1.0, about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, or about 2.0 vector copy number per cell. 
     In certain embodiments, the subject lacks a human leukocyte antigen (HLA)-matched donor. In certain embodiments, the transduced cell is from the same subject. In certain embodiments, the transduced cell is from bone marrow of the same subject. Thus, administration of the transduced cells do not incur the risk of graft-versus host disease in the subject. The method does not require immune suppression to prevent graft rejection, e.g., the method does not comprise administering an immunosuppressive agent to the subject. 
     The present disclosed subject matter also provides a method of increasing the proportion of red blood cells or erythrocytes compared to white blood cells or leukocytes in a subject. In various non-limiting embodiments, the method comprises administering an effective amount of a presently disclosed transduced cell or a population of the presently disclosed transduced cells (e.g., HSCs, embryonic stem cells, or iPSCs) to the subject, wherein the proportion of red blood cell progeny cells of the hematopoietic stem cells are increased compared to white blood cell progeny cells of the hematopoietic stem cells in the subject. 
     An important advantage provided by the expression cassette, vectors and other delivery systems (e.g., nucleases and CRISPR-Cas systems), compositions, and methods of the presently disclosed subject is the high efficacy of globin gene therapy that can be achieved by administering populations of cells comprising lower percentages of transduced cells compared to existing methods. This provides important safety advantages associated with reduced chances of deleterious mutation, transformation, or oncogene activation of cellular genes in transduced cells. The transduced cells can be administered as part of a bone marrow or cord blood transplant in an individual that has or has not undergone bone marrow ablative therapy. 
     One consideration concerning the therapeutic use of the presently disclosed cells transduced with the expression cassette described herein (“transduced cells”) is the quantity of cells necessary to achieve an optimal effect. The quantity of transduced cells to be administered will vary for the subject being treated. In certain embodiments, from about 1×10 4  to about 1×10 5  cells/kg, from about 1×10 5  to about 1×10 6  cells/kg, from about 1×10 6  to about 1×10 7  cells/kg, from about 1×10 7  to about 1×10 8  cells/kg, from about 1×10 8  to about 1×10 9  cells/kg, or from about 1×10 9  to about 1×10 10  cells/kg of the presently disclosed transduced cells are administered to a subject. More effective cells may be administered in even smaller numbers. In certain embodiments, at least about 1×10 8  cells/kg, at least about 2×10 8  cells/kg, at least about 3×10 8  cells/kg, at least about 4×10 8  cells/kg, or at least about 5×10 8  cells/kg of the presently disclosed transduced cells are administered to a subject. The precise determination of what would be considered an effective dose may be based on factors individual to each subject, including their size, age, sex, weight, and condition of the particular subject. Dosages can be readily ascertained by those skilled in the art from this disclosure and the knowledge in the art. 
     In certain embodiments, the expression cassettes, vectors and other delivery systems (nucleases and CRISPR-Cas systems), compositions, and methods of the presently disclosed subject matter offer improved methods of gene therapy using ex vivo gene therapy and autologous transplantation. Transplantation of cells transduced with the expression cassette or into subjects having a hemoglobinopathy results in long-term correction of the disease. 
     One or more presently disclosed transduced cells can be administered by any methods known in the art, including, but not limited to, parenteral administration (e.g., intramuscular administration, intravenous administration, subcutaneous administration, or intraperitoneal administration), spinal administration, and epidermal administration. In certain embodiments, one or more transduced cells are delivered to a subject intravenously. One or more presently disclosed transduced cells can be administered by injection, infusion, or implantation. In certain embodiments, one or more transduced cells are administered by injection. In certain embodiments, one or more transduced cells are administered by intravenous injection. 
     The subjects can have an advanced form of disease, in which case the treatment objective can include mitigation or reversal of disease progression, and/or amelioration of side effects. The subjects can have a history of the condition, for which they have already been treated, in which case the therapeutic objective will typically include a decrease or delay in the risk of recurrence. 
     VII. Kits 
     The presently disclosed subject matter provides kits for the treatment or prevention of a hemoglobinopathy. In certain embodiments, the kit comprises a therapeutic or prophylactic composition containing an effective amount of a cell transduced with the presently disclosed expression cassette in unit dosage form. In certain embodiments, the kit comprises one or more expression cassettes disclosed herein. In certain embodiments, the kit comprises one or more vectors comprising an expression cassette disclosed herein. In certain embodiments, the kit comprises a sterile container, which can be a box, an ampule, a bottle, a vial, a tube, a bag, a pouch, a blister-pack, or other suitable container forms known in the art. Such containers can be made of plastic, glass, laminated paper, metal foil, or other materials suitable for holding medicaments. 
     If desired, the transduced cell is provided together with instructions for administering the cell to a subject having or at risk of developing a hemoglobinopathy. The instructions will generally include information about the use of the composition for the treatment or prevention of a hemoglobinopathy. In certain embodiments, the instructions include at least one of the following: description of the therapeutic agent; dosage schedule and administration for treatment or prevention of a hemoglobinopathy or symptoms thereof; precautions; warnings; indications; counter-indications; overdosage information; adverse reactions; animal pharmacology; clinical studies; and/or references. Alternatively or additionally, the kit can include instructions for transducing a cell with the one or more expression cassettes and/or vectors comprising such expression cassettes. The instructions may be printed directly on the container (when present), or as a label applied to the container, or as a separate sheet, pamphlet, card, or folder supplied in or with the container. 
     EXAMPLES 
     The practice of the presently disclosed subject matter employs, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry and immunology, which are well within the purview of the skilled artisan. Such techniques are explained fully in the literature, such as, “Molecular Cloning: A Laboratory Manual”, second edition (Sambrook, 1989); “Oligonucleotide Synthesis” (Gait, 1984); “Animal Cell Culture” (Freshney, 1987); “Methods in Enzymology” “Handbook of Experimental Immunology” (Weir, 1996); “Gene Transfer Vectors for Mammalian Cells” (Miller and Calos, 1987); “Current Protocols in Molecular Biology” (Ausubel, 1987); “PCR: The Polymerase Chain Reaction”, (Mullis, 1994); “Current Protocols in Immunology” (Coligan, 1991). These techniques are applicable to the production of the polynucleotides and polypeptides of the presently disclosed subject matter, and, as such, may be considered in making and practicing the presently disclosed subject matter. Particularly useful techniques for particular embodiments will be discussed in the sections that follow. 
     The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the expression cassettes, vectors, delivery systems, and therapeutic methods of the presently disclosed subject matter, and are not intended to limit the scope of what the inventors regard as their invention. 
     Example 1: Discovery of Novel Insulators 
     The problems created by insertional mutagenesis of viral vectors are widely known (Nienhuis (2013), Baum et al. (2006), Nienhuis et al. (2006)) as is the evidence that the risks of genotoxicity can be reduced by the use of chromatin insulators (Arumugam et al. (2007), Emery (2011), Evans-Galea et al. (2007), Rivella et al. (2000), Emery et al. (2000), Emery et al. (2002), Yannaki et al. (2002), Hino et al. (2004), Ramezani et al. (2003), Ramezani et al. (2008)). Approaches allowing the efficient identification of enhancer blocking insulators in the human genome have been developed. These new insulators are short, on the average 150 bp, and they do not affect adversely the titers of viral vectors and they are several times more powerful than the insulator cHS4. Genomic approaches were used to discover the most powerful enhancer blocker and barrier insulators of the human genome. For gene therapy of the hemoglobinopathies, powerful enhancers are required to achieve therapeutic levels of globin gene expression. Powerful insulators may thus provide one means to protect the genomic environment from the powerful enhancers of the integrating vectors. 
     Several studies have demonstrated the ability of the cHS4 insulator to reduce position-effect silencing of gammaretroviral vectors (Evans-Galea et al. (2007), Rivella et al. (2000), Emery et al. (2000), Emery et al. (2002), Yannaki et al. (2002), Hino et al. (2004), Ramezani et al. (2006), Yao et al. (2003), Nishino et al. (2006), Aker et al. (2007), Li and Emery (2008)), and lentiviral vectors (Evans-Galea et al. (2007), Ramezani et al. (2003), Puthenveetil et al. (2004), Arumugam et al. (2007), Bank et al. (2005), Aker et al. (2007), Ma et al. (2003), Chang et al. (2005), Pluta et al. (2005)). Those studies that were appropriately designed demonstrated that inclusion of the 1.2 kb version of the cHS4 insulator increased the likelihood and/or consistency of vector transgene expression in at least some settings (Arumugam et al. (2007), Evans-Galea et al. (2007), Emery et al. (2002), Yannaki et al. (2002), Hino et al. (2004), Ramezani et al. (2006), Aker et al. (2007), Li and Emery (2008), Pluta et al. (2005), Jakobsson et al. (2004)). Nevertheless, the degree of protection afforded by the cHS4 insulator is far from complete. In addition, the inclusion of the 1.2 Kb cHS4 can adversely affect vector titers while the smallest cHS4 core has been proven ineffective (Aker et al. (2007), Jakobsson et al. (2004)). 
     Effects on genotoxicity were tested using an in vivo assay based on quantitation of tumor formation in mice. Vectors insulated by insulator A1 decreased tumor formation induced by random vector integration in hematopoietic chimeras compared to mice that received uninsulated or cHS4-insulated controls. 
     To assess effects on vector titers, insulator A1 was introduced into the double-copy region of a third-generation lentiviral vector expressing GFP from a constitutive package promoter, and the viral titers and GFP expression were measured. Insulator A1 did not affect adversely vector GFP expression. 
     In the in vivo genotoxicity assay, a cell line transduced with gammaretroviral vectors produced tumors after transplantation in mice and allowed quantitation of genotoxic effects by measuring rates of tumor free survival. Effects of an insulator on genotoxicity were quantitated by the number of tumors formed in the mice and the rates of tumor free survival. Insulator A1 was inserted in the proximal portion of the 3′ LTR, from which it is copied into the 5′ LTR during reverse transcription and vector integration. The resulting topology places copies of the insulator between the genomic regions located 5′ and 3′ of the integrated provirus and enhancer activity from the 5′ viral LTR and internal Pgk promoter, but does not contain the enhancer in the 3′ LTR. This can decrease genotoxicity thus resulting in decreased tumor formation and increased survival of the animals. Gamma-retroviral reporter vectors flanked with insulator A1 or control regions were used to transduce the growth factor-dependent cell line 32D, and 10 independent sub-pools for each vector were transplanted into syngeneic C3H/HeJ mice. All 10 mice transplanted with mock-transduced cells remained free of 32D cell-derived tumors, while nearly all mice transplanted with 32D cells transduced with vectors containing no inserts or a 790 bp neutral spacer developed tumors within a median of 16 weeks ( FIG. 5B ). Flanking this vector with the cHS4 insulator delayed the onset of tumor formation by several weeks, and reduced the frequency of animals that developed tumors to 6 of 10. In contrast, only two of 10 animals developed tumors following transplantation with 32D cells transduced with the vector flanked with insulator A1 ( FIG. 5B ). The frequency of animals with tumors and the number of vector transduction events in the original sub-pools suggested that flanking the vector with insulator A1 reduced the overall rate of tumor formation 12-fold, from 46.9 tumors per 10 5  provirus to 3.9 tumors per 10 5  provirus ( FIG. 5C ). In comparison, the cHS4 insulator reduced the overall rate of tumor formation 2.8-fold (to 16.9 tumors per 10 5  provirus), while the neutral spacer had no statistically discernable effect on the rate of tumor formation. These results indicate that the discovered enhancer blocking insulators can decrease substantially the risks of insertional mutagenesis and genotoxicity. 
     Example 2: Characterization of Globin Vectors Comprising at Least One Insulator 
     A presently disclosed expression cassette (designated as “Expression Cassette 1”; as shown in  FIG. 1 ), which comprises insulator A1, and a human β A -globin gene encoding a threonine to glutamine mutation at codon 87 (β A-T87Q ) operably linked to a β-globin LCR comprising a HS2 region having the nucleotide sequence set forth in SEQ ID NO:9, a HS3 region having the nucleotide sequence set forth in SEQ ID NO:5, and a HS4 region having the nucleotide sequence set forth in SEQ ID NO:7, was generated. The rationale for using the variant β chain (β A ) is to facilitate the detection of the vector-encoded β-globin gene, distinguishing it from endogenous or transfused beta chains. The glutamine (GLN) residue at position 87 in the γ-globin chain augments the anti-sickling activity of the gamma chain relative to the β chain, while preserving adult oxygen-binding characteristics of the β chain (Nagel et al. (1979)). In Vector 1, a point mutation altering codon 87 (β A-T87Q , or β87) replaces the normal threonine with glutamine and augments anti-sickling activity of the vector-encoded β chain. This β87 chain has been safely used in a patient with HbE-thalassemia (Cavazzana-Calvo et al. (2010)). 
     Expression cassette 1 was incorporated or introduced to a lentivirus vector (designated as “Vector 1”). Vector 1 was introduced in bone marrow cells of C57BL/6-Hbb th3/+ mice and transplanted to syngeneic lethally irradiated recipients as previously described (May et al. (2000), May et al. (2002), Lisowski et al. (2007)). The vector titer of V1 was comparable to that of a lentivirus vector comprising an expression cassette lacking insulator A1. The β-globin expression of Vector 1 was compared to that of a lentivirus vector (designated as “Vector 2”) comprising an expression cassette that lacks an insulator and comprises a wild human β-globin gene operably linked to a β-globin LCR comprising a HS2 region having the nucleotide sequence set forth in SEQ ID NO:9, a HS3 region having the nucleotide sequence set forth in SEQ ID NO:5, and a HS4 region having the nucleotide sequence set forth in SEQ ID NO:6. In comparison to Vector 2, β-globin expression of Vector 1 normalized to vector copy was equivalent or slightly increased, suggesting an added benefit for in vivo expression provided by the flanking barrier elements, as shown in  FIG. 6 . 
     Example 3: Evaluation of Enhancer Activity in Non-Erythroid K562 Cells 
     The enhancer activity of HS2 was evaluated in Non-erythroid K562 Cells. As shown in  FIG. 7 , GFP expression in K562 cells transduced with vectors driven by a minimal promoter linked to no enhancer (“Empty”, HS2, HS3-4, HS2-3-4 or the runx1 enhancer used as positive control (“RUNX1”). Background expression was on the order or 0.01% (“empty), but increased over 10-fold with HS2-3-4 (“Lcr9”, 0.17%). This enhancement was mostly due to HS2 (0.15%) but not HS3-4 (0.05%). All cell lines were comparably transduced (mean vector copy number 2.5). The results support that HS2 but not HS3-HS4 may pose an oncogenic risk in non-erythroid hematopoietic stem and progenitor cells. 
     Example 4: Novel Erythroid-Specific Enhancers 
     As shown in  FIGS. 8 and 9 , five erythroid-specific enhancers were substituted for HS2: ALAS Intron 1, ALAS Intron 8, BLVRB, PPDX, and Spectrin-alpha. The inventors have shown that all these five enhancers are powerful enhancers, and lack enhancer activity in non-erythroid tissues, and do not reduce the vector titer. 
     Example 5: Increasing Globin Lentiviral Vector Production Through 3′ LTR Modifications 
     An essential feature of therapeutic globin vectors is to achieve a high titer, sufficient for effective transduction of patient cells. By virtue of their large cargo, comprising a gene, promoter, enhancers and/or LCR elements, globin lentiviral vectors inherently have low titer, complicating their manufacture and limiting their clinical use. This problem is further compounded by the incorporation of additional genomic elements such as an insulator, which further increase the size of the vector. 
     The inventors explored different modifications of the 3′ long terminal repeat (LTR) of globin vectors to increase the titer of globin vectors. Over 62 variations were evaluated, numbered 1 through 62, modeled on a lentivirus vector comprising a human β-globin gene operably linked to a β-globin LCR comprising a HS2 region having the nucleotide sequence set forth in SEQ ID NO:9, a HS3 region having the nucleotide sequence set forth in SEQ ID NO:5, and a HS4 region having the nucleotide sequence set forth in SEQ ID NO:7. In other words, all of Vectors #1 through Vector 62 comprise a β-globin LCR comprising a HS2 region having the nucleotide sequence set forth in SEQ ID NO:9, a HS3 region having the nucleotide sequence set forth in SEQ ID NO:5, and a HS4 region having the nucleotide sequence set forth in SEQ ID NO:7. Vector #18 served as a baseline, comprising a standard U3 deletion in the 3′LTR. Vector #1 (not depicted) comprised a full, i.e., wild-type LTR, which cannot be used clinically. Modifications to the 3′LTR are depicted in  FIGS. 10A and 10B , and their titers shown in  FIGS. 11 and 12  (the Y axis shows the vector copy number of vector stocks manufactured and tested under strictly identical conditions). Titrations were measured in triple replicas, performed in parallel by two operators, and repeated in multiple experiments. 
     As shown in  FIGS. 11 and 12 , Vector #55 repeatedly showed a higher titer. This vector comprises a Woodchuck hepatitis post-regulatory element (WPRE) and a bovine growth hormone polyadenylation signal 3′ to the R region in the 3′ LTR. The WPRE element is therefore not transferred to the transduced cells. 
     Example 6: Generation of Globin Vectors 
     Methods and Materials 
     Vector Production 
     Various lentiviral vectors were produced, including TNS9.B87.A1, SNS22.2.B87.A1, SNS23.2.B87.A1, and SNS24.2.B87.A1. The nucleotide sequences of TNS9.B87.A1, SNS22.2.B87.A1, SNS23.2.B87.A1, SNS24.2.B87.A1, SNS26.B87.A1 and SNS27.2.B87.A1 are set forth in SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 76, and SEQ ID NO: 77, respectively. 
     
       
         
           
               
               
            
               
                 [SEQ ID NO: 37] 
                   
               
               
                 attggccattgcatacgttgtatccatatcataatatgtacatttatattggctcatgtccaacattaccgccatgttga 
                   
               
               
                   
               
               
                 cattgattattgactagttattaatagtaatcaattacggggtcattagttcatagcccatatatggagttccgcgttac 
               
               
                   
               
               
                 ataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccattgacgtcaataatgacgtatgttccca 
               
               
                   
               
               
                 tagtaacgccaatagggactttccattgacgtcaatgggtggagtatttacggtaaactgcccacttggcagtacatcaa 
               
               
                   
               
               
                 gtgtatcatatgccaagtacgccccctattgacgtcaatgacggtaaatggcccgcctggcattatgcccagtacatgac 
               
               
                   
               
               
                 cttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtaca 
               
               
                   
               
               
                 tcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttgg 
               
               
                   
               
               
                 caccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtg 
               
               
                   
               
               
                 ggaggtctatataagcagagctcgtttagtgaaccggggtctctctggttagaccagatctgagcctgggagctctctgg 
               
               
                   
               
               
                 ctaactagggaacccactgcttaagcctcaataaagcttgccttgagtgcttcaagtagtgtgtgcccgtctgttgtgtg 
               
               
                   
               
               
                 actctggtaactagagatccctcagacccttttagtcagtgtggaaaatctctagcagtggcgcccgaacagggacttga 
               
               
                   
               
               
                 aagcgaaagggaaaccagaggagctctctcgacgcaggactcggcttgctgaagcgcgcacggcaagaggcgaggggcgg 
               
               
                   
               
               
                 cgactggtgagtacgccaaaaattttgactagcggaggctagaaggagagagacgggtgcgagagcgtcagtattaagcg 
               
               
                   
               
               
                 ggggtgaataagatcgcgatgggaaaaaattcggttaaggccagggggaaagaaaaaatataaattaaaacatatagtat 
               
               
                   
               
               
                 gggcaagcagggagctagaacgattcgcagttaatcctggcctgttagaaacatcagaaggctgtagacaaatactggga 
               
               
                   
               
               
                 cagctacaaccatcccttcagacaggatcagaagaacttagatcattatataatacagtagcaaccctctattgtgtgca 
               
               
                   
               
               
                 tcaaaggatagagataaaagacaccaaggaagctttagacaagatagaggaagagcaaaacaaaagtaagaccaccgcac 
               
               
                   
               
               
                 agcaagcggccgctgatcttcagacctggaggaggagatatgagggacaattggagaagtgaattatataaatataaagt 
               
               
                   
               
               
                 agtaaaaattgaaccattaggagtagcacccaccaaggcaaagagaagagtggtgcagagagaaaaaagagcagtgggaa 
               
               
                   
               
               
                 taggagctttgttccttgggttcttgggagcagcaggaagcactatgggcgcagcgtcaatgacgctgacggtacaggcc 
               
               
                   
               
               
                 agacaattattgtctggtatagtgcagcagcagaacaatttgctgagggctattgaggcgcaacagcatctgttgcaact 
               
               
                   
               
               
                 cacagtctggggcatcaagcagctccaggcaagaatcctggctgtggaaagatacctaaaggatcaacagctcctgggga 
               
               
                   
               
               
                 tttggggttgctctggaaaactcatttgcaccactgctgtgccttggaatgctagttggagtaataaatctctggaacag 
               
               
                   
               
               
                 atttggaatcacacgacctggatggagtgggacagagaaattaacaattacacaagcttaatacactccttaattgaaga 
               
               
                   
               
               
                 atcgcaaaaccagcaagaaaagaatgaacaagaattattggaattagataaatgggcaagtttgtggaattggtttaaca 
               
               
                   
               
               
                 taacaaattggctgtggtatataaaattattcataatgatagtaggaggcttggtaggtttaagaatagtttttgctgta 
               
               
                   
               
               
                 ctttctatagtgaatagagttaggcagggatattcaccattatcgtttcagacccacctcccaaccccgaggggacccga 
               
               
                   
               
               
                 caggcccgaaggaatagaagaagaaggtggagagagagacagagacagatccattcgattagtgaacggatctcgacggt 
               
               
                   
               
               
                 atcgttttaaaagaaaaggggggattggggggtacagtgcaggggaaagaatagtagacataatagcaacagacatacaa 
               
               
                   
               
               
                 actaaagaattacaaaaacaaattacaaaaattcaaaattttatcggcgtgttgggggtggaccatcctctaggtattga 
               
               
                   
               
               
                 ataagaaaaatgaagttaaggtggttgatggtaacactatgctaataactgcagagccagaagcaccataagggacatga 
               
               
                   
               
               
                 taagggagccagcagacctctgatctcttcctgaatgctaatcttaaacatcctgaggaagaatgggacttccatttggg 
               
               
                   
               
               
                 gtgggcctatgatagggtaataagacagtagtgaatatcaagctacaaaaagccccctttcaaattcttctcagtcctaa 
               
               
                   
               
               
                 cttttcatactaagcccagtccttccaaagcagactgtgaaagagtgatagttccgggagactagcactgcagattccgg 
               
               
                   
               
               
                 gtcactgtgagtgggggaggcagggaagaagggctcacaggacagtcaaaccatgccccctgtttttccttcttcaagta 
               
               
                   
               
               
                 gacctctataagacaacagagacaactaaggctgagtggccaggcgaggagaaaccatctcgccgtaaaacatggaagga 
               
               
                   
               
               
                 acacttcaggggaaaggtggtatctctaagcaagagaactgagtggagtcaaggctgagagatgcaggataagcaaatgg 
               
               
                   
               
               
                 gtagtgaaaagacattcatgaggacagctaaaacaataagtaatgtaaaatacagcatagcaaaactttaacctccaaat 
               
               
                   
               
               
                 caagcctctacttgaatccttttctgagggatgaataaggcataggcatcaggggctgttgccaatgtgcattagctgtt 
               
               
                   
               
               
                 tgcagcctcaccttctttcatggagtttaagatatagtgtattttcccaaggtttgaactagctcttcatttctttatgt 
               
               
                   
               
               
                 tttaaatgcactgacctcccacattccctttttagtaaaatattcagaaataatttaaatacatcattgcaatgaaaata 
               
               
                   
               
               
                 aatgttttttattaggcagaatccagatgctcaaggcccttcataatatcccccagtttagtagttggacttagggaaca 
               
               
                   
               
               
                 aaggaacctttaatagaaattggacagcaagaaagcgagcttagtgatacttgtgggccagggcattagccacaccagcc 
               
               
                   
               
               
                 accactttctgataggcagcctgcactggtggggtgAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACGTT 
               
               
                   
               
               
                 GCCCAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAAAAGGGCCTAGCTTGGACTCAGAATAATCCAGC 
               
               
                   
               
               
                 CTTATCCCAACCATAAAATAAAAGCAGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACCTCTTACATCAGT 
               
               
                   
               
               
                 TACAATTTATATGCAGAAATATTTATATGCAGAAATATTGCTATTGCCTTAACCCAGAAATTATCACTGTTATTCTTTAG 
               
               
                   
               
               
                 AATGGTGCAAAGAGGCATGATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAGGGAAAGTATTAGAAATAAGATA 
               
               
                   
               
               
                 AACAAAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAAATTACAAATGCAAAATTACCCTGATTTGGTCAATATGTGTA 
               
               
                   
               
               
                 CCCTGTTACTTCTCCCCTTCCTATGACATGAACTTAACCATAGAAAAGAAGGGGAAAGAAAACATCAAGGGTCCCATAGA 
               
               
                   
               
               
                 CTCACCCTGAAGTTCTCAGGATCCACGTGCAGCTTGTCACAGTGCAGCTCACTCAGctgGGCAAAGGTGCCCTTGAGGTT 
               
               
                   
               
               
                 GTCCAGGTGAGCCAGGCCATCACTAAAGGCACCGAGCACTTTCTTGCCATGAGCCTTCACCTTAGGGTTGCCCATAACAG 
               
               
                   
               
               
                 CATCAGGAGTGGACAGATCCCCAAAGGACTCAAAGAACCTCTGGGTCCAAGGGTAGACCACCAGCAGCCTAAGGGTGGGA 
               
               
                   
               
               
                 AAATAGACCAATAGGCAGAGAGAGTCAGTGCCTATCAGAAACCCAAGAGTCTTCTCTGTCTCCACATGCCCAGTTTCTAT 
               
               
                   
               
               
                 TGGTCTCCTTAAACCTGTCTTGTAACCTTGATACCAACCTGCCCAGGGCCTCACCACCAACTTCATCCACGTTCACCTTG 
               
               
                   
               
               
                 CCCCACAGGGCAGTAACGGCAGACTTCTCCTCAGGAGTCAGGTGCACCATGGTGTCTGTTTGAGGTTGCTAGTGAACACA 
               
               
                   
               
               
                 GTTGTGTCAGAAGCAAATGTAAGCAATAGATGGCTCTGCCCTGACTTTTATGCCCAGCCCTGGCTCCTGCCCTCCCTGCT 
               
               
                   
               
               
                 CCTGGGAGTAGATTGGCCAACCctagggtgtggctccacagggtgaggtctaagtgatgacagccgtacctgtccttggc 
               
               
                   
               
               
                 tcttctggcactggcttaggagttggacttcaaaccctcagccctccctctaagatatatctcttggccccataccatca 
               
               
                   
               
               
                 gtacaaattgctactaaaaacatcctcctttgcaagtgtatttacgtaatatttggaatcacagcttggtaagcatattg 
               
               
                   
               
               
                 aagatcgttttcccaattttcttattacacaaataagaaattgatgcactaaaagtggaagagttttgtctaccataatt 
               
               
                   
               
               
                 cagctttgggatatgtagatggatctcttcctgcgtctccagaatatgcaaaatacttacaggacagaatggatgaaaac 
               
               
                   
               
               
                 tctacctcagttctaagcatatcttctccttatttggattaaaaccttctggtaagaaaagaaaaaaaatatatatatat 
               
               
                   
               
               
                 atgtgtatatatacacacatacatatacatatatatgcattcatttgttgttgtttttcttaatttgctcatggtatatg 
               
               
                   
               
               
                 tgtatatatatatatatatattcaggaaataatatattctagaatatgtcacattctgtctcaggcatccattttcttta 
               
               
                   
               
               
                 tgatgccgtttgaggtggagttttagtcaggtggtcagcttctccttttttttgccatctgccctgtaagcatcctgctg 
               
               
                   
               
               
                 gggacccagataggagtcatcactctaggctgagaacatctgggcacacaccctaagcctcagcatgactcatcatgact 
               
               
                   
               
               
                 cagcattgctgtgcttgagccagaaggtttgcttagaaggttacacagaaccagaaggcgggggtggggcactgaccccg 
               
               
                   
               
               
                 acaggggcctggccagaactgctcatgcttggactatgggaggtcactaatggagacacacagaaatgtaacaggaacta 
               
               
                   
               
               
                 aggaaaaactgaagcttatttaatcagagatgagatgctggaagggatagagggagctgagcttgtaaaaagtatagtaa 
               
               
                   
               
               
                 tcattcagcaaatggttttgaagcacctgctggatgctaaacactattttcagtgcttgaatcataaataagaataaaac 
               
               
                   
               
               
                 atgtatcttattccccacaagagtccaagtaaaaaataacagttaattataatgtgctctgtcccccaggctggagtgca 
               
               
                   
               
               
                 gtggcacgatctcagctcactgcaacctccgcctcccgggttcaagcaattctcctgcctcagccaccctaatagctggg 
               
               
                   
               
               
                 attacaggtgcacaccaccatgccaggctaatttttgtactttttgtagaggcagggtatcaccatgttgtccaagatgg 
               
               
                   
               
               
                 tcttgaactcctgagctccaagcagtccacccacctcagcctcccaaagtgctatctgcggccgcctatctgtaccacta 
               
               
                   
               
               
                 gtctcgagaagctttcattaaaaaaagtctaaccagctgcattcgactttgactgcagcagctggttagaaggttctact 
               
               
                   
               
               
                 ggaggagggtcccagcccattgctaaattaacatcaggctctgagactggcagtatatctctaacagtggttgatgctat 
               
               
                   
               
               
                 cttctggaacttgcctgctacattgagaccactgacccatacataggaagcccatagctctgtcctgaactgttaggcca 
               
               
                   
               
               
                 ctggtccagagagtgtgcatctcctttgatcctcataataaccctatgagatagacacaattattactcttactttatag 
               
               
                   
               
               
                 atgatgatcctgaaaacataggagtcaaggcacttgcccctagctgggggtataggggagcagtcccatgtagtagtaga 
               
               
                   
               
               
                 atgaaaaatgctgctatgctgtgcctcccccacctttcccatgtctgccctctactcatggtctatctctcctggctcct 
               
               
                   
               
               
                 gggagtcatggactccacccagcaccaccaacctgacctaaccacctatctgagcctgccagcctataacccatctgggc 
               
               
                   
               
               
                 cctgatagctggtggccagccctgaccccaccccaccctccctggaacctctgatagacacatctggcacaccagctcgc 
               
               
                   
               
               
                 aaagtcaccgtgagggtcttgtgtttgctgagtcaaaattccttgaaatccaagtccttagagactcctgctcccaaatt 
               
               
                   
               
               
                 tacagtcatagacttcttcatggctgtctcctttatccacagaatgattcctttgcttcattgccccatccatctgatcc 
               
               
                   
               
               
                 tcctcatcagtgcagcacagggcccatgagcagtagctgcagagtctcacataggtctggcactgcctctgacatgtccg 
               
               
                   
               
               
                 accttaggcaaatgcttgactcttctgagctcagtcttgtcatggcaaaataaagataataatagtgtttttttatggag 
               
               
                   
               
               
                 ttagcgtgaggatggaaaacaatagcaaaattgattagactataaaaggtctcaacaaatagtagtagattttatcatcc 
               
               
                   
               
               
                 attaatccttccctctcctctcttactcatcccatcacgtatgcctcttaattttcccttacctataataagagttattc 
               
               
                   
               
               
                 ctcttattatattcttcttatagtgattctggatattaaagtgggaatgaggggcaggccactaacgaagaagatgtttc 
               
               
                   
               
               
                 tcaaagaagccattctccccacatagatcatctcagcagggttcaggaagataaaggaggatcaaggtcgaaggtaggaa 
               
               
                   
               
               
                 ctaaggaagaacactgggcaagtggatcctgagccccttttcctctaactgaaagaaggaaaaaaaaaatggaacccaaa 
               
               
                   
               
               
                 atattctacatagtttccatgtcacagccagggctgggcagtctcctgttatttcttttaaaataaatatatcatttaaa 
               
               
                   
               
               
                 tgcataaataagcaaaccctgctcgggaatgggagggagagtctctggagtccaccccttctcggccctggctctgcaga 
               
               
                   
               
               
                 tagtgctatcaaagccctgacagagccctgcccattgctgggccttggagtgagtcagcctagtagagaggcagggcaag 
               
               
                   
               
               
                 ccatctcatagctgctgagtgggagagagaaaagggctcattgtctataaactcaggtcatggctattcttattctcaca 
               
               
                   
               
               
                 ctaagaaaaagaatgagatgtctacatataccctgcgtcccctcttgtgtactggggcccccaagagctctctaaaagtg 
               
               
                   
               
               
                 atggcaaagtcattgcgctagatgccatcccatctattataaacctgcatttgtctccacacaccagtcatggacaataa 
               
               
                   
               
               
                 ccctcctcccaggtccacgtgcttgtctttgtataatactcaagtaatttcggaaaatgtattctttcaatcttgttctg 
               
               
                   
               
               
                 ttattcctgtttcaatggcttagtagaaaaagtacatacttgttttcccataaattgacaatagacaatttcacatcaat 
               
               
                   
               
               
                 gtctatatgggtcgttgtgtttgctgtgtttgcaaaaactcacaataactttatattgttactactctaagaaagttaca 
               
               
                   
               
               
                 acatggtgaatacaagagaaagctattacaagtccagaaaataaaagttatcatcttgaggcctcagctttctaggaata 
               
               
                   
               
               
                 atatcaatattacaaaatttaatctaacaattatgaacagcaatgagataatatgtacaaagtacccagacctatgtggt 
               
               
                   
               
               
                 agagcatcaaggaagcgcattgcggagcagttttttgtttgtttgtttttgtattctgtttcgtgaggcaaggtttcact 
               
               
                   
               
               
                 ctgctgtccaggctggagtgcagtggcaagatcatgtctcactgcagccttgacacgcgtcgacggtaccgttaacgatc 
               
               
                   
               
               
                 ttagccactttttaaaagaaaaggggggactggaagggctaattcactcccaacgaagacaagatatcctgCTAGTCCTT 
               
               
                   
               
               
                 CCTTTCTAAATGACGAGAGAGACAGAAGAATTCTTCAAGGTTAGTGTGTCCAGCATGCAACCTTTCCTTCCTGGATGAGC 
               
               
                   
               
               
                 ATCCCTGGAGTAGGAGAGCCAGCCTGCCTCCTGCGCTGGCACAGAGCCCGGTTCCCTAGACAACTGCCTCTCCAAATCTG 
               
               
                   
               
               
                 ATGTCCAGCGCCACCTGGTGTCCACATCAAGCAGACACAATTAATAGTCAACCTGTTCAGGAAAACTGTGAGGGGGAAAA 
               
               
                   
               
               
                 AAAAGAAAGAGGATTTATGAAGGGAAAAGAAAGTTTAGAGGATATGCCACGATTGGctagcagctgctttttgcctgtac 
               
               
                   
               
               
                 tgggtctctctggttagaccagatctgagcctgggagctctctggctaactagggaacccactgcttaagcctcaataaa 
               
               
                   
               
               
                 gcttgccttgagtgcttcatccggAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGT 
               
               
                   
               
               
                 TGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCT 
               
               
                   
               
               
                 CCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACT 
               
               
                   
               
               
                 GTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCT 
               
               
                   
               
               
                 CCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATT 
               
               
                   
               
               
                 CCGTGGTGTTGTCGGGGAAGCTGACGTCCTTTCCATGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCC 
               
               
                   
               
               
                 TTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCG 
               
               
                   
               
               
                 TCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCCTGtccggtAGCTTGCCAGCCTCGACT 
               
               
                   
               
               
                 GTGCCTTCTAGTTGCCAGCCGTCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGT 
               
               
                   
               
               
                 CCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGG 
               
               
                   
               
               
                 ACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGcaagcttggcgtaatcatggtcatagctgtttcctgtgtga 
               
               
                   
               
               
                 aattgttatccgctcacaattccacacaacatacgagccggaagcataaagtgtaaagcctggggtgcctaatgagtgag 
               
               
                   
               
               
                 ctaactcacattaattgcgttgcgctcactgcccgctttccagtcgggaaacctgtcgtgccagctgcattaatgaatcg 
               
               
                   
               
               
                 gccaacgcgcggggagaggcggtttgcgtattgggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgtt 
               
               
                   
               
               
                 cggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaa 
               
               
                   
               
               
                 catgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgccccc 
               
               
                   
               
               
                 ctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccc 
               
               
                   
               
               
                 cctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaag 
               
               
                   
               
               
                 cgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacg 
               
               
                   
               
               
                 aaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcg 
               
               
                   
               
               
                 ccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcc 
               
               
                   
               
               
                 taactacggctacactagaagaacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggta 
               
               
                   
               
               
                 gctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaa 
               
               
                   
               
               
                 ggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggt 
               
               
                   
               
               
                 catgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatg 
               
               
                   
               
               
                 agtaaacttggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgtctatttcgttcatccata 
               
               
                   
               
               
                 gttgcctgactccccgtcgtgtagataactacgatacgggagggcttaccatctggccccagtgctgcaatgataccgcg 
               
               
                   
               
               
                 agacccacgctcaccggctccagatttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaa 
               
               
                   
               
               
                 ctttatccgcctccatccagtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgcaac 
               
               
                   
               
               
                 gttgttgccattgctacaggcatcgtggtgtcacgctcgtcgtttggtatggcttcattcagctccggttcccaacgatc 
               
               
                   
               
               
                 aaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctccttcggtcctccgatcgttgtcagaagtaagt 
               
               
                   
               
               
                 tggccgcagtgttatcactcatggttatggcagcactgcataattctcttactgtcatgccatccgtaagatgcttttct 
               
               
                   
               
               
                 gtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcggcgaccgagttgctcttgcccggcgtcaatacg 
               
               
                   
               
               
                 ggataataccgcgccacatagcagaactttaaaagtgctcatcattggaaaacgttcttcggggcaaaactctcaaggat 
               
               
                   
               
               
                 cttaccgctgttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatcttttactttcaccagcg 
               
               
                   
               
               
                 tttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaagggaataagggcgacacggaaatgttgaatactcata 
               
               
                   
               
               
                 ctcttcctttttcaatattattgaagcatttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaa 
               
               
                   
               
               
                 aaataaacaaataggggttccgcgcacatttccccgaaaagtgccacctgacgtctaagaaaccattattatcatgacat 
               
               
                   
               
               
                 taacctataaaaataggcgtatcacgaggccctttcgtctcgcgcgtttcggtgatgacggtgaaaacctctgacacatg 
               
               
                   
               
               
                 cagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgt 
               
               
                   
               
               
                 tggcgggtgtcggggctggcttaactatgcggcatcagagcagattgtactgagagtgcacc 
               
               
                   
               
               
                 [SEQ ID NO: 38] 
                   
               
               
                 ATTGGCCATTGCATACGTTGTATCCATATCATAATATGTACATTTATATTGGCTCATGTCCAACATTACCGCCATGTTGA 
                   
               
               
                   
               
               
                 CATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTAC 
               
               
                   
               
               
                 ATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCA 
               
               
                   
               
               
                 TAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAA 
               
               
                   
               
               
                 GTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGAC 
               
               
                   
               
               
                 CTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACA 
               
               
                   
               
               
                 TCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGG 
               
               
                   
               
               
                 CACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTG 
               
               
                   
               
               
                 GGAGGTCTATATAAGCAGAGCTCGTTTAGTGAACCGGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGG 
               
               
                   
               
               
                 CTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGTGTGCCCGTCTGTTGTGTG 
               
               
                   
               
               
                 ACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTCAGTGTGGAAAATCTCTAGCAGTGGCGCCCGAACAGGGACTTGA 
               
               
                   
               
               
                 AAGCGAAAGGGAAACCAGAGGAGCTCTCTCGACGCAGGACTCGGCTTGCTGAAGCGCGCACGGCAAGAGGCGAGGGGCGG 
               
               
                   
               
               
                 CGACTGGTGAGTACGCCAAAAATTTTGACTAGCGGAGGCTAGAAGGAGAGAGACGGGTGCGAGAGCGTCAGTATTAAGCG 
               
               
                   
               
               
                 GGGGTGAATAAGATCGCGATGGGAAAAAATTCGGTTAAGGCCAGGGGGAAAGAAAAAATATAAATTAAAACATATAGTAT 
               
               
                   
               
               
                 GGGCAAGCAGGGAGCTAGAACGATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGTAGACAAATACTGGGA 
               
               
                   
               
               
                 CAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATCATTATATAATACAGTAGCAACCCTCTATTGTGTGCA 
               
               
                   
               
               
                 TCAAAGGATAGAGATAAAAGACACCAAGGAAGCTTTAGACAAGATAGAGGAAGAGCAAAACAAAAGTAAGACCACCGCAC 
               
               
                   
               
               
                 AGCAAGCGGCCGCTGATCTTCAGACCTGGAGGAGGAGATATGAGGGACAATTGGAGAAGTGAATTATATAAATATAAAGT 
               
               
                   
               
               
                 AGTAAAAATTGAACCATTAGGAGTAGCACCCACCAAGGCAAAGAGAAGAGTGGTGCAGAGAGAAAAAAGAGCAGTGGGAA 
               
               
                   
               
               
                 TAGGAGCTTTGTTCCTTGGGTTCTTGGGAGCAGCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACAGGCC 
               
               
                   
               
               
                 AGACAATTATTGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTGAGGCGCAACAGCATCTGTTGCAACT 
               
               
                   
               
               
                 CACAGTCTGGGGCATCAAGCAGCTCCAGGCAAGAATCCTGGCTGTGGAAAGATACCTAAAGGATCAACAGCTCCTGGGGA 
               
               
                   
               
               
                 TTTGGGGTTGCTCTGGAAAACTCATTTGCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTCTGGAACAG 
               
               
                   
               
               
                 ATTTGGAATCACACGACCTGGATGGAGTGGGACAGAGAAATTAACAATTACACAAGCTTAATACACTCCTTAATTGAAGA 
               
               
                   
               
               
                 ATCGCAAAACCAGCAAGAAAAGAATGAACAAGAATTATTGGAATTAGATAAATGGGCAAGTTTGTGGAATTGGTTTAACA 
               
               
                   
               
               
                 TAACAAATTGGCTGTGGTATATAAAATTATTCATAATGATAGTAGGAGGCTTGGTAGGTTTAAGAATAGTTTTTGCTGTA 
               
               
                   
               
               
                 CTTTCTATAGTGAATAGAGTTAGGCAGGGATATTCACCATTATCGTTTCAGACCCACCTCCCAACCCCGAGGGGACCCGA 
               
               
                   
               
               
                 CAGGCCCGAAGGAATAGAAGAAGAAGGTGGAGAGAGAGACAGAGACAGATCCATTCGATTAGTGAACGGATCTCGACGGT 
               
               
                   
               
               
                 ATCGTTTTAAAAGAAAAGGGGGGATTGGGGGGTACAGTGCAGGGGAAAGAATAGTAGACATAATAGCAACAGACATACAA 
               
               
                   
               
               
                 ACTAAAGAATTACAAAAACAAATTACAAAAATTCAAAATTTTATCGGCGTGTTGGGGGTGGACCATCCTCTAGGTATTGA 
               
               
                   
               
               
                 ATAAGAAAAATGAAGTTAAGGTGGTTGATGGTAACACTATGCTAATAACTGCAGAGCCAGAAGCACCATAAGGGACATGA 
               
               
                   
               
               
                 TAAGGGAGCCAGCAGACCTCTGATCTCTTCCTGAATGCTAATCTTAAACATCCTGAGGAAGAATGGGACTTCCATTTGGG 
               
               
                   
               
               
                 GTGGGCCTATGATAGGGTAATAAGACAGTAGTGAATATCAAGCTACAAAAAGCCCCCTTTCAAATTCTTCTCAGTCCTAA 
               
               
                   
               
               
                 CTTTTCATACTAAGCCCAGTCCTTCCAAAGCAGACTGTGAAAGAGTGATAGTTCCGGGAGACTAGCACTGCAGATTCCGG 
               
               
                   
               
               
                 GTCACTGTGAGTGGGGGAGGCAGGGAAGAAGGGCTCACAGGACAGTCAAACCATGCCCCCTGTTTTTCCTTCTTCAAGTA 
               
               
                   
               
               
                 GACCTCTATAAGACAACAGAGACAACTAAGGCTGAGTGGCCAGGCGAGGAGAAACCATCTCGCCGTAAAACATGGAAGGA 
               
               
                   
               
               
                 ACACTTCAGGGGAAAGGTGGTATCTCTAAGCAAGAGAACTGAGTGGAGTCAAGGCTGAGAGATGCAGGATAAGCAAATGG 
               
               
                   
               
               
                 GTAGTGAAAAGACATTCATGAGGACAGCTAAAACAATAAGTAATGTAAAATACAGCATAGCAAAACTTTAACCTCCAAAT 
               
               
                   
               
               
                 CAAGCCTCTACTTGAATCCTTTTCTGAGGGATGAATAAGGCATAGGCATCAGGGGCTGTTGCCAATGTGCATTAGCTGTT 
               
               
                   
               
               
                 TGCAGCCTCACCTTCTTTCATGGAGTTTAAGATATAGTGTATTTTCCCAAGGTTTGAACTAGCTCTTCATTTCTTTATGT 
               
               
                   
               
               
                 TTTAAATGCACTGACCTCCCACATTCCCTTTTTAGTAAAATATTCAGAAATAATTTAAATACATCATTGCAATGAAAATA 
               
               
                   
               
               
                 AATGTTTTTTATTAGGCAGAATCCAGATGCTCAAGGCCCTTCATAATATCCCCCAGTTTAGTAGTTGGACTTAGGGAACA 
               
               
                   
               
               
                 AAGGAACCTTTAATAGAAATTGGACAGCAAGAAAGCGAGCTTAGTGATACTTGTGGGCCAGGGCATTAGCCACACCAGCC 
               
               
                   
               
               
                 ACCACTTTCTGATAGGCAGCCTGCACTGGTGGGGTGAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACGTT 
               
               
                   
               
               
                 GCCCAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAAAAGGGCCTAGCTTGGACTCAGAATAATCCAGC 
               
               
                   
               
               
                 CTTATCCCAACCATAAAATAAAAGCAGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACCTCTTACATCAGT 
               
               
                   
               
               
                 TACAATTTATATGCAGAAATATTTATATGCAGAAATATTGCTATTGCCTTAACCCAGAAATTATCACTGTTATTCTTTAG 
               
               
                   
               
               
                 AATGGTGCAAAGAGGCATGATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAGGGAAAGTATTAGAAATAAGATA 
               
               
                   
               
               
                 AACAAAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAAATTACAAATGCAAAATTACCCTGATTTGGTCAATATGTGTA 
               
               
                   
               
               
                 CCCTGTTACTTCTCCCCTTCCTATGACATGAACTTAACCATAGAAAAGAAGGGGAAAGAAAACATCAAGGGTCCCATAGA 
               
               
                   
               
               
                 CTCACCCTGAAGTTCTCAGGATCCACGTGCAGCTTGTCACAGTGCAGCTCACTCAGCTGGGCAAAGGTGCCCTTGAGGTT 
               
               
                   
               
               
                 GTCCAGGTGAGCCAGGCCATCACTAAAGGCACCGAGCACTTTCTTGCCATGAGCCTTCACCTTAGGGTTGCCCATAACAG 
               
               
                   
               
               
                 CATCAGGAGTGGACAGATCCCCAAAGGACTCAAAGAACCTCTGGGTCCAAGGGTAGACCACCAGCAGCCTAAGGGTGGGA 
               
               
                   
               
               
                 AAATAGACCAATAGGCAGAGAGAGTCAGTGCCTATCAGAAACCCAAGAGTCTTCTCTGTCTCCACATGCCCAGTTTCTAT 
               
               
                   
               
               
                 TGGTCTCCTTAAACCTGTCTTGTAACCTTGATACCAACCTGCCCAGGGCCTCACCACCAACTTCATCCACGTTCACCTTG 
               
               
                   
               
               
                 CCCCACAGGGCAGTAACGGCAGACTTCTCCTCAGGAGTCAGGTGCACCATGGTGTCTGTTTGAGGTTGCTAGTGAACACA 
               
               
                   
               
               
                 GTTGTGTCAGAAGCAAATGTAAGCAATAGATGGCTCTGCCCTGACTTTTATGCCCAGCCCTGGCTCCTGCCCTCCCTGCT 
               
               
                   
               
               
                 CCTGGGAGTAGATTGGCCAACCCTAGGGTGTGGCTCCACAGGGTGAGGTCTAAGTGATGACAGCCGTACCTGTCCTTGGC 
               
               
                   
               
               
                 TCTTCTGGCACTGGCTTAGGAGTTGGACTTCAAACCCTCAGCCCTCCCTCTAAGATATATCTCTTGGCCCCATACCATCA 
               
               
                   
               
               
                 GTACAAATTGCTACTAAAAACATCCTCCTTTGCAAGTGTATTTACGGCATCTGTGAAGGAAAGAAACATCTCCTCTAAAC 
               
               
                   
               
               
                 CACTATGCTGCTAGAGCCTCTTTTCTGTACTCAAGCCTCATTCAGACACTAGTGTCACCAGTCTCCTCATATACCTATTG 
               
               
                   
               
               
                 TATTTTCTTCTTCTTGCTGGTTTAGTCATGTTTTCTGGGAGCTTAGGGGCTTATTTTATTTTGTTTTGTTTTCTAATCAA 
               
               
                   
               
               
                 CAGAGATGGGCAAACCCATTATTTTTTTCTTTAGACTTGGGATGGTGATAGCTGGGCAGCGTCAGAAACTGTGTGTGGAT 
               
               
                   
               
               
                 ATAGATAAGAGCTCGGACTATGCTGAGCTGTGATGAGGGAGGGACCTAGCCAAAGGCAGTGAGAGTCAGAATGCTCCTGC 
               
               
                   
               
               
                 TATTGCCTTCTCAGTCCCCACGCTTGGTTTCTACACAAGTAGATACATAGAAAAGGCTATAGGTTAGTGTTTGAGAGTCC 
               
               
                   
               
               
                 TGCATGAGTTAGTTGCTCAGAAATGCCCGATAAATATGTTATGTGTGTTTATGTATATATATGTTTTATATATATATATG 
               
               
                   
               
               
                 TGTGTGTGTGTGTGTGTGTGTGTTGTGTTTACAAATATGTGATTATCATCAAAACGTGAGGGCTAAAGTGACCAGATAAC 
               
               
                   
               
               
                 TTGCAGGTCTAGAATATGTCACATTCTGTCTCAGGCATCCATTTTCTTTATGATGCCGTTTGAGGTGGAGTTTTAGTCAG 
               
               
                   
               
               
                 GTGGTCAGCTTCTCCTTTTTTTTGCCATCTGCCCTGTAAGCATCCTGCTGGGGACCCAGATAGGAGTCATCACTCTAGGC 
               
               
                   
               
               
                 TGAGAACATCTGGGCACACACCCTAAGCCTCAGCATGACTCATCATGACTCAGCATTGCTGTGCTTGAGCCAGAAGGTTT 
               
               
                   
               
               
                 GCTTAGAAGGTTACACAGAACCAGAAGGCGGGGGTGGGGCACTGACCCCGACAGGGGCCTGGCCAGAACTGCTCATGCTT 
               
               
                   
               
               
                 GGACTATGGGAGGTCACTAATGGAGACACACAGAAATGTAACAGGAACTAAGGAAAAACTGAAGCTTATTTAATCAGAGA 
               
               
                   
               
               
                 TGAGATGCTGGAAGGGATAGAGGGAGCTGAGCTTGTAAAAAGTATAGTAATCATTCAGCAAATGGTTTTGAAGCACCTGC 
               
               
                   
               
               
                 TGGATGCTAAACACTATTTTCAGTGCTTGAATCATAAATAAGAACAAAACATGTATCTTATTCCCCACAAGAGTCCAAGT 
               
               
                   
               
               
                 AAAAAATAACAGTTAATTATAATGTGCTCTGTCCCCCAGGCTGGAGTGCAGTGGCACGATCTCAGCTCACTGCAACCTCC 
               
               
                   
               
               
                 GCCTCCCGGGTTCAAGCAATTCTCCTGCCTCAGCCACCCTAATAGCTGGGATTACAGGTGCACACCACCATGCCAGGCTA 
               
               
                   
               
               
                 ATTTTTGTACTTTTTGTAGAGGCAGGGTATCACCATGTTGTCCAAGATGGTCTTGAACTCCTGAGCTCCAAGCAGTCCAC 
               
               
                   
               
               
                 CCACCTCAGCCTCCCAAAGTGCTATCTGCGGCCGCCTATCTGTACCACTAGTCTCGAGAAGCTTTCATCAAAAAAAGTCT 
               
               
                   
               
               
                 AACCAGCTGCATTCGACTTTGACTGCAGCAGCTGGTTAGAAGGTTCTACTGGAGGAGGGTCCCAGCCCATTGCTAAATTA 
               
               
                   
               
               
                 ACATCAGGCTCTGAGACTGGCAGTATATCTCTAACAGTGGTTGATGCTATCTTCTGGAACTTGCCTGCTACATTGAGACC 
               
               
                   
               
               
                 ACTGACCCATACATAGGAAGCCCATAGCTCTGTCCTGAACTGTTAGGCCACTGGTCCAGAGAGTGTGCATCTCCTTTGAT 
               
               
                   
               
               
                 CCTCATAATAACCCTATGAGATAGACACAATTATTACTCTTACTTTATAGATGATGATCCTGAAAACATAGGAGTCAAGG 
               
               
                   
               
               
                 CACTTGCCCCTAGCTGGGGGTATAGGGGAGCAGTCCCATGTAGTAGTAGAATGAAAAATGCTGCTATGCTGTGCCTCCCC 
               
               
                   
               
               
                 CACCTTTCCCATGTCTGCCCTCTACTCATGGTCTATCTCTCCTGGCTCCTGGGAGTCATGGACTCCACCCAGCACCACCA 
               
               
                   
               
               
                 ACCTGACCTAACCACCTATCTGAGCCTGCCAGCCTATAACCCATCTGGGCCCTGATAGCTGGTGGCCAGCCCTGACCCCA 
               
               
                   
               
               
                 CCCCACCCTCCCTGGAACCTCTGATAGACACATCTGGCACACCAGCTCGCAAAGTCACCGTGAGGGTCTTGTGTTTGCTG 
               
               
                   
               
               
                 AGTCAAAATTCCTTGAAATCCAAGTCCTTAGAGACTCCTGCTCCCAAATTTACAGTCATAGACTTCTTCATGGCTGTCTC 
               
               
                   
               
               
                 CTTTATCCACAGAATGATTCCTTTGCTTCATTGCCCCATCCATCTGATCCTCCTCATCAGTGCAGCACAGGGCCCATGAG 
               
               
                   
               
               
                 CAGTAGCTGCAGAGTCTCACATAGGTCTGGCACTGCCTCTGACATGTCCGACCTTAGGCAAATGCTTGACTCTTCTGAGC 
               
               
                   
               
               
                 TCAGTCTTGTCATGGCAAAACAAAGATAATAATAGTGTTTTTTTATGGAGTTAGCGTGAGGATGGAAAACAATAGCAAAA 
               
               
                   
               
               
                 TTGATTAGACTATAAAAGGTCTCAACAAATAGTAGTAGATTTTATCATCCATTAATCCTTCCCTCTCCTCTCTTACTCAT 
               
               
                   
               
               
                 CCCATCACGTATGCCTCTTAATTTTCCCTTACCTATAATAAGAGTTATTCCTCTTATTATATTCTTCTTATAGTGATTCT 
               
               
                   
               
               
                 GGATATCAAAGTGGGAATGAGGGGCAGGCCACTAACGAAGAAGATGTTTCTCAAAGAAGCCATTCTCCCCACATAGATCA 
               
               
                   
               
               
                 TCTCAGCAGGGTTCAGGAAGATAAAGGAGGATCAAGGTCGAAGGTAGGAACTAAGGAAGAACACTGGGCAAGTGGATCCT 
               
               
                   
               
               
                 AAATATATCATTTAAATGCATAAATAAGCAAACCCTGCTCGGGAATGGGAGGGAGAGTCTCTGGAGTCCACCCCTTCTCG 
               
               
                   
               
               
                 GCCCTGGCTCTGCAGATAGTGCTATCAAAGCCCTGACAGAGCCCTGCCCATTGCTGGGCCTTGGAGTGAGTCAGCCTAGT 
               
               
                   
               
               
                 AGAGAGGCAGGGCAAGCCATCTCATAGCTGCTGAGTGGGAGAGAGAAAAGGGCTCATTGTCTATAAACTCAGGTCATGGC 
               
               
                   
               
               
                 TATTCTTATTCTCACACTAAGAAAAAGAATGAGATGTCTACATATACCCTGCGTCCCCTCTTGTGTACTGGGGCCCCCAA 
               
               
                   
               
               
                 GAGCTCTCTAAAAGTGATGGCAAAGTCATTGCGCTAGATGCCATCCCATCTATTATAAACCTGCATTTGTCTCCACACAC 
               
               
                   
               
               
                 CAGTCATGGACAATAACCCTCCTCCCAGGTCCACGTGCTTGTCTTTGTATAATACTCAAGTAATTTCGGAAAATGTATTC 
               
               
                   
               
               
                 TTTCAATCTTGTTCTGTTATTCCTGTTTCAATGGCTTAGTAGAAAAAGTACATACTTGTTTTCCCATAAATTGACAATAG 
               
               
                   
               
               
                 ACAATTTCACATCAATGTCTATATGGGTCGTTGTGTTTGCTGTGTTTGCAAAAACTCACAATAACTTTATATTGTTACTA 
               
               
                   
               
               
                 CTCTAAGAAAGTTACAACATGGTGAATACAAGAGAAAGCTATTACAAGTCCAGAAAACAAAAGTTATCATCTTGAGGCCT 
               
               
                   
               
               
                 CAGCTTTCTAGGAATAATATCAATATTACAAAACGCGTCGACGGTACCGTTAACGATCTTAGCCACTTTTTAAAAGAAAA 
               
               
                   
               
               
                 GGGGGGACTGGAAGGGCTAATTCACTCCCAACGAAGACAAGATATCCTGCTAGTCCTTCCTTTCTAAATGACGAGAGAGA 
               
               
                   
               
               
                 CAGAAGAATTCTTCAAGGTTAGTGTGTCCAGCATGCAACCTTTCCTTCCTGGATGAGCATCCCTGGAGTAGGAGAGCCAG 
               
               
                   
               
               
                 CCTGCCTCCTGCGCTGGCACAGAGCCCGGTTCCCTAGACAACTGCCTCTCCAAATCTGATGTCCAGCGCCACCTGGTGTC 
               
               
                   
               
               
                 CACATCAAGCAGACACAATTAATAGTCAACCTGTTCAGGAAAACTGTGAGGGGGAAAAAAAAGAAAGAGGATTTATGAAG 
               
               
                   
               
               
                 GGAAAAGAAAGTTTAGAGGATATGCCACGATTGGCTAGCAGCTGCTTTTTGCCTGTACTGGGTCTCTCTGGTTAGACCAG 
               
               
                   
               
               
                 ATCTGAGCCTGGGAGCTCTCTGGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCATCC 
               
               
                   
               
               
                 GGAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGA 
               
               
                   
               
               
                 TACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTT 
               
               
                   
               
               
                 GCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCA 
               
               
                   
               
               
                 CTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTC 
               
               
                   
               
               
                 ATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAGCT 
               
               
                   
               
               
                 GACGTCCTTTCCATGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCC 
               
               
                   
               
               
                 TCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACG 
               
               
                   
               
               
                 AGTCGGATCTCCCTTTGGGCCGCCTCCCCGCCTGTCCGGTAGCTTGCCAGCCTCGACTGTGCCTTCTAGTTGCCAGCCGT 
               
               
                   
               
               
                 CTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAA 
               
               
                   
               
               
                 ATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGA 
               
               
                   
               
               
                 AGACAATAGCAGGCATGCAAGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAATTGTTATCCGCTCACAATTC 
               
               
                   
               
               
                 CACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTG 
               
               
                   
               
               
                 CGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGG 
               
               
                   
               
               
                 TTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAG 
               
               
                   
               
               
                 CTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAA 
               
               
                   
               
               
                 AAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCG 
               
               
                   
               
               
                 ACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCT 
               
               
                   
               
               
                 CTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCA 
               
               
                   
               
               
                 CGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCG 
               
               
                   
               
               
                 CTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTA 
               
               
                   
               
               
                 ACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGA 
               
               
                   
               
               
                 ACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAAC 
               
               
                   
               
               
                 CACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGA 
               
               
                   
               
               
                 TCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATC 
               
               
                   
               
               
                 TTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTA 
               
               
                   
               
               
                 CCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGT 
               
               
                   
               
               
                 AGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCA 
               
               
                   
               
               
                 GATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTC 
               
               
                   
               
               
                 TATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCA 
               
               
                   
               
               
                 TCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCC 
               
               
                   
               
               
                 ATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCAT 
               
               
                   
               
               
                 GGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCA 
               
               
                   
               
               
                 AGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGC 
               
               
                   
               
               
                 AGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGT 
               
               
                   
               
               
                 TCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGG 
               
               
                   
               
               
                 AAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATT 
               
               
                   
               
               
                 GAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCG 
               
               
                   
               
               
                 CGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTAT 
               
               
                   
               
               
                 CACGAGGCCCTTTCGTCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCGGAGACGGTCACAG 
               
               
                   
               
               
                 CTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGCCCGTCAGGGCGCGTCAGCGGGTGTTGGCGGGTGTCGGGGCTGGCTT 
               
               
                   
               
               
                 AACTATGCGGCATCAGAGCAGATTGTACTGAGAGTGCACC 
               
               
                   
               
               
                 [SEQ ID NO: 39] 
                   
               
               
                 ATTGGCCATTGCATACGTTGTATCCATATCATAATATGTACATTTATATTGGCTCATGTCCAACATTACCGCCATGTTGA 
                   
               
               
                   
               
               
                 CATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTAC 
               
               
                   
               
               
                 ATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCA 
               
               
                   
               
               
                 TAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAA 
               
               
                   
               
               
                 GTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGAC 
               
               
                   
               
               
                 CTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACA 
               
               
                   
               
               
                 TCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGG 
               
               
                   
               
               
                 CACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTG 
               
               
                   
               
               
                 GGAGGTCTATATAAGCAGAGCTCGTTTAGTGAACCGGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGG 
               
               
                   
               
               
                 CTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGTGTGCCCGTCTGTTGTGTG 
               
               
                   
               
               
                 ACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTCAGTGTGGAAAATCTCTAGCAGTGGCGCCCGAACAGGGACTTGA 
               
               
                   
               
               
                 AAGCGAAAGGGAAACCAGAGGAGCTCTCTCGACGCAGGACTCGGCTTGCTGAAGCGCGCACGGCAAGAGGCGAGGGGCGG 
               
               
                   
               
               
                 CGACTGGTGAGTACGCCAAAAATTTTGACTAGCGGAGGCTAGAAGGAGAGAGACGGGTGCGAGAGCGTCAGTATTAAGCG 
               
               
                   
               
               
                 GGGGTGAATAAGATCGCGATGGGAAAAAATTCGGTTAAGGCCAGGGGGAAAGAAAAAATATAAATTAAAACATATAGTAT 
               
               
                   
               
               
                 GGGCAAGCAGGGAGCTAGAACGATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGTAGACAAATACTGGGA 
               
               
                   
               
               
                 CAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATCATTATATAATACAGTAGCAACCCTCTATTGTGTGCA 
               
               
                   
               
               
                 TCAAAGGATAGAGATAAAAGACACCAAGGAAGCTTTAGACAAGATAGAGGAAGAGCAAAACAAAAGTAAGACCACCGCAC 
               
               
                   
               
               
                 AGCAAGCGGCCGCTGATCTTCAGACCTGGAGGAGGAGATATGAGGGACAATTGGAGAAGTGAATTATATAAATATAAAGT 
               
               
                   
               
               
                 AGTAAAAATTGAACCATTAGGAGTAGCACCCACCAAGGCAAAGAGAAGAGTGGTGCAGAGAGAAAAAAGAGCAGTGGGAA 
               
               
                   
               
               
                 TAGGAGCTTTGTTCCTTGGGTTCTTGGGAGCAGCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACAGGCC 
               
               
                   
               
               
                 AGACAATTATTGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTGAGGCGCAACAGCATCTGTTGCAACT 
               
               
                   
               
               
                 CACAGTCTGGGGCATCAAGCAGCTCCAGGCAAGAATCCTGGCTGTGGAAAGATACCTAAAGGATCAACAGCTCCTGGGGA 
               
               
                   
               
               
                 TTTGGGGTTGCTCTGGAAAACTCATTTGCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTCTGGAACAG 
               
               
                   
               
               
                 ATTTGGAATCACACGACCTGGATGGAGTGGGACAGAGAAATTAACAATTACACAAGCTTAATACACTCCTTAATTGAAGA 
               
               
                   
               
               
                 ATCGCAAAACCAGCAAGAAAAGAATGAACAAGAATTATTGGAATTAGATAAATGGGCAAGTTTGTGGAATTGGTTTAACA 
               
               
                   
               
               
                 TAACAAATTGGCTGTGGTATATAAAATTATTCATAATGATAGTAGGAGGCTTGGTAGGTTTAAGAATAGTTTTTGCTGTA 
               
               
                   
               
               
                 CTTTCTATAGTGAATAGAGTTAGGCAGGGATATTCACCATTATCGTTTCAGACCCACCTCCCAACCCCGAGGGGACCCGA 
               
               
                   
               
               
                 CAGGCCCGAAGGAATAGAAGAAGAAGGTGGAGAGAGAGACAGAGACAGATCCATTCGATTAGTGAACGGATCTCGACGGT 
               
               
                   
               
               
                 ATCGTTTTAAAAGAAAAGGGGGGATTGGGGGGTACAGTGCAGGGGAAAGAATAGTAGACATAATAGCAACAGACATACAA 
               
               
                   
               
               
                 ACTAAAGAATTACAAAAACAAATTACAAAAATTCAAAATTTTATCGGCGTGTTGGGGGTGGACCATCCTCTAGGTATTGA 
               
               
                   
               
               
                 ATAAGAAAAATGAAGTTAAGGTGGTTGATGGTAACACTATGCTAATAACTGCAGAGCCAGAAGCACCATAAGGGACATGA 
               
               
                   
               
               
                 TAAGGGAGCCAGCAGACCTCTGATCTCTTCCTGAATGCTAATCTTAAACATCCTGAGGAAGAATGGGACTTCCATTTGGG 
               
               
                   
               
               
                 GTGGGCCTATGATAGGGTAATAAGACAGTAGTGAATATCAAGCTACAAAAAGCCCCCTTTCAAATTCTTCTCAGTCCTAA 
               
               
                   
               
               
                 CTTTTCATACTAAGCCCAGTCCTTCCAAAGCAGACTGTGAAAGAGTGATAGTTCCGGGAGACTAGCACTGCAGATTCCGG 
               
               
                   
               
               
                 GTCACTGTGAGTGGGGGAGGCAGGGAAGAAGGGCTCACAGGACAGTCAAACCATGCCCCCTGTTTTTCCTTCTTCAAGTA 
               
               
                   
               
               
                 GACCTCTATAAGACAACAGAGACAACTAAGGCTGAGTGGCCAGGCGAGGAGAAACCATCTCGCCGTAAAACATGGAAGGA 
               
               
                   
               
               
                 ACACTTCAGGGGAAAGGTGGTATCTCTAAGCAAGAGAACTGAGTGGAGTCAAGGCTGAGAGATGCAGGATAAGCAAATGG 
               
               
                   
               
               
                 GTAGTGAAAAGACATTCATGAGGACAGCTAAAACAATAAGTAATGTAAAATACAGCATAGCAAAACTTTAACCTCCAAAT 
               
               
                   
               
               
                 CAAGCCTCTACTTGAATCCTTTTCTGAGGGATGAATAAGGCATAGGCATCAGGGGCTGTTGCCAATGTGCATTAGCTGTT 
               
               
                   
               
               
                 TGCAGCCTCACCTTCTTTCATGGAGTTTAAGATATAGTGTATTTTCCCAAGGTTTGAACTAGCTCTTCATTTCTTTATGT 
               
               
                   
               
               
                 TTTAAATGCACTGACCTCCCACATTCCCTTTTTAGTAAAATATTCAGAAATAATTTAAATACATCATTGCAATGAAAATA 
               
               
                   
               
               
                 AATGTTTTTTATTAGGCAGAATCCAGATGCTCAAGGCCCTTCATAATATCCCCCAGTTTAGTAGTTGGACTTAGGGAACA 
               
               
                   
               
               
                 AAGGAACCTTTAATAGAAATTGGACAGCAAGAAAGCGAGCTTAGTGATACTTGTGGGCCAGGGCATTAGCCACACCAGCC 
               
               
                   
               
               
                 ACCACTTTCTGATAGGCAGCCTGCACTGGTGGGGTGAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACGTT 
               
               
                   
               
               
                 GCCCAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAAAAGGGCCTAGCTTGGACTCAGAATAATCCAGC 
               
               
                   
               
               
                 CTTATCCCAACCATAAAATAAAAGCAGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACCTCTTACATCAGT 
               
               
                   
               
               
                 TACAATTTATATGCAGAAATATTTATATGCAGAAATATTGCTATTGCCTTAACCCAGAAATTATCACTGTTATTCTTTAG 
               
               
                   
               
               
                 AATGGTGCAAAGAGGCATGATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAGGGAAAGTATTAGAAATAAGATA 
               
               
                   
               
               
                 AACAAAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAAATTACAAATGCAAAATTACCCTGATTTGGTCAATATGTGTA 
               
               
                   
               
               
                 CCCTGTTACTTCTCCCCTTCCTATGACATGAACTTAACCATAGAAAAGAAGGGGAAAGAAAACATCAAGGGTCCCATAGA 
               
               
                   
               
               
                 CTCACCCTGAAGTTCTCAGGATCCACGTGCAGCTTGTCACAGTGCAGCTCACTCAGCTGGGCAAAGGTGCCCTTGAGGTT 
               
               
                   
               
               
                 GTCCAGGTGAGCCAGGCCATCACTAAAGGCACCGAGCACTTTCTTGCCATGAGCCTTCACCTTAGGGTTGCCCATAACAG 
               
               
                   
               
               
                 CATCAGGAGTGGACAGATCCCCAAAGGACTCAAAGAACCTCTGGGTCCAAGGGTAGACCACCAGCAGCCTAAGGGTGGGA 
               
               
                   
               
               
                 AAATAGACCAATAGGCAGAGAGAGTCAGTGCCTATCAGAAACCCAAGAGTCTTCTCTGTCTCCACATGCCCAGTTTCTAT 
               
               
                   
               
               
                 TGGTCTCCTTAAACCTGTCTTGTAACCTTGATACCAACCTGCCCAGGGCCTCACCACCAACTTCATCCACGTTCACCTTG 
               
               
                   
               
               
                 CCCCACAGGGCAGTAACGGCAGACTTCTCCTCAGGAGTCAGGTGCACCATGGTGTCTGTTTGAGGTTGCTAGTGAACACA 
               
               
                   
               
               
                 GTTGTGTCAGAAGCAAATGTAAGCAATAGATGGCTCTGCCCTGACTTTTATGCCCAGCCCTGGCTCCTGCCCTCCCTGCT 
               
               
                   
               
               
                 CCTGGGAGTAGATTGGCCAACCCTAGGGTGTGGCTCCACAGGGTGAGGTCTAAGTGATGACAGCCGTACCTGTCCTTGGC 
               
               
                   
               
               
                 TCTTCTGGCACTGGCTTAGGAGTTGGACTTCAAACCCTCAGCCCTCCCTCTAAGATATATCTCTTGGCCCCATACCATCA 
               
               
                   
               
               
                 GTACAAATTGCTACTAAAAACATCCTCCTTTGCAAGTGTATTTACTCTAGAATATGTCACATTCTGTCTCAGGCATCCAT 
               
               
                   
               
               
                 TTTCTTTATGATGCCGTTTGAGGTGGAGTTTTAGTCAGGTGGTCAGCTTCTCCTTTTTTTTGCCATCTGCCCTGTAAGCA 
               
               
                   
               
               
                 TCCTGCTGGGGACCCAGATAGGAGTCATCACTCTAGGCTGAGAACATCTGGGCACACACCCTAAGCCTCAGCATGACTCA 
               
               
                   
               
               
                 TCATGACTCAGCATTGCTGTGCTTGAGCCAGAAGGTTTGCTTAGAAGGTTACACAGAACCAGAAGGCGGGGGTGGGGCAC 
               
               
                   
               
               
                 TGACCCCGACAGGGGCCTGGCCAGAACTGCTCATGCTTGGACTATGGGAGGTCACTAATGGAGACACACAGAAATGTAAC 
               
               
                   
               
               
                 AGGAACTAAGGAAAAACTGAAGCTTATTTAATCAGAGATGAGATGCTGGAAGGGATAGAGGGAGCTGAGCTTGTAAAAAG 
               
               
                   
               
               
                 TATAGTAATCATTCAGCAAATGGTTTTGAAGCACCTGCTGGATGCTAAACACTATTTTCAGTGCTTGAATCATAAATAAG 
               
               
                   
               
               
                 AACAAAACATGTATCTTATTCCCCACAAGAGTCCAAGTAAAAAATAACAGTTAATTATAATGTGCTCTGTCCCCCAGGCT 
               
               
                   
               
               
                 GGAGTGCAGTGGCACGATCTCAGCTCACTGCAACCTCCGCCTCCCGGGTTCAAGCAATTCTCCTGCCTCAGCCACCCTAA 
               
               
                   
               
               
                 TAGCTGGGATTACAGGTGCACACCACCATGCCAGGCTAATTTTTGTACTTTTTGTAGAGGCAGGGTATCACCATGTTGTC 
               
               
                   
               
               
                 CAAGATGGTCTTGAACTCCTGAGCTCCAAGCAGTCCACCCACCTCAGCCTCCCAAAGTGCTATCTGCGGCCGCCTATCTG 
               
               
                   
               
               
                 TACCACTAGTCTCGAGAAGCTTTCATCAAAAAAAGTCTAACCAGCTGCATTCGACTTTGACTGCAGCAGCTGGTTAGAAG 
               
               
                   
               
               
                 GTTCTACTGGAGGAGGGTCCCAGCCCATTGCTAAATTAACATCAGGCTCTGAGACTGGCAGTATATCTCTAACAGTGGTT 
               
               
                   
               
               
                 GATGCTATCTTCTGGAACTTGCCTGCTACATTGAGACCACTGACCCATACATAGGAAGCCCATAGCTCTGTCCTGAACTG 
               
               
                   
               
               
                 TTAGGCCACTGGTCCAGAGAGTGTGCATCTCCTTTGATCCTCATAATAACCCTATGAGATAGACACAATTATTACTCTTA 
               
               
                   
               
               
                 CTTTATAGATGATGATCCTGAAAACATAGGAGTCAAGGCACTTGCCCCTAGCTGGGGGTATAGGGGAGCAGTCCCATGTA 
               
               
                   
               
               
                 GTAGTAGAATGAAAAATGCTGCTATGCTGTGCCTCCCCCACCTTTCCCATGTCTGCCCTCTACTCATGGTCTATCTCTCC 
               
               
                   
               
               
                 TGGCTCCTGGGAGTCATGGACTCCACCCAGCACCACCAACCTGACCTAACCACCTATCTGAGCCTGCCAGCCTATAACCC 
               
               
                   
               
               
                 ATCTGGGCCCTGATAGCTGGTGGCCAGCCCTGACCCCACCCCACCCTCCCTGGAACCTCTGATAGACACATCTGGCACAC 
               
               
                   
               
               
                 CAGCTCGCAAAGTCACCGTGAGGGTCTTGTGTTTGCTGAGTCAAAATTCCTTGAAATCCAAGTCCTTAGAGACTCCTGCT 
               
               
                   
               
               
                 CCCAAATTTACAGTCATAGACTTCTTCATGGCTGTCTCCTTTATCCACAGAATGATTCCTTTGCTTCATTGCCCCATCCA 
               
               
                   
               
               
                 TCTGATCCTCCTCATCAGTGCAGCACAGGGCCCATGAGCAGTAGCTGCAGAGTCTCACATAGGTCTGGCACTGCCTCTGA 
               
               
                   
               
               
                 CATGTCCGACCTTAGGCAAATGCTTGACTCTTCTGAGCTCAGTCTTGTCATGGCAAAACAAAGATAATAATAGTGTTTTT 
               
               
                   
               
               
                 TTATGGAGTTAGCGTGAGGATGGAAAACAATAGCAAAATTGATTAGACTATAAAAGGTCTCAACAAATAGTAGTAGATTT 
               
               
                   
               
               
                 TATCATCCATTAATCCTTCCCTCTCCTCTCTTACTCATCCCATCACGTATGCCTCTTAATTTTCCCTTACCTATAATAAG 
               
               
                   
               
               
                 AGTTATTCCTCTTATTATATTCTTCTTATAGTGATTCTGGATATCAAAGTGGGAATGAGGGGCAGGCCACTAACGAAGAA 
               
               
                   
               
               
                 GATGTTTCTCAAAGAAGCCATTCTCCCCACATAGATCATCTCAGCAGGGTTCAGGAAGATAAAGGAGGATCAAGGTCGAA 
               
               
                   
               
               
                 GGTAGGAACTAAGGAAGAACACTGGGCAAGTGGATCCTAAATATATCATTTAAATGCATAAATAAGCAAACCCTGCTCGG 
               
               
                   
               
               
                 GAATGGGAGGGAGAGTCTCTGGAGTCCACCCCTTCTCGGCCCTGGCTCTGCAGATAGTGCTATCAAAGCCCTGACAGAGC 
               
               
                   
               
               
                 CCTGCCCATTGCTGGGCCTTGGAGTGAGTCAGCCTAGTAGAGAGGCAGGGCAAGCCATCTCATAGCTGCTGAGTGGGAGA 
               
               
                   
               
               
                 GAGAAAAGGGCTCATTGTCTATAAACTCAGGTCATGGCTATTCTTATTCTCACACTAAGAAAAAGAATGAGATGTCTACA 
               
               
                   
               
               
                 TATACCCTGCGTCCCCTCTTGTGTACTGGGGCCCCCAAGAGCTCTCTAAAAGTGATGGCAAAGTCATTGCGCTAGATGCC 
               
               
                   
               
               
                 ATCCCATCTATTATAAACCTGCATTTGTCTCCACACACCAGTCATGGACAATAACCCTCCTCCCAGGTCCACGTGCTTGT 
               
               
                   
               
               
                 CTTTGTATAATACTCAAGTAATTTCGGAAAATGTATTCTTTCAATCTTGTTCTGTTATTCCTGTTTCAATGGCTTAGTAG 
               
               
                   
               
               
                 AAAAAGTACATACTTGTTTTCCCATAAATTGACAATAGACAATTTCACATCAATGTCTATATGGGTCGTTGTGTTTGCTG 
               
               
                   
               
               
                 TGTTTGCAAAAACTCACAATAACTTTATATTGTTACTACTCTAAGAAAGTTACAACATGGTGAATACAAGAGAAAGCTAT 
               
               
                   
               
               
                 TACAAGTCCAGAAAACAAAAGTTATCATCTTGAGGCCTCAGCTTTCTAGGAATAATATCAATATTACAAAACGCGTCGAC 
               
               
                   
               
               
                 GGTACCGTTAACGATCTTAGCCACTTTTTAAAAGAAAAGGGGGGACTGGAAGGGCTAATTCACTCCCAACGAAGACAAGA 
               
               
                   
               
               
                 TATCCTGCTAGTCCTTCCTTTCTAAATGACGAGAGAGACAGAAGAATTCTTCAAGGTTAGTGTGTCCAGCATGCAACCTT 
               
               
                   
               
               
                 TCCTTCCTGGATGAGCATCCCTGGAGTAGGAGAGCCAGCCTGCCTCCTGCGCTGGCACAGAGCCCGGTTCCCTAGACAAC 
               
               
                   
               
               
                 TGCCTCTCCAAATCTGATGTCCAGCGCCACCTGGTGTCCACATCAAGCAGACACAATTAATAGTCAACCTGTTCAGGAAA 
               
               
                   
               
               
                 ACTGTGAGGGGGAAAAAAAAGAAAGAGGATTTATGAAGGGAAAAGAAAGTTTAGAGGATATGCCACGATTGGCTAGCAGC 
               
               
                   
               
               
                 TGCTTTTTGCCTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGGCTAACTAGGGAACCCACTG 
               
               
                   
               
               
                 CTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCATCCGGAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTG 
               
               
                   
               
               
                 GTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGT 
               
               
                   
               
               
                 ATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACG 
               
               
                   
               
               
                 TGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGA 
               
               
                   
               
               
                 CTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTG 
               
               
                   
               
               
                 TTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAGCTGACGTCCTTTCCATGGCTGCTCGCCTGTGTTGCCACCTGGAT 
               
               
                   
               
               
                 TCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTC 
               
               
                   
               
               
                 TGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCCTGTCCGGTAG 
               
               
                   
               
               
                 CTTGCCAGCCTCGACTGTGCCTTCTAGTTGCCAGCCGTCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAG 
               
               
                   
               
               
                 GTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGG 
               
               
                   
               
               
                 GGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCAAGCTTGGCGTAATCATGGTCATA 
               
               
                   
               
               
                 GCTGTTTCCTGTGTGAAATTGTTATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGG 
               
               
                   
               
               
                 GTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAG 
               
               
                   
               
               
                 CTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTC 
               
               
                   
               
               
                 GCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGG 
               
               
                   
               
               
                 ATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTC 
               
               
                   
               
               
                 CATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAG 
               
               
                   
               
               
                 ATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCT 
               
               
                   
               
               
                 TTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAG 
               
               
                   
               
               
                 CTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGT 
               
               
                   
               
               
                 AAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGT 
               
               
                   
               
               
                 TCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTC 
               
               
                   
               
               
                 GGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGAT 
               
               
                   
               
               
                 TACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCAC 
               
               
                   
               
               
                 GTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCA 
               
               
                   
               
               
                 ATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCT 
               
               
                   
               
               
                 ATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTG 
               
               
                   
               
               
                 CTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGC 
               
               
                   
               
               
                 AGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGT 
               
               
                   
               
               
                 TAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCT 
               
               
                   
               
               
                 CCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATC 
               
               
                   
               
               
                 GTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATC 
               
               
                   
               
               
                 CGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTT 
               
               
                   
               
               
                 GCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGG 
               
               
                   
               
               
                 CAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCT 
               
               
                   
               
               
                 TTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAA 
               
               
                   
               
               
                 ATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATAT 
               
               
                   
               
               
                 TTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACC 
               
               
                   
               
               
                 ATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTCTCGCGCGTTTCGGTGATGACGGTGA 
               
               
                   
               
               
                 AAACCTCTGACACATGCAGCTCCCGGAGACGGTCACAGCTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGCCCGTCAGG 
               
               
                   
               
               
                 GCGCGTCAGCGGGTGTTGGCGGGTGTCGGGGCTGGCTTAACTATGCGGCATCAGAGCAGATTGTACTGAGAGTGCACC 
               
               
                   
               
               
                 [SEQ ID NO: 40] 
                   
               
               
                 ATTGGCCATTGCATACGTTGTATCCATATCATAATATGTACATTTATATTGGCTCATGTCCAACATTACCGCCATGTTGA 
                   
               
               
                   
               
               
                 CATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTAC 
               
               
                   
               
               
                 ATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCA 
               
               
                   
               
               
                 TAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAA 
               
               
                   
               
               
                 GTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGAC 
               
               
                   
               
               
                 CTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACA 
               
               
                   
               
               
                 TCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGG 
               
               
                   
               
               
                 CACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTG 
               
               
                   
               
               
                 GGAGGTCTATATAAGCAGAGCTCGTTTAGTGAACCGGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGG 
               
               
                   
               
               
                 CTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGTGTGCCCGTCTGTTGTGTG 
               
               
                   
               
               
                 ACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTCAGTGTGGAAAATCTCTAGCAGTGGCGCCCGAACAGGGACTTGA 
               
               
                   
               
               
                 AAGCGAAAGGGAAACCAGAGGAGCTCTCTCGACGCAGGACTCGGCTTGCTGAAGCGCGCACGGCAAGAGGCGAGGGGCGG 
               
               
                   
               
               
                 CGACTGGTGAGTACGCCAAAAATTTTGACTAGCGGAGGCTAGAAGGAGAGAGACGGGTGCGAGAGCGTCAGTATTAAGCG 
               
               
                   
               
               
                 GGGGTGAATAAGATCGCGATGGGAAAAAATTCGGTTAAGGCCAGGGGGAAAGAAAAAATATAAATTAAAACATATAGTAT 
               
               
                   
               
               
                 GGGCAAGCAGGGAGCTAGAACGATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGTAGACAAATACTGGGA 
               
               
                   
               
               
                 CAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATCATTATATAATACAGTAGCAACCCTCTATTGTGTGCA 
               
               
                   
               
               
                 TCAAAGGATAGAGATAAAAGACACCAAGGAAGCTTTAGACAAGATAGAGGAAGAGCAAAACAAAAGTAAGACCACCGCAC 
               
               
                   
               
               
                 AGCAAGCGGCCGCTGATCTTCAGACCTGGAGGAGGAGATATGAGGGACAATTGGAGAAGTGAATTATATAAATATAAAGT 
               
               
                   
               
               
                 AGTAAAAATTGAACCATTAGGAGTAGCACCCACCAAGGCAAAGAGAAGAGTGGTGCAGAGAGAAAAAAGAGCAGTGGGAA 
               
               
                   
               
               
                 TAGGAGCTTTGTTCCTTGGGTTCTTGGGAGCAGCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACAGGCC 
               
               
                   
               
               
                 AGACAATTATTGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTGAGGCGCAACAGCATCTGTTGCAACT 
               
               
                   
               
               
                 CACAGTCTGGGGCATCAAGCAGCTCCAGGCAAGAATCCTGGCTGTGGAAAGATACCTAAAGGATCAACAGCTCCTGGGGA 
               
               
                   
               
               
                 TTTGGGGTTGCTCTGGAAAACTCATTTGCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTCTGGAACAG 
               
               
                   
               
               
                 ATTTGGAATCACACGACCTGGATGGAGTGGGACAGAGAAATTAACAATTACACAAGCTTAATACACTCCTTAATTGAAGA 
               
               
                   
               
               
                 ATCGCAAAACCAGCAAGAAAAGAATGAACAAGAATTATTGGAATTAGATAAATGGGCAAGTTTGTGGAATTGGTTTAACA 
               
               
                   
               
               
                 TAACAAATTGGCTGTGGTATATAAAATTATTCATAATGATAGTAGGAGGCTTGGTAGGTTTAAGAATAGTTTTTGCTGTA 
               
               
                   
               
               
                 CTTTCTATAGTGAATAGAGTTAGGCAGGGATATTCACCATTATCGTTTCAGACCCACCTCCCAACCCCGAGGGGACCCGA 
               
               
                   
               
               
                 CAGGCCCGAAGGAATAGAAGAAGAAGGTGGAGAGAGAGACAGAGACAGATCCATTCGATTAGTGAACGGATCTCGACGGT 
               
               
                   
               
               
                 ATCGTTTTAAAAGAAAAGGGGGGATTGGGGGGTACAGTGCAGGGGAAAGAATAGTAGACATAATAGCAACAGACATACAA 
               
               
                   
               
               
                 ACTAAAGAATTACAAAAACAAATTACAAAAATTCAAAATTTTATCGGCGTGTTGGGGGTGGACCATCCTCTAGGTATTGA 
               
               
                   
               
               
                 ATAAGAAAAATGAAGTTAAGGTGGTTGATGGTAACACTATGCTAATAACTGCAGAGCCAGAAGCACCATAAGGGACATGA 
               
               
                   
               
               
                 TAAGGGAGCCAGCAGACCTCTGATCTCTTCCTGAATGCTAATCTTAAACATCCTGAGGAAGAATGGGACTTCCATTTGGG 
               
               
                   
               
               
                 GTGGGCCTATGATAGGGTAATAAGACAGTAGTGAATATCAAGCTACAAAAAGCCCCCTTTCAAATTCTTCTCAGTCCTAA 
               
               
                   
               
               
                 CTTTTCATACTAAGCCCAGTCCTTCCAAAGCAGACTGTGAAAGAGTGATAGTTCCGGGAGACTAGCACTGCAGATTCCGG 
               
               
                   
               
               
                 GTCACTGTGAGTGGGGGAGGCAGGGAAGAAGGGCTCACAGGACAGTCAAACCATGCCCCCTGTTTTTCCTTCTTCAAGTA 
               
               
                   
               
               
                 GACCTCTATAAGACAACAGAGACAACTAAGGCTGAGTGGCCAGGCGAGGAGAAACCATCTCGCCGTAAAACATGGAAGGA 
               
               
                   
               
               
                 ACACTTCAGGGGAAAGGTGGTATCTCTAAGCAAGAGAACTGAGTGGAGTCAAGGCTGAGAGATGCAGGATAAGCAAATGG 
               
               
                   
               
               
                 GTAGTGAAAAGACATTCATGAGGACAGCTAAAACAATAAGTAATGTAAAATACAGCATAGCAAAACTTTAACCTCCAAAT 
               
               
                   
               
               
                 CAAGCCTCTACTTGAATCCTTTTCTGAGGGATGAATAAGGCATAGGCATCAGGGGCTGTTGCCAATGTGCATTAGCTGTT 
               
               
                   
               
               
                 TGCAGCCTCACCTTCTTTCATGGAGTTTAAGATATAGTGTATTTTCCCAAGGTTTGAACTAGCTCTTCATTTCTTTATGT 
               
               
                   
               
               
                 TTTAAATGCACTGACCTCCCACATTCCCTTTTTAGTAAAATATTCAGAAATAATTTAAATACATCATTGCAATGAAAATA 
               
               
                   
               
               
                 AATGTTTTTTATTAGGCAGAATCCAGATGCTCAAGGCCCTTCATAATATCCCCCAGTTTAGTAGTTGGACTTAGGGAACA 
               
               
                   
               
               
                 AAGGAACCTTTAATAGAAATTGGACAGCAAGAAAGCGAGCTTAGTGATACTTGTGGGCCAGGGCATTAGCCACACCAGCC 
               
               
                   
               
               
                 ACCACTTTCTGATAGGCAGCCTGCACTGGTGGGGTGAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACGTT 
               
               
                   
               
               
                 GCCCAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAAAAGGGCCTAGCTTGGACTCAGAATAATCCAGC 
               
               
                   
               
               
                 CTTATCCCAACCATAAAATAAAAGCAGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACCTCTTACATCAGT 
               
               
                   
               
               
                 TACAATTTATATGCAGAAATATTTATATGCAGAAATATTGCTATTGCCTTAACCCAGAAATTATCACTGTTATTCTTTAG 
               
               
                   
               
               
                 AATGGTGCAAAGAGGCATGATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAGGGAAAGTATTAGAAATAAGATA 
               
               
                   
               
               
                 AACAAAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAAATTACAAATGCAAAATTACCCTGATTTGGTCAATATGTGTA 
               
               
                   
               
               
                 CCCTGTTACTTCTCCCCTTCCTATGACATGAACTTAACCATAGAAAAGAAGGGGAAAGAAAACATCAAGGGTCCCATAGA 
               
               
                   
               
               
                 CTCACCCTGAAGTTCTCAGGATCCACGTGCAGCTTGTCACAGTGCAGCTCACTCAGCTGGGCAAAGGTGCCCTTGAGGTT 
               
               
                   
               
               
                 GTCCAGGTGAGCCAGGCCATCACTAAAGGCACCGAGCACTTTCTTGCCATGAGCCTTCACCTTAGGGTTGCCCATAACAG 
               
               
                   
               
               
                 CATCAGGAGTGGACAGATCCCCAAAGGACTCAAAGAACCTCTGGGTCCAAGGGTAGACCACCAGCAGCCTAAGGGTGGGA 
               
               
                   
               
               
                 AAATAGACCAATAGGCAGAGAGAGTCAGTGCCTATCAGAAACCCAAGAGTCTTCTCTGTCTCCACATGCCCAGTTTCTAT 
               
               
                   
               
               
                 TGGTCTCCTTAAACCTGTCTTGTAACCTTGATACCAACCTGCCCAGGGCCTCACCACCAACTTCATCCACGTTCACCTTG 
               
               
                   
               
               
                 CCCCACAGGGCAGTAACGGCAGACTTCTCCTCAGGAGTCAGGTGCACCATGGTGTCTGTTTGAGGTTGCTAGTGAACACA 
               
               
                   
               
               
                 GTTGTGTCAGAAGCAAATGTAAGCAATAGATGGCTCTGCCCTGACTTTTATGCCCAGCCCTGGCTCCTGCCCTCCCTGCT 
               
               
                   
               
               
                 CCTGGGAGTAGATTGGCCAACCCTAGGGTGTGGCTCCACAGGGTGAGGTCTAAGTGATGACAGCCGTACCTGTCCTTGGC 
               
               
                   
               
               
                 TCTTCTGGCACTGGCTTAGGAGTTGGACTTCAAACCCTCAGCCCTCCCTCTAAGATATATCTCTTGGCCCCATACCATCA 
               
               
                   
               
               
                 GTACAAATTGCTACTAAAAACATCCTCCTTTGCAAGTGTATTTACGGCATCTGTGAAGGAAAGAAACATCTCCTCTAAAC 
               
               
                   
               
               
                 CACTATGCTGCTAGAGCCTCTTTTCTGTACTCAAGCCTCATTCAGACACTAGTGTCACCAGTCTCCTCATATACCTATTG 
               
               
                   
               
               
                 TATTTTCTTCTTCTTGCTGGTTTAGTCATGTTTTCTGGGAGCTTAGGGGCTTATTTTATTTTGTTTTGTTTTCTAATCAA 
               
               
                   
               
               
                 CAGAGATGGGCAAACCCATTATTTTTTTCTTTAGACTTGGGATGGTGATAGCTGGGCAGCGTCAGAAACTGTGTGTGGAT 
               
               
                   
               
               
                 ATAGATAAGAGCTCGGACTATGCTGAGCTGTGATGAGGGAGGGACCTAGCCAAAGGCAGTGAGAGTCAGAATGCTCCTGC 
               
               
                   
               
               
                 TATTGCCTTCTCAGTCCCCACGCTTGGTTTCTACACAAGTAGATACATAGAAAAGGCTATAGGTTAGTGTTTGAGAGTCC 
               
               
                   
               
               
                 TGCATGAGTTAGTTGCTCAGAAATGCCCGATAAATATGTTATGTGTGTTTATGTATATATATGTTTTATATATATATATG 
               
               
                   
               
               
                 TGTGTGTGTGTGTGTGTGTGTGTTGTGTTTACAAATATGTGATTATCATCAAAACGTGAGGGCTAAAGTGACCAGATAAC 
               
               
                   
               
               
                 TTGCAGGTCTAGACACCCTTTTCCGGCACGCAGATAGTCAATATCTTCAGCGTCCCCAAGGCCTGCAAGGGTGGGGCCCC 
               
               
                   
               
               
                 ATATCTGGAAGTCCCAGGCGGAGCTGGGAGTTGGTCAAGTCTGGGCTGTGGGGGCAGGGAGTGCTGGGGGATGGCTCGAG 
               
               
                   
               
               
                 AAGCTTTCATCAAAAAAAGTCTAACCAGCTGCATTCGACTTTGACTGCAGCAGCTGGTTAGAAGGTTCTACTGGAGGAGG 
               
               
                   
               
               
                 GTCCCAGCCCATTGCTAAATTAACATCAGGCTCTGAGACTGGCAGTATATCTCTAACAGTGGTTGATGCTATCTTCTGGA 
               
               
                   
               
               
                 ACTTGCCTGCTACATTGAGACCACTGACCCATACATAGGAAGCCCATAGCTCTGTCCTGAACTGTTAGGCCACTGGTCCA 
               
               
                   
               
               
                 GAGAGTGTGCATCTCCTTTGATCCTCATAATAACCCTATGAGATAGACACAATTATTACTCTTACTTTATAGATGATGAT 
               
               
                   
               
               
                 CCTGAAAACATAGGAGTCAAGGCACTTGCCCCTAGCTGGGGGTATAGGGGAGCAGTCCCATGTAGTAGTAGAATGAAAAA 
               
               
                   
               
               
                 TGCTGCTATGCTGTGCCTCCCCCACCTTTCCCATGTCTGCCCTCTACTCATGGTCTATCTCTCCTGGCTCCTGGGAGTCA 
               
               
                   
               
               
                 TGGACTCCACCCAGCACCACCAACCTGACCTAACCACCTATCTGAGCCTGCCAGCCTATAACCCATCTGGGCCCTGATAG 
               
               
                   
               
               
                 CTGGTGGCCAGCCCTGACCCCACCCCACCCTCCCTGGAACCTCTGATAGACACATCTGGCACACCAGCTCGCAAAGTCAC 
               
               
                   
               
               
                 CGTGAGGGTCTTGTGTTTGCTGAGTCAAAATTCCTTGAAATCCAAGTCCTTAGAGACTCCTGCTCCCAAATTTACAGTCA 
               
               
                   
               
               
                 TAGACTTCTTCATGGCTGTCTCCTTTATCCACAGAATGATTCCTTTGCTTCATTGCCCCATCCATCTGATCCTCCTCATC 
               
               
                   
               
               
                 AGTGCAGCACAGGGCCCATGAGCAGTAGCTGCAGAGTCTCACATAGGTCTGGCACTGCCTCTGACATGTCCGACCTTAGG 
               
               
                   
               
               
                 CAAATGCTTGACTCTTCTGAGCTCAGTCTTGTCATGGCAAAACAAAGATAATAATAGTGTTTTTTTATGGAGTTAGCGTG 
               
               
                   
               
               
                 AGGATGGAAAACAATAGCAAAATTGATTAGACTATAAAAGGTCTCAACAAATAGTAGTAGATTTTATCATCCATTAATCC 
               
               
                   
               
               
                 TTCCCTCTCCTCTCTTACTCATCCCATCACGTATGCCTCTTAATTTTCCCTTACCTATAATAAGAGTTATTCCTCTTATT 
               
               
                   
               
               
                 ATATTCTTCTTATAGTGATTCTGGATATCAAAGTGGGAATGAGGGGCAGGCCACTAACGAAGAAGATGTTTCTCAAAGAA 
               
               
                   
               
               
                 GCCATTCTCCCCACATAGATCATCTCAGCAGGGTTCAGGAAGATAAAGGAGGATCAAGGTCGAAGGTAGGAACTAAGGAA 
               
               
                   
               
               
                 GAACACTGGGCAAGTGGATCCTAAATATATCATTTAAATGCATAAATAAGCAAACCCTGCTCGGGAATGGGAGGGAGAGT 
               
               
                   
               
               
                 CTCTGGAGTCCACCCCTTCTCGGCCCTGGCTCTGCAGATAGTGCTATCAAAGCCCTGACAGAGCCCTGCCCATTGCTGGG 
               
               
                   
               
               
                 CCTTGGAGTGAGTCAGCCTAGTAGAGAGGCAGGGCAAGCCATCTCATAGCTGCTGAGTGGGAGAGAGAAAAGGGCTCATT 
               
               
                   
               
               
                 GTCTATAAACTCAGGTCATGGCTATTCTTATTCTCACACTAAGAAAAAGAATGAGATGTCTACATATACCCTGCGTCCCC 
               
               
                   
               
               
                 TCTTGTGTACTGGGGCCCCCAAGAGCTCTCTAAAAGTGATGGCAAAGTCATTGCGCTAGATGCCATCCCATCTATTATAA 
               
               
                   
               
               
                 ACCTGCATTTGTCTCCACACACCAGTCATGGACAATAACCCTCCTCCCAGGTCCACGTGCTTGTCTTTGTATAATACTCA 
               
               
                   
               
               
                 AGTAATTTCGGAAAATGTATTCTTTCAATCTTGTTCTGTTATTCCTGTTTCAATGGCTTAGTAGAAAAAGTACATACTTG 
               
               
                   
               
               
                 TTTTCCCATAAATTGACAATAGACAATTTCACATCAATGTCTATATGGGTCGTTGTGTTTGCTGTGTTTGCAAAAACTCA 
               
               
                   
               
               
                 CAATAACTTTATATTGTTACTACTCTAAGAAAGTTACAACATGGTGAATACAAGAGAAAGCTATTACAAGTCCAGAAAAC 
               
               
                   
               
               
                 AAAAGTTATCATCTTGAGGCCTCAGCTTTCTAGGAATAATATCAATATTACAAAACGCGTCGACGGTACCGTTAACGATC 
               
               
                   
               
               
                 TTAGCCACTTTTTAAAAGAAAAGGGGGGACTGGAAGGGCTAATTCACTCCCAACGAAGACAAGATATCCTGCTAGTCCTT 
               
               
                   
               
               
                 CCTTTCTAAATGACGAGAGAGACAGAAGAATTCTTCAAGGTTAGTGTGTCCAGCATGCAACCTTTCCTTCCTGGATGAGC 
               
               
                   
               
               
                 ATCCCTGGAGTAGGAGAGCCAGCCTGCCTCCTGCGCTGGCACAGAGCCCGGTTCCCTAGACAACTGCCTCTCCAAATCTG 
               
               
                   
               
               
                 ATGTCCAGCGCCACCTGGTGTCCACATCAAGCAGACACAATTAATAGTCAACCTGTTCAGGAAAACTGTGAGGGGGAAAA 
               
               
                   
               
               
                 AAAAGAAAGAGGATTTATGAAGGGAAAAGAAAGTTTAGAGGATATGCCACGATTGGCTAGCAGCTGCTTTTTGCCTGTAC 
               
               
                   
               
               
                 TGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAA 
               
               
                   
               
               
                 GCTTGCCTTGAGTGCTTCATCCGGAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGT 
               
               
                   
               
               
                 TGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCT 
               
               
                   
               
               
                 CCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACT 
               
               
                   
               
               
                 GTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCT 
               
               
                   
               
               
                 CCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATT 
               
               
                   
               
               
                 CCGTGGTGTTGTCGGGGAAGCTGACGTCCTTTCCATGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCC 
               
               
                   
               
               
                 TTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCG 
               
               
                   
               
               
                 TCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCCTGTCCGGTAGCTTGCCAGCCTCGACT 
               
               
                   
               
               
                 GTGCCTTCTAGTTGCCAGCCGTCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGT 
               
               
                   
               
               
                 CCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGG 
               
               
                   
               
               
                 ACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCAAGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTGTGA 
               
               
                   
               
               
                 AATTGTTATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAG 
               
               
                   
               
               
                 CTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCG 
               
               
                   
               
               
                 GCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTT 
               
               
                   
               
               
                 CGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAA 
               
               
                   
               
               
                 CATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCC 
               
               
                   
               
               
                 CTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCC 
               
               
                   
               
               
                 CCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAG 
               
               
                   
               
               
                 CGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACG 
               
               
                   
               
               
                 AACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCG 
               
               
                   
               
               
                 CCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCC 
               
               
                   
               
               
                 TAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTA 
               
               
                   
               
               
                 GCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAA 
               
               
                   
               
               
                 GGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGT 
               
               
                   
               
               
                 CATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATG 
               
               
                   
               
               
                 AGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATA 
               
               
                   
               
               
                 GTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCG 
               
               
                   
               
               
                 AGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAA 
               
               
                   
               
               
                 CTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAAC 
               
               
                   
               
               
                 GTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATC 
               
               
                   
               
               
                 AAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGT 
               
               
                   
               
               
                 TGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCT 
               
               
                   
               
               
                 GTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACG 
               
               
                   
               
               
                 GGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCAAAACTCTCAAGGAT 
               
               
                   
               
               
                 CTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCG 
               
               
                   
               
               
                 TTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATA 
               
               
                   
               
               
                 CTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAA 
               
               
                   
               
               
                 AAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACAT 
               
               
                   
               
               
                 TAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATG 
               
               
                   
               
               
                 CAGCTCCCGGAGACGGTCACAGCTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGCCCGTCAGGGCGCGTCAGCGGGTGT 
               
               
                   
               
               
                 TGGCGGGTGTCGGGGCTGGCTTAACTATGCGGCATCAGAGCAGATTGTACTGAGAGTGCACC 
               
               
                   
               
               
                 [SEQ ID NO: 76] 
                   
               
               
                 attggccattgcatacgttgtatccatatcataatatgtacatttatattggctcatgtccaacattaccgccatgttga 
                   
               
               
                   
               
               
                 cattgattattgactagttattaatagtaatcaattacggggtcattagttcatagcccatatatggagttccgcgttac 
               
               
                   
               
               
                 ataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccattgacgtcaataatgacgtatgttccca 
               
               
                   
               
               
                 tagtaacgccaatagggactttccattgacgtcaatgggtggagtatttacggtaaactgcccacttggcagtacatcaa 
               
               
                   
               
               
                 gtgtatcatatgccaagtacgccccctattgacgtcaatgacggtaaatggcccgcctggcattatgcccagtacatgac 
               
               
                   
               
               
                 cttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtaca 
               
               
                   
               
               
                 tcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttgg 
               
               
                   
               
               
                 caccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtg 
               
               
                   
               
               
                 ggaggtctatataagcagagctcgtttagtgaaccggggtctctctggttagaccagatctgagcctgggagctctctgg 
               
               
                   
               
               
                 ctaactagggaacccactgcttaagcctcaataaagcttgccttgagtgcttcaagtagtgtgtgcccgtctgttgtgtg 
               
               
                   
               
               
                 actctggtaactagagatccctcagacccttttagtcagtgtggaaaatctctagcagtggcgcccgaacagggacttga 
               
               
                   
               
               
                 aagcgaaagggaaaccagaggagctctctcgacgcaggactcggcttgctgaagcgcgcacggcaagaggcgaggggcgg 
               
               
                   
               
               
                 cgactggtgagtacgccaaaaattttgactagcggaggctagaaggagagagacgggtgcgagagcgtcagtattaagcg 
               
               
                   
               
               
                 ggggtgaataagatcgcgatgggaaaaaattcggttaaggccagggggaaagaaaaaatataaattaaaacatatagtat 
               
               
                   
               
               
                 gggcaagcagggagctagaacgattcgcagttaatcctggcctgttagaaacatcagaaggctgtagacaaatactggga 
               
               
                   
               
               
                 cagctacaaccatcccttcagacaggatcagaagaacttagatcattatataatacagtagcaaccctctattgtgtgca 
               
               
                   
               
               
                 tcaaaggatagagataaaagacaccaaggaagctttagacaagatagaggaagagcaaaacaaaagtaagaccaccgcac 
               
               
                   
               
               
                 agcaagcggccgctgatcttcagacctggaggaggagatatgagggacaattggagaagtgaattatataaatataaagt 
               
               
                   
               
               
                 agtaaaaattgaaccattaggagtagcacccaccaaggcaaagagaagagtggtgcagagagaaaaaagagcagtgggaa 
               
               
                   
               
               
                 taggagctttgttccttgggttcttgggagcagcaggaagcactatgggcgcagcgtcaatgacgctgacggtacaggcc 
               
               
                   
               
               
                 agacaattattgtctggtatagtgcagcagcagaacaatttgctgagggctattgaggcgcaacagcatctgttgcaact 
               
               
                   
               
               
                 cacagtctggggcatcaagcagctccaggcaagaatcctggctgtggaaagatacctaaaggatcaacagctcctgggga 
               
               
                   
               
               
                 tttggggttgctctggaaaactcatttgcaccactgctgtgccttggaatgctagttggagtaataaatctctggaacag 
               
               
                   
               
               
                 atttggaatcacacgacctggatggagtgggacagagaaattaacaattacacaagcttaatacactccttaattgaaga 
               
               
                   
               
               
                 atcgcaaaaccagcaagaaaagaatgaacaagaattattggaattagataaatgggcaagtttgtggaattggtttaaca 
               
               
                   
               
               
                 taacaaattggctgtggtatataaaattattcataatgatagtaggaggcttggtaggtttaagaatagtttttgctgta 
               
               
                   
               
               
                 ctttctatagtgaatagagttaggcagggatattcaccattatcgtttcagacccacctcccaaccccgaggggacccga 
               
               
                   
               
               
                 caggcccgaaggaatagaagaagaaggtggagagagagacagagacagatccattcgattagtgaacggatctcgacggt 
               
               
                   
               
               
                 atcgttttaaaagaaaaggggggattggggggtacagtgcaggggaaagaatagtagacataatagcaacagacatacaa 
               
               
                   
               
               
                 actaaagaattacaaaaacaaattacaaaaattcaaaattttatcggcgtgttgggggtggaccatcctctaggtattga 
               
               
                   
               
               
                 ataagaaaaatgaagttaaggtggttgatggtaacactatgctaataactgcagagccagaagcaccataagggacatga 
               
               
                   
               
               
                 taagggagccagcagacctctgatctcttcctgaatgctaatcttaaacatcctgaggaagaatgggacttccatttggg 
               
               
                   
               
               
                 gtgggcctatgatagggtaataagacagtagtgaatatcaagctacaaaaagccccctttcaaattcttctcagtcctaa 
               
               
                   
               
               
                 cttttcatactaagcccagtccttccaaagcagactgtgaaagagtgatagttccgggagactagcactgcagattccgg 
               
               
                   
               
               
                 gtcactgtgagtgggggaggcagggaagaagggctcacaggacagtcaaaccatgccccctgtttttccttcttcaagta 
               
               
                   
               
               
                 gacctctataagacaacagagacaactaaggctgagtggccaggcgaggagaaaccatctcgccgtaaaacatggaagga 
               
               
                   
               
               
                 acacttcaggggaaaggtggtatctctaagcaagagaactgagtggagtcaaggctgagagatgcaggataagcaaatgg 
               
               
                   
               
               
                 gtagtgaaaagacattcatgaggacagctaaaacaataagtaatgtaaaatacagcatagcaaaactttaacctccaaat 
               
               
                   
               
               
                 caagcctctacttgaatccttttctgagggatgaataaggcataggcatcaggggctgttgccaatgtgcattagctgtt 
               
               
                   
               
               
                 tgcagcctcaccttctttcatggagtttaagatatagtgtattttcccaaggtttgaactagctcttcatttctttatgt 
               
               
                   
               
               
                 tttaaatgcactgacctcccacattccctttttagtaaaatattcagaaataatttaaatacatcattgcaatgaaaata 
               
               
                   
               
               
                 aatgttttttattaggcagaatccagatgctcaaggcccttcataatatcccccagtttagtagttggacttagggaaca 
               
               
                   
               
               
                 aaggaacctttaatagaaattggacagcaagaaagcgagcttagtgatacttgtgggccagggcattagccacaccagcc 
               
               
                   
               
               
                 accactttctgataggcagcctgcactggtggggtgAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACGTT 
               
               
                   
               
               
                 GCCCAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAAAAGGGCCTAGCTTGGACTCAGAATAATCCAGC 
               
               
                   
               
               
                 CTTATCCCAACCATAAAATAAAAGCAGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACCTCTTACATCAGT 
               
               
                   
               
               
                 TACAATTTATATGCAGAAATATTTATATGCAGAAATATTGCTATTGCCTTAACCCAGAAATTATCACTGTTATTCTTTAG 
               
               
                   
               
               
                 AATGGTGCAAAGAGGCATGATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAGGGAAAGTATTAGAAATAAGATA 
               
               
                   
               
               
                 AACAAAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAAATTACAAATGCAAAATTACCCTGATTTGGTCAATATGTGTA 
               
               
                   
               
               
                 CCCTGTTACTTCTCCCCTTCCTATGACATGAACTTAACCATAGAAAAGAAGGGGAAAGAAAACATCAAGGGTCCCATAGA 
               
               
                   
               
               
                 CTCACCCTGAAGTTCTCAGGATCCACGTGCAGCTTGTCACAGTGCAGCTCACTCAGctgGGCAAAGGTGCCCTTGAGGTT 
               
               
                   
               
               
                 GTCCAGGTGAGCCAGGCCATCACTAAAGGCACCGAGCACTTTCTTGCCATGAGCCTTCACCTTAGGGTTGCCCATAACAG 
               
               
                   
               
               
                 CATCAGGAGTGGACAGATCCCCAAAGGACTCAAAGAACCTCTGGGTCCAAGGGTAGACCACCAGCAGCCTAAGGGTGGGA 
               
               
                   
               
               
                 AAATAGACCAATAGGCAGAGAGAGTCAGTGCCTATCAGAAACCCAAGAGTCTTCTCTGTCTCCACATGCCCAGTTTCTAT 
               
               
                   
               
               
                 TGGTCTCCTTAAACCTGTCTTGTAACCTTGATACCAACCTGCCCAGGGCCTCACCACCAACTTCATCCACGTTCACCTTG 
               
               
                   
               
               
                 CCCCACAGGGCAGTAACGGCAGACTTCTCCTCAGGAGTCAGGTGCACCATGGTGTCTGTTTGAGGTTGCTAGTGAACACA 
               
               
                   
               
               
                 GTTGTGTCAGAAGCAAATGTAAGCAATAGATGGCTCTGCCCTGACTTTTATGCCCAGCCCTGGCTCCTGCCCTCCCTGCT 
               
               
                   
               
               
                 CCTGGGAGTAGATTGGCCAACCctagggtgtggctccacagggtgaggtctaagtgatgacagccgtacctgtccttggc 
               
               
                   
               
               
                 tcttctggcactggcttaggagttggacttcaaaccctcagccctccctctaagatatatctcttggccccataccatca 
               
               
                   
               
               
                 gtacaaattgctactaaaaacatcctcctttgcaagtgtatttacCACCCTTTTCCGGCACGCAGATAGTCAATATCTTC 
               
               
                   
               
               
                 AGCGTCCCCAAGGCCTGCAAGGGTGGGGCCCCATATCTGGAAGTCCCAGGCGGAGCTGGGAGTTGGTCAAGTCTGGGCTG 
               
               
                   
               
               
                 TGGGGGCAGGGAGTGCTGGGGGATGGacgcgtcgacggtaccgttaacgatcttagccactttttaaaagaaaagggggg 
               
               
                   
               
               
                 actggaagggctaattcactcccaacgaagacaagatatcctgCTAGTCCTTCCTTTCTAAATGACGAGAGAGACAGAAG 
               
               
                   
               
               
                 AATTCTTCAAGGTTAGTGTGTCCAGCATGCAACCTTTCCTTCCTGGATGAGCATCCCTGGAGTAGGAGAGCCAGCCTGCC 
               
               
                   
               
               
                 TCCTGCGCTGGCACAGAGCCCGGTTCCCTAGACAACTGCCTCTCCAAATCTGATGTCCAGCGCCACCTGGTGTCCACATC 
               
               
                   
               
               
                 AAGCAGACACAATTAATAGTCAACCTGTTCAGGAAAACTGTGAGGGGGAAAAAAAAGAAAGAGGATTTATGAAGGGAAAA 
               
               
                   
               
               
                 GAAAGTTTAGAGGATATGCCACGATTGGctagcagctgctttttgcctgtactgggtctctctggttagaccagatctga 
               
               
                   
               
               
                 gcctgggagctctctggctaactagggaacccactgcttaagcctcaataaagcttgccttgagtgcttcatccggAATC 
               
               
                   
               
               
                 AACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCT 
               
               
                   
               
               
                 GCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTC 
               
               
                   
               
               
                 TCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTT 
               
               
                   
               
               
                 GGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCC 
               
               
                   
               
               
                 GCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAGCTGACGTC 
               
               
                   
               
               
                 CTTTCCATGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATC 
               
               
                   
               
               
                 CAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGG 
               
               
                   
               
               
                 ATCTCCCTTTGGGCCGCCTCCCCGCCTGtccggtAGCTTGCCAGCCTCGACTGTGCCTTCTAGTTGCCAGCCGTCTGTTG 
               
               
                   
               
               
                 TTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCA 
               
               
                   
               
               
                 TCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAA 
               
               
                   
               
               
                 TAGCAGGCATGcaagcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctcacaattccacaca 
               
               
                   
               
               
                 acatacgagccggaagcataaagtgtaaagcctggggtgcctaatgagtgagctaactcacattaattgcgttgcgctca 
               
               
                   
               
               
                 ctgcccgctttccagtcgggaaacctgtcgtgccagctgcattaatgaatcggccaacgcgcggggagaggcggtttgcg 
               
               
                   
               
               
                 tattgggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcact 
               
               
                   
               
               
                 caaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggcc 
               
               
                   
               
               
                 aggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctc 
               
               
                   
               
               
                 aagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctg 
               
               
                   
               
               
                 ttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgt 
               
               
                   
               
               
                 aggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgc 
               
               
                   
               
               
                 cttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacagga 
               
               
                   
               
               
                 ttagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaagaacagta 
               
               
                   
               
               
                 tttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgc 
               
               
                   
               
               
                 tggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatctttt 
               
               
                   
               
               
                 ctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacc 
               
               
                   
               
               
                 tagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatg 
               
               
                   
               
               
                 cttaatcagtgaggcacctatctcagcgatctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataa 
               
               
                   
               
               
                 ctacgatacgggagggcttaccatctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccagattta 
               
               
                   
               
               
                 tcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaactttatccgcctccatccagtctattaa 
               
               
                   
               
               
                 ttgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgcaacgttgttgccattgctacaggcatcgtgg 
               
               
                   
               
               
                 tgtcacgctcgtcgtttggtatggcttcattcagctccggttcccaacgatcaaggcgagttacatgatcccccatgttg 
               
               
                   
               
               
                 tgcaaaaaagcggttagctccttcggtcctccgatcgttgtcagaagtaagttggccgcagtgttatcactcatggttat 
               
               
                   
               
               
                 ggcagcactgcataattctcttactgtcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcat 
               
               
                   
               
               
                 tctgagaatagtgtatgcggcgaccgagttgctcttgcccggcgtcaatacgggataataccgcgccacatagcagaact 
               
               
                   
               
               
                 ttaaaagtgctcatcattggaaaacgttcttcggggcaaaactctcaaggatcttaccgctgttgagatccagttcgatg 
               
               
                   
               
               
                 taacccactcgtgcacccaactgatcttcagcatcttttactttcaccagcgtttctgggtgagcaaaaacaggaaggca 
               
               
                   
               
               
                 aaatgccgcaaaaaagggaataagggcgacacggaaatgttgaatactcatactcttcctttttcaatattattgaagca 
               
               
                   
               
               
                 tttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaataaacaaataggggttccgcgcaca 
               
               
                   
               
               
                 tttccccgaaaagtgccacctgacgtctaagaaaccattattatcatgacattaacctataaaaataggcgtatcacgag 
               
               
                   
               
               
                 gccctttcgtctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtc 
               
               
                   
               
               
                 tgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggctggcttaactat 
               
               
                   
               
               
                 gcggcatcagagcagattgtactgagagtgcacc 
               
               
                   
               
               
                 [SEQ ID NO: 77] 
                   
               
               
                 attggccattgcatacgttgtatccatatcataatatgtacatttatattggctcatgtccaacattaccgccatgttga 
                   
               
               
                   
               
               
                 cattgattattgactagttattaatagtaatcaattacggggtcattagttcatagcccatatatggagttccgcgttac 
               
               
                   
               
               
                 ataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccattgacgtcaataatgacgtatgttccca 
               
               
                   
               
               
                 tagtaacgccaatagggactttccattgacgtcaatgggtggagtatttacggtaaactgcccacttggcagtacatcaa 
               
               
                   
               
               
                 gtgtatcatatgccaagtacgccccctattgacgtcaatgacggtaaatggcccgcctggcattatgcccagtacatgac 
               
               
                   
               
               
                 cttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtaca 
               
               
                   
               
               
                 tcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggagtttgttttgg 
               
               
                   
               
               
                 caccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacggtg 
               
               
                   
               
               
                 ggaggtctatataagcagagctcgtttagtgaaccggggtctctctggttagaccagatctgagcctgggagctctctgg 
               
               
                   
               
               
                 ctaactagggaacccactgcttaagcctcaataaagcttgccttgagtgcttcaagtagtgtgtgcccgtctgttgtgtg 
               
               
                   
               
               
                 actctggtaactagagatccctcagacccttttagtcagtgtggaaaatctctagcagtggcgcccgaacagggacttga 
               
               
                   
               
               
                 aagcgaaagggaaaccagaggagctctctcgacgcaggactcggcttgctgaagcgcgcacggcaagaggcgaggggcgg 
               
               
                   
               
               
                 cgactggtgagtacgccaaaaattttgactagcggaggctagaaggagagagacgggtgcgagagcgtcagtattaagcg 
               
               
                   
               
               
                 ggggtgaataagatcgcgatgggaaaaaattcggttaaggccagggggaaagaaaaaatataaattaaaacatatagtat 
               
               
                   
               
               
                 gggcaagcagggagctagaacgattcgcagttaatcctggcctgttagaaacatcagaaggctgtagacaaatactggga 
               
               
                   
               
               
                 cagctacaaccatcccttcagacaggatcagaagaacttagatcattatataatacagtagcaaccctctattgtgtgca 
               
               
                   
               
               
                 tcaaaggatagagataaaagacaccaaggaagctttagacaagatagaggaagagcaaaacaaaagtaagaccaccgcac 
               
               
                   
               
               
                 agcaagcggccgctgatcttcagacctggaggaggagatatgagggacaattggagaagtgaattatataaatataaagt 
               
               
                   
               
               
                 agtaaaaattgaaccattaggagtagcacccaccaaggcaaagagaagagtggtgcagagagaaaaaagagcagtgggaa 
               
               
                   
               
               
                 taggagctttgttccttgggttcttgggagcagcaggaagcactatgggcgcagcgtcaatgacgctgacggtacaggcc 
               
               
                   
               
               
                 agacaattattgtctggtatagtgcagcagcagaacaatttgctgagggctattgaggcgcaacagcatctgttgcaact 
               
               
                   
               
               
                 cacagtctggggcatcaagcagctccaggcaagaatcctggctgtggaaagatacctaaaggatcaacagctcctgggga 
               
               
                   
               
               
                 tttggggttgctctggaaaactcatttgcaccactgctgtgccttggaatgctagttggagtaataaatctctggaacag 
               
               
                   
               
               
                 atttggaatcacacgacctggatggagtgggacagagaaattaacaattacacaagcttaatacactccttaattgaaga 
               
               
                   
               
               
                 atcgcaaaaccagcaagaaaagaatgaacaagaattattggaattagataaatgggcaagtttgtggaattggtttaaca 
               
               
                   
               
               
                 taacaaattggctgtggtatataaaattattcataatgatagtaggaggcttggtaggtttaagaatagtttttgctgta 
               
               
                   
               
               
                 ctttctatagtgaatagagttaggcagggatattcaccattatcgtttcagacccacctcccaaccccgaggggacccga 
               
               
                   
               
               
                 caggcccgaaggaatagaagaagaaggtggagagagagacagagacagatccattcgattagtgaacggatctcgacggt 
               
               
                   
               
               
                 atcgttttaaaagaaaaggggggattggggggtacagtgcaggggaaagaatagtagacataatagcaacagacatacaa 
               
               
                   
               
               
                 actaaagaattacaaaaacaaattacaaaaattcaaaattttatcggcgtgttgggggtggaccatcctctaggtattga 
               
               
                   
               
               
                 ataagaaaaatgaagttaaggtggttgatggtaacactatgctaataactgcagagccagaagcaccataagggacatga 
               
               
                   
               
               
                 taagggagccagcagacctctgatctcttcctgaatgctaatcttaaacatcctgaggaagaatgggacttccatttggg 
               
               
                   
               
               
                 gtgggcctatgatagggtaataagacagtagtgaatatcaagctacaaaaagccccctttcaaattcttctcagtcctaa 
               
               
                   
               
               
                 cttttcatactaagcccagtccttccaaagcagactgtgaaagagtgatagttccgggagactagcactgcagattccgg 
               
               
                   
               
               
                 gtcactgtgagtgggggaggcagggaagaagggctcacaggacagtcaaaccatgccccctgtttttccttcttcaagta 
               
               
                   
               
               
                 gacctctataagacaacagagacaactaaggctgagtggccaggcgaggagaaaccatctcgccgtaaaacatggaagga 
               
               
                   
               
               
                 acacttcaggggaaaggtggtatctctaagcaagagaactgagtggagtcaaggctgagagatgcaggataagcaaatgg 
               
               
                   
               
               
                 gtagtgaaaagacattcatgaggacagctaaaacaataagtaatgtaaaatacagcatagcaaaactttaacctccaaat 
               
               
                   
               
               
                 caagcctctacttgaatccttttctgagggatgaataaggcataggcatcaggggctgttgccaatgtgcattagctgtt 
               
               
                   
               
               
                 tgcagcctcaccttctttcatggagtttaagatatagtgtattttcccaaggtttgaactagctcttcatttctttatgt 
               
               
                   
               
               
                 tttaaatgcactgacctcccacattccctttttagtaaaatattcagaaataatttaaatacatcattgcaatgaaaata 
               
               
                   
               
               
                 aatgttttttattaggcagaatccagatgctcaaggcccttcataatatcccccagtttagtagttggacttagggaaca 
               
               
                   
               
               
                 aaggaacctttaatagaaattggacagcaagaaagcgagcttagtgatacttgtgggccagggcattagccacaccagcc 
               
               
                   
               
               
                 accactttctgataggcagcctgcactggtggggtgAATTCTTTGCCAAAGTGATGGGCCAGCACACAGACCAGCACGTT 
               
               
                   
               
               
                 GCCCAGGAGCTGTGGGAGGAAGATAAGAGGTATGAACATGATTAGCAAAAGGGCCTAGCTTGGACTCAGAATAATCCAGC 
               
               
                   
               
               
                 CTTATCCCAACCATAAAATAAAAGCAGAATGGTAGCTGGATTGTAGCTGCTATTAGCAATATGAAACCTCTTACATCAGT 
               
               
                   
               
               
                 TACAATTTATATGCAGAAATATTTATATGCAGAAATATTGCTATTGCCTTAACCCAGAAATTATCACTGTTATTCTTTAG 
               
               
                   
               
               
                 AATGGTGCAAAGAGGCATGATACATTGTATCATTATTGCCCTGAAAGAAAGAGATTAGGGAAAGTATTAGAAATAAGATA 
               
               
                   
               
               
                 AACAAAAAAGTATATTAAAAGAAGAAAGCATTTTTTAAAATTACAAATGCAAAATTACCCTGATTTGGTCAATATGTGTA 
               
               
                   
               
               
                 CCCTGTTACTTCTCCCCTTCCTATGACATGAACTTAACCATAGAAAAGAAGGGGAAAGAAAACATCAAGGGTCCCATAGA 
               
               
                   
               
               
                 CTCACCCTGAAGTTCTCAGGATCCACGTGCAGCTTGTCACAGTGCAGCTCACTCAGctgGGCAAAGGTGCCCTTGAGGTT 
               
               
                   
               
               
                 GTCCAGGTGAGCCAGGCCATCACTAAAGGCACCGAGCACTTTCTTGCCATGAGCCTTCACCTTAGGGTTGCCCATAACAG 
               
               
                   
               
               
                 CATCAGGAGTGGACAGATCCCCAAAGGACTCAAAGAACCTCTGGGTCCAAGGGTAGACCACCAGCAGCCTAAGGGTGGGA 
               
               
                   
               
               
                 AAATAGACCAATAGGCAGAGAGAGTCAGTGCCTATCAGAAACCCAAGAGTCTTCTCTGTCTCCACATGCCCAGTTTCTAT 
               
               
                   
               
               
                 TGGTCTCCTTAAACCTGTCTTGTAACCTTGATACCAACCTGCCCAGGGCCTCACCACCAACTTCATCCACGTTCACCTTG 
               
               
                   
               
               
                 CCCCACAGGGCAGTAACGGCAGACTTCTCCTCAGGAGTCAGGTGCACCATGGTGTCTGTTTGAGGTTGCTAGTGAACACA 
               
               
                   
               
               
                 GTTGTGTCAGAAGCAAATGTAAGCAATAGATGGCTCTGCCCTGACTTTTATGCCCAGCCCTGGCTCCTGCCCTCCCTGCT 
               
               
                   
               
               
                 CCTGGGAGTAGATTGGCCAACCctagggtgtggctccacagggtgaggtctaagtgatgacagccgtacctgtccttggc 
               
               
                   
               
               
                 tcttctggcactggcttaggagttggacttcaaaccctcagccctccctctaagatatatctcttggccccataccatca 
               
               
                   
               
               
                 gtacaaattgctactaaaaacatcctcctttgcaagtgtatttacCACCCTTTTCCGGCACGCAGATAGTCAATATCTTC 
               
               
                   
               
               
                 AGCGTCCCCAAGGCCTGCAAGGGTGGGGCCCCATATCTGGAAGTCCCAGGCGGAGCTGGGAGTTGGTCAAGTCTGGGCTG 
               
               
                   
               
               
                 TGGGGGCAGGGAGTGCTGGGGGATGGacgcgtatctgcggccgcctatctgtaccactagtctcgagaagctttcatcaa 
               
               
                   
               
               
                 aaaaagtctaaccagctgcattcgactttgactgcagcagctggttagaaggttctactggaggagggtcccagcccatt 
               
               
                   
               
               
                 gctaaattaacatcaggctctgagactggcagtatatctctaacagtggttgatgctatcttctggaacttgcctgctac 
               
               
                   
               
               
                 attgagaccactgacccatacataggaagcccatagctctgtcctgaactgttaggccactggtccagagagtgtgcatc 
               
               
                   
               
               
                 tcctttgatcctcataataaccctatgagatagacacaattattactcttactttatagatgatgatcctgaaaacatag 
               
               
                   
               
               
                 gagtcaaggcacttgcccctagctgggggtataggggagcagtcccatgtagtagtagaatgaaaaatgctgctatgctg 
               
               
                   
               
               
                 tgcctcccccacctttcccatgtctgccctctactcatggtctatctctcctggctcctgggagtcatggactccaccca 
               
               
                   
               
               
                 gcaccaccaacctgacctaaccacctatctgagcctgccagcctataacccatctgggccctgatagctggtggccagcc 
               
               
                   
               
               
                 ctgaccccaccccaccctccctggaacctctgatagacacatctggcacaccagctcgcaaagtcaccgtgagggtcttg 
               
               
                   
               
               
                 tgtttgctgagtcaaaattccttgaaatccaagtccttagagactcctgctcccaaatttacagtcatagacttcttcat 
               
               
                   
               
               
                 ggctgtctcctttatccacagaatgattcctttgcttcattgccccatccatctgatcctcctcatcagtgcagcacagg 
               
               
                   
               
               
                 gcccatgagcagtagctgcagagtctcacataggtctggcactgcctctgacatgtccgaccttaggcaaatgcttgact 
               
               
                   
               
               
                 cttctgagctcagtcttgtcatggcaaaacaaagataataatagtgtttttttatggagttagcgtgaggatggaaaaca 
               
               
                   
               
               
                 atagcaaaattgattagactataaaaggtctcaacaaatagtagtagattttatcatccattaatccttccctctcctct 
               
               
                   
               
               
                 cttactcatcccatcacgtatgcctcttaattttcccttacctataataagagttattcctcttattatattcttcttat 
               
               
                   
               
               
                 agtgattctggatatcaaagtgggaatgaggggcaggccactaacgaagaagatgtttctcaaagaagccattctcccca 
               
               
                   
               
               
                 catagatcatctcagcagggttcaggaagataaaggaggatcaaggtcgaaggtaggaactaaggaagaacactgggcaa 
               
               
                   
               
               
                 gtgacgcgtcgacggtaccgttaacgatcttagccactttttaaaagaaaaggggggactggaagggctaattcactccc 
               
               
                   
               
               
                 aacgaagacaagatatcctgCTAGTCCTTCCTTTCTAAATGACGAGAGAGACAGAAGAATTCTTCAAGGTTAGTGTGTCC 
               
               
                   
               
               
                 AGCATGCAACCTTTCCTTCCTGGATGAGCATCCCTGGAGTAGGAGAGCCAGCCTGCCTCCTGCGCTGGCACAGAGCCCGG 
               
               
                   
               
               
                 TTCCCTAGACAACTGCCTCTCCAAATCTGATGTCCAGCGCCACCTGGTGTCCACATCAAGCAGACACAATTAATAGTCAA 
               
               
                   
               
               
                 CCTGTTCAGGAAAACTGTGAGGGGGAAAAAAAAGAAAGAGGATTTATGAAGGGAAAAGAAAGTTTAGAGGATATGCCACG 
               
               
                   
               
               
                 ATTGGctagcagctgctttttgcctgtactgggtctctctggttagaccagatctgagcctgggagctctctggctaact 
               
               
                   
               
               
                 agggaacccactgcttaagcctcaataaagcttgccttgagtgcttcatccggAATCAACCTCTGGATTACAAAATTTGT 
               
               
                   
               
               
                 GAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGC 
               
               
                   
               
               
                 TATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCG 
               
               
                   
               
               
                 TTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAG 
               
               
                   
               
               
                 CTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGAC 
               
               
                   
               
               
                 AGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAGCTGACGTCCTTTCCATGGCTGCTCGCCTGTG 
               
               
                   
               
               
                 TTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGC 
               
               
                   
               
               
                 CTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCC 
               
               
                   
               
               
                 GCCTGtccggtAGCTTGCCAGCCTCGACTGTGCCTTCTAGTTGCCAGCCGTCTGTTGTTTGCCCCTCCCCCGTGCCTTCC 
               
               
                   
               
               
                 TTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCA 
               
               
                   
               
               
                 TTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGcaagcttggcgt 
               
               
                   
               
               
                 aatcatggtcatagctgtttcctgtgtgaaattgttatccgctcacaattccacacaacatacgagccggaagcataaag 
               
               
                   
               
               
                 tgtaaagcctggggtgcctaatgagtgagctaactcacattaattgcgttgcgctcactgcccgctttccagtcgggaaa 
               
               
                   
               
               
                 cctgtcgtgccagctgcattaatgaatcggccaacgcgcggggagaggcggtttgcgtattgggcgctcttccgcttcct 
               
               
                   
               
               
                 cgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatcc 
               
               
                   
               
               
                 acagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgtt 
               
               
                   
               
               
                 gctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccga 
               
               
                   
               
               
                 caggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccgga 
               
               
                   
               
               
                 tacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggt 
               
               
                   
               
               
                 cgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttg 
               
               
                   
               
               
                 agtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggc 
               
               
                   
               
               
                 ggtgctacagagttcttgaagtggtggcctaactacggctacactagaagaacagtatttggtatctgcgctctgctgaa 
               
               
                   
               
               
                 gccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgttt 
               
               
                   
               
               
                 gcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtgg 
               
               
                   
               
               
                 aacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatg 
               
               
                   
               
               
                 aagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaatcagtgaggcacctatct 
               
               
                   
               
               
                 cagcgatctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagggcttacca 
               
               
                   
               
               
                 tctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccagatttatcagcaataaaccagccagccgg 
               
               
                   
               
               
                 aagggccgagcgcagaagtggtcctgcaactttatccgcctccatccagtctattaattgttgccgggaagctagagtaa 
               
               
                   
               
               
                 gtagttcgccagttaatagtttgcgcaacgttgttgccattgctacaggcatcgtggtgtcacgctcgtcgtttggtatg 
               
               
                   
               
               
                 gcttcattcagctccggttcccaacgatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctcctt 
               
               
                   
               
               
                 cggtcctccgatcgttgtcagaagtaagttggccgcagtgttatcactcatggttatggcagcactgcataattctctta 
               
               
                   
               
               
                 ctgtcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcggcga 
               
               
                   
               
               
                 ccgagttgctcttgcccggcgtcaatacgggataataccgcgccacatagcagaactttaaaagtgctcatcattggaaa 
               
               
                   
               
               
                 acgttcttcggggcaaaactctcaaggatcttaccgctgttgagatccagttcgatgtaacccactcgtgcacccaactg 
               
               
                   
               
               
                 atcttcagcatcttttactttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaagggaataa 
               
               
                   
               
               
                 gggcgacacggaaatgttgaatactcatactcttcctttttcaatattattgaagcatttatcagggttattgtctcatg 
               
               
                   
               
               
                 agcggatacatatttgaatgtatttagaaaaataaacaaataggggttccgcgcacatttccccgaaaagtgccacctga 
               
               
                   
               
               
                 cgtctaagaaaccattattatcatgacattaacctataaaaataggcgtatcacgaggccctttcgtctcgcgcgtttcg 
               
               
                   
               
               
                 gtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcaga 
               
               
                   
               
               
                 caagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggctggcttaactatgcggcatcagagcagattgtact 
               
               
                   
               
               
                 gagagtgcacc 
               
            
           
         
       
     
     These vectors were produced by transient triple transfection of the recombinant vector, pCMVΔR8.9 and pMD.G into 293T cells. A total of 5×10 6  293 T cells were seeded in 10-cm-diameter dishes 24 h prior to transfection in DMEM with 10% fetal bovine serum, penicillin (100 IU/ml), and streptomycin (100 μg/ml) in a 5% CO 2  incubator. A total of 20 μg of plasmid DNA was used for the transfection of one 10 cm dish: 3.5 μg of the envelope plasmid pMD.G, 6.5 μs of packaging plasmid, and 10 μg of transfer vector plasmid. The medium (10 ml) was replaced after 14 to 16 h; the conditioned medium was collected after another 24 h, cleared by low-speed centrifugation, and filtered through 0.45-μm-pore-size cellulose acetate filters. The pseudotyped virions were than concentrated by ultracentrifugation and resuspended for frozen down in aliquots for subsequent titration and stem cell transduction. 
     Titration Assay 
     These vectors were produced by transient triple transfection of the recombinant vector, pCMVΔR8.9 and pMD.G into 293T cells. A total of 5×10 6  293 T cells were seeded in 10-cm-diameter dishes 24 h prior to transfection in DMEM with 10% fetal bovine serum, penicillin (100 IU/ml), and streptomycin (100 μg/ml) in a 5% CO 2  incubator. A total of 20 μg of plasmid DNA was used for the transfection of one 10 cm dish: 3.5 μg of the envelope plasmid pMD.G, 6.5 μs of packaging plasmid, and 10 μg of transfer vector plasmid. The medium (10 ml) was replaced after 14 to 16 h; the conditioned medium was collected after another 24 h, cleared by low-speed centrifugation, and filtered through 0.45-μm-pore-size cellulose acetate filters. The pseudotyped virions were than used directly for titration assays in Hela cells. Serial dilutions of un-concentrated virus were used to infect 20 5  cells in a six-well plate in the presence of Polybrene (8 μg/ml). each dilution was tested in triplicate. The cells were kept in culture for no less than 15 days, harvested and genomic DNA were extracted, quantified and used to Vector Copy Number (VCN) quantification. Transducing activity was expressed in transducing units (TU). VCN analysis was performed by TaqMan PCR analysis using: 
     Gag-specific probe—5′-acagccttctgatgtttctaacaggccagg-3′ (SEQ ID NO: 41)
 
Gag-primer-Forward:—5′-ggagctagaacgattcgcagtt-3′ (SEQ ID NO: 42)
 
Gag-primer-Reverse:—5′-gttgtagctgtcccagtatttgtc-3′ (SEQ ID NO: 43)
 
Human Albumin Probe—5′-tgctgaaacattcaccttccatgcagt-3′ (SEQ ID NO: 44)
 
Human Albumin Forward—5′tgaaacatacgttcccaaagagttt-3′ (SEQ ID NO: 45)
 
Human Albumin Reverse—5′ ctctccttctcagaaagtgtgcatat-3′ (SEQ ID NO: 46)
 
     Hematopoietic Stem Cell Transduction and Transplantation 
     Donor bone marrow was flushed from the femurs of 8- to 16-week-old male Hbb th3/+  thalassemic mice that had been injected intravenously (i.v.) 6 days earlier with 5-flurouracil (5-FU, Pharmacia; 150 mg kg −1  body weight). Bone marrow cells were resuspended in serum-free medium, and supplemented with rmSCF (100 ng ml −1 ), rmTPO (100 ng ml −1 ), β-mercaptoethanol (55 μM; Sigma), L-glutamine (2 mM), pen/strep (10 IU ml −1 ), and cultured for 18 h. Bone marrow cells were pelleted and resuspended in serum-free medium containing concentrated lentiviral supernatant at a multiplicity of infection (MOI) of 25 and supplemented with polybrene (8 μs ml −1 ), L-glutamine (2 mM), pen/strep (100 IU ml −1 ) and incubated for 8 h. Transduced bone marrow cells (5×10 5 ) were then i.v. injected into each of the irradiated female recipients to establish bone marrow chimaeras. Recipient mice (11- to 14-week-old C57/Hbb th3/+  mice) were irradiated with 10.5 Gy (split dose 2×5.25 Gy) on the day of transplantation. 
     Post-Transplant Vector Studies 
     Peripheral Blood was collected periodically starting 5-6 weeks after hematopoietic stem cell transplantation. Blood samples were obtained by retro-orbital puncture under ether anesthesia, according to MSKCC animal protocol. Total hemoglobin levels, red cell counts, hematocrit levels, neutrophil counts, platelet counts, and reticulocyte counts were measured on a blood cell count analyzer. 
     VCN analysis: To evaluate the integrated vector copy number in peripheral blood, mouse genomic DNA was isolated, quantified and analyzed with multiplex real-time TaqMan PCR using following probes and primer: 
     Gag-specific probe—5′-acagccttctgatgtactaacaggccagg-3′ (SEQ ID NO: 41)
 
Gag-primer-Forward—5′-ggagctagaacgattcgcagtt-3′ (SEQ ID NO: 42)
 
Gag-primer-Reverse—5′-gttgtagctgtcccagtatttgtc-3′ (SEQ ID NO: 43)
 
Mouse β-actin Probe—5′-tacgagggctatgctctccctcacgc-3′ (SEQ ID NO: 47)
 
Mouse β-actin-primer-Forward—5′-tcacccacactgtgcccat-3′ (SEQ ID NO: 48)
 
Mouse β-actin-primer-reverse—5′-agccaggtccagacgcag-3′ (SEQ ID NO: 49)
 
     Hemoglobin assays: automated hemoglobin (Hb) quantification; based line anemia in Hbb th3/+  mice is 7.5-8.5 g/dL. Vector performance, i.e., protein output per vector copy, was determined as the ratio of gain in hemoglobin expression (i.e., correction of anemia), ΔHb g/dL, per vector copy number (VCN) measured in circulating blood cells. 
     Results and Discussion 
     As shown in  FIG. 17 , the SNS22.2.B87.A1, SNS23.2.B87.A1, SNS24.2.B87.A1, SNS26.B87.A1 and SNS27.2.B87.A1 vectors achieved higher titer than the TNS9.B87.A1 vector. Moreover, as shown in  FIG. 16  and Table 1, the multiple mutations and/or deletions introduced into the promoter, HS2, HS3, and HS4 regions of the β-globin LCR of SNS23.2.B87.A1 did not affect (e.g., did not reduce) globin expression in mice. Furthermore, vectors lacking HS2 or all LCR elements but including an BLV enhancer and flanked by the A1 insulator, e.g., SNS26.B87.A1 and SNS27.2.B87.A1, can effectively treat anemia in thalassemic mice. While the globin protein output per copy of the SNS26.B87.A1 vector or the SNS27.2.B87.A1 vector was less than that of the SNS23.2.B87.A1 vector (see  FIG. 18  and Table 1), SNS26.B87.A1 and SNS27.2.B87.A1 vectors were able to correct the disease with a higher vector copy number (see  FIG. 16 ). The higher titer of the SNS26.B87.A1 and SNS27.2.B87.A1 vectors facilitated higher gene transfer, resulting in a higher vector copy number (VCN) and therapeutic activity. 
     As shown in  FIG. 16 , the expression of an incomplete or lacking LCR required 0.8 or higher copies of vector per cell, while in the case of SNS23.2.B87.A1, a VCN of 0.3 copies was sufficient to achieve correction of anemia 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 Vector copy number (VCN) and Hb levels in long-term  
               
               
                 hematopoietic chimeras. Total Hb level [g/dL] in peripheral  
               
               
                 blood (PB) of chimeric mice. Representative data for week 6  
               
               
                 after transplant is shown. ΔHb level was obtained by  
               
               
                 subtracting Th3/+ hemoglobin (value = 9 g/dL) from  
               
               
                 total Hb level for each animal tested. ΔHb/copy = 
               
               
                 Correlation between delta(Δ)Hb and vector copy number.  
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                 HGB 
                   
                 ΔHGB/ 
               
               
                 mouse ID 
                 VCN 
                 (g/dL) 
                 Δ(HGB-9) 
                 copy 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 TNS9.B87.A1 #1 
                 0.2 
                 11.3 
                 2.3 
                 11.5 
               
               
                 TNS9.B87.A1 #2 
                 0.2 
                 12.0 
                 3.0 
                 16.7 
               
               
                 TNS9.B87.A1 #3 
                 0.2 
                 8.5 
                 −0.5 
                 −2.5 
               
               
                 TNS9.B87.A1 #4 
                 0.2 
                 13.5 
                 4.5 
                 30.0 
               
               
                 SNS23.2.B87.A1 #1 
                 0.3 
                 13.1 
                 4.1 
                 15.8 
               
               
                 SNS23.2.B87.A1 #2 
                 0.3 
                 13.3 
                 4.3 
                 13.0 
               
               
                 SNS23.2.B87.A1 #3 
                 0.3 
                 13.2 
                 4.2 
                 14.5 
               
               
                 SNS23.2.B87.A1 #4 
                 0.3 
                 13.0 
                 4.0 
                 13.8 
               
               
                 SNS23.2.B87.A1 #5 
                 0.3 
                 13.2 
                 4.2 
                 13.5 
               
               
                 SNS26.B87.A1 #1 
                 1.2 
                 11.4 
                 2.4 
                 2.0 
               
               
                 SNS26.B87.A1 #2 
                 1.2 
                 11.9 
                 2.9 
                 2.3 
               
               
                 SNS26.B87.A1 #3 
                 1.3 
                 9.7 
                 0.7 
                 0.6 
               
               
                 SNS26.B87.A1 #4 
                 1.7 
                 11.9 
                 2.9 
                 1.8 
               
               
                 SNS26.B87.A1 #5 
                 1.3 
                 11.3 
                 2.3 
                 1.8 
               
               
                 SNS27.2.B87.A1 #1 
                 0.8 
                 11.7 
                 2.7 
                 3.3 
               
               
                 SNS27.2.B87.A1 #2 
                 0.8 
                 13.0 
                 4.0 
                 5.3 
               
               
                 SNS27.2.B87.A1 #3 
                 0.9 
                 11.5 
                 2.5 
                 2.7 
               
               
                 SNS27.2.B87.A1 #4 
                 0.9 
                 11.5 
                 2.5 
                 2.7 
               
               
                 SNS27.2.B87.A1 #5 
                 0.7 
                 11.3 
                 2.3 
                 3.2 
               
               
                 TH3/+ MOCK #1 
                 — 
                 7.5 
                 −1.5 
                   
               
               
                   
               
            
           
         
       
     
     Example 7: Evaluation of Globin Production at Different Time Points 
     Additional experiments were conducted to measure the globin production at different time points in thalassemic mice transfected with vectors disclosed herein. The methods used in Example 6 were applied in this Example. Average total Hb levels and average gains in Hb levels (ΔHb) in peripheral blood were measured in thalassemic mice transfected with vectors for 6-week, 5-month, or 9-month. ΔHb was normalized to vector copy (VCN), and ΔHb=Hb level−7.5 (baseline level in thalassemic mice, as all the time points consistently show an HGB value of 7.5 g/dL in the Thalassemic mice used as controls, this value was used). As shown in  FIG. 19  and Table 2, globin productions from all vectors, including SNS23.2.B87A1, were stable over time. As shown in  FIG. 20 , SNS23.2.B87.A1 exhibited equal or better output of B-globin relative to TNS9.B87.A1, especially at late time points. 
                     TABLE 2                  Representative data for three different time points after transplant                                         HGB        ΔHGB/       Vectors   VCN   (g/dL)   Δ(HGB-7.5)   copy                                         TNS9.B87.A1 (6 weeks)   0.5   10.6   3.8   8.6       TNS9.B87.A1 (5 months)   0.5   10.7   3.2   5.1       TNS9.B87.A1 (9 months)   0.4   9.8   2.3   6.4       SNS23.2.B87.A1 (6 weeks)   1.1   13.2   5.7   6.0       SNS23.2.B87.A1 (5 months)   1.1   11.3   3.8   4.3       SNS23.2.B87.A1 (9 months)   0.5   11.6   4.1   8.3       SNS26.B87.A1 (6 weeks)   2.0   11.2   3.7   2.0       SNS26.B87.A1 (5 months)   3.0   11.1   3.6   1.5       SNS26.B87.A1 (9 months)   3.1   11.1   3.6   1.4       SNS27.2.B87.A1 (6 weeks)   0.8   11.8   4.3   5.3       SNS27.2.B87.A1 (5 months)   1.5   12.1   4.6   3.5       SNS27.2.B87.A1 (9 months)   1.1   10.4   2.7   2.4       TH3/+ MOCK (6 weeks)   0   7.5   0   0       TH3/+ MOCK (5 months)   0   7.5   0   0       TH3/+ MOCK (9 months)   0   7.5   0   0                    
Average of Vector copy number (VCN) in long-term hematopoietic chimeras. Average Hb level [g/dL] in peripheral blood (PB) of chimeric mice. ΔHGb level was obtained by subtracting Th3/+ hemoglobin (value=7.5 g/dL) from total Hb level for each animal tested. ΔHGb/copy=Correlation between delta(Δ)Hb and vector copy number.
 
All the ΔHGb calculations are made using TH3/+ MOCK (7.5 g/dL) as basal HGB, value that is consistent in all the time points.
 
     Example 8: Human Primary CD34+ Cells Transduced with Vectors Disclosed Herein 
     Human primary CD34 +  cells were isolated by centrifugation on a gradient of Ficoll-Hypaque Plus density. CD34+ cells were purified by positive selection using separation columns and beads. After one day of cytokine stimulation the CD34 +  cells were transduced with SNS.23.2.B87.A1 or TNS9.B87.A1 vectors disclosed herein using 4 different MOI. 10 and 15 days after transduction, the cells were harvested and the Vector copy number (VCN) was measured using the methods disclosed in Example 6. As shown in Table 3, at MOIs from 20×1 to 100×1, a linearly increased VCN response was observed in cells transduced with SNS.23.2.B87.A1 vector, with maximum VCN&gt;2. By contrast, cells transduced with TNS9.B87.A1 vector did not exhibit proportional increases in VCN at MOIs from 20×1 to 100×1. Unlike TNS9.B87.A1 vector, the SNS23.2.B87.A1 vector resulted in an increased globin gene transduction in CD34 +  cells up to &gt;2 VCN per cell (2.33 at MOI 100, day 15, Table 3). 
     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                 Transduction of human primary CD34+ cells 
               
            
           
           
               
               
               
               
            
               
                 Groups 
                 MOI 
                 VCN day10 
                 VCN day15 
               
               
                   
               
               
                 SNS23.2.B87.A1 
                  10 × 1 
                 1   
                 0.21 
               
               
                 Concentrate virus 
                  20 × 1 
                 1.78 
                 0.56 
               
               
                   
                  40 × 1 
                 3.35 
                 0.86 
               
               
                   
                 100 × 1 
                 7.33 
                 2.33 
               
               
                 TNS9.B87.A1  
                  10 × 1 
                 0.47 
                 0.38 
               
               
                 Concentrate virus 
                  20 × 1 
                 0.52 
                 0.44 
               
               
                   
                  40 × 1 
                 0.48 
                 0.40 
               
               
                   
                 100 × 1 
                 0.60 
                 0.44 
               
               
                 UT 
                 NA 
                 0.00 
                 0.00 
               
               
                   
               
            
           
         
       
     
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     From the foregoing description, it will be apparent that variations and modifications may be made to the presently disclosed subject matter described herein to adopt it to various usages and conditions. Such embodiments are also within the scope of the following claims. 
     All patents and publications and sequences referred to by accession or reference number mentioned in this specification are herein incorporated by reference to the same extent as if each independent patent and publication and sequence was specifically and individually indicated to be incorporated by reference.