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pRetroX-Tight-Pur载体

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  • ¥3500
  • ZYbscience
  • 中国/美国
  • ZY 632104
  • 2025年07月14日
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    • 详细信息
    • 文献和实验
    • 技术资料
    • 保存条件

      -20℃低温保存

    • 保质期

      三年

    • 英文名

      pRetroX-Tight-Pur

    • 库存

      20

    • 供应商

      泽叶生物

    载体基本信息

    出品公司: ZYbscience
    载体名称: pRetroX-Tight-Pur
    质粒类型: 逆病毒载体;四环素调控载体
    高拷贝/低拷贝: 高拷贝
    克隆方法: 限制性内切酶,多克隆位点
    启动子: Tight
    载体大小: 6568 bp
    5' 测序引物及序列: --
    3' 测序引物及序列: --
    载体标签: --
    载体抗性: 氨苄青霉素
    筛选标记: 嘌呤霉素(Puromycin)
    克隆菌株: DH5α
    宿主细胞(系): 常规细胞系(293、CV-1、CHO等)
    备注: 逆病毒载体pRetroX-Tight-Pur是四环素调控系统载体
    产品目录号: ZY632104
    稳定性: 稳表达
    组成型/诱导型: 诱导型
    病毒/非病毒: 逆转录病毒

    载体质粒图谱和多克隆位点信息

    pRetroX-Tight-Pur载体图谱



    pRetroX-Tight-Pur 多克隆位点

    pRetroX-Tight-Pur 载体特征

    载体简介

    pRetroX-Tight-Pur载体描述
    
    pRetroX-Tight-Pur is an inducible, self-inactivating, retroviral expression vector designed to express a gene of interest under the control of PTight, a modified Tet-responsive promoter. PTight consists of seven direct repeats of a 36 bp regulatory sequence that contains the 19 bp tet operator sequence (tetO), and a modified minimal CMV promoter (1). This vector is supplied with our Retro-XTM Tet-On Advanced and Tet-Off Advanced Inducible Expression Systems (Cat. Nos. 632104 and 632105). These systems provide ready access to the inducible gene expression strategy of Gossen & Bujard, and incorporate major improvements described by Urlinger, et al. (2-6).
    pRetroX-Tight-Pur is optimized to eliminate promoter interference through LTR self-inactivation. The hybrid 5’ LTR consists of the cytomegalovirus (CMV) type I enhancer and the mouse sarcoma virus (MSV) promoter. This promoter drives high levels of viral genome transcription in HEK 293-based packaging cell lines due, in part, to the presence of adenoviral E1A (7-8) in these cells. The self-inactivating feature of the vector is provided by a deletion in the 3’ LTR enhancer region (U3). During reverse transcription of the retroviral RNA, a copy of the inactivated 3’ LTR U3 region replaces the corresponding region of the 5’ LTR, resulting in the inactivation of the 5’ LTR CMV enhancer sequence. This mechanism can reduce the phenomenon known as promoter interference (9-10) and allow more efficient expression. In this way, pRetroX-Tight-Pur supports high viral titers, yet eliminates potential downstream interference between the inducible PTight promoter and its adjacent viral LTR.
    
    pRetroX-Tight-Pur contains all of the necessary viral RNA processing elements; these include the 5’ and 3’ LTRs, the packaging signal (Ψ+), and the tRNA primer binding site. For safety reasons, however, the vector lacks the structural genes (gag, pol, and env) necessary for retroviral particle formation and replication. pRetroX-Tight-Pur contains a puromycin resistance gene (Purr) under the control of the murine phosphoglycerate kinase (PGK) promoter (PPGK) for the selection of stable transfectants. In addition, the vector contains a ColE1 origin of replication and an E. coli Ampr gene for propagation and selection in bacteria.
    
    
    pRetroX-Tight-Pur can be used as either a plasmid or retroviral expression vector. When used as a retroviral expression vector, it must be transfected into a packaging cell line, such as GP2-293; we recommend the Retro-X Universal Packaging System (Cat. No. 631530). Packaging cell lines allow you to produce infectious, replication-incompetent retroviral particles. These retroviral particles can infect a wide range of target cells and transmit your gene of interest, but they cannot replicate within these cells due to the absence of viral structural genes. The separate introduction and integration of the structural genes into the packaging cell line minimizes the chance of producing replication-competent virus due to recombination events during cell proliferation.
    
    Propagation in E. coli
     Suitable host strains: DH5α, DH10B and other general purpose strains.
     Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) in E. coli hosts.
     E. coli replication origin: ColE1
     Copy number: high
    
    Notes:
    The vector sequence was compiled from information in the sequence databases, published literature, and other sources, together with partial sequences obtained by Clontech. This vector has not been completely sequenced.
    The viral supernatants produced by this retroviral vector could contain potentially hazardous recombinant virus. Due caution must be exercised in the production and handling of recombinant retrovirus. Appropriate NIH, regional, and institutional guidelines apply. 
    
    

    载体序列

    LOCUS       pRetroX-Tight-Pur	6567 bp 	DNA	SYN
    DEFINITION  pRetroX-Tight-Pur
    ACCESSION   
    KEYWORDS    
    SOURCE      
      ORGANISM  other sequences; artificial sequences; vectors.
    FEATURES             Location/Qualifiers
         source          1..6567
                         /organism="pRetroX-Tight-Pur"
                         /mol_type="other DNA"
         promoter        2..506
                         /label="CMV_immearly_promoter"
         misc_feature    6..688
                         /label="5_LTR"
         misc_feature    9..296
                         /label="CAG_enhancer"
         misc_feature    463..483
                         /label="CMV_fwd_primer"
         misc_feature    758..1567
                         /label="psi_plus_pack"
         misc_feature    1154..1561
                         /label="gag"
         misc_feature    1497..1519
                         /label="MSCV_primer"
         misc_feature    1497..1519
                         /label="pLXSN_5_primer"
         misc_feature    1535..1551
                         /label="pBABE_5_primer"
         CDS             1669..2196
                         /label="ORF frame 1"
                         /translation="MSSLLPISDRERMSSLLPISDRERMSSLLPISDRERMSSLSLSV
                         IENVCRVYSLSVIENVCRGRRVRWEAYISRARLVNRQIAWRRIRGRAGSRSRTREFYR
                         VGEALFPRQSGACALAAPLGTWRYTSGLWPRTHSTSTGRRQPAPFFGGPFAPPSTPPL
                         VRKFPPAPQLASCRT*"
         misc_feature    1888..1907
                         /label="pCEP_fwd_primer"
                         /translation="MSSLLPISDRERMSSLLPISDRERMSSLLPISDRERMSSLSLSV
                         IENVCRVYSLSVIENVCRGRRVRWEAYISRARLVNRQIAWRRIRGRAGSRSRTREFYR
                         VGEALFPRQSGACALAAPLGTWRYTSGLWPRTHSTSTGRRQPAPFFGGPFAPPSTPPL
                         VRKFPPAPQLASCRT*"
         misc_feature    1890..1914
                         /label="LNCX_primer"
                         /translation="MSSLLPISDRERMSSLLPISDRERMSSLLPISDRERMSSLSLSV
                         IENVCRVYSLSVIENVCRGRRVRWEAYISRARLVNRQIAWRRIRGRAGSRSRTREFYR
                         VGEALFPRQSGACALAAPLGTWRYTSGLWPRTHSTSTGRRQPAPFFGGPFAPPSTPPL
                         VRKFPPAPQLASCRT*"
         misc_feature    complement(2008..2031)
                         /label="MSCV_rev_primer"
                         /translation="MSSLLPISDRERMSSLLPISDRERMSSLLPISDRERMSSLSLSV
                         IENVCRVYSLSVIENVCRGRRVRWEAYISRARLVNRQIAWRRIRGRAGSRSRTREFYR
                         VGEALFPRQSGACALAAPLGTWRYTSGLWPRTHSTSTGRRQPAPFFGGPFAPPSTPPL
                         VRKFPPAPQLASCRT*"
         CDS             2253..3092
                         /label="ORF frame 3"
                         /translation="MEAGRPLGQRPIAALLLRFLGSEAGKGWVRGRAQGRAQGRGGRP
                         KVLRRPGILHASKAHVCRAVLLFLISGPFDLQPKLTMTEYKPTVRLATRDDVPRAVRT
                         LAAAFADYPATRHTVDPDRHIERVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPE
                         SVEAGAVFAEIGPRMAELSGSRLAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKG
                         LGSAVVLPGVEAAERAGVPAFLETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRK
                         PGA*"
         misc_feature    2405..2424
                         /label="mPGK_F_primer"
                         /translation="MEAGRPLGQRPIAALLLRFLGSEAGKGWVRGRAQGRAQGRGGRP
                         KVLRRPGILHASKAHVCRAVLLFLISGPFDLQPKLTMTEYKPTVRLATRDDVPRAVRT
                         LAAAFADYPATRHTVDPDRHIERVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPE
                         SVEAGAVFAEIGPRMAELSGSRLAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKG
                         LGSAVVLPGVEAAERAGVPAFLETSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRK
                         PGA*"
         CDS             complement(2418..3143)
                         /label="ORF frame 2"
                         /translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
                         PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
                         RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
                         DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
                         KLGLQVERPGDEEEENSAADVRF*"
         gene            2493..3092
                         /label="puro (variant)"
                         /gene="puro (variant)"
                         /translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
                         PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
                         RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
                         DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
                         KLGLQVERPGDEEEENSAADVRF*"
         misc_feature    complement(3801..3823)
                         /label="pGEX_3_primer"
                         /translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
                         PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
                         RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
                         DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
                         KLGLQVERPGDEEEENSAADVRF*"
         misc_feature    complement(3959..3979)
                         /label="pBABE_3_primer"
                         /translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
                         PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
                         RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
                         DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
                         KLGLQVERPGDEEEENSAADVRF*"
         misc_feature    complement(3965..4180)
                         /label="SV40_enhancer"
                         /translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
                         PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
                         RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
                         DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
                         KLGLQVERPGDEEEENSAADVRF*"
         promoter        3977..4245
                         /label="SV40_promoter"
                         /translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
                         PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
                         RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
                         DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
                         KLGLQVERPGDEEEENSAADVRF*"
         rep_origin      4144..4221
                         /label="SV40_origin"
                         /translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
                         PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
                         RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
                         DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
                         KLGLQVERPGDEEEENSAADVRF*"
         misc_feature    4206..4225
                         /label="SV40pro_F_primer"
                         /translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
                         PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
                         RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
                         DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
                         KLGLQVERPGDEEEENSAADVRF*"
         gene            complement(5345..6205)
                         /label="Ampicillin"
                         /gene="Ampicillin"
                         /translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
                         PLVEGEVAGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
                         RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
                         DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
                         KLGLQVERPGDEEEENSAADVRF*"
         CDS             complement(5345..6205)
                         /label="ORF frame 3"
                         /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                         IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                         YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                         DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                         LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                         EIGASLIKHW*"
         promoter        complement(6247..6275)
                         /label="AmpR_promoter"
                         /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                         IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                         YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                         DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                         LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                         EIGASLIKHW*"
         promoter        6497..506
                         /label="CMV_immearly_promoter"
                         /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                         IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                         YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                         DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                         LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                         EIGASLIKHW*"
         misc_feature    6561..688
                         /label="5_LTR"
                         /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                         IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                         YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                         DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                         LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                         EIGASLIKHW*"
    ORIGIN
        1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
       61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      301 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      361 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      421 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      481 gtgtacggtg ggaggtctat ataagcagag ctcaataaaa gagcccacaa cccctcactc
      541 ggcgcgccag tcttccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc
      601 ttgctgtttg catccgaatc gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
      661 tgactaccca cgacgggggt ctttcatttg ggggctcgtc cgggatttgg agacccctgc
      721 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc
      781 cgattgtcta gtgtctatgt ttgatgttat gcgcctgcgt ctgtactagt tagctaacta
      841 gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc
      901 tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc
      961 gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca
     1021 gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct
     1081 gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc
     1141 tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga
     1201 tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt
     1261 ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg
     1321 agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga
     1381 ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa
     1441 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct
     1501 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct
     1561 aggcgccgga attgaagatc tgaggccctt tcgtcttcac tcgagtttac tccctatcag
     1621 tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgat gtcgagttta
     1681 ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacgt
     1741 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttatc cctatcagtg
     1801 atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgaggtagg
     1861 cgtgtacggt gggaggccta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
     1921 agaaggatcc gcggccgcgc cggctctaga tcgcgaacgc gtgaattcta ccgggtaggg
     1981 gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg
     2041 cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg
     2101 gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc
     2161 ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact
     2221 agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc
     2281 ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc
     2341 gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc
     2401 ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct
     2461 ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc
     2521 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     2581 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     2641 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     2701 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     2761 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     2821 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     2881 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     2941 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     3001 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     3061 cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga
     3121 ccgaaaggag cgcacgaccc catgcatcgg cgtctcgaga tatcagtggt ccaggctcta
     3181 gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa
     3241 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     3301 aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag
     3361 aagttcagat caaggtcagg aacagatgga acagggtcga ccctagagaa ccatcagatg
     3421 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     3481 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     3541 cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca
     3601 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     3661 ctgagtgatt gactacccgt cagcgggggt ctttcatttg ggggctcgtc cgggatcggg
     3721 agacccctgc ccagggacca ccgacccacc accgggaggt aagctggctg cctcgcgcgt
     3781 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     3841 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     3901 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtagat ccggctgtgg
     3961 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4021 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4081 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4141 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4201 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4261 ggaggctttt ttggaggcct aggcttttgc aaaaagctta ctggcttaac tatgcggcat
     4321 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     4381 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4441 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4501 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4561 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4621 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4681 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4741 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4801 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4861 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4921 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4981 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     5041 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5101 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5161 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5221 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc

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