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  • pRetroX-Tet-On Advanced

    價格:3500元

    聯系方式:I47-825O-882O

    買家導航

    載體基本信息

    出品公司: Clontech
    載體名稱: pRetroX-Tet-On Advanced
    質粒類型: 逆病毒載體;四環素調控載體
    高拷貝/低拷貝: 高拷貝
    克隆方法: 限制性內切酶,多克隆位點
    啟動子: CMV IE
    載體大小: 8130 bp
    5' 測序引物及序列: --
    3' 測序引物及序列: --
    載體標簽: --
    載體抗性: 氨芐青霉素
    篩選標記: 新霉素(Neomycin)
    克隆菌株: DH5α
    宿主細胞(系): 常規細胞系(293、CV-1、CHO等)
    備注: 逆病毒載體pRetroX-Tet-On Advanced是四環素調控系統載體,組成型表達rtTA Advanced。
    產品目錄號: 632104
    穩定性: 穩表達
    組成型/誘導型: 組成型
    病毒/非病毒: 逆轉錄病毒

    載體質粒圖譜和多克隆位點信息

    pRetroX-Tet-On Advanced載體圖譜



    pRetroX-Tet-On Advanced 載體特征

    載體簡介

    pRetroX-Tet-On Advanced載體描述
    
    pRetroX-Tet-On Advanced is a Retro-X Q retroviral vector that expresses rtTA-Advanced, an improved version of the reverse tetracycline(Tet)-controlled transactivator protein rtTA (1–4). rtTA-Advanced is more sensitive to doxycycline (Dox) and yields lower background expression than the original rtTA. The rtTA-Advanced protein is a fusion of the Tet repressor (TetR) DNA-binding domain, and three minimal “F”-type transcriptional activation domains from the herpes simplex virus VP16 protein. The gene encoding rtTA-Advanced is completely synthetic, lacks cryptic splice sites, and utilizes human codon preferences for stable expression in mammalian cells. Expression of rtTA-Advanced is driven by the powerful, constitutively active CMV promoter.
    As with all of our Retro-X Q vectors, pRetroX-Tet-On Advanced uses LTR self-inactivation to eliminate promoter interference. In LTR self-inactivation, the mechanism of viral integration is used to introduce a deletion into the 5’ LTR. As a result of this mutation, the 5’ LTR CMV/MSV promoter, which drives high expression of the complete viral genome in packaging cells, is inactive in stably transduced target cells. This allows the expression of rtTA-Advanced (from PCMV IE) to proceed unimpeded (5, 6). The vector also contains a neomycin resistance gene (Neor) that allows G418 selection of stably transduced cells. rtTA-Advanced and the Neor marker are coexpressed from a bicistronic transcript containing an internal ribosome entry site (IRES).This ensures that a high frequency of G418 resistant clones express the Tet-On Advanced transactivator.
    
    pRetroX-Tet-On Advanced contains all of the necessary viral RNA processing elements; these include the 5’ and 3’ LTRs, a packaging signal (Ψ+), and a tRNA primer binding site. The vector also contains an SV40 origin of replication for plasmid propagation in mammalian cells that express SV40 T antigen. In addition, a pUC origin of replication and an E. coli Ampr gene allow for propagation and selection in bacteria.
    
    The pRetroX-Tet-On Advanced vector is used in conjunction with Tet response vectors to create double stable Retro-X Tet-On Advanced cell lines. Such cell lines are essentially Doxycycline (Dox)-controlled gene expression systems in which the Tet response vector expresses a gene of interest under the control of a Tet-responsive element (TRE; e.g. PTight; 7, 8). In the presence of Dox, rtTA-Advanced binds to the TRE and activates expression of the gene of interest. In the absence of Dox, rtTA-Advanced is unable to bind to the TRE, and the system is inactive. Additional information on TRE-containing vectors and protocols describing how to construct a Retro-X Tet-On Advanced cell line can be found in the Retro-X Tet-On Advanced Inducible Gene Expression System User manual (PT3958-1).
    
    Selection of Stable Transfectants
     Selectable marker: plasmid confers resistance to G418.
    
    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: pUC
    
    

    載體序列

    LOCUS       pRetroX-Tet-On Advanced	8152 bp 	DNA	SYN
    DEFINITION  pRetroX-Tet-On Advanced
    ACCESSION   
    KEYWORDS    
    SOURCE      
      ORGANISM  other sequences; artificial sequences; vectors.
    FEATURES             Location/Qualifiers
         source          1..8152
                         /organism="pRetroX-Tet-On Advanced"
                         /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"
         misc_feature    1608..1895
                         /label="CAG_enhancer"
         promoter        1613..2105
                         /label="CMV_immearly_promoter"
         misc_feature    2062..2082
                         /label="CMV_fwd_primer"
         promoter        2063..2132
                         /label="CMV_promoter"
         promoter        2075..2194
                         /label="CMV2_promoter"
         misc_feature    2106..2125
                         /label="pCEP_fwd_primer"
         misc_feature    2108..2132
                         /label="LNCX_primer"
         CDS             2287..3033
                         /label="ORF frame 1"
                         /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWH
                         VKNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLG
                         TRPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERET
                         PTTDSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLD
                         MLPADALDDFDLDMLPADALDDFDLDMLPG*"
         misc_feature    3071..3622
                         /label="IRES"
                         /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWH
                         VKNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLG
                         TRPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERET
                         PTTDSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLD
                         MLPADALDDFDLDMLPADALDDFDLDMLPG*"
         CDS             3647..4441
                         /label="ORF frame 2"
                         /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                         PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                         LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                         EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                         QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
         gene            3662..4438
                         /label="NeoR/KanR"
                         /gene="NeoR/KanR"
                         /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                         PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                         LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                         EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                         QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
         misc_feature    complement(5386..5408)
                         /label="pGEX_3_primer"
                         /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                         PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                         LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                         EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                         QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
         misc_feature    complement(5544..5564)
                         /label="pBABE_3_primer"
                         /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                         PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                         LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                         EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                         QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
         misc_feature    complement(5550..5765)
                         /label="SV40_enhancer"
                         /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                         PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                         LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                         EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                         QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
         promoter        5562..5830
                         /label="SV40_promoter"
                         /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                         PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                         LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                         EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                         QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
         rep_origin      5729..5806
                         /label="SV40_origin"
                         /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                         PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                         LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                         EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                         QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
         misc_feature    5791..5810
                         /label="SV40pro_F_primer"
                         /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                         PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                         LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                         EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                         QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
         gene            complement(6930..7790)
                         /label="Ampicillin"
                         /gene="Ampicillin"
                         /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                         PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                         LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                         EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                         QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
         CDS             complement(6930..7790)
                         /label="ORF frame 3"
                         /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                         IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                         YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                         DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                         LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                         EIGASLIKHW*"
         promoter        complement(7832..7860)
                         /label="AmpR_promoter"
                         /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                         IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                         YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                         DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                         LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                         EIGASLIKHW*"
         promoter        8082..506
                         /label="CMV_immearly_promoter"
                         /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                         IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                         YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                         DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                         LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                         EIGASLIKHW*"
         misc_feature    8146..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 TGGGGGATCG ATCCTCTAGA GTCCGTTACA TAACTTACGG
     1621 TAAATGGCCC GCCTGGCTGA CCGCCCAACG ACCCCCGCCC ATTGACGTCA ATAATGACGT
     1681 ATGTTCCCAT AGTAACGCCA ATAGGGACTT TCCATTGACG TCAATGGGTG GAGTATTTAC
     1741 GGTAAACTGC CCACTTGGCA GTACATCAAG TGTATCATAT GCCAAGTACG CCCCCTATTG
     1801 ACGTCAATGA CGGTAAATGG CCCGCCTGGC ATTATGCCCA GTACATGACC TTATGGGACT
     1861 TTCCTACTTG GCAGTACATC TACGTATTAG TCATCGCTAT TACCATGGTG ATGCGGTTTT
     1921 GGCAGTACAT CAATGGGCGT GGATAGCGGT TTGACTCACG GGGATTTCCA AGTCTCCACC
     1981 CCATTGACGT CAATGGGAGT TTGTTTTGGC ACCAAAATCA ACGGGACTTT CCAAAATGTC
     2041 GTAACAACTC CGCCCCATTG ACGCAAATGG GCGGTAGGCG TGTACGGTGG GAGGTCTATA
     2101 TAAGCAGAGC TCGTTTAGTG AACCGTCAGA TCGCCTGGAG ACGCCATCCA CGCTGTTTTG
     2161 ACCTCCATAG AAGACACCGG GACCGATCCA GCCTCCGCGG CCGGGAACGG TGCATTGGAA
     2221 CGGACCTGCA GGAATTGATC CGCGGCCGCA CCGGTAGGCC TCGTACGCTT AATTAACGGA
     2281 TCCACCATGT CTAGACTGGA CAAGAGCAAA GTCATAAACG GCGCTCTGGA ATTACTCAAT
     2341 GGAGTCGGTA TCGAAGGCCT GACGACAAGG AAACTCGCTC AAAAGCTGGG AGTTGAGCAG
     2401 CCTACCCTGT ACTGGCACGT GAAGAACAAG CGGGCCCTGC TCGATGCCCT GCCAATCGAG
     2461 ATGCTGGACA GGCATCATAC CCACTTCTGC CCCCTGGAAG GCGAGTCATG GCAAGACTTT
     2521 CTGCGGAACA ACGCCAAGTC ATTCCGCTGT GCTCTCCTCT CACATCGCGA CGGGGCTAAA
     2581 GTGCATCTCG GCACCCGCCC AACAGAGAAA CAGTACGAAA CCCTGGAAAA TCAGCTCGCG
     2641 TTCCTGTGTC AGCAAGGCTT CTCCCTGGAG AACGCACTGT ACGCTCTGTC CGCCGTGGGC
     2701 CACTTTACAC TGGGCTGCGT ATTGGAGGAA CAGGAGCATC AAGTAGCAAA AGAGGAAAGA
     2761 GAGACACCTA CCACCGATTC TATGCCCCCA CTTCTGAGAC AAGCAATTGA GCTGTTCGAC
     2821 CGGCAGGGAG CCGAACCTGC CTTCCTTTTC GGCCTGGAAC TAATCATATG TGGCCTGGAG
     2881 AAACAGCTAA AGTGCGAAAG CGGCGGGCCG GCCGACGCCC TTGACGATTT TGACTTAGAC
     2941 ATGCTCCCAG CCGATGCCCT TGACGACTTT GACCTTGATA TGCTGCCTGC TGACGCTCTT
     3001 GACGATTTTG ACCTTGACAT GCTCCCCGGG TAACTAGAAT TCCGCCCCCC CCCCCCCCCC
     3061 CCCTCTCCCT CCCCCCCCCC TAACGTTACT GGCCGAAGCC GCTTGGAATA AGGCCGGTGT
     3121 GCGTTTGTCT ATATGTTATT TTCCACCATA TTGCCGTCTT TTGGCAATGT GAGGGCCCGG
     3181 AAACCTGGCC CTGTCTTCTT GACGAGCATT CCTAGGGGTC TTTCCCCTCT CGCCAAAGGA
     3241 ATGCAAGGTC TGTTGAATGT CGTGAAGGAA GCAGTTCCTC TGGAAGCTTC TTGAAGACAA
     3301 ACAACGTCTG TAGCGACCCT TTGCAGGCAG CGGAACCCCC CACCTGGCGA CAGGTGCCTC
     3361 TGCGGCCAAA AGCCACGTGT ATAAGATACA CCTGCAAAGG CGGCACAACC CCAGTGCCAC
     3421 GTTGTGAGTT GGATAGTTGT GGAAAGAGTC AAATGGCTCT CCTCAAGCGT ATTCAACAAG
     3481 GGGCTGAAGG ATGCCCAGAA GGTACCCCAT TGTATGGGAT CTGATCTGGG GCCTCGGTGC
     3541 ACATGCTTTA CATGTGTTTA GTCGAGGTTA AAAAAACGTC TAGGCCCCCC GAACCACGGG
     3601 GACGTGGTTT TCCTTTGAAA AACACGATGA TAAGCTTGCC ACAACCATGG CTGAACAAGA
     3661 TGGATTGCAC GCAGGTTCTC CGGCCGCTTG GGTGGAGAGG CTATTCGGCT ATGACTGGGC
     3721 ACAACAGACA ATCGGCTGCT CTGATGCCGC CGTGTTCCGG CTGTCAGCGC AGGGGCGCCC
     3781 GGTTCTTTTT GTCAAGACCG ACCTGTCCGG TGCCCTGAAT GAACTGCAGG ACGAGGCAGC
     3841 GCGGCTATCG TGGCTGGCCA CGACGGGCGT TCCTTGCGCA GCTGTGCTCG ACGTTGTCAC
     3901 TGAAGCGGGA AGGGACTGGC TGCTATTGGG CGAAGTGCCG GGGCAGGATC TCCTGTCATC
     3961 TCACCTTGCT CCTGCCGAGA AAGTATCCAT CATGGCTGAT GCAATGCGGC GGCTGCATAC
     4021 GCTTGATCCG GCTACCTGCC CATTCGACCA CCAAGCGAAA CATCGCATCG AGCGAGCACG
     4081 TACTCGGATG GAAGCCGGTC TTGTCGATCA GGATGATCTG GACGAAGAGC ATCAGGGGCT
     4141 CGCGCCAGCC GAACTGTTCG CCAGGCTCAA GGCGCGCATG CCCGACGGCG AGGATCTCGT
     4201 CGTGACCCAT GGCGATGCCT GCTTGCCGAA TATCATGGTG GAAAATGGCC GCTTTTCTGG
     4261 ATTCATCGAC TGTGGCCGGC TGGGTGTGGC GGACCGCTAT CAGGACATAG CGTTGGCTAC
     4321 CCGTGATATT GCTGAAGAGC TTGGCGGCGA ATGGGCTGAC CGCTTCCTCG TGCTTTACGG
     4381 TATCGCCGCT CCCGATTCGC AGCGCATCGC CTTCTATCGC CTTCTTGACG AGTTCTTCTG
     4441 AGCGGGATCG GCTAGAGTCG ACCCGGACGG GACGCTCCTG CGCCTGATAC AGAACGAATT
     4501 GCTTGCAGGC ATCTCATGAG TGTGTCTTCC CGTTTTCCGC CTGAGGTCAC TGCGTGGATG
     4561 GAGCGCTGGC GCCTGCTGCG CGACGGCGAG CTGCTCACCA CCCACTCGAG GGCTGCAGCG
     4621 CTGCAGAGGC CGAGTGCAGA ACTGCTCCAA AGGGACCTCA AGGCTTTCCG AGGGACACTA
     4681 GGCTGACTCC ATCGAGCCAG TGTAGAGATA AGCTTATCGA TTAGTCCAAT TTGTTAAAGA
     4741 CAGGATATCA GTGGTCCAGG CTCTAGTTTT GACTCAACAA TATCACCAGC TGAAGCCTAT
     4801 AGAGTACGAG CCATAGATAA AATAAAAGAT TTTATTTAGT CTCCAGAAAA AGGGGGGAAT
     4861 GAAAGACCCC ACCTGTAGGT TTGGCAAGCT AGCTTAAGTA ACGCCATTTT GCAAGGCATG
     4921 GAAAAATACA TAACTGAGAA TAGAGAAGTT CAGATCAAGG TCAGGAACAG ATGGAACAGG
     4981 GTCGACCCTA GAGAACCATC AGATGTTTCC AGGGTGCCCC AAGGACCTGA AATGACCCTG
     5041 TGCCTTATTT GAACTAACCA ATCAGTTCGC TTCTCGCTTC TGTTCGCGCG CTTCTGCTCC
     5101 CCGAGCTCAA TAAAAGAGCC CACAACCCCT CACTCGGGGC GCCAGTCCTC CGATTGACTG
     5161 AGTCGCCCGG GTACCCGTGT ATCCAATAAA CCCTCTTGCA GTTGCATCCG ACTTGTGGTC
     5221 TCGCTGTTCC TTGGGAGGGT CTCCTCTGAG TGATTGACTA CCCGTCAGCG GGGGTCTTTC
     5281 ATTTGGGGGC TCGTCCGGGA TCGGGAGACC CCTGCCCAGG GACCACCGAC CCACCACCGG
     5341 GAGGTAAGCT GGCTGCCTCG CGCGTTTCGG TGATGACGGT GAAAACCTCT GACACATGCA
     5401 GCTCCCGGAG ACGGTCACAG CTTGTCTGTA AGCGGATGCC GGGAGCAGAC AAGCCCGTCA
     5461 GGGCGCGTCA GCGGGTGTTG GCGGGTGTCG GGGCGCAGCC ATGACCCAGT CACGTAGCGA
     5521 TAGCGGAGTG TAGATCCGGC TGTGGAATGT GTGTCAGTTA GGGTGTGGAA AGTCCCCAGG
     5581 CTCCCCAGCA GGCAGAAGTA TGCAAAGCAT GCATCTCAAT TAGTCAGCAA CCAGGTGTGG
     5641 AAAGTCCCCA GGCTCCCCAG CAGGCAGAAG TATGCAAAGC ATGCATCTCA ATTAGTCAGC
     5701 AACCATAGTC CCGCCCCTAA CTCCGCCCAT CCCGCCCCTA ACTCCGCCCA GTTCCGCCCA
     5761 TTCTCCGCCC CATGGCTGAC TAATTTTTTT TATTTATGCA GAGGCCGAGG CCGCCTCGGC
     5821 CTCTGAGCTA TTCCAGAAGT AGTGAGGAGG CTTTTTTGGA GGCCTAGGCT TTTGCAAAAA
     5881 GCTTACTGGC TTAACTATGC GGCATCAGAG CAGATTGTAC TGAGAGTGCA CCATATGCGG
     5941 TGTGAAATAC CGCACAGATG CGTAAGGAGA AAATACCGCA TCAGGCGCTC TTCCGCTTCC
     6001 TCGCTCACTG ACTCGCTGCG CTCGGTCGTT CGGCTGCGGC GAGCGGTATC AGCTCACTCA
     6061 AAGGCGGTAA TACGGTTATC CACAGAATCA GGGGATAACG CAGGAAAGAA CATGTGAGCA
     6121 AAAGGCCAGC AAAAGGCCAG GAACCGTAAA AAGGCCGCGT TGCTGGCGTT TTTCCATAGG
     6181 CTCCGCCCCC CTGACGAGCA TCACAAAAAT CGACGCTCAA GTCAGAGGTG GCGAAACCCG
     6241 ACAGGACTAT AAAGATACCA GGCGTTTCCC CCTGGAAGCT CCCTCGTGCG CTCTCCTGTT
     6301 CCGACCCTGC CGCTTACCGG ATACCTGTCC GCCTTTCTCC CTTCGGGAAG CGTGGCGCTT
     6361 TCTCATAGCT CACGCTGTAG GTATCTCAGT TCGGTGTAGG TCGTTCGCTC CAAGCTGGGC
     6421 TGTGTGCACG AACCCCCCGT TCAGCCCGAC CGCTGCGCCT TATCCGGTAA CTATCGTCTT
     6481 GAGTCCAACC CGGTAAGACA CGACTTATCG CCACTGGCAG CAGCCACTGG TAACAGGATT
     6541 AGCAGAGCGA GGTATGTAGG CGGTGCTACA GAGTTCTTGA AGTGGTGGCC TAACTACGGC
     6601 TACACTAGAA GGACAGTATT TGGTATCTGC GCTCTGCTGA AGCCAGTTAC CTTCGGAAAA
     6661 AGAGTTGGTA GCTCTTGATC CGGCAAACAA ACCACCGCTG GTAGCGGTGG TTTTTTTGTT
     6721 TGCAAGCAGC AGATTACGCG CAGAAAAAAA GGATCTCAAG AAGATCCTTT GATCTTTTCT
     6781 ACGGGGTCTG ACGCTCAGTG GAACGAAAAC TCACGTTAAG GGATTTTGGT CATGAGATTA
     6841 TCAAAAAGGA TCTTCACCTA GATCCTTTTA AATTAAAAAT GAAGTTTTAA ATCAATCTAA
     6901 AGTATATATG AGTAAACTTG GTCTGACAGT TACCAATGCT TAATCAGTGA GGCACCTATC
     6961 TCAGCGATCT GTCTATTTCG TTCATCCATA GTTGCCTGAC TCCCCGTCGT GTAGATAACT
     7021 ACGATACGGG AGGGCTTACC ATCTGGCCCC AGTGCTGCAA TGATACCGCG AGACCCACGC
     7081 TCACCGGCTC CAGATTTATC AGCAATAAAC CAGCCAGCCG GAAGGGCCGA GCGCAGAAGT
     7141 GGTCCTGCAA CTTTATCCGC CTCCATCCAG TCTATTAATT GTTGCCGGGA AGCTAGAGTA
     7201 AGTAGTTCGC CAGTTAATAG TTTGCGCAAC GTTGTTGCCA TTGCTGCAGG CATCGTGGTG
     7261 TCACGCTCGT CGTTTGGTAT GGCTTCATTC AGCTCCGGTT CCCAACGATC AAGGCGAGTT
     7321 ACATGATCCC CCATGTTGTG CAAAAAAGCG GTTAGCTCCT TCGGTCCTCC GATCGTTGTC
     7381 AGAAGTAAGT TGGCCGCAGT GTTATCACTC ATGGTTATGG CAGCACTGCA TAATTCTCTT
     7441 ACTGTCATGC CATCCGTAAG ATGCTTTTCT GTGACTGGTG AGTACTCAAC CAAGTCATTC
     7501 TGAGAATAGT GTATGCGGCG ACCGAGTTGC TCTTGCCCGG CGTCAACACG GGATAATACC
     7561 GCGCCACATA GCAGAACTTT AAAAGTGCTC ATCATTGGAA AACGTTCTTC GGGGCGAAAA
     7621 CTCTCAAGGA TCTTACCGCT GTTGAGATCC AGTTCGATGT AACCCACTCG TGCACCCAAC
     7681 TGATCTTCAG CATCTTTTAC TTTCACCAGC GTTTCTGGGT GAGCAAAAAC AGGAAGGCAA
     7741 AATGCCGCAA AAAAGGGAAT AAGGGCGACA CGGAAATGTT GAATACTCAT ACTCTTCCTT
     7801 TTTCAATATT ATTGAAGCAT TTATCAGGGT TATTGTCTCA TGAGCGGATA CATATTTGAA
     7861 TGTATTTAGA AAAATAAACA AATAGGGGTT CCGCGCACAT TTCCCCGAAA AGTGCCACCT
     7921 GACGTCTAAG AAACCATTAT TATCATGACA TTAACCTATA AAAATAGGCG TATCACGAGG
     7981 CCCTTTCGTC TTCAAGAATT AGCTTGGCCA TTGCATACGT TGTATCCATA TCATAATATG
     8041 TACATTTATA TTGGCTCATG TCCAACATTA CCGCCATGTT GACATTGATT ATTGACTAGT
     8101 TATTAATAGT AATCAATTAC GGGGTCATTA GTTCATAGCC CATATATGGA GT
    
    //
    

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