LOCUS Exported 12394 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Tet-regulated (Tet-on) lentiviral vector for transgene (hEF-1alpha promoter) - OR - shRNA (H1 promoter when subcloned from pLVTHM (Addgene#12247)) - 2nd generation. ACCESSION . VERSION . KEYWORDS pLVET-tTR-KRAB SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12394) AUTHORS Szulc J, Wiznerowicz M, Sauvain MO, Trono D, Aebischer P TITLE A versatile tool for conditional gene expression and knockdown. JOURNAL Nat Methods. 2006 Feb . 3(2):109-16. PUBMED 16432520 REFERENCE 2 (bases 1 to 12394) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12394 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 2..635 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 682..807 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1300..1533 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1718..1762 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" promoter 2120..3298 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2351..3289 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3246..3266 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" misc_feature 3346..3463 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" regulatory 3540..3549 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3546..4262 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3546..4262 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3591..3612) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3852..3871) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4199..4220 /label=EGFP-C /note="EGFP, forward primer" misc_feature 4341..4892 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4485..4502) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4712..4731 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4895..5512 /codon_start=1 /gene="tetR from transposon Tn10" /product="tetracycline repressor TetR" /label=TetR /note="TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline." /translation="MARLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESG" primer_bind complement(4968..4987) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" CDS 5516..5536 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 5639..5833 /codon_start=1 /product="Kruppel-associated box (KRAB) transcriptional repression domain from the human zinc finger protein ZNF10 (Margolin et al., 1994)" /label=KRAB /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY QLTKPDVILRLEKGEEPWLV" misc_feature 5975..6563 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(6028..6048) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6446..6457) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" protein_bind 6701..6971 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" LTR 7010..7190 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" promoter complement(7215..7233) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(7216..7233) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(7523..7542) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind complement(7756..7775) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7875..7897 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(7935..7953) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 8021..8125 /gene="bla" /label=AmpR promoter CDS 8126..8986 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(8344..8363) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 9157..9745 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9646..9665 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 9899..9916 /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(9986..10006) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 9991..10320 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 10171..10306 /label=SV40 ori /note="SV40 origin of replication" primer_bind 10233..10252 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" intron 11454..11519 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 11649..11669 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 12094..12228 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(12131..12150) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 12185..12204 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" ORIGIN 1 ttggaagggc taattcactc ccaaagaaga caagatatcc ttgatctgtg gatctaccac 61 acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca 121 ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc 181 aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg 241 gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga 301 gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc 361 gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga 421 tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg 481 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc 541 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct 601 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa 661 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg 721 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag 781 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg 841 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg 901 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc 961 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga 1021 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac 1081 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag 1141 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga 1201 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa 1261 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt 1321 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag 1381 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca 1441 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc 1501 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact 1561 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat 1621 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat 1681 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga 1741 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat 1801 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact 1861 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc 1921 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag 1981 agacagatcc awtcgatyas krrackrryc tcgacsnnat cgatswcgac gataagcttt 2041 gcaaagatgg ataaagtttt aaacagagag gaatctttgc agctaatgga ccttctaggt 2101 cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac 2161 agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg 2221 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg 2281 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc 2341 cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg 2401 cccttgcgtg ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct 2461 tcgggttgga agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg 2521 tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct 2581 tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc 2641 tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg 2701 tatttcggtt tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc 2761 ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg 2821 ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag 2881 gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc 2941 agggagctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca 3001 aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc 3061 gccgtccagg cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg 3121 ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc 3181 agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt 3241 cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgagg 3301 aatttcgaca tttaaattta attaatctcg acggtatcgg ttaactttta aaagaaaagg 3361 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca 3421 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttatcgatc acgagactag 3481 cctcgaggtt taaactacgg gatccaggcc taagcttacg cgtcctagcg ctaccggtcg 3541 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc 3601 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca 3661 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc 3721 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca 3781 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca 3841 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca 3901 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg 3961 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga 4021 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc 4081 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca 4141 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca 4201 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca 4261 agtccggact cagatctcga ctagctagta gctagctagc tagtcgagct caagcttcga 4321 attccgcccc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc 4381 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa 4441 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat 4501 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac 4561 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg 4621 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt 4681 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg 4741 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac 4801 atgctttaca tgtgtttagt cgaggttaaa aaacgtctag gccccccgaa ccacggggac 4861 gtggttttcc tttgaaaaac acgatgataa taccatggct agattagata aaagtaaagt 4921 gattaacagc gcattagagc tgcttaatga ggtcggaatc gaaggtttaa caacccgtaa 4981 actcgcccag aagctaggtg tagagcagcc tacattgtat tggcatgtaa aaaataagcg 5041 ggctttgctc gacgccttag ccattgagat gttagatagg caccatactc acttttgccc 5101 tttagaaggg gaaagctggc aagatttttt acgtaataac gctaaaagtt ttagatgtgc 5161 tttactaagt catcgcgatg gagcaaaagt acatttaggt acacggccta cagaaaaaca 5221 gtatgaaact ctcgaaaatc aattagcctt tttatgccaa caaggttttt cactagagaa 5281 tgcattatat gcactcagcg ctgtggggca ttttacttta ggttgcgtat tggaagatca 5341 agagcatcaa gtcgctaaag aagaaaggga aacacctact actgatagta tgccgccatt 5401 attacgacaa gctatcgaat tatttgatca ccaaggtgca gagccagcct tcttattcgg 5461 ccttgaattg atcatatgcg gattagaaaa acaacttaaa tgtgaaagtg ggtcgccaaa 5521 aaagaagaga aaggtcgacg gcggtggtgc tttgtctcct cagcactctg ctgtcactca 5581 aggaagtatc atcaagaaca aggagggcat ggatgctaag tcactaactg cctggtcccg 5641 gacactggtg accttcaagg atgtatttgt ggacttcacc agggaggagt ggaagctgct 5701 ggacactgct cagcagatcg tgtacagaaa tgtgatgctg gagaactata agaacctggt 5761 ttccttgggt tatcagctta ctaagccaga tgtgatcctc cggttggaga agggagaaga 5821 gccctggctg gtggagagag aaattcacca agagacccat cctgattcag agactgcatt 5881 tgaaatcaaa tcatcagttt aaggatctcg actagctagt agctagctag ctagtcgagc 5941 tcaagcttcg aattcccggg actagtcata tgataatcaa cctctggatt acaaaatttg 6001 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc 6061 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta 6121 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt 6181 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca 6241 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc 6301 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt 6361 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg 6421 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg 6481 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat 6541 ctccctttgg gccgcctccc cgcatcggta cgtatggcca ggtaccttta agaccaatga 6601 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc 6661 taattcactc ccaacgaaga caagatctcg agtttaccac tccctatcag tgatagagaa 6721 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta 6781 ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat 6841 agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg 6901 agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc 6961 agtgatagag aaaagtgaaa gtcggatcga tctgcttttt gcttgtactg ggtctctctg 7021 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 7081 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 7141 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gcatctagaa 7201 ttaattccgt gtattctata gtgtcaccta aatcgtatgt gtatgataca taaggttatg 7261 tattaattgt agccgcgttc taacgacaat atgtacaagc ctaattgtgt agcatctggc 7321 ttactgaagc agaccctatc atctctctcg taaactgccg tcagagtcgg tttggttgga 7381 cgaaccttct gagtttctgg taacgccgtc ccgcacccgg aaatggtcag cgaaccaatc 7441 agcagggtca tcgctagcca gatcctctac gccggacgca tcgtggccgg catcaccggc 7501 gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct 7561 cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc 7621 gggggactgt tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac 7681 ggcctcaacc tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga 7741 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc 7801 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca 7861 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg 7921 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat 7981 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt 8041 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct 8101 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc 8161 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa 8221 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg 8281 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt 8341 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg 8401 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac 8461 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc 8521 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa 8581 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc 8641 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt 8701 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga 8761 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa 8821 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa 8881 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa 8941 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt 9001 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt 9061 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg 9121 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt 9181 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca 9241 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac 9301 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac 9361 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct 9421 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg 9481 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca 9541 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt 9601 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta 9661 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc 9721 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc 9781 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa 9841 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag 9901 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg 9961 ttggccgatt cattaatgca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca 10021 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt 10081 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 10141 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 10201 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 10261 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa 10321 aagcttggac acaagacagg cttgcgagat atgtttgaga ataccacttt atcccgcgtc 10381 agggagaggc agtgcgtaaa aagacgcgga ctcatgtgaa atactggttt ttagtgcgcc 10441 agatctctat aatctcgcgc aacctatttt cccctcgaac actttttaag ccgtagataa 10501 acaggctggg acacttcaca tgagcgaaaa atacatcgtc acctgggaca tgttgcagat 10561 ccatgcacgt aaactcgcaa gccgactgat gccttctgaa caatggaaag gcattattgc 10621 cgtaagccgt ggcggtctgt accgggtgcg ttactggcgc gtgaactggg tattcgtcat 10681 gtcgataccg tttgtatttc cagctacgat cacgacaacc agcgcgagct taaagtgctg 10741 aaacgcgcag aaggcgatgg cgaaggcttc atcgttattg atgacctggt ggataccggt 10801 ggtactgcgg ttgcgattcg tgaaatgtat ccaaaagcgc actttgtcac catcttcgca 10861 aaaccggctg gtcgtccgct ggttgatgac tatgttgttg atatcccgca agatacctgg 10921 attgaacagc cgtgggatat gggcgtcgta ttcgtcccgc caatctccgg tcgctaatct 10981 tttcaacgcc tggcactgcc gggcgttgtt ctttttaact tcaggcgggt tacaatagtt 11041 tccagtaagt attctggagg ctgcatccat gacacaggca aacctgagcg aaaccctgtt 11101 caaaccccgc tttaaacatc ctgaaacctc gacgctagtc cgccgcttta atcacggcgc 11161 acaaccgcct gtgcagtcgg cccttgatgg taaaaccatc cctcactggt atcgcatgat 11221 taaccgtctg atgtggatct ggcgcggcat tgacccacgc gaaatcctcg acgtccaggc 11281 acgtattgtg atgagcgatg ccgaacgtac cgacgatgat ttatacgata cggtgattgg 11341 ctaccgtggc ggcaactgga tttatgagtg ggccccggat ctttgtgaag gaaccttact 11401 tctgtggtgt gacataattg gacaaactac ctacagagat ttaaagctct aaggtaaata 11461 taaaattttt aagtgtataa tgtgttaaac tactgattct aattgtttgt gtattttaga 11521 ttccaaccta tggaactgat gaatgggagc agtggtggaa tgcctttaat gaggaaaacc 11581 tgttttgctc agaagaaatg ccatctagtg atgatgaggc tactgctgac tctcaacatt 11641 ctactcctcc aaaaaagaag agaaaggtag aagaccccaa ggactttcct tcagaattgc 11701 taagtttttt gagtcatgct gtgtttagta atagaactct tgcttgcttt gctatttaca 11761 ccacaaagga aaaagctgca ctgctataca agaaaattat ggaaaaatat tctgtaacct 11821 ttataagtag gcataacagt tataatcata acatactgtt ttttcttact ccacacaggc 11881 atagagtgtc tgctattaat aactatgctc aaaaattgtg tacctttagc tttttaattt 11941 gtaaaggggt taataaggaa tatttgatgt atagtgcctt gactagagat cataatcagc 12001 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac 12061 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt 12121 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 12181 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact 12241 caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac 12301 ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat 12361 tgtatttgtt aaatatgtac tacaaactta gtag //