LOCUS Exported 11634 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Tet-regulated (Tet-on) lentiviral vector for transgene (hPGK promoter) - AND/OR - shRNA (H1 promoter when subcloned from pLVTHM (Addgene#12247)) - 2nd generation. ACCESSION . VERSION . KEYWORDS pLVPT-tTR-KRAB SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11634) 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 11634) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11634 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 294..411 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 450..949 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(485..508) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 882..901 /label=mPGK-F /note="Mouse PGK promoter, forward primer" regulatory 1000..1009 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1006..1722 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 1006..1722 /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(1051..1072) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1312..1331) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1659..1680 /label=EGFP-C /note="EGFP, forward primer" misc_feature 1818..2369 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(1962..1979) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 2189..2208 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 2372..2989 /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(2445..2464) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" CDS 2993..3013 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 3116..3310 /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" primer_bind complement(3449..3465) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3450..3466) /label=pBluescriptKS /note="For pBluescript vector" misc_feature 3480..4068 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3533..3553) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3951..3962) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" protein_bind 4206..4476 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" LTR 4515..4695 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" promoter complement(4720..4738) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(4721..4738) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(5028..5047) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind complement(5261..5280) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5380..5402 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5440..5458) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5526..5630 /gene="bla" /label=AmpR promoter CDS 5631..6491 /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(5849..5868) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6662..7250 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7151..7170 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7404..7421 /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(7491..7511) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 7496..7825 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 7676..7811 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7738..7757 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" intron 8959..9024 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 9154..9174 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 9599..9733 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(9636..9655) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 9690..9709 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" LTR 9901..10534 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 10581..10706 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 11199..11432 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 11617..27 /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" ORIGIN 1 ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg 61 tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct 121 gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac 181 ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga 241 gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt aacttttaaa 301 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca 361 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tatcgatcac 421 gagactagcc tcgacgatgg tcgagtaccg ggtaggggag gcgcttttcc caaggcagtc 481 tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc 541 tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt 601 cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc 661 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac 721 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt 781 cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct 841 caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa 901 gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctctagcgg 961 gatccaggcc taagcttacg cgtcctagcg ctaccggtcg ccaccatggt gagcaagggc 1021 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc 1081 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg 1141 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg 1201 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc 1261 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc 1321 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag 1381 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac 1441 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac 1501 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag 1561 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag 1621 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg 1681 accgccgccg ggatcactct cggcatggac gagctgtaca agtccggact cagatctcga 1741 ctagctagta gctagctagc tagtcgagct caagcttcga attcccgggc tgcaggaatt 1801 ccgccccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt 1861 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc 1921 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca 1981 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac 2041 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg 2101 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt 2161 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct 2221 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg 2281 ctttacatgt gtttagtcga ggttaaaaaa cgtctaggcc ccccgaacca cggggacgtg 2341 gttttccttt gaaaaacacg atgataatac catggctaga ttagataaaa gtaaagtgat 2401 taacagcgca ttagagctgc ttaatgaggt cggaatcgaa ggtttaacaa cccgtaaact 2461 cgcccagaag ctaggtgtag agcagcctac attgtattgg catgtaaaaa ataagcgggc 2521 tttgctcgac gccttagcca ttgagatgtt agataggcac catactcact tttgcccttt 2581 agaaggggaa agctggcaag attttttacg taataacgct aaaagtttta gatgtgcttt 2641 actaagtcat cgcgatggag caaaagtaca tttaggtaca cggcctacag aaaaacagta 2701 tgaaactctc gaaaatcaat tagccttttt atgccaacaa ggtttttcac tagagaatgc 2761 attatatgca ctcagcgctg tggggcattt tactttaggt tgcgtattgg aagatcaaga 2821 gcatcaagtc gctaaagaag aaagggaaac acctactact gatagtatgc cgccattatt 2881 acgacaagct atcgaattat ttgatcacca aggtgcagag ccagccttct tattcggcct 2941 tgaattgatc atatgcggat tagaaaaaca acttaaatgt gaaagtgggt cgccaaaaaa 3001 gaagagaaag gtcgacggcg gtggtgcttt gtctcctcag cactctgctg tcactcaagg 3061 aagtatcatc aagaacaagg agggcatgga tgctaagtca ctaactgcct ggtcccggac 3121 actggtgacc ttcaaggatg tatttgtgga cttcaccagg gaggagtgga agctgctgga 3181 cactgctcag cagatcgtgt acagaaatgt gatgctggag aactataaga acctggtttc 3241 cttgggttat cagcttacta agccagatgt gatcctccgg ttggagaagg gagaagagcc 3301 ctggctggtg gagagagaaa ttcaccaaga gacccatcct gattcagaga ctgcatttga 3361 aatcaaatca tcagtttaag gatctcgact agctagtagc tagctagcta gtcgagctca 3421 agcttcgaat tcgatatcaa gcttatcgcg ataccgtcga cctcgactag tcatatgata 3481 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 3541 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 3601 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 3661 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 3721 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 3781 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 3841 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg 3901 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 3961 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc 4021 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cggtacgtat 4081 ggccaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta 4141 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctcgagttt 4201 accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga 4261 tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc 4321 gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat 4381 cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg 4441 aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcgg atcgatctgc 4501 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct 4561 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt 4621 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt 4681 ggaaaatctc tagcagcatc tagaattaat tccgtgtatt ctatagtgtc acctaaatcg 4741 tatgtgtatg atacataagg ttatgtatta attgtagccg cgttctaacg acaatatgta 4801 caagcctaat tgtgtagcat ctggcttact gaagcagacc ctatcatctc tctcgtaaac 4861 tgccgtcaga gtcggtttgg ttggacgaac cttctgagtt tctggtaacg ccgtcccgca 4921 cccggaaatg gtcagcgaac caatcagcag ggtcatcgct agccagatcc tctacgccgg 4981 acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga 5041 catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt 5101 gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc 5161 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca 5221 ggagtcgcat aagggagagc gtcgaatggt gcactctcag tacaatctgc tctgatgccg 5281 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc 5341 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga 5401 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt 5461 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa 5521 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca 5581 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc 5641 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc 5701 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt 5761 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt 5821 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg 5881 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact 5941 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg 6001 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga 6061 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg 6121 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa 6181 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac 6241 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc 6301 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca 6361 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga 6421 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta 6481 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc 6541 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc 6601 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 6661 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 6721 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 6781 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact 6841 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 6901 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 6961 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 7021 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 7081 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 7141 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 7201 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 7261 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg 7321 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc 7381 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa 7441 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgt ggaatgtgtg 7501 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 7561 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 7621 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc 7681 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 7741 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt 7801 ttttggaggc ctaggctttt gcaaaaagct tggacacaag acaggcttgc gagatatgtt 7861 tgagaatacc actttatccc gcgtcaggga gaggcagtgc gtaaaaagac gcggactcat 7921 gtgaaatact ggtttttagt gcgccagatc tctataatct cgcgcaacct attttcccct 7981 cgaacacttt ttaagccgta gataaacagg ctgggacact tcacatgagc gaaaaataca 8041 tcgtcacctg ggacatgttg cagatccatg cacgtaaact cgcaagccga ctgatgcctt 8101 ctgaacaatg gaaaggcatt attgccgtaa gccgtggcgg tctgtaccgg gtgcgttact 8161 ggcgcgtgaa ctgggtattc gtcatgtcga taccgtttgt atttccagct acgatcacga 8221 caaccagcgc gagcttaaag tgctgaaacg cgcagaaggc gatggcgaag gcttcatcgt 8281 tattgatgac ctggtggata ccggtggtac tgcggttgcg attcgtgaaa tgtatccaaa 8341 agcgcacttt gtcaccatct tcgcaaaacc ggctggtcgt ccgctggttg atgactatgt 8401 tgttgatatc ccgcaagata cctggattga acagccgtgg gatatgggcg tcgtattcgt 8461 cccgccaatc tccggtcgct aatcttttca acgcctggca ctgccgggcg ttgttctttt 8521 taacttcagg cgggttacaa tagtttccag taagtattct ggaggctgca tccatgacac 8581 aggcaaacct gagcgaaacc ctgttcaaac cccgctttaa acatcctgaa acctcgacgc 8641 tagtccgccg ctttaatcac ggcgcacaac cgcctgtgca gtcggccctt gatggtaaaa 8701 ccatccctca ctggtatcgc atgattaacc gtctgatgtg gatctggcgc ggcattgacc 8761 cacgcgaaat cctcgacgtc caggcacgta ttgtgatgag cgatgccgaa cgtaccgacg 8821 atgatttata cgatacggtg attggctacc gtggcggcaa ctggatttat gagtgggccc 8881 cggatctttg tgaaggaacc ttacttctgt ggtgtgacat aattggacaa actacctaca 8941 gagatttaaa gctctaaggt aaatataaaa tttttaagtg tataatgtgt taaactactg 9001 attctaattg tttgtgtatt ttagattcca acctatggaa ctgatgaatg ggagcagtgg 9061 tggaatgcct ttaatgagga aaacctgttt tgctcagaag aaatgccatc tagtgatgat 9121 gaggctactg ctgactctca acattctact cctccaaaaa agaagagaaa ggtagaagac 9181 cccaaggact ttccttcaga attgctaagt tttttgagtc atgctgtgtt tagtaataga 9241 actcttgctt gctttgctat ttacaccaca aaggaaaaag ctgcactgct atacaagaaa 9301 attatggaaa aatattctgt aacctttata agtaggcata acagttataa tcataacata 9361 ctgttttttc ttactccaca caggcataga gtgtctgcta ttaataacta tgctcaaaaa 9421 ttgtgtacct ttagcttttt aatttgtaaa ggggttaata aggaatattt gatgtatagt 9481 gccttgacta gagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa 9541 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa 9601 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 9661 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 9721 tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc 9781 ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc 9841 tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt 9901 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 9961 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 10021 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 10081 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 10141 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 10201 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 10261 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 10321 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 10381 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 10441 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 10501 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 10561 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 10621 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 10681 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 10741 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 10801 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 10861 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 10921 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 10981 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 11041 aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa 11101 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag 11161 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc 11221 ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga 11281 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa 11341 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct 11401 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc 11461 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt 11521 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata 11581 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga atta //