LOCUS Exported 15398 bp ds-DNA circular SYN 15-NOV-2022 DEFINITION Bipartite dCas9 repressor (dCas9-KRAB-MeCP2) in a UCOE-SFFV-dCas9 lentiviral vector.. ACCESSION . VERSION . KEYWORDS lenti_dCas9-KRAB-MeCP2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 15398) TITLE Validation of VIPER-inferred master regulators by pooled CRISPR screening JOURNAL Unpublished REFERENCE 2 (bases 1 to 15398) AUTHORS . TITLE Direct Submission JOURNAL Exported Nov 15, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..15398 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 103..510 /label=SFFV promoter /note="spleen focus-forming virus long terminal repeat (LTR) promoter" regulatory 579..588 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 585..4688 /codon_start=1 /product="catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=dCas9 /note="RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain" /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 4692..4718 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 4737..4757 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 4764..4784 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 4824..5009 /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 QLTKPDVILRLEKGEEP" CDS 5043..5063 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 5073..5930 /codon_start=1 /product="transcriptional repression domain from rat methyl-CpG-binding protein 2 (Nan et al., 1997)" /label=MeCP2 /translation="VQVKRVLEKSPGKLLVKMPFQASPGGKGEGGGATTSAQVMVIKRP GRKRKAEADPQAIPKKRGRKPGSVVAAAAAEAKKKAVKESSIRSVQETVLPIKKRKTRE TVSIEVKEVVKPLLVSTLGEKSGKGLKTCKSPGRKSKESSPKGRSSSASSPPKKEHHHH HHHAESPKAPMPLLPPPPPPEPQSSEDPISPPEPQDLSSSICKEEKMPRAGSLESDGCP KEPAKTQPMVAAAATTTTTTTTTVAEKYKHRGEGERKDIVSSSMPRPNREEPVDSRTPV TERVS" CDS 5940..5996 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 5997..6395 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" misc_feature 6456..7044 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(6509..6529) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6927..6938) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(7047..7063) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7048..7064) /label=pBluescriptKS /note="For pBluescript vector" LTR 7154..7387 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" promoter complement(7412..7430) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(7413..7430) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(7720..7739) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind complement(7953..7972) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 8072..8094 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(8132..8150) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 8218..8322 /gene="bla" /label=AmpR promoter CDS 8323..9183 /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(8541..8560) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 9354..9942 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9843..9862 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 10096..10113 /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(10183..10203) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 10188..10517 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 10368..10503 /label=SV40 ori /note="SV40 origin of replication" primer_bind 10430..10449 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" intron 11651..11716 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 11846..11866 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 12291..12425 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(12328..12347) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 12382..12401 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" LTR 12593..13226 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 13273..13398 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 13895..14128 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 14313..14357 /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" misc_feature 14642..14759 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" ORIGIN 1 gcgcggagtc ctgcagcccc gataaaataa aagattttat ttagtctcca gaaaaagggg 61 ggaatgaaag accccacctg taggtttggc aagctagctg cagtaacgcc attttgcaag 121 gcatggaaaa ataccaaacc aagaatagag aagttcagat caagggcggg tacatgaaaa 181 tagctaacgt tgggccaaac aggatatctg cggtgagcag tttcggcccc ggcccggggc 241 caagaacaga tggtcaccgc agtttcggcc ccggcccgag gccaagaaca gatggtcccc 301 agatatggcc caaccctcag cagtttctta agacccatca gatgtttcca ggctccccca 361 aggacctgaa atgaccctgc gccttatttg aattaaccaa tcagcctgct tctcgcttct 421 gttcgcgcgc ttctgcttcc cgagctctat aaaagagctc acaacccctc actcggcgcg 481 ccagtcctcc gacagactga gtcgcccggg ggggatctgg agctctcgag aattctcacg 541 cgtctgcagg atatcaagct tgcggtaccg cgggcccggc caccatggac aagaagtaca 601 gcatcggcct ggccatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca 661 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga 721 acctgatcgg cgccctgctg ttcgacagcg gagaaacagc cgaggccacc cggctgaaga 781 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct 841 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc 901 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg 961 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca 1021 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg 1081 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca 1141 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg 1201 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga 1261 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg caacctgatt gccctgagcc 1321 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc 1381 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt 1441 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc 1501 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg 1561 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga 1621 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac atcgatggcg 1681 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca 1741 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg 1801 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc 1861 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct 1921 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga 1981 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg 2041 ccagcgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga 2101 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtacaac gagctgacca 2161 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa 2221 aagccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag 2281 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc 2341 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact 2401 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt 2461 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca 2521 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc 2581 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg 2641 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag 2701 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca 2761 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg 2821 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag 2881 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag 2941 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc 3001 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc 3061 aggaactgga catcaaccgg ctgtccgact acgatgtgga cgctatcgtg cctcagagct 3121 ttctgaagga cgactccatc gataacaaag tgctgactcg gagcgacaag aaccggggca 3181 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcgccagc 3241 tgctgaatgc caagctgatt acccagagga agttcgacaa tctgaccaag gccgagagag 3301 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc 3361 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga 3421 acgacaaact gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt 3481 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg 3541 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa 3601 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg 3661 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt 3721 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa 3781 acggcgaaac aggcgagatc gtgtgggata agggccggga ctttgccacc gtgcggaaag 3841 tgctgtctat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca 3901 gcaaagagtc tatcctgccc aagaggaaca gcgacaagct gatcgccaga aagaaggact 3961 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg 4021 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga 4081 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg 4141 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc 4201 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac 4261 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga 4321 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aaacactacc 4381 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta 4441 atctggacaa ggtgctgagc gcctacaaca agcacagaga caagcctatc agagagcagg 4501 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt 4561 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca 4621 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg 4681 gaggcgacgc ctatccctat gacgtgcccg attatgccag cctgggcagc ggctccccca 4741 agaaaaaacg caaggtggaa gatcctaaga aaaagcggaa agtggacggc attggtagtg 4801 ggagcaacgg cagcagcgga tcccggacac tggtgacctt caaggatgta tttgtggact 4861 tcaccaggga ggagtggaag ctgctggaca ctgctcagca gatcgtgtac agaaatgtga 4921 tgctggagaa ctataagaac ctggtttcct tgggttatca gcttactaag ccagatgtga 4981 tcctccggtt ggagaaggga gaagagccct cgggaggtgg ttcgggtggc tctggatcaa 5041 gcccaaagaa gaaacggaag gtggaagcct cagtgcaggt gaaaagggtg ctggaaaaat 5101 cccccggcaa actcctcgtg aagatgccct tccaggcttc ccctggcgga aaaggtgaag 5161 ggggtggcgc aaccacatct gcccaggtca tggtcatcaa gcgacctgga aggaaaagaa 5221 aggccgaggc tgaccctcag gccattccaa agaaacgggg acgcaagcca gggtccgtgg 5281 tcgcagctgc agcagctgag gctaagaaaa aggcagtgaa ggaaagctcc atccgcagtg 5341 tgcaggagac tgtcctgccc atcaagaaga ggaagactag ggagaccgtg tccatcgagg 5401 tcaaagaagt ggtcaagccc ctgctcgtgt ccaccctggg cgaaaaatct ggaaaggggc 5461 tcaaaacatg caagtcacct ggacggaaaa gcaaggagtc tagtccaaag gggcgctcaa 5521 gctccgcttc tagtccccct aaaaaggaac accatcacca tcaccatcac gccgagtctc 5581 ctaaggctcc tatgccactg ctcccaccac ctccaccacc tgagccacag tcaagcgaag 5641 accccatcag cccacccgag cctcaggatc tgtcctctag tatttgcaaa gaggaaaaga 5701 tgcccagagc aggcagcctg gagagtgatg gctgtccaaa agaacccgcc aagacccagc 5761 ctatggtggc agccgctgca actaccacca caaccacaac taccacagtg gccgaaaaat 5821 acaagcatcg cggcgagggc gaacgaaagg acattgtgtc aagctccatg cccagaccta 5881 accgggagga accagtcgat agtaggacac ccgtgactga gagagtctca gcggccgccg 5941 caacaaactt ctctctgctg aaacaagccg gagatgtcga agagaatcct ggaccgatgg 6001 ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca 6061 gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca 6121 tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc 6181 tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg 6241 agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc 6301 ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg 6361 aattgctgcc ctctggttat gtgtgggagg gctaataagg ccgcgactct agagtcgacc 6421 tgcaggcatg caagcttgat atcaagctta tcgataatca acctctggat tacaaaattt 6481 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg 6541 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt 6601 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg 6661 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc 6721 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg 6781 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt 6841 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc 6901 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg 6961 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga 7021 tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgagggaat taattcgagc 7081 tcggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa 7141 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct 7201 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg 7261 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt 7321 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc 7381 tctagcagca tctagaatta attccgtgta ttctatagtg tcacctaaat cgtatgtgta 7441 tgatacataa ggttatgtat taattgtagc cgcgttctaa cgacaatatg tacaagccta 7501 attgtgtagc atctggctta ctgaagcaga ccctatcatc tctctcgtaa actgccgtca 7561 gagtcggttt ggttggacga accttctgag tttctggtaa cgccgtcccg cacccggaaa 7621 tggtcagcga accaatcagc agggtcatcg ctagccagat cctctacgcc ggacgcatcg 7681 tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg 7741 atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg 7801 tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca ccattccttg 7861 cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc 7921 ataagggaga gcgtcgaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta 7981 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg 8041 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca 8101 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt 8161 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc 8221 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa 8281 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc 8341 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa 8401 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa 8461 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg 8521 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa 8581 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc 8641 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc 8701 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta 8761 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag 8821 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca 8881 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata 8941 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc 9001 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca 9061 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca 9121 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg 9181 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa 9241 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt 9301 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat 9361 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 9421 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga 9481 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac 9541 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt 9601 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag 9661 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc 9721 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag 9781 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca 9841 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt 9901 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc 9961 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc 10021 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc 10081 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa 10141 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct gtggaatgtg tgtcagttag 10201 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 10261 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 10321 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 10381 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 10441 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag 10501 gcctaggctt ttgcaaaaag cttggacaca agacaggctt gcgagatatg tttgagaata 10561 ccactttatc ccgcgtcagg gagaggcagt gcgtaaaaag acgcggactc atgtgaaata 10621 ctggttttta gtgcgccaga tctctataat ctcgcgcaac ctattttccc ctcgaacact 10681 ttttaagccg tagataaaca ggctgggaca cttcacatga gcgaaaaata catcgtcacc 10741 tgggacatgt tgcagatcca tgcacgtaaa ctcgcaagcc gactgatgcc ttctgaacaa 10801 tggaaaggca ttattgccgt aagccgtggc ggtctgtacc gggtgcgtta ctggcgcgtg 10861 aactgggtat tcgtcatgtc gataccgttt gtatttccag ctacgatcac gacaaccagc 10921 gcgagcttaa agtgctgaaa cgcgcagaag gcgatggcga aggcttcatc gttattgatg 10981 acctggtgga taccggtggt actgcggttg cgattcgtga aatgtatcca aaagcgcact 11041 ttgtcaccat cttcgcaaaa ccggctggtc gtccgctggt tgatgactat gttgttgata 11101 tcccgcaaga tacctggatt gaacagccgt gggatatggg cgtcgtattc gtcccgccaa 11161 tctccggtcg ctaatctttt caacgcctgg cactgccggg cgttgttctt tttaacttca 11221 ggcgggttac aatagtttcc agtaagtatt ctggaggctg catccatgac acaggcaaac 11281 ctgagcgaaa ccctgttcaa accccgcttt aaacatcctg aaacctcgac gctagtccgc 11341 cgctttaatc acggcgcaca accgcctgtg cagtcggccc ttgatggtaa aaccatccct 11401 cactggtatc gcatgattaa ccgtctgatg tggatctggc gcggcattga cccacgcgaa 11461 atcctcgacg tccaggcacg tattgtgatg agcgatgccg aacgtaccga cgatgattta 11521 tacgatacgg tgattggcta ccgtggcggc aactggattt atgagtgggc cccggatctt 11581 tgtgaaggaa ccttacttct gtggtgtgac ataattggac aaactaccta cagagattta 11641 aagctctaag gtaaatataa aatttttaag tgtataatgt gttaaactac tgattctaat 11701 tgtttgtgta ttttagattc caacctatgg aactgatgaa tgggagcagt ggtggaatgc 11761 ctttaatgag gaaaacctgt tttgctcaga agaaatgcca tctagtgatg atgaggctac 11821 tgctgactct caacattcta ctcctccaaa aaagaagaga aaggtagaag accccaagga 11881 ctttccttca gaattgctaa gttttttgag tcatgctgtg tttagtaata gaactcttgc 11941 ttgctttgct atttacacca caaaggaaaa agctgcactg ctatacaaga aaattatgga 12001 aaaatattct gtaaccttta taagtaggca taacagttat aatcataaca tactgttttt 12061 tcttactcca cacaggcata gagtgtctgc tattaataac tatgctcaaa aattgtgtac 12121 ctttagcttt ttaatttgta aaggggttaa taaggaatat ttgatgtata gtgccttgac 12181 tagagatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc 12241 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta 12301 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 12361 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 12421 ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta 12481 ttaccactgc caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat 12541 tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta gttggaaggg 12601 ctaattcact cccaaagaag acaagatatc cttgatctgt ggatctacca cacacaaggc 12661 tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc actgaccttt 12721 ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc caataaagga 12781 gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa 12841 gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat 12901 ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc cgctggggac 12961 tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata 13021 taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga 13081 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct 13141 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt 13201 ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg 13261 aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc 13321 gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga 13381 gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt 13441 cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg 13501 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa 13561 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat 13621 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa 13681 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc 13741 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat 13801 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa 13861 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg 13921 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat 13981 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc 14041 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg 14101 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt 14161 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga 14221 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact 14281 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag 14341 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat 14401 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta 14461 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc 14521 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca 14581 gatccattcg attagtgaac ggatctcgac ggtatcgcca aatggcagta ttcatccaca 14641 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa 14701 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc 14761 gggtttatta cagggacagc agagatccag tttggatcga taagcttgat atcgaattcg 14821 ggaggtggtc cctgcagtta cgccaatgat aacccccgcc agaaaaatct tagtagcctt 14881 ccctttttgt tttccgtgcc ccaactcggc ggattgactc ggccccttcc ggaaacaccc 14941 gaatcaactt ctagtcaaat tattgttcac gccgcaatga cccacccctg gcccgcgtct 15001 gtggaactga cccctggtgt acaggagagt tcgctgctga aagtggtccc aaaggggtac 15061 tagtttttaa gctcccaact ccccctcccc cagcgtctgg aggattccac accctcgcac 15121 cgcaggggcg aggaagtggg cggagtccgg ttttggcgcc agccgctgag gctgccaagc 15181 agaaaagcca ccgctgagga gactccggtc actgtcctcg ccccgcctcc cccttccctc 15241 cccttgggga ccaccgggcg ccacgccgcg aacggtaagt gccgcggtcg tcggcgcctc 15301 cgccctcccc ctagggcccc aattcccagc gggcgcggcg cgcggccccg cgcgcagctc 15361 ccggctccct cccccttcgg atgtggcttg agctgtag //