LOCUS Exported 14820 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral expression of dCas9-HA-BFP-KRAB-NLS for CRISPRi driven by a hEF1alpha promoter. ACCESSION . VERSION . KEYWORDS pHR-EF1a-dCas9-HA-BFP-KRAB-NLS SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14820) TITLE Corn Lab Cas9 plasmids REFERENCE 2 (bases 1 to 14820) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..14820 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 1..634 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 681..806 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1303..1536 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1721..1765 /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 2050..2167 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2241..2452 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" intron 2453..3391 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3348..3368 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 3449..3458 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3455..7558 /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 7562..7588 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 7595..7615 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 7655..8350 /codon_start=1 /product="monomeric blue fluorescent protein" /label=TagBFP /note="mammalian codon-optimized" /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD LPSKLGHKLN" CDS 8396..8581 /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 8600..8620 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" misc_feature 8684..9272 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(8737..8757) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(9155..9166) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(9275..9291) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(9276..9292) /label=pBluescriptKS /note="For pBluescript vector" LTR 9382..9615 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" promoter complement(9640..9658) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(9641..9658) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(9948..9967) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind complement(10181..10200) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 10300..10322 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(10360..10378) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 10446..10550 /gene="bla" /label=AmpR promoter CDS 10551..11411 /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(10769..10788) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 11582..12170 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 12071..12090 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 12324..12341 /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(12411..12431) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 12416..12745 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 12596..12731 /label=SV40 ori /note="SV40 origin of replication" primer_bind 12658..12677 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" intron 13879..13944 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 14074..14094 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 14519..14653 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(14556..14575) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 14610..14629 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccaaa tggcagtatt 2041 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt 2101 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca 2161 aaattttcgg gtttattaca gggacagcag agatccagtt tggatcgata agcttgatat 2221 cgaattctgt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg 2281 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag 2341 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc 2401 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc 2461 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat 2521 tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg 2581 ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg 2641 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc 2701 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg 2761 gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc 2821 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg 2881 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc 2941 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc 3001 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag 3061 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc 3121 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga 3181 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat 3241 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta 3301 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac 3361 agtggttcaa agtttttttc ttccatttca ggtgtcgtgg aattctcacg cgtctgcagg 3421 atatcaagct tgcggtaccg cgggcccggc caccatggac aagaagtaca gcatcggcct 3481 ggccatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag 3541 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg 3601 cgccctgctg ttcgacagcg gagaaacagc cgaggccacc cggctgaaga gaaccgccag 3661 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga 3721 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga 3781 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca 3841 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc 3901 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct 3961 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt 4021 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa 4081 ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct 4141 gcccggcgag aagaagaatg gcctgttcgg caacctgatt gccctgagcc tgggcctgac 4201 ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga 4261 cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct 4321 gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa 4381 caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca 4441 ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga 4501 gattttcttc gaccagagca agaacggcta cgccggctac atcgatggcg gagccagcca 4561 ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact 4621 gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag 4681 catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt 4741 ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc 4801 ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag 4861 cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg ccagcgccca 4921 gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc 4981 caagcacagc ctgctgtacg agtacttcac cgtgtacaac gagctgacca aagtgaaata 5041 cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aagccatcgt 5101 ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt 5161 caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc 5221 ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa 5281 tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag 5341 agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa 5401 gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg 5461 catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc 5521 caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca 5581 gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg 5641 cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa 5701 agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac 5761 cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa 5821 agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga 5881 gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga 5941 catcaaccgg ctgtccgact acgatgtgga cgctatcgtg cctcagagct ttctgaagga 6001 cgactccatc gataacaaag tgctgactcg gagcgacaag aaccggggca agagcgacaa 6061 cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcgccagc tgctgaatgc 6121 caagctgatt acccagagga agttcgacaa tctgaccaag gccgagagag gcggcctgag 6181 cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa 6241 gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga acgacaaact 6301 gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga 6361 tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct 6421 gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt 6481 gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat 6541 cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga 6601 gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac 6661 aggcgagatc gtgtgggata agggccggga ctttgccacc gtgcggaaag tgctgtctat 6721 gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc 6781 tatcctgccc aagaggaaca gcgacaagct gatcgccaga aagaaggact gggaccctaa 6841 gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt 6901 ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat 6961 ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga 7021 agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg 7081 ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc 7141 ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc 7201 cgaggataat gagcagaaac agctgtttgt ggaacagcac aaacactacc tggacgagat 7261 catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa 7321 ggtgctgagc gcctacaaca agcacagaga caagcctatc agagagcagg ccgagaatat 7381 catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac 7441 caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca 7501 ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacgc 7561 ctatccctat gacgtgcccg attatgccga agatcctaag aaaaagcgta aagtggacgg 7621 cattggtagt gggagcaacg gcagcagcgg atccagcgag ctgattaagg agaacatgca 7681 catgaagctg tacatggagg gcaccgtgga caaccatcac ttcaagtgca catccgaggg 7741 cgaaggcaag ccctacgagg gcacccagac catgagaatc aaggtggtcg agggcggccc 7801 tctccccttc gccttcgaca tcctggctac tagcttcctc tacggcagca agaccttcat 7861 caaccacacc cagggcatcc ccgacttctt caagcagtcc ttccctgagg gcttcacatg 7921 ggagagagtc accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct 7981 ccaggacggc tgcctcatct acaacgtcaa gatcagaggg gtgaacttca catccaacgg 8041 ccctgtgatg cagaagaaaa cactcggctg ggaggccttc accgagacgc tgtaccccgc 8101 tgacggcggc ctggaaggca gaaacgacat ggccctgaag ctcgtgggcg ggagccatct 8161 gatcgcaaac atcaagacca catatagatc caagaaaccc gctaagaacc tcaagatgcc 8221 tggcgtctac tatgtggact acagactgga aagaatcaag gaggccaaca acgagaccta 8281 cgtcgagcag cacgaggtgg cagtggccag atactgcgac ctccctagca aactggggca 8341 caagcttaat ggcggtggcg gagggatgga tgctaagtca ctaactgcct ggtcccggac 8401 actggtgacc ttcaaggatg tatttgtgga cttcaccagg gaggagtgga agctgctgga 8461 cactgctcag cagatcgtgt acagaaatgt gatgctggag aactataaga acctggtttc 8521 cttgggttat cagcttacta agccagatgt gatcctccgg ttggagaagg gagaagagcc 8581 cagcctgggc agcggctccc ccaagaaaaa acgcaaggtg tagtaaggcc gcgactctag 8641 agtcgacctg caggcatgca agcttgatat caagcttatc gataatcaac ctctggatta 8701 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg 8761 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc 8821 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca 8881 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac 8941 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact 9001 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc 9061 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg 9121 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc 9181 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac 9241 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gagggaatta 9301 attcgagctc ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact 9361 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc 9421 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct 9481 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt 9541 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt 9601 ggaaaatctc tagcagcatc tagaattaat tccgtgtatt ctatagtgtc acctaaatcg 9661 tatgtgtatg atacataagg ttatgtatta attgtagccg cgttctaacg acaatatgta 9721 caagcctaat tgtgtagcat ctggcttact gaagcagacc ctatcatctc tctcgtaaac 9781 tgccgtcaga gtcggtttgg ttggacgaac cttctgagtt tctggtaacg ccgtcccgca 9841 cccggaaatg gtcagcgaac caatcagcag ggtcatcgct agccagatcc tctacgccgg 9901 acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga 9961 catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt 10021 gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc 10081 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca 10141 ggagtcgcat aagggagagc gtcgaatggt gcactctcag tacaatctgc tctgatgccg 10201 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc 10261 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga 10321 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt 10381 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa 10441 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca 10501 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc 10561 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc 10621 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt 10681 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt 10741 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg 10801 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact 10861 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg 10921 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga 10981 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg 11041 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa 11101 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac 11161 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc 11221 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca 11281 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga 11341 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta 11401 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc 11461 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc 11521 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 11581 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 11641 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 11701 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact 11761 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 11821 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 11881 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 11941 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 12001 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 12061 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 12121 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 12181 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg 12241 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc 12301 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa 12361 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgt ggaatgtgtg 12421 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 12481 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 12541 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc 12601 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 12661 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt 12721 ttttggaggc ctaggctttt gcaaaaagct tggacacaag acaggcttgc gagatatgtt 12781 tgagaatacc actttatccc gcgtcaggga gaggcagtgc gtaaaaagac gcggactcat 12841 gtgaaatact ggtttttagt gcgccagatc tctataatct cgcgcaacct attttcccct 12901 cgaacacttt ttaagccgta gataaacagg ctgggacact tcacatgagc gaaaaataca 12961 tcgtcacctg ggacatgttg cagatccatg cacgtaaact cgcaagccga ctgatgcctt 13021 ctgaacaatg gaaaggcatt attgccgtaa gccgtggcgg tctgtaccgg gtgcgttact 13081 ggcgcgtgaa ctgggtattc gtcatgtcga taccgtttgt atttccagct acgatcacga 13141 caaccagcgc gagcttaaag tgctgaaacg cgcagaaggc gatggcgaag gcttcatcgt 13201 tattgatgac ctggtggata ccggtggtac tgcggttgcg attcgtgaaa tgtatccaaa 13261 agcgcacttt gtcaccatct tcgcaaaacc ggctggtcgt ccgctggttg atgactatgt 13321 tgttgatatc ccgcaagata cctggattga acagccgtgg gatatgggcg tcgtattcgt 13381 cccgccaatc tccggtcgct aatcttttca acgcctggca ctgccgggcg ttgttctttt 13441 taacttcagg cgggttacaa tagtttccag taagtattct ggaggctgca tccatgacac 13501 aggcaaacct gagcgaaacc ctgttcaaac cccgctttaa acatcctgaa acctcgacgc 13561 tagtccgccg ctttaatcac ggcgcacaac cgcctgtgca gtcggccctt gatggtaaaa 13621 ccatccctca ctggtatcgc atgattaacc gtctgatgtg gatctggcgc ggcattgacc 13681 cacgcgaaat cctcgacgtc caggcacgta ttgtgatgag cgatgccgaa cgtaccgacg 13741 atgatttata cgatacggtg attggctacc gtggcggcaa ctggatttat gagtgggccc 13801 cggatctttg tgaaggaacc ttacttctgt ggtgtgacat aattggacaa actacctaca 13861 gagatttaaa gctctaaggt aaatataaaa tttttaagtg tataatgtgt taaactactg 13921 attctaattg tttgtgtatt ttagattcca acctatggaa ctgatgaatg ggagcagtgg 13981 tggaatgcct ttaatgagga aaacctgttt tgctcagaag aaatgccatc tagtgatgat 14041 gaggctactg ctgactctca acattctact cctccaaaaa agaagagaaa ggtagaagac 14101 cccaaggact ttccttcaga attgctaagt tttttgagtc atgctgtgtt tagtaataga 14161 actcttgctt gctttgctat ttacaccaca aaggaaaaag ctgcactgct atacaagaaa 14221 attatggaaa aatattctgt aacctttata agtaggcata acagttataa tcataacata 14281 ctgttttttc ttactccaca caggcataga gtgtctgcta ttaataacta tgctcaaaaa 14341 ttgtgtacct ttagcttttt aatttgtaaa ggggttaata aggaatattt gatgtatagt 14401 gccttgacta gagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa 14461 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa 14521 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 14581 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 14641 tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc 14701 ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc 14761 tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt //