LOCUS Exported 11170 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of a catalytically inactive, human codon-optimized Cas9-BFP fusion under the control of Murine Stem Cell retroVirus LTR promoter for mammalian gene knockdown. ACCESSION . VERSION . KEYWORDS pdCas9::BFP-humanized SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11170) AUTHORS Qi LS, Larson MH, Gilbert LA, Doudna JA, Weissman JS, Arkin AP, Lim WA TITLE Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression. JOURNAL Cell. 2013 Feb 28;152(5):1173-83. doi: 10.1016/j.cell.2013.02.022. PUBMED 23452860 REFERENCE 2 (bases 1 to 11170) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11170 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 579..920 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 987..1403 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1333..1355 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1371..1387 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" regulatory 1417..1426 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1423..5526 /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 5536..5556 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 5566..6261 /codon_start=1 /product="monomeric blue fluorescent protein" /label=TagBFP /note="mammalian codon-optimized" /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD LPSKLGHKLN" CDS 6271..6291 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" promoter 6313..6812 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(6348..6371) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 6745..6764 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 6833..7432 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(6833..6852) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 7329..7349 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 7532..8046 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" primer_bind complement(8215..8231) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8215..8231) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(8228..8250) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 8239..8255 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(8263..8293) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8308..8329 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(8446..8463) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8617..9205) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8697..8716) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(9376..10236) /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 9999..10018 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(10237..10341) /gene="bla" /label=AmpR promoter primer_bind 10409..10427 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(10465..10487) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 10587..10606 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 10800..10822 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 10950..10969 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" LTR 11169..515 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" ORIGIN 1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat 61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca 121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga 181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt 241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc 301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc 361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata 421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca 481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga 541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg 601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt 661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa 721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga 781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag 841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa 901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg 961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct 1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga 1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag 1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc 1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc 1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg 1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag 1381 ccctcactcc ttctctaggc gccggaatta gatctcgcca ccatggacaa gaagtattct 1441 atcggactgg ccatcgggac taatagcgtc gggtgggccg tgatcactga cgagtacaag 1501 gtgccctcta agaagttcaa ggtgctcggg aacaccgacc ggcattccat caagaaaaat 1561 ctgatcggag ctctcctctt tgattcaggg gagaccgctg aagcaacccg cctcaagcgg 1621 actgctagac ggcggtacac caggaggaag aaccggattt gttaccttca agagatattc 1681 tccaacgaaa tggcaaaggt cgacgacagc ttcttccata ggctggaaga atcattcctc 1741 gtggaagagg ataagaagca tgaacggcat cccatcttcg gtaatatcgt cgacgaggtg 1801 gcctatcacg agaaataccc aaccatctac catcttcgca aaaagctggt ggactcaacc 1861 gacaaggcag acctccggct tatctacctg gccctggccc acatgatcaa gttcagaggc 1921 cacttcctga tcgagggcga cctcaatcct gacaatagcg atgtggataa actgttcatc 1981 cagctggtgc agacttacaa ccagctcttt gaagagaacc ccatcaatgc aagcggagtc 2041 gatgccaagg ccattctgtc agcccggctg tcaaagagcc gcagacttga gaatcttatc 2101 gctcagctgc cgggtgaaaa gaaaaatgga ctgttcggga acctgattgc tctttcactt 2161 gggctgactc ccaatttcaa gtctaatttc gacctggcag aggatgccaa gctgcaactg 2221 tccaaggaca cctatgatga cgatctcgac aacctcctgg cccagatcgg tgaccaatac 2281 gccgaccttt tccttgctgc taagaatctt tctgacgcca tcctgctgtc tgacattctc 2341 cgcgtgaaca ctgaaatcac caaggcccct ctttcagctt caatgattaa gcggtatgat 2401 gagcaccacc aggacctgac cctgcttaag gcactcgtcc ggcagcagct tccggagaag 2461 tacaaggaaa tcttctttga ccagtcaaag aatggatacg ccggctacat cgacggaggt 2521 gcctcccaag aggaatttta taagtttatc aaacctatcc ttgagaagat ggacggcacc 2581 gaagagctcc tcgtgaaact gaatcgggag gatctgctgc ggaagcagcg cactttcgac 2641 aatgggagca ttccccacca gatccatctt ggggagcttc acgccatcct tcggcgccaa 2701 gaggacttct acccctttct taaggacaac agggagaaga ttgagaaaat tctcactttc 2761 cgcatcccct actacgtggg acccctcgcc agaggaaata gccggtttgc ttggatgacc 2821 agaaagtcag aagaaactat cactccctgg aacttcgaag aggtggtgga caagggagcc 2881 agcgctcagt cattcatcga acggatgact aacttcgata agaacctccc caatgagaag 2941 gtcctgccga aacattccct gctctacgag tactttaccg tgtacaacga gctgaccaag 3001 gtgaaatatg tcaccgaagg gatgaggaag cccgcattcc tgtcaggcga acaaaagaag 3061 gcaattgtgg accttctgtt caagaccaat agaaaggtga ccgtgaagca gctgaaggag 3121 gactatttca agaaaattga atgcttcgac tctgtggaga ttagcggggt cgaagatcgg 3181 ttcaacgcaa gcctgggtac ctaccatgat ctgcttaaga tcatcaagga caaggatttt 3241 ctggacaatg aggagaacga ggacatcctt gaggacattg tcctgactct cactctgttc 3301 gaggaccggg aaatgatcga ggagaggctt aagacctacg cccatctgtt cgacgataaa 3361 gtgatgaagc aacttaaacg gagaagatat accggatggg gacgccttag ccgcaaactc 3421 atcaacggaa tccgggacaa acagagcgga aagaccattc ttgatttcct taagagcgac 3481 ggattcgcta atcgcaactt catgcaactt atccatgatg attccctgac ctttaaggag 3541 gacatccaga aggcccaagt gtctggacaa ggtgactcac tgcacgagca tatcgcaaat 3601 ctggctggtt cacccgctat taagaagggt attctccaga ccgtgaaagt cgtggacgag 3661 ctggtcaagg tgatgggtcg ccataaacca gagaacattg tcatcgagat ggccagggaa 3721 aaccagacta cccagaaggg acagaagaac agcagggagc ggatgaaaag aattgaggaa 3781 gggattaagg agctcgggtc acagatcctt aaagagcacc cggtggaaaa cacccagctt 3841 cagaatgaga agctctatct gtactacctt caaaatggac gcgatatgta tgtggaccaa 3901 gagcttgata tcaacaggct ctcagactac gacgtggacg ccatcgtccc tcagagcttc 3961 ctcaaagacg actcaattga caataaggtg ctgactcgct cagacaagaa ccggggaaag 4021 tcagataacg tgccctcaga ggaagtcgtg aaaaagatga agaactattg gcgccagctt 4081 ctgaacgcaa agctgatcac tcagcggaag ttcgacaatc tcactaaggc tgagaggggc 4141 ggactgagcg aactggacaa agcaggattc attaaacggc aacttgtgga gactcggcag 4201 attactaaac atgtcgccca aatccttgac tcacgcatga ataccaagta cgacgaaaac 4261 gacaaactta tccgcgaggt gaaggtgatt accctgaagt ccaagctggt cagcgatttc 4321 agaaaggact ttcaattcta caaagtgcgg gagatcaata actatcatca tgctcatgac 4381 gcatatctga atgccgtggt gggaaccgcc ctgatcaaga agtacccaaa gctggaaagc 4441 gagttcgtgt acggagacta caaggtctac gacgtgcgca agatgattgc caaatctgag 4501 caggagatcg gaaaggccac cgcaaagtac ttcttctaca gcaacatcat gaatttcttc 4561 aagaccgaaa tcacccttgc aaacggtgag atccggaaga ggccgctcat cgagactaat 4621 ggggagactg gcgaaatcgt gtgggacaag ggcagagatt tcgctaccgt gcgcaaagtg 4681 ctttctatgc ctcaagtgaa catcgtgaag aaaaccgagg tgcaaaccgg aggcttttct 4741 aaggaatcaa tcctccccaa gcgcaactcc gacaagctca ttgcaaggaa gaaggattgg 4801 gaccctaaga agtacggcgg attcgattca ccaactgtgg cttattctgt cctggtcgtg 4861 gctaaggtgg aaaaaggaaa gtctaagaag ctcaagagcg tgaaggaact gctgggtatc 4921 accattatgg agcgcagctc cttcgagaag aacccaattg actttctcga agccaaaggt 4981 tacaaggaag tcaagaagga ccttatcatc aagctcccaa agtatagcct gttcgaactg 5041 gagaatgggc ggaagcggat gctcgcctcc gctggcgaac ttcagaaggg taatgagctg 5101 gctctcccct ccaagtacgt gaatttcctc taccttgcaa gccattacga gaagctgaag 5161 gggagccccg aggacaacga gcaaaagcaa ctgtttgtgg agcagcataa gcattatctg 5221 gacgagatca ttgagcagat ttccgagttt tctaaacgcg tcattctcgc tgatgccaac 5281 ctcgataaag tccttagcgc atacaataag cacagagaca aaccaattcg ggagcaggct 5341 gagaatatca tccacctgtt caccctcacc aatcttggtg cccctgccgc attcaagtac 5401 ttcgacacca ccatcgaccg gaaacgctat acctccacca aagaagtgct ggacgccacc 5461 ctcatccacc agagcatcac cggactttac gaaactcgga ttgacctctc acagctcgga 5521 ggggatgagg gagctcccaa gaaaaagcgc aaggtaggta gttccagcga gctgattaag 5581 gagaacatgc acatgaagct gtacatggag ggcaccgtgg acaaccatca cttcaagtgc 5641 acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggtggtc 5701 gagggcggcc ctctcccctt cgccttcgac atcctggcta ctagcttcct ctacggcagc 5761 aagaccttca tcaaccacac ccagggcatc cccgacttct tcaagcagtc cttccctgag 5821 ggcttcacat gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag 5881 gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc 5941 acatccaacg gccctgtgat gcagaagaaa acactcggct gggaggcctt caccgagacg 6001 ctgtaccccg ctgacggcgg cctggaaggc agaaacgaca tggccctgaa gctcgtgggc 6061 gggagccatc tgatcgcaaa catcaagacc acatatagat ccaagaaacc cgctaagaac 6121 ctcaagatgc ctggcgtcta ctatgtggac tacagactgg aaagaatcaa ggaggccaac 6181 aacgagacct acgtcgagca gcacgaggtg gcagtggcca gatactgcga cctccctagc 6241 aaactggggc acaagcttaa tgagggagct ccaaagaaga agcgtaaggt aggtagttcc 6301 tagaattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag 6361 cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat 6421 ccaccggtag gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc 6481 ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa 6541 tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc 6601 gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga 6661 ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg 6721 cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc 6781 tgttctcctc ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga 6841 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct 6901 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat 6961 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 7021 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 7081 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 7141 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 7201 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 7261 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 7321 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 7381 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc 7441 ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcga taaaataaaa 7501 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa 7561 gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag 7621 ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg 7681 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc 7741 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac 7801 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg 7861 ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag 7921 actgcgtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt 7981 ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc 8041 tttcatgggt aacagtttct tgaagttgga gaacaacatt ctgagggtag gagtcgaata 8101 ttaagtaatc ctgactcaat tagccactgt tttgaatcca catactccaa tactcctgaa 8161 atagttcatt atggacagcg cagaagagct ggggagaatt aattcgtaat catggtcata 8221 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 8281 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 8341 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 8401 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 8461 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 8521 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 8581 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 8641 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 8701 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 8761 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 8821 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 8881 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 8941 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 9001 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac 9061 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 9121 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat 9181 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 9241 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 9301 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 9361 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 9421 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg 9481 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga 9541 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt 9601 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 9661 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 9721 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 9781 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc 9841 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 9901 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 9961 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 10021 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt 10081 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 10141 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 10201 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg 10261 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 10321 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac 10381 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc 10441 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc 10501 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg 10561 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca 10621 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc 10681 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg 10741 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc 10801 ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg catgcaagga gatggcgccc 10861 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc 10921 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc 10981 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggcg attagtccaa 11041 tttgttaaag acaggatatc agtggtccag gctctagttt tgactcaaca atatcaccag 11101 ctgaagccta tagagtacga gccatagata aaataaaaga ttttatttag tctccagaaa 11161 aaggggggaa //