LOCUS Exported 14004 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Genedrive entry vector for Candida albicans. ACCESSION . VERSION . KEYWORDS pV1539 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14004) AUTHORS Vyas VK, Bushkin GG, Bernstein DA, Getz MA, Sewastianik M, Barrasa MI, Bartel DP, Fink GR TITLE New CRISPR Mutagenesis Strategies Reveal Variation in Repair Mechanisms among Fungi. JOURNAL mSphere. 2018 Apr 25;3(2). pii: 3/2/e00154-18. doi: 10.1128/mSphere.00154-18. Print 2018 Apr 25. PUBMED 29695624 REFERENCE 2 (bases 1 to 14004) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..14004 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(26..481) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(163..184) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 375..394 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 608..630 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 622..639 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 623..639 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 649..668 /label=T7 /note="T7 promoter, forward primer" promoter 649..667 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 2254..6357 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 6370..6390 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 6394..6414 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 6418..6438 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 6439..6504 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" terminator 6569..6816 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(6585..6603) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS 7376..8647 /codon_start=1 /product="site-specific recombinase" /label=FLP /note="FLP is a site-specific recombinase from Saccharomyces cerevisiae. Recombination occurs at FRT sequences." /translation="MPQFDILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR YPAWNGIISQEVLDYLSSYINRRI" terminator 9074..9271 /label=TEF terminator /note="Ashbya gossypii TEF terminator" promoter complement(9872..10215) /label=TEF promoter /note="Ashbya gossypii TEF promoter" misc_RNA 11284..11359 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" protein_bind complement(11439..11472) /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" promoter complement(11839..11857) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(11878..11894) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(11878..11894) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(11891..11913) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 11902..11918 /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(11926..11956) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 11971..11992 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(12109..12126) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(12280..12868) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(12360..12379) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(13039..13899) /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 13662..13681 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(13900..14004) /gene="bla" /label=AmpR promoter ORIGIN 1 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg 61 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct 121 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt 181 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat 241 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca 301 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac 361 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta 421 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg 481 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg 541 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca 601 gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca 661 ctatagggcg aattgaggcc tgcatgcggt acctgccact actaccactg ggagtttcgt 721 tcttctcgat actattagct ttacttcctg cactagcagt ggttggatca acagaatctt 781 cataatcatc aaaatcgtct tttgaagacc ccccgtttga tgtatggccc tgtcttttca 841 tcaaactttt tatatagttg actgaactga ggctaaatat gtgatcatct tcactataga 901 caatctttct cttatttgca ccaccgccac caccagtctt tgagaaattc tcaaaacctt 961 ttacgatatt accaagcggg ctctcttcga aataatctat ctctttttga tatatcgaat 1021 cctctagcgt ggttagcttt ctagttagtt cttgcttctt aagaatttgc tggattagtt 1081 tatttttcaa ttcaacgtat ttctcagagt catctttaga ttttgatgaa gatgtgcgtt 1141 cattcgctat atccttcttg gtcgtgtctt ttcgatcctc cttggctggc actgaactcg 1201 tcttttttgg cgttgctgtt ccagacagac ttatctcatt agatttggaa cttgtgggtt 1261 taacatcatt tgcatcttta gtagacatga ttgtgtaata ccgtgattgt ttgttttgaa 1321 aggtctgtca tatttctatc aatttcaaaa caaaatgttc atcagaaaaa agccaaaaat 1381 gtctcttcta gcttcttagt ggtgtcgcaa aatacacaat gtcgctcaac aatccacatt 1441 cccggcgcat agctcaaatc acatgactac agctaacaat tacacaaaaa aattctcttt 1501 tcgatgtagc aactatcttc aactaaaaca ttttctcctt cggcccatga ttgtcctccg 1561 ggtcgacagc aagccgttac aattgagatg gaaagcgacc taccttcact cgataaggtg 1621 cttaattgta cttcatataa atctggcccg gatctaaaca aatgagttcc attaagccgt 1681 gggttctcaa ttagggtttt tgtttttgat ttagaaaaaa gagatcaaga tttgtttaca 1741 ggtgatgcct ttttttagaa cttatgcgtt gcaaaagttg actaacgatt tctataaggt 1801 gatccacact aattatacaa acgtacaaac agacatactt ttcctgcgtt cacctgatgt 1861 tggccagatt tctctcttca ttgcatagaa cataaccaca ctagggcaac agnaaaaaaa 1921 aaaaaaaagt gcatcgggaa gttgtgttcc attcattata tgtctactac tgcatatgag 1981 tagcccaccc accaccacca tagtaagttt ttgtgtatgc gcgccgtcag gttatttcat 2041 ttctgaattt ttcaaccacc ttactccctt tattgttgat tgacaatttt gctcacagta 2101 agatctttta gactccaatt aatataaaat aagtctgatt ttccaattcc tgttttttct 2161 ttttttttct gtttctattt ctttcctttt ctcccttttt tttaattctt cattcaatca 2221 tcaattgata attcaggaat attacaaccc gggatggata aaaagtatag tattggttta 2281 gatattggta ctaactctgt gggttgggca gttatcactg acgaatataa agttccatca 2341 aagaaattta aggtgttagg taacactgac agacactcaa taaaaaagaa tcttatcggt 2401 gctcttttgt tcgactccgg tgaaactgcc gaggctacac gtttaaaaag aacagcaaga 2461 agaagatata cccgtagaaa aaatagaata tgttatttac aagaaatctt ttctaatgaa 2521 atggctaaag ttgatgattc ctttttccat agattggaag agtcattttt ggttgaagaa 2581 gacaaaaagc atgagagaca tccaatcttt gggaatatag ttgatgaagt ggcttaccat 2641 gaaaaatatc ctaccattta tcatttaaga aagaaattgg tagattcaac tgataaagct 2701 gaccttagat taatctattt agcacttgcc catatgatta aatttagagg tcattttttg 2761 attgaaggtg atttgaaccc agataattct gacgtggata aattatttat tcaattagtc 2821 caaacctaca accaattatt tgaggaaaat ccaattaatg ctagtggtgt cgatgccaaa 2881 gctatattat cagccagatt atcaaaatct agacgtttgg aaaatttgat tgcccaattg 2941 ccaggagaaa aaaagaatgg attatttgga aacttgatcg cattatcatt gggtttgaca 3001 ccaaatttta aatctaattt tgatttagct gaagatgcta aattacaatt atcaaaagac 3061 acctatgacg acgatttgga caatttactt gctcaaattg gtgatcaata tgcagatttg 3121 ttcttagctg ctaaaaactt atctgatgct attttgttgt ctgatatttt gagagtgaac 3181 acagaaataa ccaaagctcc attatcagca tctatgatca aacgttatga tgaacaccat 3241 caggatttga ctttattgaa agctttggtg agacaacaat tgccagagaa gtataaagaa 3301 atctttttcg atcaatctaa aaacgggtat gcaggttata ttgatggggg tgcctcccaa 3361 gaggaatttt acaaatttat aaaacctatt ttagaaaaga tggatgggac tgaggaactt 3421 ttggtcaaat tgaacagaga agatttgtta cgtaaacaga gaacttttga taatggtagt 3481 atacctcacc aaattcattt gggtgagttg catgcaattt taagaagaca agaagatttt 3541 tatccatttt taaaagataa tagagaaaaa atcgagaaaa ttttaacctt tagaattcca 3601 tactatgttg ggcctttggc tagaggtaat tcaagatttg cctggatgac acgtaaatca 3661 gaagaaacta ttaccccttg gaattttgaa gaggttgttg ataaaggagc atcagcacag 3721 agttttattg aaagaatgac caatttcgat aaaaacttac caaatgaaaa agttttacca 3781 aaacattcct tgttatacga atattttact gtttacaatg aacttacaaa ggttaaatat 3841 gttactgaag gtatgcgtaa gccagccttt ttatctggag aacagaaaaa ggcaatagtt 3901 gatttattgt ttaaaacaaa tagaaaagtt actgttaaac aattaaaaga agattacttt 3961 aagaaaattg aatgttttga ttcagttgaa atcagtggtg ttgaagacag atttaatgct 4021 agtttaggaa cttaccatga tttacttaaa attatcaaag ataaagattt cttggataac 4081 gaagaaaatg aagacatttt agaagacatt gttttaacct taactttatt cgaagataga 4141 gagatgattg aagaacgttt gaagacttat gcacatttgt ttgacgataa agtgatgaaa 4201 cagttgaaaa gaagacgtta tactggatgg ggtagattgt ctcgtaaatt gatcaatgga 4261 attagagata aacaaagtgg taaaactatc ttggactttt tgaaatctga cggatttgct 4321 aatagaaatt tcatgcaatt gatccacgac gatagtttga catttaaaga agacatccaa 4381 aaggcccaag tgagtgggca aggtgattca ttacatgaac atattgcaaa tttagccgga 4441 tctcctgcta ttaagaaagg gatattacaa actgttaaag ttgtggatga attagtgaaa 4501 gtaatgggaa gacataaacc tgaaaacatt gtcattgaga tggcaagaga aaatcaaact 4561 acacaaaaag gacagaaaaa tagtagagaa cgtatgaaaa gaatagaaga gggtattaaa 4621 gaattgggta gtcaaatatt gaaagaacac ccagtggaaa atacccagtt gcaaaatgaa 4681 aaattatatc tttactacct tcaaaatgga cgtgatatgt atgttgatca ggaattagat 4741 ataaatagac tttcagatta tgatgtagat catatagttc cacaatcttt cttgaaagat 4801 gattccatag acaataaagt attaactaga agtgataaaa atagaggtaa aagtgataat 4861 gtcccaagtg aggaagtcgt caaaaagatg aaaaattact ggcgtcaact tttgaatgct 4921 aaattaatta ctcaaagaaa atttgataat ttgactaaag cagaaagagg tgggctttct 4981 gaattagata aagccgggtt cattaaaaga caattggtcg aaactagaca aattactaaa 5041 catgttgccc aaattttaga ttcccgtatg aacactaagt atgacgaaaa tgataagtta 5101 atacgtgagg ttaaagtcat tactttaaaa tcaaaacttg tctctgattt cagaaaggat 5161 ttccaattct ataaagttag agaaattaat aattatcatc atgctcatga tgcatatttg 5221 aatgctgtag ttggaactgc tttaatcaag aaatacccta aattagaatc tgaatttgta 5281 tatggtgatt acaaagtcta tgatgttaga aagatgattg ctaaatcaga acaagaaatt 5341 ggtaaagcta cagctaaata cttcttttac tctaacatta tgaatttctt taaaacagaa 5401 attactttgg caaacggtga aattagaaaa agacctctta ttgaaacaaa tggtgagact 5461 ggagagatag tttgggacaa agggcgtgat ttcgctactg ttagaaaagt tttatcaatg 5521 ccacaagtta acattgtaaa gaaaacagag gttcaaactg gtggtttctc aaaagaaagt 5581 attttgccta aaagaaatag tgataaatta attgccagaa aaaaggattg ggatccaaag 5641 aaatatggtg gtttcgactc accaaccgta gcctattctg ttttggttgt ggcaaaggtt 5701 gaaaagggta aaagtaaaaa gcttaaatca gtaaaagaac ttttgggtat tacaataatg 5761 gaaagaagtt cctttgaaaa gaaccctatt gattttttgg aagctaaagg ttataaggaa 5821 gtaaagaagg acttaataat caaattgcct aaatattctt tatttgaatt agaaaatggg 5881 agaaaaagaa tgttggcttc tgctggagaa ttgcaaaagg gtaatgaatt agcattgcct 5941 tccaaatatg ttaacttctt gtatttagct tcacactatg aaaagttgaa agggtcacca 6001 gaagataacg agcaaaaaca attatttgtt gaacaacaca aacactactt agatgagatt 6061 atagaacaaa ttagtgaatt cagtaaaaga gtgatattag ctgatgcaaa tttagataaa 6121 gttttgtcag cctataacaa acatagagat aagccaatta gagaacaagc agaaaacatt 6181 attcacttat ttacccttac caatttagga gcacctgctg ctttcaagta ttttgataca 6241 acaattgatc gtaaaagata tacctcaaca aaagaagtct tagacgccac cttaattcat 6301 caatcaatca ctggattgta tgagacaaga attgatttgt ctcaattggg tggtgatgaa 6361 ggggctgatc ctaagaagaa aagaaaagtt gatccaaaga aaaagcgtaa ggtggatcct 6421 aagaaaaaga gaaaggttga ctacaaagac catgacggtg attataaaga tcatgacatc 6481 gactacaagg atgacgatga caagtgataa tgactgcaga gatccatcga cctgccgcca 6541 agctaattcc gggcgaattt ctgtcgagtc atgtaattag ttatgtcacg cttacattca 6601 cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag 6661 gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt 6721 tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt 6781 gagaaggttt tgggacgctc gaaggcttta atttgcggcc gggccccccc gtttgatatt 6841 tttgtctagt accatctgta ccattacact taaattatct ttatatctgt ctaactcgac 6901 tgtctggatt tcattgatgt agtcgtatgc atcgttagtt ccaaaaaata ttgtcatcaa 6961 tttgatattg gtttccgact ctaaaatttt tggaagaatt tgtctagcgt gctctgagtt 7021 gtagccactg aaaccacggt taataacatc caattttcgg atatacacat tctgtaatgc 7081 tggatgaaag ccatactggg tacaactaaa ctgggtgatg gagtcaccga acaacacaaa 7141 tttaccgtat tccatgattg ctatggttga gaattttttt tttttcttgt cccacgccat 7201 ttttcaaatt atgcagttga gaatgttagt ttttgtgtac accccgttcg ctgaatattt 7261 cggaataatt caaagattgg ggagtggggg aggcgataga cgaagacacg gtataaaaat 7321 gggcaaaatt ttccccaact ttttgcagtg gtttaactaa taatcaacta caatgatgcc 7381 acaatttgat atattatgta aaacaccacc taaggtgctt gttcgtcagt ttgtggaaag 7441 gtttgaaaga ccttcaggtg agaaaatagc attatgtgct gctgaactaa cctatttatg 7501 ttggatgatt acacataacg gaacagcaat caagagagcc acattcatga gctataatac 7561 tatcataagc aattcgttga gtttcgatat tgtcaataaa tcactccagt ttaaatacaa 7621 gacgcaaaaa gcaacaattt tggaagcctc attaaagaaa ttgattcctg cttgggaatt 7681 tacaattatt ccttactatg gacaaaaaca tcaatctgat atcactgata ttgtaagtag 7741 tttgcaatta cagttcgaat catcggaaga agcagataag gggaatagcc acagtaaaaa 7801 aatgcttaaa gcacttctaa gtgagggtga aagcatctgg gagatcactg agaaaatact 7861 aaattcgttt gagtatactt cgagatttac aaaaacaaaa actttatacc aattcctctt 7921 cctagctact ttcatcaact gtggaagatt cagcgatatt aagaacgttg atccgaaatc 7981 atttaaatta gtccaaaata agtatttggg agtaataatc cagtgtttag tgacagagac 8041 aaagacaagc gttagtaggc acatatactt ctttagcgca aggggtagga tcgatccact 8101 tgtatatttg gatgaatttt tgaggaattc tgaaccagtc ctaaaacgag taaataggac 8161 cggcaattct tcaagcaata aacaggaata ccaattatta aaagataact tagtcagatc 8221 gtacaataaa gctttgaaga aaaatgcgcc ttattcaatc tttgctataa aaaatggccc 8281 aaaatctcac attggaagac atttgatgac ctcatttctt tcaatgaagg gcctaacgga 8341 gttgactaat gttgtgggaa attggagcga taagcgtgct tctgccgtgg ccaggacaac 8401 gtatactcat cagataacag caatacctga tcactacttc gcactagttt ctcggtacta 8461 tgcatatgat ccaatatcaa aggaaatgat agcattgaag gatgagacta atccaattga 8521 ggagtggcag catatagaac agctaaaggg tagtgctgaa ggaagcatac gataccccgc 8581 atggaatggg ataatatcac aggaggtact agactacctt tcatcctaca taaatagacg 8641 catataagag tgaaattctg gaaatctgga aatctggttt tgtattcttg ttattcttct 8701 ttttgttatt acatatataa cttgttactt ttttaaaaaa atctttgtat attttataaa 8761 tatataaaac taaatttaag aaaaagagaa aaatgtttta tttgagagat tgaaatttta 8821 cttgaattta gcttagcttt tataaagtat tattatgtaa aaaaacaaaa caaatataca 8881 tcaaaaagtt aagactataa aatagccacc caaggcattt ctatatcttg ttgttgttgt 8941 tttcatcttc tgtatcagag gaacttattt tattattttc gtcacgggta ttttctcttg 9001 tttgatgatt catcccattc attccatcat aaaatgtcga cactggatgg cggcgttagt 9061 atcgaatcga cagcagtata gcgaccagca ttcacatacg attgacgcat gatattactt 9121 tctgcgcact taacttcgca tctgggcaga tgatgtcgag gcgaaaaaaa atataaatca 9181 cgctaacatt tgattaaaat agaacaacta caatataaaa aaactataca aatgacaagt 9241 tcttgaaaac aagaatcttt ttattgtcag tactgactcg agttattatg gacatggcat 9301 agacatatac aaagcttgtt caccatcgga agcagtacca tcgtataaag cagtatccaa 9361 accacacaaa gtgaaaccca ttcctctata agcatgaata gctggagcat taacattggt 9421 aacttccaac cacaaatgac cagcacctct ttctctggcg aattcagtag ccaaacccat 9481 caaagctcta ccaacaccat gacctctatg ttctggagca acttcaatat cttcaacagt 9541 caatcttctg ttccaaccag aataagaaac aacaacgaaa ccagccaaat caccatcatc 9601 accataagca acgaaagttc tagaatctgg atcaccatct tcaccagcat cggattcatc 9661 atcggattca tcatctggga aaaccttagt caatggtgga tcaactggaa cttctctcaa 9721 agtgaaacca tcaccagtag cagtaactct aaaaacagta tcggtagtga aagaaccatc 9781 caaagcttca atagcttcag catcacctgg aacagaagtt ctgtatctat aagcagtatc 9841 atccaaagta gtagacataa ttgtaggatc cggttgttta tgttcggatg tgatgtgaga 9901 actgtatcct agcaagattt taaaaggaag tatatgaaag aagaacctca gtggcaaacc 9961 ctaacctttt atatttctct acaggggcgc ggcgtgggga caattcaacg cgtctgtgag 10021 gggagcgttt ccctgctcgc aggtctgcag cgaggagccg taatttttgc ttcgcgccgt 10081 gcggccatca aaatgtatgg atgcaaatga ttatacatgg ggatgtatgg gctaaatgta 10141 cgggcgacag tcacatcatg cccctgagct gcgcacgtca agactgtcaa ggagggtatt 10201 ctgggcctcc atgtcgctgg ccgggtgacc cggcggggac gaggcaagct tgatgtgcgg 10261 ccgcaagtga ttagacttag tccgttcaaa tcaagcacaa ctctgttcat tgtttcaaca 10321 agaattaatt caaaaacagg ttcggtgcat aatttgcaaa aaaatattgc agcttctgtg 10381 gctcgaacac agtacctcca gatttcaggt ttgaaatact tcagtctgac gctctcccag 10441 atgagctaaa gctgcaataa gaaaacccac gccgggattc gaacccggaa tcctttgatt 10501 agaagtcaaa agcgataacc atttcgccac gcaggcctac ttgatgggtt tgtaaatggt 10561 ctactttttc agacctaaca gaaattttaa tgaaagtcat attcttatac aataaaactg 10621 tgtcataaaa gcagatattc gactttcgta gattatatag gacccaagaa ctaaaattta 10681 atgccatatt atgcattttt aatctgtaaa agtgttgttt ccaacctatc acaagtacgt 10741 tcttgtaact tgtgtttgta gggttgcaaa tgaatcataa caacatctca acagaacatg 10801 tatagcaaag cttagtataa aatcagtgtt ttgagaggca atccaagaat gtttacatca 10861 aagtttcaat aaatatcgac cgaaactgaa aatcttttta ggttattgtt cacttttttg 10921 taaatattta aacatttttt ggacctaaaa aaatacaagc accaattacg taccaagaag 10981 catctaatca actcccagat caccactata catttaaaag tcattggtca ataactatac 11041 tcgagtattg cctcatcaaa gaaacaatca aatattatag atactcactc catcacgtga 11101 taatttcact ggtatggaaa agtggaaaat tttataaaaa aaaatttgat gcctttggca 11161 tagctgaaac ttcggcccaa taggattgga gaatatgttt tcgcagcgtt cttacaatta 11221 aattgtggtg gaagttcgag acttgcgtaa actattttta atttggagac ggaattccgt 11281 ctcgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa 11341 agtggcaccg agtcggtgct ttttttctcg agttttttta tcgagtgttt aaggataatg 11401 ataactgaag agaagaatta gttttgccgc caccgcggga agttcctata ctttctagag 11461 aataggaact tcacgccggt aaacaagtgg tattcaagca caattcagtt ttctggatct 11521 ctaataattg caattgcaat tgcttcacat actctgttta taattcagtt tctttctctt 11581 gatgttactg atatattaag ttaataaata ccaataactt tttttcaaaa atcttctatt 11641 ttcctttcaa ttgattattt aaacacgaat cgttaataag ctgtgattgc tttttctttt 11701 tctttttctt ttctttgttc aagtttgaaa aaagtaaaaa aaaaggtttc tcttgttcga 11761 tgtagaattc agtgggttat agttgtaatg atatatggaa taaaaagaaa gaaagaaaga 11821 gagctccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt aatcatggtc 11881 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 11941 gagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 12001 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 12061 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 12121 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 12181 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 12241 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 12301 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 12361 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 12421 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 12481 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 12541 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 12601 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 12661 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 12721 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 12781 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 12841 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 12901 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 12961 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 13021 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 13081 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 13141 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 13201 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 13261 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 13321 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 13381 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 13441 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 13501 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 13561 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 13621 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 13681 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 13741 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 13801 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 13861 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 13921 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 13981 aaataaacaa ataggggttc cgcg //