LOCUS Exported 13957 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION N. castellii Solo CRISPR vector, marked with URA3 and NatR. ACCESSION . VERSION . KEYWORDS pV1464 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13957) 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 13957) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13957 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 43..60 /label=L4440 /note="L4440 vector, forward primer" protein_bind 177..198 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 213..243 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 251..267 /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)." primer_bind 256..278 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 275..291 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 275..291 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 310..330 /label=T3 /note="T3 promoter, forward primer" promoter 312..330 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" misc_RNA complement(2450..2525) /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" promoter 2942..3285 /label=TEF promoter /note="Ashbya gossypii TEF promoter" terminator 3886..4083 /label=TEF terminator /note="Ashbya gossypii TEF terminator" primer_bind 4241..4259 /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" CDS complement(4340..4405) /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS complement(4406..4426) /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS complement(4430..4450) /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS complement(4454..4474) /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS complement(4487..8590) /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" primer_bind complement(11135..11154) /label=T7 /note="T7 promoter, forward primer" promoter complement(11136..11154) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(11164..11181) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(11164..11180) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(11173..11195) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 11321..11776 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(11408..11427) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 11618..11639 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(11907..12710) /codon_start=1 /gene="S. cerevisiae URA3" /product="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /label=URA3 /note="yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" promoter complement(12711..12926) /gene="S. cerevisiae URA3" /label=URA3 promoter primer_bind complement(12950..12969) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 13072..13094 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(13136..13154) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" misc_feature 13195..13698 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" promoter 13735..13839 /gene="bla" /label=AmpR promoter ORIGIN 1 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag 61 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa 121 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat 181 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg 241 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac 301 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctccg aacactcttg 361 tgctgatcta gtagatatca acaggtgcac ttgttgcaag agcaatttct gtgttggaat 421 atcgactttg gtatagaatt tcattagtct cggaagacca tcattattga ctatccaatg 481 attatgttag tactaaagta gaccacaaat aaaattcacg agtttactaa gtacatacat 541 attaaaacgg aatgaatcat agtttctcga ttgtaaagct ctcaagctgt tagaattgtt 601 gttgaggccc aaaggcgtct tttttttttc caccattcgc aattatctcc cacataatac 661 tgggtatcaa ctttttttgt tgtttccgat ttttttcttc aattctatct ctccggttat 721 accaccaact actccaaccc cattttatta tcagatgtca ttagccgagg caaagaaatt 781 ggacaaaaag ttcccaactt ttaaaaatga gtttgccata ccaacttttg gatcattagg 841 tatcaaaaat aataaatatg agtcatcaac tgagtctatt tatttatgtg gaaactctct 901 tgggttgatg cctaagaata caaagaaagc tatcaatgat gaattgaatg catgggttga 961 acgaggggtt gagtctcatt tcaatcatcc tgacaaactg ttaacaccgt gggtagacat 1021 cgatttaccc ttattaccat taatagcccc cattgttggt gcaaaggaaa acgaagttgc 1081 agttatgggt tctttaacag caaacttgaa tgcattgttg atccattttt acaaacctga 1141 ggggaaaaga accaaaattt tatttgaaaa acaagcattt ccttctgact actatgcatt 1201 tttgaacatt gttaaattgt tcggatacga cgagaaacat ttgattcaat tggaagttca 1261 acctggtgaa acttatatta aaacagaaag aataatcaag gctatagatg agaacctgga 1321 tgaattggct ctagtgtgtt ttcctggtat ccaatactac actggtcaat ttttcaagat 1381 agaggaaata actaaatacg ctaaggaaaa gagccaacaa attaaagttg ggtgggatct 1441 tgcccatgca gttgggaatg ttccattaaa tttacatgat tggggagtag attttgctgc 1501 ttggtgttcc tacaaatatt taaactcggg acctggagcc ataggtggaa tttttgtgca 1561 tgaaaaatac actatagaga acaagaagag cagtttcgtt cctagactag ctggttggtg 1621 gggcaataac tcccaagaac gattcaaaat gttagaagag tttgatccaa taaactcggc 1681 tctttcttac agacagtcaa atccttctgt tcttgatgtt gttgctgtga agtcgtcatt 1741 ggaagtgtat gctaaagtgg gtggagtatc taaacttcgt gaaaaaagtg ttgctttgac 1801 tcaattttta caggatctat tgacgaattc caagtattac ataccacaat ccagtacatc 1861 aaactccaaa tttgggttta aaattttaac tcctttgaat ccagctgaaa gaggatgtca 1921 attgtccatt atgtttcaac cgcatttcga tgaaaagggc aagaacgtca tggaacgagt 1981 taatgcgtac ttgcatgatc atgctataat ctgtgacgaa agaagaccag atgtaatcag 2041 attagcccca ttgccgttat acaatacttt tgaagagact tttattgctg tccagagatt 2101 atttgaagcc ctagataata tctctaaaga atatatgtaa atagacgtta gcatcgatta 2161 ataaaatgtg caattgaaaa gaatcaacgc gtctaagtta gtctccgttc atctcttcac 2221 caggtgctgt atcccttttt ctttgactgc agctcagtga ttaagagtaa agatgggtaa 2281 aaaattatca tttaattagt tcatatattc aagatgttcc tataaaagaa aaaaaagcac 2341 cagctttttt ttatttaatc agaggcaaac ccgcggtggc ggcaaaacta attcttctct 2401 tcagttatca ttatccttaa acactcgata aaaaaactcg agaaaaaaag caccgactcg 2461 gtgccacttt ttcaagttga taacggacta gccttatttt aacttgctat ttctagctct 2521 aaaacgagac ggaattccgt ctcctattac tgttcacaaa ggtagtttcg aacccaaacc 2581 cttttaggaa ggaaattacc acttgaccaa atcaagaaac aataaaattt cggagaaact 2641 ttgaattgat aagcaaaact taaggtcaca agattaatcc gggtaatgat cgcttcttac 2701 gatttttttc ctatttcgtt ctggaaaaaa aaaacaccac cagtttcagt tcagtttgtt 2761 gtcgttgttc ctttccatcc cattcatctc tttcgttatt atcctcaaaa attagtacat 2821 cagtgttttt attcaacttg atcttagtta gtccctcttt aatctcattt attagctgct 2881 actacctcct caacgcggcc gcatcaagct tgcctcgtcc ccgccgggtc acccggccag 2941 cgacatggag gcccagaata ccctccttga cagtcttgac gtgcgcagct caggggcatg 3001 atgtgactgt cgcccgtaca tttagcccat acatccccat gtataatcat ttgcatccat 3061 acattttgat ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc agacctgcga 3121 gcagggaaac gctcccctca cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag 3181 aaatataaaa ggttaggatt tgccactgag gttcttcttt catatacttc cttttaaaat 3241 cttgctagga tacagttctc acatcacatc cgaacataaa caaccggatc ctacaattat 3301 gtctactact ttggatgata ctgcttatag atacagaact tctgctccag gtgatgctga 3361 agctattgaa gctttggatg gttctttcac taccgatact gtttttagag ttactgctac 3421 tggtgatggt ttcactttga gagaagttcc agttgatcca ccattgacta aggttttccc 3481 agatgatgaa tccgatgatg aatccgatgc tggtgaagat ggcgatccag attctagaac 3541 tttcgttgct tatggtgatg atggtgattt ggctggtttc gttgttgttt cttattctgg 3601 ttggaacaga agattgactg ttgaagatat tgaagttgct ccagaacata gaggtcatgg 3661 tgttggtaga gctttgatgg gtttggctac tgaattcgcc agagaaagag gtgctggtca 3721 tttgtggttg gaagttacca atgttaatgc tccagctatt catgcttata gaagaatggg 3781 tttcactttg tgtggtttgg atactgcttt atacgatggt actgcttccg atggtgaaca 3841 agctttgtat atgtctatgc catgtccata ataactcgag tcagtactga caataaaaag 3901 atccttgttt tcaagaactt gtcatttgta tagttttttt atattgtagt tgttctattt 3961 taatcaaatg ttagcgtgat ttatattttt tttcgcctcg acatcatctg cccagatgcg 4021 aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata 4081 ctgctgtcga ttcgatacta acgccgccaa gaaaaaaaag aaaaatttga aatataaata 4141 acgttcttaa tacaaacata actataaaaa aataaatagg gacctagact tcaggttgtc 4201 taactccttc cttttcggtt agagcggatg tggggggagg gcgtgaatgt aagcgtgaca 4261 taactaatta catgactcga cagaaattcg cccggaatta gcttggcggc aggtcgatgg 4321 atctctgcag tcattatcac ttgtcatcgt catccttgta gtcgatgtca tgatctttat 4381 aatcaccgtc atggtctttg tagtcaacct ttctcttttt cttaggatcc accttacgct 4441 ttttctttgg atcaactttt cttttcttct taggatcagc cccttcatca ccacccaatt 4501 gagacaaatc aattcttgtc tcatacaatc cagtgattga ttgatgaatt aaggtggcgt 4561 ctaagacttc ttttgttgag gtatatcttt tacgatcaat tgttgtatca aaatacttga 4621 aagcagcagg tgctcctaaa ttggtaaggg taaataagtg aataatgttt tctgcttgtt 4681 ctctaattgg cttatctcta tgtttgttat aggctgacaa aactttatct aaatttgcat 4741 cagctaatat cactctttta ctgaattcac taatttgttc tataatctca tctaagtagt 4801 gtttgtgttg ttcaacaaat aattgttttt gctcgttatc ttctggtgac cctttcaact 4861 tttcatagtg tgaagctaaa tacaagaagt taacatattt ggaaggcaat gctaattcat 4921 tacccttttg caattctcca gcagaagcca acattctttt tctcccattt tctaattcaa 4981 ataaagaata tttaggcaat ttgattatta agtccttctt tacttcctta taacctttag 5041 cttccaaaaa atcaataggg ttcttttcaa aggaacttct ttccattatt gtaataccca 5101 aaagttcttt tactgattta agctttttac ttttaccctt ttcaaccttt gccacaacca 5161 aaacagaata ggctacggtt ggtgagtcga aaccaccata tttctttgga tcccaatcct 5221 tttttctggc aatcaattta tcactatttc ttttaggcaa aatactttct tttgagaaac 5281 caccagtttg aacctctgtt ttctttacaa tgttaacttg tggcattgat aaaacttttc 5341 taacagtagc gaaatcacgc cctttgtccc aaactatctc tccagtctca ccatttgttt 5401 caataagagg tctttttcta atttcaccgt ttgccaaagt aatttctgtt ttaaagaaat 5461 tcataatgtt agagtaaaag aagtatttag ctgtagcttt accaatttct tgttctgatt 5521 tagcaatcat ctttctaaca tcatagactt tgtaatcacc atatacaaat tcagattcta 5581 atttagggta tttcttgatt aaagcagttc caactacagc attcaaatat gcatcatgag 5641 catgatgata attattaatt tctctaactt tatagaattg gaaatccttt ctgaaatcag 5701 agacaagttt tgattttaaa gtaatgactt taacctcacg tattaactta tcattttcgt 5761 catacttagt gttcatacgg gaatctaaaa tttgggcaac atgtttagta atttgtctag 5821 tttcgaccaa ttgtctttta atgaacccgg ctttatctaa ttcagaaagc ccacctcttt 5881 ctgctttagt caaattatca aattttcttt gagtaattaa tttagcattc aaaagttgac 5941 gccagtaatt tttcatcttt ttgacgactt cctcacttgg gacattatca cttttacctc 6001 tatttttatc acttctagtt aatactttat tgtctatgga atcatctttc aagaaagatt 6061 gtggaactat atgatctaca tcataatctg aaagtctatt tatatctaat tcctgatcaa 6121 catacatatc acgtccattt tgaaggtagt aaagatataa tttttcattt tgcaactggg 6181 tattttccac tgggtgttct ttcaatattt gactacccaa ttctttaata ccctcttcta 6241 ttcttttcat acgttctcta ctatttttct gtcctttttg tgtagtttga ttttctcttg 6301 ccatctcaat gacaatgttt tcaggtttat gtcttcccat tactttcact aattcatcca 6361 caactttaac agtttgtaat atccctttct taatagcagg agatccggct aaatttgcaa 6421 tatgttcatg taatgaatca ccttgcccac tcacttgggc cttttggatg tcttctttaa 6481 atgtcaaact atcgtcgtgg atcaattgca tgaaatttct attagcaaat ccgtcagatt 6541 tcaaaaagtc caagatagtt ttaccacttt gtttatctct aattccattg atcaatttac 6601 gagacaatct accccatcca gtataacgtc ttcttttcaa ctgtttcatc actttatcgt 6661 caaacaaatg tgcataagtc ttcaaacgtt cttcaatcat ctctctatct tcgaataaag 6721 ttaaggttaa aacaatgtct tctaaaatgt cttcattttc ttcgttatcc aagaaatctt 6781 tatctttgat aattttaagt aaatcatggt aagttcctaa actagcatta aatctgtctt 6841 caacaccact gatttcaact gaatcaaaac attcaatttt cttaaagtaa tcttctttta 6901 attgtttaac agtaactttt ctatttgttt taaacaataa atcaactatt gcctttttct 6961 gttctccaga taaaaaggct ggcttacgca taccttcagt aacatattta acctttgtaa 7021 gttcattgta aacagtaaaa tattcgtata acaaggaatg ttttggtaaa actttttcat 7081 ttggtaagtt tttatcgaaa ttggtcattc tttcaataaa actctgtgct gatgctcctt 7141 tatcaacaac ctcttcaaaa ttccaagggg taatagtttc ttctgattta cgtgtcatcc 7201 aggcaaatct tgaattacct ctagccaaag gcccaacata gtatggaatt ctaaaggtta 7261 aaattttctc gattttttct ctattatctt ttaaaaatgg ataaaaatct tcttgtcttc 7321 ttaaaattgc atgcaactca cccaaatgaa tttggtgagg tatactacca ttatcaaaag 7381 ttctctgttt acgtaacaaa tcttctctgt tcaatttgac caaaagttcc tcagtcccat 7441 ccatcttttc taaaataggt tttataaatt tgtaaaattc ctcttgggag gcacccccat 7501 caatataacc tgcatacccg tttttagatt gatcgaaaaa gatttcttta tacttctctg 7561 gcaattgttg tctcaccaaa gctttcaata aagtcaaatc ctgatggtgt tcatcataac 7621 gtttgatcat agatgctgat aatggagctt tggttatttc tgtgttcact ctcaaaatat 7681 cagacaacaa aatagcatca gataagtttt tagcagctaa gaacaaatct gcatattgat 7741 caccaatttg agcaagtaaa ttgtccaaat cgtcgtcata ggtgtctttt gataattgta 7801 atttagcatc ttcagctaaa tcaaaattag atttaaaatt tggtgtcaaa cccaatgata 7861 atgcgatcaa gtttccaaat aatccattct ttttttctcc tggcaattgg gcaatcaaat 7921 tttccaaacg tctagatttt gataatctgg ctgataatat agctttggca tcgacaccac 7981 tagcattaat tggattttcc tcaaataatt ggttgtaggt ttggactaat tgaataaata 8041 atttatccac gtcagaatta tctgggttca aatcaccttc aatcaaaaaa tgacctctaa 8101 atttaatcat atgggcaagt gctaaataga ttaatctaag gtcagcttta tcagttgaat 8161 ctaccaattt ctttcttaaa tgataaatgg taggatattt ttcatggtaa gccacttcat 8221 caactatatt cccaaagatt ggatgtctct catgcttttt gtcttcttca accaaaaatg 8281 actcttccaa tctatggaaa aaggaatcat caactttagc catttcatta gaaaagattt 8341 cttgtaaata acatattcta ttttttctac gggtatatct tcttcttgct gttcttttta 8401 aacgtgtagc ctcggcagtt tcaccggagt cgaacaaaag agcaccgata agattctttt 8461 ttattgagtg tctgtcagtg ttacctaaca ccttaaattt ctttgatgga actttatatt 8521 cgtcagtgat aactgcccaa cccacagagt tagtaccaat atctaaacca atactatact 8581 ttttatccat cccgggtttt atgtatgtga attagtttgg attgctatga ggttttaaac 8641 taagaacaag attgtaataa gaaataacag aagaaaaaag ggttttcact gaaagttgag 8701 acgagatgat cttaaatata aatgtttaat gggtaagaaa aaaattttct aaggatggga 8761 atgaaaattt tagtgaaaaa aaaaagagaa aaagaaaaag gaaaggaaga gatgcccctc 8821 gctcatttct tttttctttc gtttctggct cctaatcctc tcgagcgttg tgcggttatt 8881 tctggggaaa gtgcctacgg ggcgcttgcc aaaccaaaac tgtaaaaata taccctgctg 8941 ggtacagtga aaatgccgta gcccacatgc ggtttgtctt ttcaaatttt ttcttcggtt 9001 ttttcttagt aaacttttgt tggtggcaaa gtggcggacg ttccaaccgt gcggccattt 9061 cttaggaggg aatgtctctg atgggggaca ttcaatacat cttgcctccc ttgttagtta 9121 tgcggtgtct gggtggctaa aacactctga aagatgaacg ggtgtttgtt gacaaacagt 9181 caaacttttt gtgagtgtga gcactactga taacagatgt cagatatgaa tttaacacca 9241 acattctttt aatgagcaat gtggatgtaa aaatataggg acgtcactag gtaacgttaa 9301 aaagaatttt tttgatcttg aggttatggt aaacgtgttt tcttggttat taatttcaac 9361 ttttgctggg tgttttgtct tcacaagtag cattgacaca ttgtgtgcca cactttacgc 9421 aaaactacaa aatcacagaa cagagtctat atttaactac aactacagat tgaagtgact 9481 ttactttgtg ttggagactg gaaacaacca gagatatttt gataagtaaa atactacaac 9541 taataagaat tgcgatagac catggttgtt gtaatattcc tgaattatca attgatgatc 9601 gaatgaagaa ttaaaaaaaa gggagaaaag gaaagaaata gaaacagaaa aaaaaagaaa 9661 aaacaggaat tggaaaatca gacttatttt atattaattg gagtctaaaa gatcttactg 9721 tgagcaaaat tgtcaatcaa caataaaggg agtaaggtgg ttgaaaaatt cagaaatgaa 9781 ataacctgac ggcgcgcata cacaaaaact tactatggtg gtggtgggtg ggctactcat 9841 atgcagtagt agacatataa tgaatggaac acaacttccc gatgcacttt tttttttttt 9901 ttctgttgcc ctagtgtggt tatgttctat gcaatgaaga gagaaatctg gccaacatca 9961 ggtgaacgca ggaaaagtat gtctgtttgt acgtttgtat aattagtgtg gatcacctta 10021 tagaaatcgt tagtcaactt ttgcaacgca taagttctaa aaaaaggcat cacctgtaaa 10081 caaatcttga tctctttttt ctaaatcaaa aacaaaaacc ctaattgaga acccacggct 10141 taatggaact catttgttta gatccgggcc agatttatat gaagtacaat taagcacctt 10201 atcgagtgaa ggtaggtcgc tttccatctc aattgtaacg gcttgctgtc gacccggagg 10261 acaatcatgg gccgaaggag aaaatgtttt agttgaagat agttgctaca tcgaaaagag 10321 aatttttttg tgtaattgtt agctgtagtc atgtgatttg agctatgcgc cgggaatgtg 10381 gattgttgag cgacattgtg tattttgcga caccactaag aagctagaag agacattttt 10441 ggcttttttc tgatgaacat tttgttttga aattgataga aatatgacag acctttcaaa 10501 acaaacaatc acggtattac acaatcatgt ctactaaaga tgcaaatgat gttaaaccca 10561 caagttccaa atctaatgag ataagtctgt ctggaacagc aacgccaaaa aagacgagtt 10621 cagtgccagc caaggaggat cgaaaagaca cgaccaagaa ggatatagcg aatgaacgca 10681 catcttcatc aaaatctaaa gatgactctg agaaatacgt tgaattgaaa aataaactaa 10741 tccagcaaat tcttaagaag caagaactaa ctagaaagct aaccacgcta gaggattcga 10801 tatatcaaaa agagatagat tatttcgaag agagcccgct tggtaatatc gtaaaaggtt 10861 ttgagaattt ctcaaagact ggtggtggcg gtggtgcaaa taagagaaag attgtctata 10921 gtgaagatga tcacatattt agcctcagtt cagtcaacta tataaaaagt ttgatgaaaa 10981 gacagggcca tacatcaaac ggggggtctt caaaagacga ttttgatgat tatgaagatt 11041 ctgttgatcc aaccactgct agtgcaggaa gtaaagctaa tagtatcgag aagaacgaaa 11101 ctcccagtgg tagtagtggc aggtacccaa ttcgccctat agtgagtcgt attacgcgcg 11161 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa 11221 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga 11281 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca 11341 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 11401 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 11461 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 11521 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 11581 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 11641 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 11701 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata 11761 ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 11821 accgcatagg gtaataactg atataattaa attgaagctc taatttgtga gtttagtata 11881 catgcattta cttataatac agttttttag ttttgctggc cgcatcttct caaatatgct 11941 tcccagcctg cttttctgta acgttcaccc tctaccttag catcccttcc ctttgcaaat 12001 agtcctcttc caacaataat aatgtcagat cctgtagaga ccacatcatc cacggttcta 12061 tactgttgac ccaatgcatc acccttgtca tctaaaccca caccgggtgt cataatcaac 12121 caatcgtaac cttcatctct tccacccatg tctctttgag caataaagcc gataacaaaa 12181 tctttgtcgc tcttcgcaat gtcaacagta cccttagtat attctccagt agatagggag 12241 cccttgcatg acaattctgc taacatcaaa aggcctctag gttcctttgt tacttcttct 12301 gccgcctgct tcaaaccgct aacaatacct gggcccacca caccgtgtgc attcgtaatg 12361 tctgcccatt ctgctattct gtatacaccc gcagagtact gcaatttgac tgtattacca 12421 atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt acttggcgga taatgccttt 12481 agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga tatccacatg tgtttttagt 12541 aaacaaattt tgggacctaa tgcttcaact aactccagta attccttggt ggtacgaaca 12601 tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga tattaaatag cttggcagca 12661 acaggactag gatgagtagc agcacgttcc ttatatgtag ctttcgacat gatttatctt 12721 cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga atactgggca atttcatgtt 12781 tcttcaacac tacatatgcg tatatatacc aatctaagtc tgtgctcctt ccttcgttct 12841 tccttctgtt cggagattac cgaatcaaaa aaatttcaag gaaaccgaaa tcaaaaaaaa 12901 gaataaaaaa aaaatgatga attgaaaagg tggtatggtg cactctcggt acaatctgct 12961 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac 13021 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgagatc tccgggagct 13081 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaattcgacg aaagggcctc 13141 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacggatcgc 13201 ttgcctgtaa cttacacgcg cctcgtatct tttaatgatg gaataatttg ggaatttact 13261 ctgtgtttat ttatttttat gttttgtatt tggattttag aaagtaaata aagaaggtag 13321 aagagttacg gaatgaagaa aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc 13381 gtactttaca tatatattta ttagacaaga aaagcagatt aaatagatat acattcgatt 13441 aacgataagt aaaatgtaaa atcacaggat tttcgtgtgt ggtcttctac acagacaaga 13501 tgaaacaatt cggcattaat acctgagagc aggaagagca agataaaagg tagtatttgt 13561 tggcgatccc cctagagtct tttacatctt cggaaaacaa aaactatttt ttctttaatt 13621 tcttttttta ctttctattt ttaatttata tatttatatt aaaaaattta aattataatt 13681 atttttatag cacgtgatga aaaggaccca ggtggcactt ttcggggaaa tgtgcgcgga 13741 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 13801 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt 13861 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg 13921 ctggtgaaag taaaagatgc tgaagatcag ttgggtg //