LOCUS Exported 16192 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of codon-optimized Cas9-eGFP fusion protein with Ppdc promoter in Trichoderma reesei . ACCESSION . VERSION . KEYWORDS pDHt/sk-PE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 16192) AUTHORS Liu R, Chen L, Jiang Y, Zhou Z, Zou G TITLE Efficient genome editing in filamentous fungus Trichoderma reesei using the CRISPR/Cas9 system. JOURNAL Cell Discov. 2015 May 12;1:15007. doi: 10.1038/celldisc.2015.7. eCollection 2015. PUBMED 27462408 REFERENCE 2 (bases 1 to 16192) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..16192 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(1446..2075) /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" misc_feature 3376..3400 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" primer_bind 3588..3610 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 3602..3619 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 3603..3619 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS 5163..9266 /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" regulatory 9288..9297 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 9294..10010 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQKTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(9339..9360) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(9600..9619) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 9947..9968 /label=EGFP-C /note="EGFP, forward primer" CDS 10011..10031 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind 11079..11099 /label=T3 /note="T3 promoter, forward primer" promoter 11081..11099 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" CDS 11503..12528 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIDKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" polyA_signal 13036..13210 /label=CaMV poly(A) signal /note="cauliflower mosaic virus polyadenylation signal" misc_feature 13288..13312 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" CDS 13737..14531 /codon_start=1 /gene="aphA-3" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF" rep_origin 14618..15206 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 15107..15126 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 15360..15377 /label=L4440 /note="L4440 vector, forward primer" misc_feature 15392..15532 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(15458..15477) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 15618..15640 /label=pGEX 3' /note="pGEX vectors, reverse primer" rep_origin 15876..16070 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS complement(16136..1017) /codon_start=1 /product="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 RepA /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI LVMRYRNLIEGEASAGS" ORIGIN 1 cgggccagcc tgccccgcct cctccttcaa atcgtactcc ggcaggtcat ttgacccgat 61 cagcttgcgc acggtgaaac agaacttctt gaactctccg gcgctgccac tgcgttcgta 121 gatcgtcttg aacaaccatc tggcttctgc cttgcctgcg gcgcggcgtg ccaggcggta 181 gagaaaacgg ccgatgccgg gatcgatcaa aaagtaatcg gggtgaaccg tcagcacgtc 241 cgggttcttg ccttctgtga tctcgcggta catccaatca gctagctcga tctcgatgta 301 ctccggccgc ccggtttcgc tctttacgat cttgtagcgg ctaatcaagg cttcaccctc 361 ggataccgtc accaggcggc cgttcttggc cttcttcgta cgctgcatgg caacgtgcgt 421 ggtgtttaac cgaatgcagg tttctaccag gtcgtctttc tgctttccgc catcggctcg 481 ccggcagaac ttgagtacgt ccgcaacgtg tggacggaac acgcggccgg gcttgtctcc 541 cttcccttcc cggtatcggt tcatggattc ggttagatgg gaaaccgcca tcagtaccag 601 gtcgtaatcc cacacactgg ccatgccggc cggccctgcg gaaacctcta cgtgcccgtc 661 tggaagctcg tagcggatca cctcgccagc tcgtcggtca cgcttcgaca gacggaaaac 721 ggccacgtcc atgatgctgc gactatcgcg ggtgcccacg tcatagagca tcggaacgaa 781 aaaatctggt tgctcgtcgc ccttgggcgg cttcctaatc gacggcgcac cggctgccgg 841 cggttgccgg gattctttgc ggattcgatc agcggccgct tgccacgatt caccggggcg 901 tgcttctgcc tcgatgcgtt gccgctgggc ggcctgcgcg gccttcaact tctccaccag 961 gtcatcaccc agcgccgcgc cgatttgtac cgggccggat ggtttgcgac cgctcacgcc 1021 gattcctcgg gcttgggggt tccagtgcca ttgcagggcc ggcaggcaac ccagccgctt 1081 acgcctggcc aaccgcccgt tcctccacac atggggcatt ccacggcgtc ggtgcctggt 1141 tgttcttgat tttccatgcc gcctccttta gccgctaaaa ttcatctact catttattca 1201 tttgctcatt tactctggta gctgcgcgat gtattcagat agcagctcgg taatggtctt 1261 gccttggcgt accgcgtaca tcttcagctt ggtgtgatcc tccgccggca actgaaagtt 1321 gacccgcttc atggctggcg tgtctgccag gctggccaac gttgcagcct tgctgctgcg 1381 tgcgctcgga cggccggcac ttagcgtgtt tgtgcttttg ctcattttct ctttacctca 1441 ttaactcaaa tgagttttga tttaatttca gcggccagcg cctggacctc gcgggcagcg 1501 tcgccctcgg gttctgattc aagaacggtt gtgccggcgg cggcagtgcc tgggtagctc 1561 acgcgctgcg tgatacggga ctcaagaatg ggcagctcgt acccggccag cgcctcggca 1621 acctcaccgc cgatgcgcgt gcctttgatc gcccgcgaca cgacaaaggc cgcttgtagc 1681 cttccatccg tgacctcaat gcgctgctta accagctcca ccaggtcggc ggtggcccat 1741 atgtcgtaag ggcttggctg caccggaatc agcacgaagt cggctgcctt gatcgcggac 1801 acagccaagt ccgccgcctg gggcgctccg tcgatcacta cgaagtcgcg ccggccgatg 1861 gccttcacgt cgcggtcaat cgtcgggcgg tcgatgccga caacggttag cggttgatct 1921 tcccgcacgg ccgcccaatc gcgggcactg ccctggggat cggaatcgac taacagaaca 1981 tcggccccgg cgagttgcag ggcgcgggct agatgggttg cgatggtcgt cttgcctgac 2041 ccgcctttct ggttaagtac agcgataacc ttcatgcgtt ccccttgcgt atttgtttat 2101 ttactcatcg catcatatac gcagcgaccg catgacgcaa gctgttttac tcaaatacac 2161 atcacctttt tagacggcgg cgctcggttt cttcagcggc caagctggcc ggccaggccg 2221 ccagcttggc atcagacaaa ccggccagga tttcatgcag ccgcacggtt gagacgtgcg 2281 cgggcggctc gaacacgtac ccggccgcga tcatctccgc ctcgatctct tcggtaatga 2341 aaaacggttc gtcctggccg tcctggtgcg gtttcatgct tgttcctctt ggcgttcatt 2401 ctcggcggcc gccagggcgt cggcctcggt caatgcgtcc tcacggaagg caccgcgccg 2461 cctggcctcg gtgggcgtca cttcctcgct gcgctcaagt gcgcggtaca gggtcgagcg 2521 atgcacgcca agcagtgcag ccgcctcttt cacggtgcgg ccttcctggt cgatcagctc 2581 gcgggcgtgc gcgatctgtg ccggggtgag ggtagggcgg gggccaaact tcacgcctcg 2641 ggccttggcg gcctcgcgcc cgctccgggt gcggtcgatg attagggaac gctcgaactc 2701 ggcaatgccg gcgaacacgg tcaacaccat gcggccggcc ggcgtggtgg tgtcggccca 2761 cggctctgcc aggctacgca ggcccgcgcc ggcctcctgg atgcgctcgg caatgtccag 2821 taggtcgcgg gtgctgcggg ccaggcggtc tagcctggtc actgtcacaa cgtcgccagg 2881 gcgtaggtgg tcaagcatcc tggccagctc cgggcggtcg cgcctggtgc cggtgatctt 2941 ctcggaaaac agcttggtgc agccggccgc gtgcagttcg gcccgttggt tggtcaagtc 3001 ctggtcgtcg gtgctgacgc gggcatagcc cagcaggcca gcggcggcgc tcttgttcat 3061 ggcgtaatgt ctccggttct agtcgcaagt attctacttt atgcgactaa aacacgcgac 3121 aagaaaacgc caggaaaagg gcagggcggc agcctgtcgc gtaacttagg acttgtgcga 3181 catgtcgttt tcagaagacg gctgcactga acgtcagaag ccgactgcac tatagcagcg 3241 gaggggttgg atcaaagtac tttgatcccg aggggaaccc tgtggttggc atgcacatac 3301 aaatggacga acggataaac cttttcacgc ccttttaaat atccgattat tctaataaac 3361 gctcttttct cttaggttta cccgccaata tatcctgtca aacactgata gtttaaactg 3421 aaggcgggaa acgacaatct gatccaagct caagctgctc tagcattcgc cattcaggct 3481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa 3541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg 3601 ttgtaaaacg acggccagtg ccaagcttag gacttccagg gctacttggc gcggcagaag 3661 ccgcaggtgt cgagccggga ggagttttcg cagtggctgt gccgcgcgca caacgatgtc 3721 aaccggaagc tgggcaagcc cgagtttgac tgctcgcggg tggacgagcg gtggaggacg 3781 gggtggaagg acggtcggtg cgactgattg gccatgaatt ccatctttcg aggacggacg 3841 aagatactgt acgattaatg aaaggaggga gcattcttcg acttgcggca attgcatgca 3901 catgtacgat tggaagcgcg ggcgatgtat tcgcaatcat gtttagaagg acggcgtttg 3961 gaaacgttgg gatgctgttg aagcgttgga aacaggggca attagaaaca ccgagccaga 4021 cagagtcaat ggtacgaggt cagccagtat catgacctgt gtgcgcatgg tggcgagaga 4081 ttccgagcca tgccacggga gacgagcaat gaaaaaactc ttcactcact tgtcgaggct 4141 ctctcaacct atcgacttat caagtagacg atgaaagcct tgcaactgtg gtgatgtggc 4201 tcatcaatgt gcgacgtcgt atccatgtct gaggccattc gatatcgtga tgcgactacc 4261 tagtaaagcc cggccagagg gcaaaccggg gcgacagggg caggcaattg accggatggc 4321 tgcatgtgcc gaagcagccc cgatggaatc gagatgtctg tcggatggac cgctgagcgg 4381 cctggcaagg tgtcccagat acgaagatgg aagtgaagtc agaggtggtc gttaattgtc 4441 cgacgagcga atcggccgct ccttcggatt gccggctctg ctgtatgtac cgtgcatgaa 4501 gccacccggg atccatgtta cgatggatag gttccaactc tctagtagct atagtggacc 4561 tgaggctatc tagtatcact ggaggagcag ccgtccacta tcgtcgagcg ctgtagaagc 4621 agctgcatta gcggctgccc acccgcgcag aaatggcccc attacatcac tatcatgaca 4681 gcggcgcgtc caaaagtgag ctcatgcttg ccgatggcac gagcagctgc aactggcggg 4741 gctcctgcct gccgtctccg gtgccgctgc ccatttgagt ttgtccgagc tgttgatggt 4801 tgaaaccgag accgatggat gattcaacac ttcgaagtct aggtagataa aaaacatcta 4861 tatatcctca ttcattgccc tgtcagtgtg ttggctcacg tctccaatcc tccgcccctc 4921 ctcctgcaaa gtaaatacct tctcaaaaca cgtctggaat cctgcaagtc tccatcacaa 4981 ggagcttctt catcaaccac cttatacgag caacatcatt tgcatcatcg ttgatccaca 5041 tctcctcgcg cctcagagtg tcgtcaccag tataaataac cgcatcaagc tctcgtcctt 5101 cttcgttcca caatccaaga agcacctcaa aacgatcaaa gcagcgcagc tacagcacaa 5161 tcatggacaa gaagtacagc attggcctgg acattggcac gaactcggtc ggctgggccg 5221 tcatcacgga cgagtacaag gtcccctcca agaagtttaa ggtcctgggc aacaccgacc 5281 gccactccat caagaagaac ctcattggcg ccctgctctt cgactccggc gagaccgccg 5341 aggccacccg cctcaagcgc accgcccgcc gccgatacac gcgccgcaag aaccgcatct 5401 gctacctgca ggagattttc tccaacgaga tggccaaggt cgacgactcc ttctttcacc 5461 gcctggagga gtcgttcctc gtcgaggaag acaagaagca cgagcgccac cccatctttg 5521 gcaacattgt cgacgaggtc gcctaccacg agaagtaccc cacgatctac cacctgcgca 5581 agaagctcgt cgactccacc gacaaggccg acctccgcct gatctacctc gccctggccc 5641 acatgattaa gttccgcggc cactttctga tcgagggcga cctcaacccc gacaacagcg 5701 acgtcgacaa gctgttcatc cagctcgtcc agacctacaa ccagctcttt gaggagaacc 5761 ccattaacgc ctccggcgtc gacgccaagg ccatcctctc ggcccgcctc tccaagagcc 5821 gccgactcga gaacctgatc gcccagctgc ccggcgagaa gaagaacggc ctgttcggca 5881 acctcatcgc cctctccctg ggcctcaccc ccaacttcaa gtcgaacttt gacctcgccg 5941 aggacgccaa gctgcagctc tccaaggaca cctacgacga cgacctggac aacctcctgg 6001 cccagatcgg cgaccagtac gccgacctgt tcctcgccgc caagaacctg tccgacgcca 6061 tcctcctgtc ggacattctc cgcgtcaaca ccgagattac gaaggcccct ctctccgcct 6121 cgatgatcaa gcgctacgac gagcaccacc aggacctgac cctgctcaag gccctggtcc 6181 gccagcagct ccccgagaag tacaaggaga tcttctttga ccagagcaag aacggctacg 6241 ccggctacat cgacggcggc gctagccaag aggagttcta caagtttatc aagcccattc 6301 tggagaagat ggacggcacg gaggagctcc tggtcaagct caaccgcgag gacctcctgc 6361 gcaagcagcg caccttcgac aacggcagca tcccccacca gattcacctc ggcgagctgc 6421 acgccatcct ccgccgacaa gaggacttct acccctttct caaggacaac cgcgagaaga 6481 tcgagaagat tctgacgttc cgcatcccct actacgtcgg ccccctggcc cgcggcaaca 6541 gccgctttgc ctggatgacc cgcaagtccg aggagaccat cacgccctgg aacttcgagg 6601 aagtcgtcga caagggcgcc tcggcccagt ccttcatcga gcgcatgacc aactttgaca 6661 agaacctgcc caacgagaag gtcctcccca agcactcgct cctgtacgag tacttcaccg 6721 tctacaacga gctcacgaag gtcaagtacg tcaccgaggg catgcgcaag cccgccttcc 6781 tgtcgggcga gcagaagaag gccatcgtcg acctcctgtt taagaccaac cgcaaggtca 6841 cggtcaagca gctcaaggaa gactacttca agaagattga gtgctttgac agcgtcgaga 6901 tctccggcgt cgaggaccgc tttaacgcct ccctgggcac ctaccacgac ctcctgaaga 6961 tcattaagga caaggacttc ctggacaacg aggagaacga ggacatcctc gaggacattg 7021 tcctgaccct cacgctgttt gaggaccgcg agatgatcga ggagcgcctg aagacgtacg 7081 cccacctctt cgacgacaag gtcatgaagc agctcaagcg ccgccgatac accggctggg 7141 gccgcctgag ccgcaagctc atcaacggca ttcgcgacaa gcagtcgggc aagacgatcc 7201 tcgacttcct gaagagcgac ggcttcgcca accgcaactt tatgcagctg attcacgacg 7261 actccctcac cttcaaggaa gacatccaga aggcccaggt ctccggccag ggcgactccc 7321 tgcacgagca catcgccaac ctcgccggca gccccgccat caagaagggc attctgcaga 7381 ccgtcaaggt cgtcgacgag ctcgtcaagg tcatgggccg ccacaagccc gagaacatcg 7441 tcattgagat ggcccgcgag aaccagacca cgcagaaggg ccagaagaac agccgcgagc 7501 gcatgaagcg catcgaggaa ggcatcaagg agctgggctc ccagatcctc aaggagcacc 7561 ccgtcgagaa cacccagctg cagaacgaga agctctacct gtactacctc cagaacggcc 7621 gcgacatgta cgtcgaccag gagctggaca ttaaccgcct ctcggactac gacgtcgacc 7681 acatcgtccc ccagagcttc ctgaaggacg actccatcga caacaaggtc ctcacccgca 7741 gcgacaagaa ccgcggcaag agcgacaacg tcccctccga ggaagtcgtc aagaagatga 7801 agaactactg gcgccagctc ctgaacgcca agctgatcac gcagcgcaag tttgacaacc 7861 tcaccaaggc cgagcgaggc ggcctctcgg agctggacaa ggccggcttc atcaagcgcc 7921 agctggtcga gacccgccag atcacgaagc acgtcgccca gattctcgac tcgcgcatga 7981 acacgaagta cgacgagaac gacaagctga tccgcgaggt caaggtcatt accctgaagt 8041 cgaagctcgt cagcgacttc cgcaaggact tccagtttta caaggtccgc gagatcaaca 8101 actaccacca cgcccacgac gcctacctca acgccgtcgt cggcaccgcc ctgatcaaga 8161 agtaccccaa gctcgagtcc gagttcgtct acggcgacta caaggtctac gacgtccgca 8221 agatgatcgc caagtccgag caggagattg gcaaggccac cgccaagtac ttcttttact 8281 cgaacatcat gaacttcttt aagaccgaga tcaccctcgc caacggcgag atccgcaagc 8341 gccccctcat tgagaccaac ggcgagaccg gcgagatcgt ctgggacaag ggccgcgact 8401 tcgccaccgt ccgcaaggtc ctcagcatgc cccaggtcaa catcgtcaag aagaccgagg 8461 tccagacggg cggcttctcg aaggagagca ttctgcccaa gcgcaactcc gacaagctca 8521 tcgcccgcaa gaaggactgg gaccccaaga agtacggtgg cttcgactcc cccaccgtcg 8581 cctactcggt cctggtcgtc gccaaggtcg agaagggcaa gtcgaagaag ctcaagagcg 8641 tcaaggagct cctgggcatc accattatgg agcgcagctc cttcgagaag aaccccatcg 8701 actttctcga ggccaagggc tacaaggaag tcaagaagga cctgatcatt aagctcccca 8761 agtactccct cttcgagctg gagaacggcc gcaagcgcat gctcgcctcc gccggcgagc 8821 tccagaaggg caacgagctc gccctgccca gcaagtacgt caacttcctc tacctggcca 8881 gccactacga gaagctcaag ggctcccccg aggacaacga gcagaagcag ctgtttgtcg 8941 agcagcacaa gcactacctc gacgagatca ttgagcagat ttccgagttc tcgaagcgcg 9001 tcatcctggc cgacgccaac ctggacaagg tcctcagcgc ctacaacaag caccgcgaca 9061 agcccatccg cgagcaggcc gagaacatca ttcacctctt caccctgacc aacctcggcg 9121 cccccgccgc cttcaagtac tttgacacca cgatcgaccg caagcgctac acctcgacga 9181 aggaagtcct ggacgccacc ctcatccacc agagcattac cggcctctac gagacgcgca 9241 tcgacctcag ccagctcggc ggcgactccc gcgccgacgt accggtcgcc accatggtga 9301 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg 9361 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc 9421 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga 9481 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg 9541 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg 9601 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc 9661 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg 9721 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca 9781 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact 9841 accagcagaa aacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga 9901 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg 9961 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag cccaagaaga 10021 agaggaaggt gtaacccggc atgaagtctg accgggtagt atgagggttc atcgtcacct 10081 tgatagaata atagacgata aagcaggcca cgggcaggta ccgattgtca atccggcagg 10141 ttcggaggcg tgttggaaat gagtttatgg gttatggtca aatcggatag tatgaggtac 10201 atagtttgta aatctcaaga ttattttctt ccttaatctt gcacgtcgca tgagagggac 10261 cgagaagaga attgatgaag ggctcttgaa gatgagatga atcacgtggt tgctgaagct 10321 tcagtagtct cgggtacctg ttctttccca caaacagtag ccaggctaga ggtactgagt 10381 acccgctcac cgtatctaat catccgacct gaaatcttca agctgtttta ttgacacttc 10441 gagtccatct tcattcacgt aaggagaact tctaggacat cacttatccc gccatattta 10501 gctgcaagga gtcaattgca atgtcagatt ccgctcctaa gaggaaacag ggccctggcg 10561 gctcagatgg ctcggcattg aagaagagaa aggtatgatg acaagaatgc ttcttttaca 10621 tatttacgaa gaatgccacc acccttctct ctggctacct atgtccatgg acaacgagct 10681 tgctgacaat gggactatga atgggtacct aggagggcac gggaggaaga tggaagacgc 10741 cagctcaaaa ggcaaagcag gcctcgtggg tcgagatggg caaggctctc gaggtgggcg 10801 acgaaggcat ctgggtcacg tatgcgagag gcatgaagaa caaggccgtt cgggagttta 10861 cggccatgtg ccatgaggta aagctgcata atcccatgcc cacccacggg gagactattt 10921 tagatgttta ttggttgcgg cggggatgat aataacgatg ttggcagtac ggcgaaaagc 10981 tctacgggat cacgcccccg gccgacgagt cagaagctca ccacccggcc gacggagccg 11041 aagaggaaga catcgaggcg tccatctaga ggatccgcgc aattaaccct cactaaaggg 11101 aacaaaagct ggagctccac cgcggtggcg gccgcgacgt taactgatat tgaaggagca 11161 ctttttgggc ttggctggag ctagtggagg tcaacaatga atgcctattt tggtttagtc 11221 gtccaggcgg tgagcacaaa atttgtgtcg tttgacaaga tggttcattt aggcaactgg 11281 tcagatcagc cccacttgta gcagtagcgg cggcgctcga agtgtgactc ttattagcag 11341 acaggaacta ggacattatc atcatctgct gcttggtgca cgataacttg gtgcgtttgt 11401 caagcaaggt aagtgaacga cccggtcata ccttcttaag ttcgcccttc ctccctttgt 11461 ttcagattca atctgactta cctattctac ccaagcatcg atatgaaaaa gcctgaactc 11521 accgcgacgt ctgtcgagaa gtttctgatc gacaagttcg acagcgtctc cgacctgatg 11581 cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat 11641 gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac 11701 tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt cagcgagagc 11761 ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc 11821 gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat 11881 cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca 11941 tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg 12001 gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag 12061 gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg 12121 gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa 12181 tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg 12241 cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg ggcgtatatg 12301 ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca 12361 gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt 12421 acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc 12481 gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagag tagatgccga 12541 ccgggatcga tccacttaac gttactgaaa tcatcaaaca gcttgacgaa tctggatata 12601 agatcgttgg tgtcgatgtc agctccggag ttgagacaaa tggtgttcag gatctcgata 12661 agatacgttc atttgtccaa gcagcaaaga gtgccttcta gtgatttaat agctccatgt 12721 caacaagaat aaaacgcgtt tcgggtttac ctcttccaga tacagctcaa ctgcaatgca 12781 ttaatgcatt ggacctcgca accctagtac gcccttcagg ctccggcgaa gcagaagaat 12841 agcttagcag agtctatttt cattttcggg agacgagatc aagcagatca acggtcgtca 12901 agagacctac gagactgagg aatccgctct tggctccacg cgactatata tttgtctcta 12961 attgtacttt gacatgctcc tcttctttac tctgatagct tgactatgaa aattccgtca 13021 ccagccctgc tcgagtttct ccataataat gtgtgagtag ttcccagata agggaattag 13081 ggttcctata gggtttcgct catgtgttga gcatataaga aacccttagt atgtatttgt 13141 atttgtaaaa tacttctatc aataaaattt ctaattccta aaaccaaaat ccagtactaa 13201 aatccagatc ccccgaatta attcggcgtt aattcagtac attaaaaacg tccgcaatgt 13261 gttattaagt tgtctaagcg tcaatttgtt tacaccacaa tatatcctgc caccagccag 13321 ccaacagctc cccgaccggc agctcggcac aaaatcacca ctcgatacag gcagcccatc 13381 agtccgggac ggcgtcagcg ggagagccgt tgtaaggcgg cagactttgc tcatgttacc 13441 gatgctattc ggaagaacgg caactaagct gccgggtttg aaacacggat gatctcgcgg 13501 agggtagcat gttgattgta acgatgacag agcgttgctg cctgtgatca ccgcggtttc 13561 aaaatcggct ccgtcgatac tatgttatac gccaactttg aaaacaactt tgaaaaagct 13621 gttttctggt atttaaggtt ttagaatgca aggaacagtg aattggagtt cgtcttgtta 13681 taattagctt cttggggtat ctttaaatac tgtagaaaag aggaaggaaa taataaatgg 13741 ctaaaatgag aatatcaccg gaattgaaaa aactgatcga aaaataccgc tgcgtaaaag 13801 atacggaagg aatgtctcct gctaaggtat ataagctggt gggagaaaat gaaaacctat 13861 atttaaaaat gacggacagc cggtataaag ggaccaccta tgatgtggaa cgggaaaagg 13921 acatgatgct atggctggaa ggaaagctgc ctgttccaaa ggtcctgcac tttgaacggc 13981 atgatggctg gagcaatctg ctcatgagtg aggccgatgg cgtcctttgc tcggaagagt 14041 atgaagatga acaaagccct gaaaagatta tcgagctgta tgcggagtgc atcaggctct 14101 ttcactccat cgacatatcg gattgtccct atacgaatag cttagacagc cgcttagccg 14161 aattggatta cttactgaat aacgatctgg ccgatgtgga ttgcgaaaac tgggaagaag 14221 acactccatt taaagatccg cgcgagctgt atgatttttt aaagacggaa aagcccgaag 14281 aggaacttgt cttttcccac ggcgacctgg gagacagcaa catctttgtg aaagatggca 14341 aagtaagtgg ctttattgat cttgggagaa gcggcagggc ggacaagtgg tatgacattg 14401 ccttctgcgt ccggtcgatc agggaggata tcggggaaga acagtatgtc gagctatttt 14461 ttgacttact ggggatcaag cctgattggg agaaaataaa atattatatt ttactggatg 14521 aattgtttta gtacctagaa tgcatgacca aaatccctta acgtgagttt tcgttccact 14581 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 14641 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 14701 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 14761 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 14821 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 14881 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 14941 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 15001 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 15061 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 15121 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 15181 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 15241 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata 15301 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca 15361 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc 15421 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat 15481 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca 15541 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag 15601 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa 15661 acgcgcgagg cagggtgcct tgatgtgggc gccggcggtc gagtggcgac ggcgcggctt 15721 gtccgcgccc tggtagattg cctggccgta ggccagccat ttttgagcgg ccagcggccg 15781 cgataggccg acgcgaagcg gcggggcgta gggagcgcag cgaccgaagg gtaggcgctt 15841 tttgcagctc ttcggctgtg cgctggccag acagttatgc acaggccagg cgggttttaa 15901 gagttttaat aagttttaaa gagttttagg cggaaaaatc gccttttttc tcttttatat 15961 cagtcactta catgtgtgac cggttcccaa tgtacggctt tgggttccca atgtacgggt 16021 tccggttccc aatgtacggc tttgggttcc caatgtacgt gctatccaca ggaaagagac 16081 cttttcgacc tttttcccct gctagggcaa tttgccctag catctgctcc gtacattagg 16141 aaccggcgga tgcttcgccc tcgatcaggt tgcggtagcg catgactagg at //