LOCUS Exported 17678 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION All-in-one genome editing vector in M. polymorpha (chlorsulfuron resistant). ACCESSION . VERSION . KEYWORDS pMpGE014 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 17678) AUTHORS Sugano SS, Nishihama R, Shirakawa M, Takagi J, Matsuda Y, Ishida S, Shimada T, Hara-Nishimura I, Osakabe K, Kohchi T TITLE Efficient CRISPR/Cas9-based genome editing and its application to conditional genetic analysis in Marchantia polymorpha. JOURNAL PLoS One. 2018 Oct 31;13(10):e0205117. doi: 10.1371/journal.pone.0205117. eCollection 2018. PUBMED 30379827 REFERENCE 2 (bases 1 to 17678) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..17678 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 54..78 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" primer_bind 266..288 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 280..297 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 281..297 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 2057..2081 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2102..6205 /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 6218..6238 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" protein_bind complement(6741..6765) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind 6794..6818 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" misc_RNA 7379..7454 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" protein_bind complement(7496..7520) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind complement(7847..7863) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7847..7863) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7860..7882) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7871..7887 /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(7895..7925) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7940..7961 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." polyA_signal 8046..8220 /label=CaMV poly(A) signal /note="cauliflower mosaic virus polyadenylation signal" promoter complement(10330..11005) /label=CaMV 35S promoter (enhanced) /note="cauliflower mosaic virus 35S promoter with a duplicated enhancer region" misc_feature 11200..11224 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" CDS 11745..12536 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 12782..13370 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 13271..13290 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 13524..13541 /label=L4440 /note="L4440 vector, forward primer" misc_feature 13556..13696 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(13622..13641) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 13782..13804 /label=pGEX 3' /note="pGEX vectors, reverse primer" rep_origin 14040..14234 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS complement(14300..15373) /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" CDS complement(15802..16431) /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" ORIGIN 1 tttcacgccc ttttaaatat ccgattattc taataaacgc tcttttctct taggtttacc 61 cgccaatata tcctgtcaaa cactgatagt ttaaactgaa ggcgggaaac gacaatctga 121 tccaagctca agctgctcta gcattcgcca ttcaggctgc gcaactgttg ggaagggcga 181 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga 241 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc 301 aagcttggcc cgggcaaatg agtcacacac attgttgaga gacatatcaa aataactcaa 361 ttgtaaggta agttgataga tttcccagtt aaatgcatta ttttaacaat atttgtatct 421 caaattttca aaatcatatc aaatttgcag atcttgatga tgattcacaa ctctgtcttc 481 gatttttgat tgttaacggg tatcgatatt tgggtaactt aaaaagaatt tgctaaagat 541 cttaaaaaac agattaaaaa aacctttcaa aatagctaat cctttaaatc cgtttcacaa 601 tataatgctc attgatgata atcgaaaaga ttccatttta aagtgcatat aaattgctta 661 attatctctt tctgtctcgt aatattttgt tctctcctat tggatcggac ccgcatcttc 721 tcgatcagtt cactatacta atatttggtc aacttcaaga gagattagaa cttcgttctc 781 caaagtgtaa aggccgacaa gcagaaaagg aaacttcaat cagaagcctt ctaggttttc 841 ccgcgtccca aaaagcgaag ggaggacggg gaggacggtg aggccagaag aagaaagcaa 901 cagtaaaatc tcatgtgatt gcgtgcatgt catggatcta tgggatgcta gtccaccgtt 961 tggaagtatg tgcagaagga gaaaggcgcg ggaggccaaa agcgacttcg gccacctctc 1021 tactttctag aaagtttgct cgcagcttgt aataagcgcg gaaaaggtac tcgaaaaacc 1081 ggagcaatcg gtgtggtatc agcagtcggg ttcgctaaac ctacgtcttc ttattttgga 1141 atgttaaatt gaaagggcgt gccggcccta agcacggacg atgtgacaga cggaggagtg 1201 tgagcaattc ggatgacgat gctggccggc agtcacgaca aagagtccga aggggcatgg 1261 acagggcaat atataaatgg aaaagaggct ttgaatattc ctaacccttc gtcgtcggcc 1321 tcccaacttg ccctcgcatg ctgaattggc cgcctatctg cgaatggccg gccgccgacc 1381 ctaagtcgca ataacttgaa cggagccgat gcaaagtcct tatcgaacgc tcgttattat 1441 aaacggggca gacctaggta tttaattata tcaaaaccct ccgcaatttc gctaatcatt 1501 ccctaatttc ttctactttg gcgtctagtg ctgtggtcgc gcttgctctc aaagctcgct 1561 gggaggcagt ggagcgtctg gcttaaaatc gatcgccgca accctagagt tcctttcact 1621 tctttctctc ttgcgtggag agcatcgaca tcgaagagct gcatcttgtc ttctggtatg 1681 cgccttgtgc cctcgtctct gtacgtcgtt acgattgctc gatattgtga ttgtctagac 1741 ctgaaacccg actggagctt ttacgtggtg tcgattgggg cctcgagcga gtgatttttt 1801 aggttcgttt tagccaagta catcccactt tggtcagtcc tgttatctcg gtaaggtgtt 1861 cagttcggtc tctggtgcgg aggtgcctcg taagtttcct gcatgataga tttgtctatc 1921 cttgacttga agaggagtcg aaggtcagcg aggttggctg tgtgctatgg acgtcggtgc 1981 tccttggttc cgtgcttcta tgggtgttgc ctcaacagta ttgatgtgcc tgcagaaagg 2041 ttgactagag ttatcaacaa gtttgtacaa aaaagcaggc tccgcggccg cccccttcac 2101 catggataag aagtactcta tcggactcga tatcggaact aactctgtgg gatgggctgt 2161 gatcaccgat gagtacaagg tgccatctaa gaagttcaag gttctcggaa acaccgatag 2221 gcactctatc aagaaaaacc ttatcggtgc tctcctcttc gattctggtg aaactgctga 2281 ggctaccaga ctcaagagaa ccgctagaag aaggtacacc agaagaaaga acaggatctg 2341 ctacctccaa gagatcttct ctaacgagat ggctaaagtg gatgattcat tcttccacag 2401 gctcgaagag tcattcctcg tggaagaaga taagaagcac gagaggcacc ctatcttcgg 2461 aaacatcgtt gatgaggtgg cataccacga gaagtaccct actatctacc acctcagaaa 2521 gaagctcgtt gattctactg ataaggctga tctcaggctc atctacctcg ctctcgctca 2581 catgatcaag ttcagaggac acttcctcat cgagggtgat ctcaaccctg ataactctga 2641 tgtggataag ttgttcatcc agctcgtgca gacctacaac cagcttttcg aagagaaccc 2701 tatcaacgct tcaggtgtgg atgctaaggc tatcctctct gctaggctct ctaagtcaag 2761 aaggcttgag aacctcattg ctcagctccc tggtgagaag aagaacggac ttttcggaaa 2821 cttgatcgct ctctctctcg gactcacccc taacttcaag tctaacttcg atctcgctga 2881 ggatgcaaag ctccagctct caaaggatac ctacgatgat gatctcgata acctcctcgc 2941 tcagatcgga gatcagtacg ctgatttgtt cctcgctgct aagaacctct ctgatgctat 3001 cctcctcagt gatatcctca gagtgaacac cgagatcacc aaggctccac tctcagcttc 3061 tatgatcaag agatacgatg agcaccacca ggatctcaca cttctcaagg ctcttgttag 3121 acagcagctc ccagagaagt acaaagagat tttcttcgat cagtctaaga acggatacgc 3181 tggttacatc gatggtggtg catctcaaga agagttctac aagttcatca agcctatcct 3241 cgagaagatg gatggaaccg aggaactcct cgtgaagctc aatagagagg atcttctcag 3301 aaagcagagg accttcgata acggatctat ccctcatcag atccacctcg gagagttgca 3361 cgctatcctt agaaggcaag aggatttcta cccattcctc aaggataaca gggaaaagat 3421 tgagaagatt ctcaccttca gaatccctta ctacgtggga cctctcgcta gaggaaactc 3481 aagattcgct tggatgacca gaaagtctga ggaaaccatc accccttgga acttcgaaga 3541 ggtggtggat aagggtgcta gtgctcagtc tttcatcgag aggatgacca acttcgataa 3601 gaaccttcca aacgagaagg tgctccctaa gcactctttg ctctacgagt acttcaccgt 3661 gtacaacgag ttgaccaagg ttaagtacgt gaccgaggga atgaggaagc ctgctttttt 3721 gtcaggtgag caaaagaagg ctatcgttga tctcttgttc aagaccaaca gaaaggtgac 3781 cgtgaagcag ctcaaagagg attacttcaa gaaaatcgag tgcttcgatt cagttgagat 3841 ttctggtgtt gaggataggt tcaacgcatc tctcggaacc taccacgatc tcctcaagat 3901 cattaaggat aaggatttct tggataacga ggaaaacgag gatatcttgg aggatatcgt 3961 tcttaccctc accctctttg aagatagaga gatgattgaa gaaaggctca agacctacgc 4021 tcatctcttc gatgataagg tgatgaagca gttgaagaga agaagataca ctggttgggg 4081 aaggctctca agaaagctca ttaacggaat cagggataag cagtctggaa agacaatcct 4141 tgatttcctc aagtctgatg gattcgctaa cagaaacttc atgcagctca tccacgatga 4201 ttctctcacc tttaaagagg atatccagaa ggctcaggtt tcaggacagg gtgatagtct 4261 ccatgagcat atcgctaacc tcgctggatc tcctgcaatc aagaagggaa tcctccagac 4321 tgtgaaggtt gtggatgagt tggtgaaggt gatgggaagg cataagcctg agaacatcgt 4381 gatcgaaatg gctagagaga accagaccac tcagaaggga cagaagaact ctagggaaag 4441 gatgaagagg atcgaggaag gtatcaaaga gcttggatct cagatcctca aagagcaccc 4501 tgttgagaac actcagctcc agaatgagaa gctctacctc tactacctcc agaacggaag 4561 ggatatgtat gtggatcaag agttggatat caacaggctc tctgattacg atgttgatca 4621 tatcgtgcca cagtcattct tgaaggatga ttctatcgat aacaaggtgc tcaccaggtc 4681 tgataagaac aggggtaaga gtgataacgt gccaagtgaa gaggttgtga agaaaatgaa 4741 gaactattgg aggcagctcc tcaacgctaa gctcatcact cagagaaagt tcgataactt 4801 gactaaggct gagaggggag gactctctga attggataag gcaggattca tcaagaggca 4861 gcttgtggaa accaggcaga tcactaagca cgttgcacag atcctcgatt ctaggatgaa 4921 caccaagtac gatgagaacg ataagttgat cagggaagtg aaggttatca ccctcaagtc 4981 aaagctcgtg tctgatttca gaaaggattt ccaattctac aaggtgaggg aaatcaacaa 5041 ctaccaccac gctcacgatg cttaccttaa cgctgttgtt ggaaccgctc tcatcaagaa 5101 gtatcctaag ctcgagtcag agttcgtgta cggtgattac aaggtgtacg atgtgaggaa 5161 gatgatcgct aagtctgagc aagagatcgg aaaggctacc gctaagtatt tcttctactc 5221 taacatcatg aatttcttca agaccgagat taccctcgct aacggtgaga tcagaaagag 5281 gccactcatc gagacaaacg gtgaaacagg tgagatcgtg tgggataagg gaagggattt 5341 cgctaccgtt agaaaggtgc tctctatgcc acaggtgaac atcgttaaga aaaccgaggt 5401 gcagaccggt ggattctcta aagagtctat cctccctaag aggaactctg ataagctcat 5461 tgctaggaag aaggattggg accctaagaa atacggtggt ttcgattctc ctaccgtggc 5521 ttactctgtt ctcgttgtgg ctaaggttga gaagggaaag agtaagaagc tcaagtctgt 5581 taaggaactt ctcggaatca ctatcatgga aaggtcatct ttcgagaaga acccaatcga 5641 tttcctcgag gctaagggat acaaagaggt taagaaggat ctcatcatca agctcccaaa 5701 gtactcactc ttcgaactcg agaacggtag aaagaggatg ctcgcttctg ctggtgagct 5761 tcaaaaggga aacgagcttg ctctcccatc taagtacgtt aactttcttt acctcgcttc 5821 tcactacgag aagttgaagg gatctccaga agataacgag cagaagcaac ttttcgttga 5881 gcagcacaag cactacttgg atgagatcat cgagcagatc tctgagttct ctaaaagggt 5941 gatcctcgct gatgcaaacc tcgataaggt gttgtctgct tacaacaagc acagagataa 6001 gcctatcagg gaacaggcag agaacatcat ccatctcttc acccttacca acctcggtgc 6061 tcctgctgct ttcaagtact tcgatacaac catcgatagg aagagataca cctctaccaa 6121 agaagtgctc gatgctaccc tcatccatca gtctatcact ggactctacg agactaggat 6181 cgatctctca cagctcggtg gtgattcaag ggctgatcct aagaagaaga ggaaggtttg 6241 aggcgcgccg agctccaggc ctcccagctt tcgtccgtat catcggtttc gacaacgttc 6301 gtcaagttca atgcatcagt ttcattgccc acacaccaga atcctactaa gtttgagtat 6361 tatggcattg gaaaagctgt tttcttctat catttgttct gcttgtaatt tactgtgttc 6421 tttcagtttt tgttttcgga catcaaaatg caaatggatg gataagagtt aataaatgat 6481 atggtccttt tgttcattct caaattatta ttatctgttg tttttacttt aatgggttga 6541 atttaagtaa gaaaggaact aacagtgtga tattaaggtg caatgttaga catataaaac 6601 agtctttcac ctctctttgg ttatgtcttg aattggtttg tttcttcact tatctgtgta 6661 atcaagttta ctatgagtct atgatcaagt aattatgcaa tcaagttaag tacagtatag 6721 gcttaagggt gggcgcgccg acccagcttt cttgtacaaa gtggttgata acagcggttg 6781 actagagtta tcaacaagtt tgtacaaaaa agcaggctcc gcggccgccc ccttgtaacg 6841 tgagactact acactcctca acgcaagttc tatctcatat ctctgcaaga gcttattcca 6901 agaaactttc ttgtgcttct atgcatgccc aggtacctta agtcgcgcgc atagtcactt 6961 cacagccttt tcagttctcc ctttgatggc tcagagtgac gcaactgcac aaaaataaga 7021 aaaaagtcga agcgtcacga gagatgtatc cggctagttt ccttggagtt cggattgctc 7081 tctttctctg tctccccgaa ccatatggaa tttatgaact gctgttcact agtcagtcac 7141 ttgtttcaga acttttgagt ggtcgattta tccaccttct tgtttcttcc tgggtccaga 7201 atctgacttc ttgagacaac aaacagtgca cagtggatcg aagaatgacc acagttgaga 7261 cacgtggcgc tggcctcatt tattgcacca tacgtttagt gaagaactga ataatggtgg 7321 atataaagca gagttgcacc cagcctctcg aaatgcaggt gatgactcac ctgcataagt 7381 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg 7441 caccgagtcg gtgctttttt tctagtatcg ggaattctaa agggtgggcg cgccgaccca 7501 gctttcttgt acaaagtggt tgataacagc gcttagaggc ttagagctcg aatttccccg 7561 atcgttcaaa catttggcaa taaagtttct taagattgaa tcctgttgcc ggtcttgcga 7621 tgattatcat ataatttctg ttgaattacg ttaagcatgt aataattaac atgtaatgca 7681 tgacgttatt tatgagatgg gtttttatga ttagagtccc gcaattatac atttaatacg 7741 cgatagaaaa caaaatatag cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta 7801 tgttactaga atcgggaaat ggttccggaa ccaattcgta atcatggtca tagctgtttc 7861 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt 7921 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aggctgaatt aggcgcgcct 7981 atttctgaat tcaacacatt gcggacgttt ttaatgtact gaattaacgc cgaattaatt 8041 cgggggatct ggattttagt actggatttt ggttttagga attagaaatt ttattgatag 8101 aagtatttta caaatacaaa tacatactaa gggtttctta tatgctcaac acatgagcga 8161 aaccctatag gaaccctaat tcccttatct gggaactact cacacattat tatggagaaa 8221 ctcgagttag tatatagtcc ttccatcacc ctctgtgata atctccttaa aacttgctcc 8281 cccaggaatc ataggcagaa cgtgctcctg gtggggaact atcacatcca gtaaataagg 8341 accgggagaa tctagcatag tttgaattgc tgctcttagt tcatccttct tcaccacacg 8401 cgcatttggc accttgaaac cgtctgcaac tgtacagaag tcggggtaga tctgctcctc 8461 gtgatctgga tcacctaagt aagtatgtgc tctgttggcc ttgtagaatc tatcctccaa 8521 ttgtacgacc attcctagat gttggttgtt taagatcatg atcttcacgg gtaacttctc 8581 cacaaaaatt gtagccaact cctgtacgtt catcacaaag ctaccatcgc catcgatgtc 8641 gatcacagga atatctggtt gagttgcggc agcccccaaa gcagaaggaa gtccgaatcc 8701 catggctcca agacctccag atgtcaacca acgccgcggt tcgttgtact tgaaccactg 8761 ggctgcccac atttgatgtt ggccaacacc agtagatatg atggcatttc caccagtgag 8821 ttcgtacagc gtctgtatgg cagattgagg agcaatggca tcctcgaata cgggaaactt 8881 cattggccat ttctgctttt gttcttctag ttctgccacc cagttggaga agtcgaaagt 8941 cctcttttga ctcttcaaga gggcattgat ttcacttagc gcaggtttca catcagcaca 9001 catggacaca tggggctgtt tgttctttcc aatttcagca gggtcaatat cgatatgaac 9061 gatgctggac ctgctagcaa aagtctccag tttaccagtc actcgatcgt cgaaacgcac 9121 tccaaatgcc aacaacagat ctgcttggtc cacggcgtag ttggcgtaaa cagttccatg 9181 cattccaagc attttcaaac ttaatggatc agactcggga tacgctccca aacccattaa 9241 agtcgatgtg ataggaattc cggtaagctc gacaaattcc ctcaactctt cgctggcgtg 9301 attgcaacct cctccagaat acactacagg cttcttagaa ctagaaatga gtcttataat 9361 ctgctcgatt aacgttggtt ccggtggccg aggcagtcgt tgtaagtagc ccttcagttg 9421 cattggctga gaccaatctg gaatagccat ttgctgttgc acgtccttgg gaatatctac 9481 caagactggg cccggcctac ctgaagctgc caaaaagaaa gcctccctga ttacgcgagg 9541 gatgtcatca atgttcataa caaggtaatt gtgcttggtg atcgatctcg taacctccac 9601 aattggagtt tcttgaaaag catcggtacc aatcatccta ctggatactt gacctgtaat 9661 ggccactagt ggcacactgt ccaagagggc atcagcgagc ccggtaacaa gattggtcgc 9721 tccaggaccg gatgtggcta tacaaacccc tactcgtcca gacgatttag cataaccttc 9781 agcagcgaaa atctccccct gctcgtggcg acagagaatg ttgcggatgc tgctggaacg 9841 tgtaagagcc tggtgaatct ccatagacgc tccgccggga taagcgaaca ccgttctaat 9901 cccttctcgc tcaagagcct ccaccaaaat gtcggctcct ttgcggggct cgtcatcctt 9961 gtagcgagac tgaaagtcgg acggggttgg tgcaacgagt gacttcccgc tgctaagccg 10021 tggtccactc acatcgactg cagactggat cttctcgcta acagcttggg ctttcacttc 10081 cagtgaacgc tcagtcttcc gagaggtgtt gtatgcatgt cccctcagag cggctcccga 10141 gaagaaacca ctcctcacct ttagctcttt agctcccttg caacaagacc ttgccgatgg 10201 ctgcaccttc tccaggctcg ctatgctcaa aggcacagct gcgcatccac taattgacgc 10261 catatctcat tgccccccgg atctgcgaaa gctcgagaga gatagatttg tagagagaga 10321 ctggtgattt cagcgtgtcc tctccaaatg aaatgaactt ccttatatag aggaaggtct 10381 tgcgaaggat agtgggattg tgcgtcatcc cttacgtcag tggagatatc acatcaatcc 10441 acttgctttg aagacgtggt tggaacgtct tctttttcca cgatgctcct cgtgggtggg 10501 ggtccatctt tgggaccact gtcggcagag gcatcttgaa cgatagcctt tcctttatcg 10561 caatgatggc atttgtaggt gccaccttcc ttttctactg tccttttgat gaagtgacag 10621 atagctgggc aatggaatcc gaggaggttt cccgatatta ccctttgttg aaaagtctca 10681 atagcccttt ggtcttctga gactgtatct ttgatattct tggagtagac gagagtgtcg 10741 tgctccacca tgttcacatc aatccacttg ctttgaagac gtggttggaa cgtcttcttt 10801 ttccacgatg ctcctcgtgg gtgggggtcc atctttggga ccactgtcgg cagaggcatc 10861 ttgaacgata gcctttcctt tatcgcaatg atggcatttg taggtgccac cttccttttc 10921 tactgtcctt ttgatgaagt gacagatagc tgggcaatgg aatccgagga ggtttcccga 10981 tattaccctt tgttgaaaag tctcaatagc cctttggtct tctgagactg tatctttgat 11041 attcttggag tagacgagag tgtcgtgctc caccatgttg gcaagctgct ctagccaata 11101 cgcaaaccga attcagaaat aaattcagcc taattcggcg ttaattcagt acattaaaaa 11161 cgtccgcaat gtgttattaa gttgtctaag cgtcaatttg tttacaccac aatatatcct 11221 gccaccagcc agccaacagc tccccgaccg gcagctcggc acaaaatcac cactcgatac 11281 aggcagccca tcagtccggg acggcgtcag cgggagagcc gttgtaaggc ggcagacttt 11341 gctcatgtta ccgatgctat tcggaagaac ggcaactaag ctgccgggtt tgaaacacgg 11401 atgatctcgc ggagggtagc atgttgattg taacgatgac agagcgttgc tgcctgtgat 11461 caattcgggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt 11521 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg 11581 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg 11641 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg 11701 atgttacgca gcagggcagt cgccctaaaa caaagttaaa catcatgggg gaagcggtga 11761 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac 11821 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca 11881 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt 11941 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg 12001 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg 12061 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca 12121 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg 12181 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc 12241 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg 12301 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg 12361 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg 12421 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt 12481 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataatgtc 12541 tagctagaaa ttcgttcaag ccgacgccgc ttcgcggcgc ggcttaactc aagcgttaga 12601 tgcactaagc acataattgc tcacagccaa actatcaggt caagtctgct tttattattt 12661 ttaagcgtgc ataataagcc ctacacaaat tgggagatat atcatgcatg accaaaatcc 12721 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 12781 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 12841 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 12901 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact 12961 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 13021 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 13081 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 13141 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 13201 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 13261 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 13321 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 13381 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg 13441 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc 13501 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga 13561 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca 13621 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga 13681 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg 13741 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca 13801 gaggttttca ccgtcatcac cgaaacgcgc gaggcagggt gccttgatgt gggcgccggc 13861 ggtcgagtgg cgacggcgcg gcttgtccgc gccctggtag attgcctggc cgtaggccag 13921 ccatttttga gcggccagcg gccgcgatag gccgacgcga agcggcgggg cgtagggagc 13981 gcagcgaccg aagggtaggc gctttttgca gctcttcggc tgtgcgctgg ccagacagtt 14041 atgcacaggc caggcgggtt ttaagagttt taataagttt taaagagttt taggcggaaa 14101 aatcgccttt tttctctttt atatcagtca cttacatgtg tgaccggttc ccaatgtacg 14161 gctttgggtt cccaatgtac gggttccggt tcccaatgta cggctttggg ttcccaatgt 14221 acgtgctatc cacaggaaag agaccttttc gacctttttc ccctgctagg gcaatttgcc 14281 ctagcatctg ctccgtacat taggaaccgg cggatgcttc gccctcgatc aggttgcggt 14341 agcgcatgac taggatcggg ccagcctgcc ccgcctcctc cttcaaatcg tactccggca 14401 ggtcatttga cccgatcagc ttgcgcacgg tgaaacagaa cttcttgaac tctccggcgc 14461 tgccactgcg ttcgtagatc gtcttgaaca accatctggc ttctgccttg cctgcggcgc 14521 ggcgtgccag gcggtagaga aaacggccga tgccgggatc gatcaaaaag taatcggggt 14581 gaaccgtcag cacgtccggg ttcttgcctt ctgtgatctc gcggtacatc caatcagcta 14641 gctcgatctc gatgtactcc ggccgcccgg tttcgctctt tacgatcttg tagcggctaa 14701 tcaaggcttc accctcggat accgtcacca ggcggccgtt cttggccttc ttcgtacgct 14761 gcatggcaac gtgcgtggtg tttaaccgaa tgcaggtttc taccaggtcg tctttctgct 14821 ttccgccatc ggctcgccgg cagaacttga gtacgtccgc aacgtgtgga cggaacacgc 14881 ggccgggctt gtctcccttc ccttcccggt atcggttcat ggattcggtt agatgggaaa 14941 ccgccatcag taccaggtcg taatcccaca cactggccat gccggccggc cctgcggaaa 15001 cctctacgtg cccgtctgga agctcgtagc ggatcacctc gccagctcgt cggtcacgct 15061 tcgacagacg gaaaacggcc acgtccatga tgctgcgact atcgcgggtg cccacgtcat 15121 agagcatcgg aacgaaaaaa tctggttgct cgtcgccctt gggcggcttc ctaatcgacg 15181 gcgcaccggc tgccggcggt tgccgggatt ctttgcggat tcgatcagcg gccgcttgcc 15241 acgattcacc ggggcgtgct tctgcctcga tgcgttgccg ctgggcggcc tgcgcggcct 15301 tcaacttctc caccaggtca tcacccagcg ccgcgccgat ttgtaccggg ccggatggtt 15361 tgcgaccgct cacgccgatt cctcgggctt gggggttcca gtgccattgc agggccggca 15421 gacaacccag ccgcttacgc ctggccaacc gcccgttcct ccacacatgg ggcattccac 15481 ggcgtcggtg cctggttgtt cttgattttc catgccgcct cctttagccg ctaaaattca 15541 tctactcatt tattcatttg ctcatttact ctggtagctg cgcgatgtat tcagatagca 15601 gctcggtaat ggtcttgcct tggcgtaccg cgtacatctt cagcttggtg tgatcctccg 15661 ccggcaactg aaagttgacc cgcttcatgg ctggcgtgtc tgccaggctg gccaacgttg 15721 cagccttgct gctgcgtgcg ctcggacggc cggcacttag cgtgtttgtg cttttgctca 15781 ttttctcttt acctcattaa ctcaaatgag ttttgattta atttcagcgg ccagcgcctg 15841 gacctcgcgg gcagcgtcgc cctcgggttc tgattcaaga acggttgtgc cggcggcggc 15901 agtgcctggg tagctcacgc gctgcgtgat acgggactca agaatgggca gctcgtaccc 15961 ggccagcgcc tcggcaacct caccgccgat gcgcgtgcct ttgatcgccc gcgacacgac 16021 aaaggccgct tgtagccttc catccgtgac ctcaatgcgc tgcttaacca gctccaccag 16081 gtcggcggtg gcccatatgt cgtaagggct tggctgcacc ggaatcagca cgaagtcggc 16141 tgccttgatc gcggacacag ccaagtccgc cgcctggggc gctccgtcga tcactacgaa 16201 gtcgcgccgg ccgatggcct tcacgtcgcg gtcaatcgtc gggcggtcga tgccgacaac 16261 ggttagcggt tgatcttccc gcacggccgc ccaatcgcgg gcactgccct ggggatcgga 16321 atcgactaac agaacatcgg ccccggcgag ttgcagggcg cgggctagat gggttgcgat 16381 ggtcgtcttg cctgacccgc ctttctggtt aagtacagcg ataaccttca tgcgttcccc 16441 ttgcgtattt gtttatttac tcatcgcatc atatacgcag cgaccgcatg acgcaagctg 16501 ttttactcaa atacacatca cctttttaga cggcggcgct cggtttcttc agcggccaag 16561 ctggccggcc aggccgccag cttggcatca gacaaaccgg ccaggatttc atgcagccgc 16621 acggttgaga cgtgcgcggg cggctcgaac acgtacccgg ccgcgatcat ctccgcctcg 16681 atctcttcgg taatgaaaaa cggttcgtcc tggccgtcct ggtgcggttt catgcttgtt 16741 cctcttggcg ttcattctcg gcggccgcca gggcgtcggc ctcggtcaat gcgtcctcac 16801 ggaaggcacc gcgccgcctg gcctcggtgg gcgtcacttc ctcgctgcgc tcaagtgcgc 16861 ggtacagggt cgagcgatgc acgccaagca gtgcagccgc ctctttcacg gtgcggcctt 16921 cctggtcgat cagctcgcgg gcgtgcgcga tctgtgccgg ggtgagggta gggcgggggc 16981 caaacttcac gcctcgggcc ttggcggcct cgcgcccgct ccgggtgcgg tcgatgatta 17041 gggaacgctc gaactcggca atgccggcga acacggtcaa caccatgcgg ccggccggcg 17101 tggtggtgtc ggcccacggc tctgccaggc tacgcaggcc cgcgccggcc tcctggatgc 17161 gctcggcaat gtccagtagg tcgcgggtgc tgcgggccag gcggtctagc ctggtcactg 17221 tcacaacgtc gccagggcgt aggtggtcaa gcatcctggc cagctccggg cggtcgcgcc 17281 tggtgccggt gatcttctcg gaaaacagct tggtgcagcc ggccgcgtgc agttcggccc 17341 gttggttggt caagtcctgg tcgtcggtgc tgacgcgggc atagcccagc aggccagcgg 17401 cggcgctctt gttcatggcg taatgtctcc ggttctagtc gcaagtattc tactttatgc 17461 gactaaaaca cgcgacaaga aaacgccagg aaaagggcag ggcggcagcc tgtcgcgtaa 17521 cttaggactt gtgcgacatg tcgttttcag aagacggctg cactgaacgt cagaagccga 17581 ctgcactata gcagcggagg ggttggatca aagtactttg atcccgaggg gaaccctgtg 17641 gttggcatgc acatacaaat ggacgaacgg ataaacct //