LOCUS Exported 15979 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION recipient plasmid [multiplexing]; pPcUbi:Cas9, nptII. ACCESSION . VERSION . KEYWORDS pDGE3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 15979) AUTHORS Ordon J, Gantner J, Kemna J, Schwalgun L, Reschke M, Streubel J, Boch J, Stuttmann J TITLE Generation of chromosomal deletions in dicotyledonous plants employing a user-friendly genome editing toolkit. JOURNAL Plant J. 2016 Aug 31. doi: 10.1111/tpj.13319. PUBMED 27579989 REFERENCE 2 (bases 1 to 15979) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..15979 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 266..290 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" terminator 333..585 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" CDS complement(975..1769) /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(1086..1105) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 1696..1715 /label=Neo-R /note="Neomycin resistance gene, reverse primer" promoter complement(1790..1973) /function="NOS promoter" /label=NOS promoter /note="nopaline synthase promoter" promoter complement(1826..2005) /label=NOS promoter /note="nopaline synthase promoter" terminator 2099..2806 /label=OCS terminator /note="octopine synthase terminator" CDS complement(2817..2837) /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS complement(2850..6953) /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" promoter 7958..8060 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 8061..8720 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(8127..8146) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" CDS 9062..9367 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 9246..9265 /label=ccdB-fwd /note="ccdB gene, forward primer" misc_feature 9709..9733 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 11034..11663 /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" CDS 12097..13164 /codon_start=1 /product="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 RepA /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV MRYRNLIEGEASAGS" rep_origin 13230..13424 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" primer_bind complement(13660..13682) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 13768..13908 /label=bom /note="basis of mobility region from pBR322" primer_bind 13823..13842 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(13923..13940) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(14094..14682) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(14174..14193) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(14926..15717) /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" ORIGIN 1 ttatgtccac tgggttcgtg cccgaattga tcacaggcag caacgctctg tcatcgttac 61 aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc ttagttgccg 121 ttcttccgaa tagcatcggt aacatgagca aagtctgccg ccttacaacg gctctcccgc 181 tgacgccgtc ccggactgat gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc 241 ggtcggggag ctgttggctg gctggtggca ggatatattg tggtgtaaac aaattgacgc 301 ttagacaact taataacaca ttgcggaatt ccgatctagt aacatagatg acaccgcgcg 361 cgataattta tcctagtttg cgcgctatat tttgttttct atcgcgtatt aaatgtataa 421 ttgcgggact ctaatcataa aaacccatct cataaataac gtcatgcatt acatgttaat 481 tattacatgc ttaacgtaat tcaacagaaa ttatatgata atcatcgcaa gaccggcaac 541 aggattcaat cttaagaaac tttattgcca aatgtttgaa cgatcgggga tcatccgggt 601 ctgtggcggg aactccacga aaatatccga acgcagcaag atatcgcggt gcatctcggt 661 cttgcctggg cagtcgccgc cgacgccgtt gatgtggacg ccgggcccga tcatattgtc 721 gctcaggatc gtggcgttgt gcttgtcggc cgttgctgtc gtaatgatat cggcaccttc 781 gaccgcctgt tccgcagaga tcccgtgggc gaagaactcc agcatgagat ccccgcgctg 841 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc 901 ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa 961 ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa 1021 tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct 1081 tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg 1141 ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca 1201 tcgccatggg tcacgacgag atcatcgccg tcgggcatgc gcgccttgag cctggcgaac 1261 agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg 1321 gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag 1381 gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg 1441 gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag 1501 tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc 1561 agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga caggtcggtc 1621 ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag 1681 ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa 1741 cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc cagatccggt gcagattatt 1801 tggattgaga gtgaatatga gactctaatt ggataccgag gggaatttat ggaacgtcag 1861 tggagcattt ttgacaagaa atatttgcta gctgatagtg accttaggcg acttttgaac 1921 gcgcaataat ggtttctgac gtatgtgctt agctcattaa actccagaaa cccgcggctg 1981 agtggctcct tcaacgttgc ggttctgtca gttccaaacg taaaacggct tgtcccgcgt 2041 catcggcggg ggtcataacg tgactccctt aattctccgc tcatgattag tagcggtcct 2101 gctgagcctc gacatgttgt cgcaaaattc gccctggacc cgcccaacga tttgtcgtca 2161 ctgtcaaggt ttgacctgca cttcatttgg ggcccacata caccaaaaaa atgctgcata 2221 attctcgggg cagcaagtcg gttacccggc cgccgtgctg gaccgggttg aatggtgccc 2281 gtaactttcg gtagagcgga cggccaatac tcaacttcaa ggaatctcac ccatgcgcgc 2341 cggcggggaa ccggagttcc cttcagtgag cgttattagt tcgccgctcg gtgtgtcgta 2401 gatactagcc cctggggcac ttttgaaatt tgaataagat ttatgtaatc agtcttttag 2461 gtttgaccgg ttctgccgct ttttttaaaa ttggatttgt aataataaaa cgcaattgtt 2521 tgttattgtg gcgctctatc atagatgtcg ctataaacct attcagcaca atatattgtt 2581 ttcattttaa tattgtacat ataagtagta gggtacaatc agtaaattga acggagaata 2641 ttattcataa aaatacgata gtaacgggtg atatattcat tagaatgaac cgaaaccggc 2701 ggtaaggatc tgagctacac atgctcaggt tttttacaac gtgcacaaca gaattgaaag 2761 caaatatcat gcgatcatag gcttctcgca tatctcatta aagcaggaca agctcacacc 2821 ttcctcttct tcttggggtc agccctgctg tctccaccga gctgagagag gtcgattctt 2881 gtttcataga gccccgtaat tgactgatga atcagtgtgg cgtccaggac ctcctttgta 2941 gaggtgtacc gctttctgtc tatggtggtg tcgaagtact tgaaggctgc aggcgcgccc 3001 aagttggtca gagtaaacaa gtggataatg ttttctgcct gctccctgat gggcttatcc 3061 ctgtgcttat tgtaagcaga aagcacctta tcgaggttag cgtcggcgag gatcactctt 3121 ttggagaatt cgcttatttg ctcgatgatc tcatcaaggt agtgtttgtg ttgttccacg 3181 aacagctgct tctgctcatt atcttcggga gatcctttga gcttttcata gtggctggcc 3241 agatacaaga aattaacgta tttagagggc agtgccagct cgttaccttt ctgcagctcg 3301 cccgcactag cgagcattcg tttccggccg ttttcaagct caaagagaga gtacttggga 3361 agcttaatga tgaggtcttt tttgacctct ttatatcctt tcgcctcgag aaagtcgatg 3421 gggttttttt cgaagcttga tcgctccatg attgtgatgc ccagcagttc cttgacgctt 3481 ttgagttttt tagacttccc tttctccact ttggccacaa ccagtacact gtaagcgact 3541 gtaggagaat cgaatccgcc gtatttcttg gggtcccaat cttttttgcg tgcgatcagc 3601 ttgtcgctgt tccttttcgg gaggatactt tccttggaga agcctccggt ctgtacttcg 3661 gtctttttaa cgatgttcac ctgcggcatg gacaggacct tccggactgt cgcgaaatcc 3721 ctacccttgt cccacacgat ttctcctgtt tctccgtttg tttcgataag tggtcgcttc 3781 cgaatctctc cattggccag tgtaatctcg gtcttgaaaa aattcataat attgctgtaa 3841 aagaagtact tagcggtggc cttgcctatt tcctgctcag actttgcgat cattttccta 3901 acatcgtaca ctttatagtc tccgtaaaca aattcagatt caagcttggg atattttttg 3961 ataagtgcag tgcctaccac tgcattcagg taggcatcat gcgcatggtg gtaattgttg 4021 atctctctca ccttataaaa ctgaaagtcc tttctgaaat ctgaaaccag cttagacttc 4081 agagtaataa ctttcacctc tcgaatcagt ttgtcatttt catcgtactt ggtgttcatg 4141 cgtgaatcga gaatttgggc cacgtgcttg gtgatctggc gtgtctcaac aagctgcctt 4201 ttgatgaagc cggctttatc caactcagac aggccacctc gttcagcctt agtcagatta 4261 tcgaacttcc gttgtgtgat cagtttggcg ttcagcagct gccgccaata atttttcatt 4321 ttcttgacaa cttcttctga ggggacgtta tcactcttcc ctctattttt atcggatctt 4381 gtcaacactt tattatcaat agaatcatct ttgagaaaag actggggcac gatatgatcc 4441 acgtcgtagt cggagagccg attgatgtcc agttcctgat ccacgtacat gtccctgccg 4501 ttctgcaggt agtacaggta gagcttctca ttctgaagct gggtgttttc aactgggtgt 4561 tccttaagga tttgggaccc cagttctttt ataccctctt caatcctctt catcctttcc 4621 ctactgttct tctgtccctt ctgggtagtt tggttctctc gggccatctc gataacgata 4681 ttctcgggct tatgccttcc cattactttg acgagttcat ccacgacctt aacggtctgc 4741 agtattccct ttttgatagc tgggctacct gcaagattag cgatgtgctc gtggagactg 4801 tccccctggc cagaaacttg tgctttctgg atgtcctcct taaaggtgag agagtcatca 4861 tggatcaact gcatgaagtt ccggttggca aatccatcgg acttaagaaa atccaggatt 4921 gtctttccac tctgcttgtc tcggatccca ttgatcagtt ttcttgacag ccgcccccat 4981 cctgtatatc ggcgcctctt gagctgtttc atgactttgt cgtcgaagag atgagcgtaa 5041 gttttcaagc gttcttcaat catctcccta tcttcaaaca acgtaagggt gaggacaatg 5101 tcctcaagaa tgtcctcgtt ctcctcattg tccaggaagt ccttgtcttt aatgattttc 5161 aggagatcgt gatacgttcc cagggatgcg ttgaagcgat cctccactcc gctgatttca 5221 acagagtcga aacattcaat ctttttgaaa taatcttctt tgagctgttt cacggtaact 5281 ttccggttcg tcttgaagag gaggtccacg atagctttct tctgctctcc agacaggaat 5341 gctggctttc tcatcccttc tgtgacgtat ttgaccttgg tgagctcgtt ataaactgtg 5401 aagtactcgt acagcagaga gtgtttagga agcacctttt cgttaggcag atttttatca 5461 aagttagtca tcctttcgat gaaggactgg gcagaggccc ccttatccac gacttcctcg 5521 aagttccagg gagtgatagt ctcttctgat ttgcgagtca tccacgcgaa tctggaattt 5581 ccccgggcga gggggcctac atagtagggt atccgaaatg tgaggatttt ctcaatcttt 5641 tccctgttat ctttcaaaaa ggggtagaaa tcctcttgcc gcctgaggat agcgtgcagt 5701 tcgcccaggt gaatctggtg ggggatgctt ccattgtcga aagtgcgctg tttgcgcaac 5761 agatcttctc tgttaagctt taccagcagc tcctcggtgc cgtccatttt ttccaagatg 5821 ggcttaataa atttgtaaaa ttcctcctgg cttgctccgc cgtcaatgta tccggcgtag 5881 ccatttttag actgatcgaa gaaaatttcc ttgtacttct caggcagttg ctgtctgaca 5941 agggccttca gcaaagtcaa gtcttggtgg tgctcatcat agcgcttgat catactagcg 6001 ctcagcggag ctttggtgat ctccgtgttc actcgcagaa tatcactcag cagaatggcg 6061 tctgacaggt tctttgccgc caaaaaaagg tctgcgtact ggtcgccgat ctgggccagc 6121 agattgtcga gatcatcatc gtaggtgtct ttgctcagtt gaagcttggc atcttcggcc 6181 aggtcgaagt tagatttaaa gttgggggtc agcccgagtg acagggcgat aagattacca 6241 aacaggccgt tcttcttctc cccagggagc tgtgcgatga ggttttcgag ccgccgggat 6301 ttggacagcc tagcgctcag gattgctttg gcgtcaactc cggatgcgtt gatcgggttc 6361 tcttcgaaaa gctgattgta agtctgaacc agttggataa agagtttgtc gacatcgctg 6421 ttgtctgggt tcaggtcccc ctcgatgagg aagtgtcccc gaaatttgat catatgcgcc 6481 agcgcgagat agatcaaccg caagtcagcc ttatcagtac tgtctacaag cttcttcctc 6541 agatgatata tggttgggta cttttcatgg tacgccacct cgtccacgat attgccaaag 6601 attgggtggc gctcgtgctt tttatcctcc tccaccaaaa aggactcctc cagcctatgg 6661 aagaaagagt catccacctt agccatctca ttactaaaga tctcctgcag gtagcagatc 6721 cgattctttc tgcgggtata tctgcgccgt gctgttcttt tgagccgcgt ggcttcggcc 6781 gtctccccgg agtcgaacag gagggcgcca atgaggttct tctttatgct gtggcgatcg 6841 gtattgccca gaactttgaa ttttttgctc ggcaccttgt actcgtccgt aatgacggcc 6901 cagccgacgc tgtttgtgcc gatatcgagc ccaatggagt acttcttgtc cattgctgca 6961 catacataac atatcaagat cagaacacac atatacacac acaaatacaa tcaagtcaac 7021 aactccaaaa agtccagatc tacatatata catacgtaaa taacaaaatc atgtaaataa 7081 tcacaatcat gtaatccaga tctatgcaca tatatatata cacaattaat aaaaaaaatg 7141 atataacaga tctatatcta tgtatgtaac aacacaatca gatgagagaa gtgatgtttt 7201 cagatctgta tacatacaaa cacaaacaga tgaacaattg atacgtagat ccatatgtat 7261 acgtacaatt agctacacga ttaaatgaaa aaaatcaacg atttcggatt ggtacacaca 7321 aacgcaacaa tatgaagaaa ttcatatctg attagatata aacataacca cgtgtagata 7381 cacagtcaaa tcaacaaatt tatagcttct aaacggatga gatgaacaag ataaagatat 7441 tcacataagg catacataag ataagcagat taacaaacta gcaataatac atacctaatt 7501 aaaacaagga ataacagaga gagagagaga gagagagatt taccttgaaa atgaagagga 7561 gaagagagga tttcttaaaa ttgggggtag agaaagaaag atgatgaatt gtgagaaagg 7621 agagatagaa gggggggttg tatatatagg ctgtagaaga ttatttttgt gtttgaggcg 7681 gtgaaggaag aggggatctg actatgacac gtttgcggtt acgtatttcg ataggagtct 7741 ttcaacgctt aacgccgtta ctctatatga ccgtttgggc cgtaacgggg ccgtttgtta 7801 acgctgatgt tgattctttt ctttctttct ttcttccttt tttaaagaag caattgtaca 7861 atcgttgcta gctgtcaaac ggataattcg gatacggata tgcctatatt catatccgta 7921 atctccaagc ttgaattctt gcagagacct gagacgttga tcggcacgta agaggttcca 7981 actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt 8041 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat 8101 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata 8161 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca 8221 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc 8281 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg 8341 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc 8401 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt 8461 tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca 8521 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg 8581 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg 8641 ccgtctgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg 8701 agtggcaggg cggggcgtaa tcgcgtggat ccggcttact aaaagccaga taacagtatg 8761 cgtatttgcg cgctgatttt tgcggtataa gaatatatac tgatatgtat acccgaagta 8821 tgtcaaaaag aggtatgcta tgaagcagcg tattacagtg acagttgaca gcgacagcta 8881 tcagttgctc aaggcatata tgatgtcaat atctccggtc tggtaagcac aaccatgcag 8941 aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga agggatggct 9001 gaggtcgccc ggtttattga aatgaacggc tcttttgctg acgagaacag gggctggtga 9061 aatgcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt 9121 acagagtgat attattgaca cgcccgggcg acggatggtg atccccctgg ccagtgcacg 9181 tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag 9241 ctggcgcatg atgaccaccg atatggccag tgtgccggtt tccgttatcg gggaagaagt 9301 ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg 9361 aatataaatg tcaggctccc ttatacacag ccagtcggtc tcagcttcca tgctagagtc 9421 cgcaaaaatc accagtctct ctctacaaat ctatctctct ctatttttct ccagaataat 9481 gtgtgagtag ttcccagata agggaattag ggttcttata gggtttcgct catgtgttga 9541 gcatataaga aacccttagt atgtatttgt atttgtaaaa tacttctatc aataaaattt 9601 ctaattccta aaaccaaaat ccagtgacct gcaggcatgc gacgtcgagc ttagcttgag 9661 cttggatcag attgtcgttt cccgccttca gtttaaacta tcagtgtttg acaggatata 9721 ttggcgggta aacctaagag aaaagagcgt ttattagaat aatcggatat ttaaaagggc 9781 gtgaaaaggt ttatccgttc gtccatttgt atgtgcatgc caaccacagg gttcccctcg 9841 ggatcaaagt actttgatcc aacccctccg ctgctatagt gcagtcggct tctgacgttc 9901 agtgcagccg tgttctgaaa acgacatgtc gcacaagtcc taagttacgc gacaggctgc 9961 cgccctgccc ttttcctggc gttttcttgt cgcgtgtttt agtcgcataa agtagaatac 10021 ttgcgactag aaccggagac attacgccat gaacaagagc gccgccgctg gcctgctggg 10081 ctatgcccgc gtcagcaccg acgaccagga cttgaccaac caacgggccg aactgcacgc 10141 ggccggctgc accaagctgt tttccgagaa gatcaccggc accaggcgcg accgcccgga 10201 gctggccagg atgcttgacc acctacgccc tggcgacgtt gtgacagtga ccaggctaga 10261 ccgcctggcc cgcagcaccc gcgacctact ggacattgcc gagcgcatcc aggaggccgg 10321 cgcgggcctg cgtagcctgg cagagccgtg ggccgacacc accacgccgg ccggccgcat 10381 ggtgttgacc gtgttcgccg gcattgccga gttcgagcgt tccctaatca tcgaccgcac 10441 ccggagcggg cgcgaggccg ccaaggcccg aggcgtgaag tttggccccc gccctaccct 10501 caccccggca cagatcgcgc acgcccgcga gctgatcgac caggaaggcc gcaccgtgaa 10561 agaggcggct gcactgcttg gcgtgcatcg ctcgaccctg taccgcgcac ttgagcgcag 10621 cgaggaagtg acgcccaccg aggccaggcg gcgcggtgcc ttccgtgagg acgcattgac 10681 cgaggccgac gccctggcgg ccgccgagaa tgaacgccaa gaggaacaag catgaaaccg 10741 caccaggacg gccaggacga accgtttttc attaccgaag agatcgaggc ggagatgatc 10801 gcggccgggt acgtgttcga gccgcccgcg cacgtctcaa ccgtgcggct gcatgaaatc 10861 ctggccggtt tgtctgatgc caagctggcg gcctggccgg ccagcttggc cgctgaagaa 10921 accgagcgcc gccgtctaaa aaggtgatgt gtatttgagt aaaacagctt gcgtcatgcg 10981 gtcgctgcgt atatgatgcg atgagtaaat aaacaaatac gcaaggggaa cgcatgaagg 11041 ttatcgctgt acttaaccag aaaggcgggt caggcaagac gaccatcgca acccatctag 11101 cccgcgccct gcaactcgcc ggggccgatg ttctgttagt cgattccgat ccccagggca 11161 gtgcccgcga ttgggcggcc gtgcgggaag atcaaccgct aaccgttgtc ggcatcgacc 11221 gcccgacgat tgaccgcgac gtgaaggcca tcggccggcg cgacttcgta gtgatcgacg 11281 gagcgcccca ggcggcggac ttggctgtgt ccgcgatcaa ggcagccgac ttcgtgctga 11341 ttccggtgca gccaagccct tacgacatat gggccaccgc cgacctggtg gagctggtta 11401 agcagcgcat tgaggtcacg gatggaaggc tacaagcggc ctttgtcgtg tcgcgggcga 11461 tcaaaggcac gcgcatcggc ggtgaggttg ccgaggcgct ggccgggtac gagctgccca 11521 ttcttgagtc ccgtatcacg cagcgcgtga gctacccagg cactgccgcc gccggcacaa 11581 ccgttcttga atcagaaccc gagggcgacg ctgcccgcga ggtccaggcg ctggccgctg 11641 aaattaaatc aaaactcatt tgagttaatg aggtaaagag aaaatgagca aaagcacaaa 11701 cacgctaagt gccggccgtc cgagcgcacg cagcagcaag gctgcaacgt tggccagcct 11761 ggcagacacg ccagccatga agcgggtcaa ctttcagttg ccggcggagg atcacaccaa 11821 gctgaagatg tacgcggtac gccaaggcaa gaccattacc gagctgctat ctgaatacat 11881 cgcgcagcta ccagagtaaa tgagcaaatg aataaatgag tagatgaatt ttagcggcta 11941 aaggaggcgg catggaaaat caagaacaac caggcaccga cgccgtggaa tgccccatgt 12001 gtggaggaac gggcggttgg ccaggcgtaa gcggctgggt tgtctgccgg ccctgcaatg 12061 gcactggaac ccccaagccc gaggaatcgg cgtgacggtc gcaaaccatc cggcccggta 12121 caaatcggcg cggcgctggg tgatgacctg gtggagaagt tgaaggccgc gcaggccgcc 12181 cagcggcaac gcatcgaggc agaagcacgc cccggtgaat cgtggcaagc ggccgctgat 12241 cgaatccgca aagaatcccg gcaaccgccg gcagccggtg cgccgtcgat taggaagccg 12301 cccaagggcg acgagcaacc agattttttc gttccgatgc tctatgacgt gggcacccgc 12361 gatagtcgca gcatcatgga cgtggccgtt ttccgtctgt cgaagcgtga ccgacgagct 12421 ggcgaggtga tccgctacga gcttccagac gggcacgtag aggtttccgc agggccggcc 12481 ggcatggcca gtgtgtggga ttacgacctg gtactgatgg cggtttccca tctaaccgaa 12541 tccatgaacc gataccggga agggaaggga gacaagcccg gccgcgtgtt ccgtccacac 12601 gttgcggacg tactcaagtt ctgccggcga gccgatggcg gaaagcagaa agacgacctg 12661 gtagaaacct gcattcggtt aaacaccacg cacgttgcca tgcagcgtac gaagaaggcc 12721 aagaacggcc gcctggtgac ggtatccgag ggtgaagcct tgattagccg ctacaagatc 12781 gtaaagagcg aaaccgggcg gccggagtac atcgagatcg agctagctga ttggatgtac 12841 cgcgagatca cagaaggcaa gaacccggac gtgctgacgg ttcaccccga ttactttttg 12901 atcgatcccg gcatcggccg ttttctctac cgcctggcac gccgcgccgc aggcaaggca 12961 gaagccagat ggttgttcaa gacgatctac gaacgcagtg gcagcgccgg agagttcaag 13021 aagttctgtt tcaccgtgcg caagctgatc gggtcaaatg acctgccgga gtacgatttg 13081 aaggaggagg cggggcaggc tggcccgatc ctagtcatgc gctaccgcaa cctgatcgag 13141 ggcgaagcat ccgccggttc ctaatgtacg gagcagatgc tagggcaaat tgccctagca 13201 ggggaaaaag gtcgaaaagg tgtctttcct gtggatagca cgtacattgg gaacccaaag 13261 ccgtacattg ggaaccggaa cccgtacatt gggaacccaa agccgtacat tgggaaccgg 13321 tcacacatgt aagtgactga tataaaagag aaaaaaggcg atttttccgc ctaaaactct 13381 ttaaaactta ttaaaactct taaaacccgc ctggcctgtg cataactgtc tggccagcgc 13441 acagccgaag agctgcaaaa agcgcctacc cttcggtcgc tgcgctccct acgccccgcc 13501 gcttcgcgtc ggcctatcgc ggccgctggc cgctcaaaaa tggctggcct acggccaggc 13561 aatctaccag ggcgcggaca agccgcgccg tcgccactcg accgccggcg cccacatcaa 13621 ggcaccctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc 13681 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc 13741 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg 13801 agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg 13861 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct 13921 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 13981 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 14041 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 14101 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 14161 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 14221 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 14281 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 14341 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 14401 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 14461 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 14521 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 14581 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 14641 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 14701 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgcat 14761 gatatatctc ccaatttgtg tagggcttat tatgcacgct taaaaataat aaaagcagac 14821 ttgacctgat agtttggctg tgagcaatta tgtgcttagt gcatctaatc gcttgagtta 14881 acgccggcga agcggcgtcg gcttgaacga atttctagct agacattatt tgccgactac 14941 cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc 15001 caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata 15061 ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc 15121 ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc 15181 ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg 15241 ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc 15301 tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc 15361 tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg 15421 ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct 15481 ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc 15541 atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc 15601 atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat 15661 gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt cccccatgat 15721 gtttaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tccataacat 15781 caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac 15841 cccaaaaaaa catgtcataa caagaagcca tgaaaaccgc cactgcgccg ttaccaccgc 15901 tgcgttcggt caaggttctg gaccagttgc gtgacggcag ttacgctact tgcattacag 15961 cttacgaacc gaacgaggc //