pTX172

LOCUS       pTX172              15416 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Express STU (Single Transcriptional Unit) Cas9 with Maize Ubiquitin1
            promoter in plants.
ACCESSION   .
VERSION     .
KEYWORDS    pTX172
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15416)
  AUTHORS   Tang X, Zheng X, Qi Y, Zhang D, Cheng Y, Tang A, Voytas DF, Zhang Y
  TITLE     A Single Transcript CRISPR-Cas9 System for Efficient Genome Editing 
            in Plants.
  JOURNAL   Mol Plant. 2016 Jul 6;9(7):1088-91. doi: 10.1016/j.molp.2016.05.001.
            Epub 2016 May 19.
  PUBMED    27212389
REFERENCE   2  (bases 1 to 15416)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..15416
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        49..2045
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             2070..6173
                     /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             6186..6206
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(6289..6594)
                     /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     complement(6391..6410)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_RNA        6921..6996
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    7073..7097
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             8398..9027
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             9456..10529
                     /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"
     rep_origin      10595..10789
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(11025..11047)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    11123..11263
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     11178..11197
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(11278..11295)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11449..12037)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11529..11548)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(12124..12918)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    13343..13367
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    13445..13619
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        complement(14709..15385)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     complement(14752..14772)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
ORIGIN
        1 tgatattctt ggagtagacg agagtgtcgt gctccaccat gggcgcgcct gcagtgcagc
       61 gtgacccggt cgtgcccctc tcttgagata aggtgagcat tgcatgtcta agttataaaa
      121 aattaccaca tatttttttt gtcacacttg tttgaagtgc agtttatcta tctttataca
      181 tatatttaaa ctttactcta cgaataatat aatctatagt actacaataa tatcagtgtt
      241 ttagagaatc atataaatga acagttagac atggtctaaa ggacaattga gtattttgac
      301 aacaggactc tacagtttta tctttttagt gtgcatgtgt tctccttttt ttttgcaaat
      361 agcttcacct atataatact tcatccattt tattagtaca tccatttagg gtttagggtt
      421 aatggttttt atagactaat ttttttagta catctatttt attctatttt agcctctaaa
      481 ttaagaaaac taaaactcta ttttagtttt tttatttaat aatttagata taaaatagaa
      541 taaaataaag tgactaaaaa ttaaacaaat accctttaag aaattaaaaa aactaaggaa
      601 acatttttct tgtttcgagt agataatgcc agcctgttaa acgccgtcga cgagtctaac
      661 ggacaccaac cagcgaacca gcagcgtcgc gtcgggccaa gcgaagcaga cggcacggca
      721 tctctgtcgc tgcctctgga cccctctcga gagttccgct ccaccgttgg acttgctccg
      781 ctgtcggcat ccagaaattg cgtggcggag cggcagacgt gagccggcac ggcaggcggc
      841 ctcctcctcc tctcacggca ccggcagcta cgggggattc ctttcccacc gctccttcgc
      901 tttcccttcc tcgcccgccg taataaatag acaccccctc cacaccctct ttccccaacc
      961 tcgtgttgtt cggagcgcac acacacacaa ccagaactcc cccaaatcca cccgtcggca
     1021 cctccgcttc aaggtacgcc gctcgtcctc cccccccccc tctctacctt ctcaagatcg
     1081 gcgttccggt ccatggttag ggcccggtag ttctacttct gttcatgttt gtgttagatc
     1141 cgtgtttgtg ttagatccgt gctactagcg ttcgtacacg gatgcgacct gtacgtcaga
     1201 cacgttctga ttgctaactt gccagtgttt ctctttgggg aatcctggga tggctctagc
     1261 cgttccgcag acgggatcga tttcatgatt ttttttgttt cgttgcatag ggtttggttt
     1321 gcccttttcc tttatttcaa tatatgccgt gcacttgttt gtcgggtcat cttttcatgc
     1381 ttttttttgt cttggttgtg atgatgtggt ctggttgggc ggtcgttcaa gatcggagta
     1441 gaattaattc tgtttcaaac tacctggtgg atttattaat tttggatctg tatgtgtgtg
     1501 ccatacatat tcatagttac gaattgaaga tgatggatgg aaatatcgat ctaggatagg
     1561 tatacatgtt gatgcgggtt ttactgatgc atatacagag atgctttttg ttcgcttggt
     1621 tgtgatgatg tggtgtggtt gggcggtcgt tcattcgttc aagatcggag tagaatactg
     1681 tttcaaacta cctggtgtat ttattaattt tggaactgta tgtgtgtgtc atacatcttc
     1741 atagttacga gtttaagatg gatggaaata tcgatctagg ataggtatac atgttgatgt
     1801 gggttttact gatgcatata catgatggca tatgcagcat ctattcatat gctctaacct
     1861 tgagtaccta tctattataa taaacaagta tgttttataa ttattttgat cttgatatac
     1921 ttggatgatg gcatatgcag cagctatatg tggatttttt tagccctgcc ttcatacgct
     1981 atttatttgc ttggtactgt ttcttttgtc gatgctcacc ctgttgtttg gtgttacttc
     2041 tgcagcctgc aggtagatcg ctcgtcgaca tggataagaa gtactctatc ggactcgata
     2101 tcggaactaa ctctgtggga tgggctgtga tcaccgatga gtacaaggtg ccatctaaga
     2161 agttcaaggt tctcggaaac accgataggc actctatcaa gaaaaacctt atcggtgctc
     2221 tcctcttcga ttctggtgaa actgctgagg ctaccagact caagagaacc gctagaagaa
     2281 ggtacaccag aagaaagaac aggatctgct acctccaaga gatcttctct aacgagatgg
     2341 ctaaagtgga tgattcattc ttccacaggc tcgaagagtc attcctcgtg gaagaagata
     2401 agaagcacga gaggcaccct atcttcggaa acatcgttga tgaggtggca taccacgaga
     2461 agtaccctac tatctaccac ctcagaaaga agctcgttga ttctactgat aaggctgatc
     2521 tcaggctcat ctacctcgct ctcgctcaca tgatcaagtt cagaggacac ttcctcatcg
     2581 agggtgatct caaccctgat aactctgatg tggataagtt gttcatccag ctcgtgcaga
     2641 cctacaacca gcttttcgaa gagaacccta tcaacgcttc aggtgtggat gctaaggcta
     2701 tcctctctgc taggctctct aagtcaagaa ggcttgagaa cctcattgct cagctccctg
     2761 gtgagaagaa gaacggactt ttcggaaact tgatcgctct ctctctcgga ctcaccccta
     2821 acttcaagtc taacttcgat ctcgctgagg atgcaaagct ccagctctca aaggatacct
     2881 acgatgatga tctcgataac ctcctcgctc agatcggaga tcagtacgct gatttgttcc
     2941 tcgctgctaa gaacctctct gatgctatcc tcctcagtga tatcctcaga gtgaacaccg
     3001 agatcaccaa ggctccactc tcagcttcta tgatcaagag atacgatgag caccaccagg
     3061 atctcacact tctcaaggct cttgttagac agcagctccc agagaagtac aaagagattt
     3121 tcttcgatca gtctaagaac ggatacgctg gttacatcga tggtggtgca tctcaagaag
     3181 agttctacaa gttcatcaag cctatcctcg agaagatgga tggaaccgag gaactcctcg
     3241 tgaagctcaa tagagaggat cttctcagaa agcagaggac cttcgataac ggatctatcc
     3301 ctcatcagat ccacctcgga gagttgcacg ctatccttag aaggcaagag gatttctacc
     3361 cattcctcaa ggataacagg gaaaagattg agaagattct caccttcaga atcccttact
     3421 acgtgggacc tctcgctaga ggaaactcaa gattcgcttg gatgaccaga aagtctgagg
     3481 aaaccatcac cccttggaac ttcgaagagg tggtggataa gggtgctagt gctcagtctt
     3541 tcatcgagag gatgaccaac ttcgataaga accttccaaa cgagaaggtg ctccctaagc
     3601 actctttgct ctacgagtac ttcaccgtgt acaacgagtt gaccaaggtt aagtacgtga
     3661 ccgagggaat gaggaagcct gcttttttgt caggtgagca aaagaaggct atcgttgatc
     3721 tcttgttcaa gaccaacaga aaggtgaccg tgaagcagct caaagaggat tacttcaaga
     3781 aaatcgagtg cttcgattca gttgagattt ctggtgttga ggataggttc aacgcatctc
     3841 tcggaaccta ccacgatctc ctcaagatca ttaaggataa ggatttcttg gataacgagg
     3901 aaaacgagga tatcttggag gatatcgttc ttaccctcac cctctttgaa gatagagaga
     3961 tgattgaaga aaggctcaag acctacgctc atctcttcga tgataaggtg atgaagcagt
     4021 tgaagagaag aagatacact ggttggggaa ggctctcaag aaagctcatt aacggaatca
     4081 gggataagca gtctggaaag acaatccttg atttcctcaa gtctgatgga ttcgctaaca
     4141 gaaacttcat gcagctcatc cacgatgatt ctctcacctt taaagaggat atccagaagg
     4201 ctcaggtttc aggacagggt gatagtctcc atgagcatat cgctaacctc gctggatctc
     4261 ctgcaatcaa gaagggaatc ctccagactg tgaaggttgt ggatgagttg gtgaaggtga
     4321 tgggaaggca taagcctgag aacatcgtga tcgaaatggc tagagagaac cagaccactc
     4381 agaagggaca gaagaactct agggaaagga tgaagaggat cgaggaaggt atcaaagagc
     4441 ttggatctca gatcctcaaa gagcaccctg ttgagaacac tcagctccag aatgagaagc
     4501 tctacctcta ctacctccag aacggaaggg atatgtatgt ggatcaagag ttggatatca
     4561 acaggctctc tgattacgat gttgatcata tcgtgccaca gtcattcttg aaggatgatt
     4621 ctatcgataa caaggtgctc accaggtctg ataagaacag gggtaagagt gataacgtgc
     4681 caagtgaaga ggttgtgaag aaaatgaaga actattggag gcagctcctc aacgctaagc
     4741 tcatcactca gagaaagttc gataacttga ctaaggctga gaggggagga ctctctgaat
     4801 tggataaggc aggattcatc aagaggcagc ttgtggaaac caggcagatc actaagcacg
     4861 ttgcacagat cctcgattct aggatgaaca ccaagtacga tgagaacgat aagttgatca
     4921 gggaagtgaa ggttatcacc ctcaagtcaa agctcgtgtc tgatttcaga aaggatttcc
     4981 aattctacaa ggtgagggaa atcaacaact accaccacgc tcacgatgct taccttaacg
     5041 ctgttgttgg aaccgctctc atcaagaagt atcctaagct cgagtcagag ttcgtgtacg
     5101 gtgattacaa ggtgtacgat gtgaggaaga tgatcgctaa gtctgagcaa gagatcggaa
     5161 aggctaccgc taagtatttc ttctactcta acatcatgaa tttcttcaag accgagatta
     5221 ccctcgctaa cggtgagatc agaaagaggc cactcatcga gacaaacggt gaaacaggtg
     5281 agatcgtgtg ggataaggga agggatttcg ctaccgttag aaaggtgctc tctatgccac
     5341 aggtgaacat cgttaagaaa accgaggtgc agaccggtgg attctctaaa gagtctatcc
     5401 tccctaagag gaactctgat aagctcattg ctaggaagaa ggattgggac cctaagaaat
     5461 acggtggttt cgattctcct accgtggctt actctgttct cgttgtggct aaggttgaga
     5521 agggaaagag taagaagctc aagtctgtta aggaacttct cggaatcact atcatggaaa
     5581 ggtcatcttt cgagaagaac ccaatcgatt tcctcgaggc taagggatac aaagaggtta
     5641 agaaggatct catcatcaag ctcccaaagt actcactctt cgaactcgag aacggtagaa
     5701 agaggatgct cgcttctgct ggtgagcttc aaaagggaaa cgagcttgct ctcccatcta
     5761 agtacgttaa ctttctttac ctcgcttctc actacgagaa gttgaaggga tctccagaag
     5821 ataacgagca gaagcaactt ttcgttgagc agcacaagca ctacttggat gagatcatcg
     5881 agcagatctc tgagttctct aaaagggtga tcctcgctga tgcaaacctc gataaggtgt
     5941 tgtctgctta caacaagcac agagataagc ctatcaggga acaggcagag aacatcatcc
     6001 atctcttcac ccttaccaac ctcggtgctc ctgctgcttt caagtacttc gatacaacca
     6061 tcgataggaa gagatacacc tctaccaaag aagtgctcga tgctaccctc atccatcagt
     6121 ctatcactgg actctacgag actaggatcg atctctcaca gctcggtggt gattcaaggg
     6181 ctgatcctaa gaagaagagg aaggtttgag gatctaaaaa aaaaaaaaaa aaaaaaaaaa
     6241 aaaaaaaaaa aaaaaaaaaa aaaaaggctt ttgtcaccgg aggagacctt atattcccca
     6301 gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg tggctgagat cagccacttc
     6361 ttccccgata acggaaaccg gcacactggc catatcggtg gtcatcatgc gccagctttc
     6421 atccccgata tgcaccaccg ggtaaagttc acgggagact ttatctgaca gcagacgtgc
     6481 actggccagg gggatcacca tccgtcgccc gggcgtgtca ataatatcac tctgtacatc
     6541 cacaaacaga cgataacggc tctctctttt ataggtgtaa accttaaact gcatttcacc
     6601 agcccctgtt ctcgtcagca aaagagccgt tcatttcaat aaaccgggcg acctcagcca
     6661 tcccttcctg attttccgct ttccagcgtt cggcacgcag acgacgggct tcattctgca
     6721 tggttgtgct taccagaccg gagatattga catcatatat gccttgagca actgatagct
     6781 gtcgctgtca actgtcactg taatacgctg cttcatagca tacctctttt tgacatactt
     6841 cgggtataca tatcagtata tattcttata ccgcaaaaat cagcgcgcaa atacgcatac
     6901 tgttatctgg cttggtctca gttttagagc tagaaatagc aagttaaaat aaggctagtc
     6961 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt gtcaccggat gtgttttccg
     7021 gtctgatgag tccgtgagga cgaaacaaaa gcgagctcaa actatcagtg tttgacagga
     7081 tatattggcg ggtaaaccta agagaaaaga gcgtttatta gaataatcgg atatttaaaa
     7141 gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc atgccaacca cagggttccc
     7201 ctcgggatca aagtactttg atccaacccc tccgctgcta tagtgcagtc ggcttctgac
     7261 gttcagtgca gccgtcttct gaaaacgaca tgtcgcacaa gtcctaagtt acgcgacagg
     7321 ctgccgccct gcccttttcc tggcgttttc ttgtcgcgtg ttttagtcgc ataaagtaga
     7381 atacttgcga ctagaaccgg agacattacg ccatgaacaa gagcgccgcc gctggcctgc
     7441 tgggctatgc ccgcgtcagc accgacgacc aggacttgac caaccaacgg gccgaactgc
     7501 acgcggccgg ctgcaccaag ctgttttccg agaagatcac cggcaccagg cgcgaccgcc
     7561 cggagctggc caggatgctt gaccacctac gccctggcga cgttgtgaca gtgaccaggc
     7621 tagaccgcct ggcccgcagc acccgcgacc tactggacat tgccgagcgc atccaggagg
     7681 ccggcgcggg cctgcgtagc ctggcagagc cgtgggccga caccaccacg ccggccggcc
     7741 gcatggtgtt gaccgtgttc gccggcattg ccgagttcga gcgttcccta atcatcgacc
     7801 gcacccggag cgggcgcgag gccgccaagg cccgaggcgt gaagtttggc ccccgcccta
     7861 ccctcacccc ggcacagatc gcgcacgccc gcgagctgat cgaccaggaa ggccgcaccg
     7921 tgaaagaggc ggctgcactg cttggcgtgc atcgctcgac cctgtaccgc gcacttgagc
     7981 gcagcgagga agtgacgccc accgaggcca ggcggcgcgg tgccttccgt gaggacgcat
     8041 tgaccgaggc cgacgccctg gcggccgccg agaatgaacg ccaagaggaa caagcatgaa
     8101 accgcaccag gacggccagg acgaaccgtt tttcattacc gaagagatcg aggcggagat
     8161 gatcgcggcc gggtacgtgt tcgagccgcc cgcgcacggc tcaaccgtgc ggctgcatga
     8221 aatcctggcc ggtttgtctg atgccaagct ggcggcctgg ccggccagct tggccgctga
     8281 agaaaccgag cgccgccgtc taaaaaggtg atgtgtattt gagtaaaaca gcttgcgtca
     8341 tgcggtcgct gcgtatatga tgcgatgagt aaataaacaa atacgcaagg ggaacgcatg
     8401 aaggttatcg ctgtacttaa ccagaaaggc gggtcaggca agacgaccat cgcaacccat
     8461 ctagcccgcg ccctgcaact cgccggggcc gatgttctgt tagtcgattc cgatccccag
     8521 ggcagtgccc gcgattgggc ggccgtgcgg gaagatcaac cgctaaccgt tgtcggcatc
     8581 gaccgcccga cgattgaccg cgacgtgaag gccatcggcc ggcgcgactt cgtagtgatc
     8641 gacggagcgc cccaggcggc ggacttggct gtgtccgcga tcaaggcagc cgacttcgtg
     8701 ctgattccgg tgcagccaag cccttacgac atatgggcca ccgccgacct ggtggagctg
     8761 gttaagcagc gcattgaggt cacggatgga aggctacaag cggcctttgt cgtgtcgcgg
     8821 gcgatcaaag gcacgcgcat cggcggtgag gttgccgagg cgctggccgg gtacgagctg
     8881 cccattcttg agtcccgtat cacgcagcgc gtgagctacc caggcactgc cgccgccggc
     8941 acaaccgttc ttgaatcaga acccgagggc gacgctgccc gcgaggtcca ggcgctggcc
     9001 gctgaaatta aatcaaaact catttgagtt aatgaggtaa agagaaaatg agcaaaagca
     9061 caaacacgct aagtgccggc cgtccgagcg cacgcagcag caaggctgca acgttggcca
     9121 gcctggcaga cacgccagcc atgaagcggg tcaactttca gttgccggcg gaggatcaca
     9181 ccaagctgaa gatgtacgcg gtacgccaag gcaagaccat taccgagctg ctatctgaat
     9241 acatcgcgca gctaccagag taaatgagca aatgaataaa tgagtagatg aattttagcg
     9301 gctaaaggag gcggcatgga aaatcaagaa caaccaggca ccgacgccgt ggaatgcccc
     9361 atgtgtggag gaacgggcgg ttggccaggc gtaagcggct gggttgcctg ccggccctgc
     9421 aatggcactg gaacccccaa gcccgaggaa tcggcgtgag cggtcgcaaa ccatccggcc
     9481 cggtacaaat cggcgcggcg ctgggtgatg acctggtgga gaagttgaag gccgcgcagg
     9541 ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg tgaatcgtgg caagcggccg
     9601 ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc cggtgcgccg tcgattagga
     9661 agccgcccaa gggcgacgag caaccagatt ttttcgttcc gatgctctat gacgtgggca
     9721 cccgcgatag tcgcagcatc atggacgtgg ccgttttccg tctgtcgaag cgtgaccgac
     9781 gagctggcga ggtgatccgc tacgagcttc cagacgggca cgtagaggtt tccgcagggc
     9841 cggccggcat ggccagtgtg tgggattacg acctggtact gatggcggtt tcccatctaa
     9901 ccgaatccat gaaccgatac cgggaaggga agggagacaa gcccggccgc gtgttccgtc
     9961 cacacgttgc ggacgtactc aagttctgcc ggcgagccga tggcggaaag cagaaagacg
    10021 acctggtaga aacctgcatt cggttaaaca ccacgcacgt tgccatgcag cgtacgaaga
    10081 aggccaagaa cggccgcctg gtgacggtat ccgagggtga agccttgatt agccgctaca
    10141 agatcgtaaa gagcgaaacc gggcggccgg agtacatcga gatcgagcta gctgattgga
    10201 tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct gacggttcac cccgattact
    10261 ttttgatcga tcccggcatc ggccgttttc tctaccgcct ggcacgccgc gccgcaggca
    10321 aggcagaagc cagatggttg ttcaagacga tctacgaacg cagtggcagc gccggagagt
    10381 tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc aaatgacctg ccggagtacg
    10441 atttgaagga ggaggcgggg caggctggcc cgatcctagt catgcgctac cgcaacctga
    10501 tcgagggcga agcatccgcc ggttcctaat gtacggagca gatgctaggg caaattgccc
    10561 tagcagggga aaaaggtcga aaaggcctct ttcctgtgga tagcacgtac attgggaacc
    10621 caaagccgta cattgggaac cggaacccgt acattgggaa cccaaagccg tacattggga
    10681 accggtcaca catgtaagtg actgatataa aagagaaaaa aggcgatttt tccgcctaaa
    10741 actctttaaa acttattaaa actcttaaaa cccgcctggc ctgtgcataa ctgtctggcc
    10801 agcgcacagc cgaagagctg caaaaagcgc ctacccttcg gtcgctgcgc tccctacgcc
    10861 ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc aaaaatggct ggcctacggc
    10921 caggcaatct accagggcgc ggacaagccg cgccgtcgcc actcgaccgc cggcgcccac
    10981 atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
    11041 ctcccggaaa cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcgcgtcag
    11101 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt
    11161 atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
    11221 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc
    11281 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
    11341 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
    11401 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
    11461 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
    11521 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
    11581 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
    11641 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
    11701 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
    11761 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
    11821 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
    11881 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
    11941 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
    12001 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
    12061 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgcattctag
    12121 gtactaaaac aattcatcca gtaaaatata atattttatt ttctcccaat caggcttgat
    12181 ccccagtaag tcaaaaaata gctcgacata ctgttcttcc ccgatatcct ccctgatcga
    12241 ccggacgcag aaggcaatgt cataccactt gtccgccctg ccgcttctcc caagatcaat
    12301 aaagccactt actttgccat ctttcacaaa gatgttgctg tctcccaggt cgccgtggga
    12361 aaagacaagt tcctcttcgg gcttttccgt ctttaaaaaa tcatacagct cgcgcggatc
    12421 tttaaatgga gtgtcttctt cccagttttc gcaatccaca tcggccagat cgttattcag
    12481 taagtaatcc aattcggcta agcggctgtc taagctattc gtatagggac aatccgatat
    12541 gtcgatggag tgaaagagcc tgatgcactc cgcatacagc tcgataatct tttcagggct
    12601 ttgttcatct tcatactctt ccgagcaaag gacgccatcg gcctcactca tgagcagatt
    12661 gctccagcca tcatgccgtt caaagtgcag gacctttgga acaggcagct ttccttccag
    12721 ccatagcatc atgtcctttt cccgttccac atcataggtg gtccctttat accggctgtc
    12781 cgtcattttt aaatataggt tttcattttc tcccaccagc ttatatacct tagcaggaga
    12841 cattccttcc gtatctttta cgcagcggta tttttcgatc agttttttca attccggtga
    12901 tattctcatt ttagccattt attatttcct tcctcttttc tacagtattt aaagataccc
    12961 caagaagcta attataacaa gacgaactcc aattcactgt tccttgcatt ctaaaacctt
    13021 aaataccaga aaacagcttt ttcaaagttg ttttcaaagt tggcgtataa catagtatcg
    13081 acggagccga ttttgaaacc gcggtgatca caggcagcaa cgctctgtca tcgttacaat
    13141 caacatgcta ccctccgcga gatcatccgt gtttcaaacc cggcagctta gttgccgttc
    13201 ttccgaatag catcggtaac atgagcaaag tctgccgcct tacaacggct ctcccgctga
    13261 cgccgtcccg gactgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt
    13321 cggggagctg ttggctggct ggtggcagga tatattgtgg tgtaaacaaa ttgacgctta
    13381 gacaacttaa taacacattg cggacgtttt taatgtactg aattaacgcc gaattaattc
    13441 gggggatctg gattttagta ctggattttg gttttaggaa ttagaaattt tattgataga
    13501 agtattttac aaatacaaat acatactaag ggtttcttat atgctcaaca catgagcgaa
    13561 accctatagg aaccctaatt cccttatctg ggaactactc acacattatt atggagaaaa
    13621 tagagcttgt cgatcgttaa ttaatcacct ccaagatccc ggacgagtgc tggggcgtcg
    13681 gtttccacta tcggcgagta cttctacaca gccatcggtc cagacggccg cgcttctgcg
    13741 ggcgatttgt gtacgcccga cagtcccggc tccggatcgg acgattgcgt cgcatcgacc
    13801 ctgcgcccaa gctgcatcat cgaaattgcc gtcaaccaag ctctgataga gttggtcaag
    13861 accaatgcgg agcatatacg cccggagccg cggcgatcct gcaagctccg gatgcctccg
    13921 ctcgaagtag cgcgtctgct gctccataca agccaaccac ggcctccaga agaagatgtt
    13981 ggcgacctcg tattgggaat ccccgaacat cgcctcgctc cagtcaatga ccgctgttat
    14041 gcggccattg tccgtcagga cattgttgga gccgaaatcc gcgtgcacga ggtgccggac
    14101 ttcggggcag tcctcggccc aaagcatcag ctcatcgaga gcctgcgcga cggacgcact
    14161 gacggtgtcg tccatcacag tttgccagtg atacacatgg ggatcagcaa tcgcgcatat
    14221 gaaatcacgc catgtagtgt attgaccgat tccttgcggt ccgaatgggc cgaacccgct
    14281 cgtctggcta agatcggccg cagcgatcgc atccatggcc tccgcgaccg gctgcagaac
    14341 agcgggcagt tcggtttcag gcaggtcttg caacgtgaca ccctgtgcac ggcgggagat
    14401 gcaataggtc aggctctcgc tgaattcccc aatgtcaagc acttccggaa tcgggagcgc
    14461 ggccgatgca aagtgccgat aaacataacg atctttgtag aaaccatcgg cgcagctatt
    14521 tacccgcagg acatatccac gccctcctac atcgaagctg aaagcacgag attcttcgcc
    14581 ctccgagagc tgcatcaggt cggacacgct gtcgaacttt tcgatcagaa acttctcgac
    14641 agacgtcgcg gtgagttcag gctttttcat cctaggagag atagatttgt agagagagac
    14701 tggtgatttc agcgtgtcct ctccaaatga aatgaacttc cttatataga ggaagggtct
    14761 tgcgaaggat agtgggattg tgcgtcatcc cttacgtcag tggagatatc acatcaatcc
    14821 acttgctttg aagacgtggt tggaacgtct tctttttcca cgatgctcct cgtgggtggg
    14881 ggtccatctt tgggaccact gtcggcagag gcatcttgaa cgatagcctt tcctttatcg
    14941 caatgatggc atttgtaggt gccaccttcc ttttctactg tccttttgat gaagtgacag
    15001 atagctgggc aatggaatcc gaggaggttt cccgatatta ccctttgttg aaaagtctca
    15061 atagcccttt ggtcttctga gactgtatct ttgatattct tggagtagac gagagtgtcg
    15121 tgctccacca tgttcacatc aatccacttg ctttgaagac gtggttggaa cgtcttcttt
    15181 ttccacgatg ctcctcgtgg gtgggggtcc atctttggga ccactgtcgg cagaggcatc
    15241 ttgaacgata gcctttcctt tatcgcaatg atggcatttg taggtgccac cttccttttc
    15301 tactgtcctt ttgatgaagt gacagatagc tgggcaatgg aatccgagga ggtttcccga
    15361 tattaccctt tgttgaaaag tctcaatagc cctttggtct tctgagactg tatctt
//