pTX179

LOCUS       pTX179               14898 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Express STU (Single Transcriptional Unit) Cas9 nickase with Maize 
            Ubiquitin1 promoter in plants.
ACCESSION   .
VERSION     .
KEYWORDS    pTX179
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14898)
  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 14898)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14898
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        40..2036
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             2061..6164
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6177..6197
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_RNA        6289..6364
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        6394..6469
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    6546..6570
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             7871..8500
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             8929..10002
                     /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      10068..10262
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(10498..10520)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    10596..10736
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     10651..10670
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(10751..10768)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10922..11510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11002..11021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11597..12391)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    12816..12840
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    12918..13092
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        complement(14182..14858)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     complement(14225..14245)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
ORIGIN
        1 tggagtagac gagagtgtcg tgctccacca tgggcgcgcc tgcagtgcag cgtgacccgg
       61 tcgtgcccct ctcttgagat aaggtgagca ttgcatgtct aagttataaa aaattaccac
      121 atattttttt tgtcacactt gtttgaagtg cagtttatct atctttatac atatatttaa
      181 actttactct acgaataata taatctatag tactacaata atatcagtgt tttagagaat
      241 catataaatg aacagttaga catggtctaa aggacaattg agtattttga caacaggact
      301 ctacagtttt atctttttag tgtgcatgtg ttctcctttt tttttgcaaa tagcttcacc
      361 tatataatac ttcatccatt ttattagtac atccatttag ggtttagggt taatggtttt
      421 tatagactaa tttttttagt acatctattt tattctattt tagcctctaa attaagaaaa
      481 ctaaaactct attttagttt ttttatttaa taatttagat ataaaataga ataaaataaa
      541 gtgactaaaa attaaacaaa taccctttaa gaaattaaaa aaactaagga aacatttttc
      601 ttgtttcgag tagataatgc cagcctgtta aacgccgtcg acgagtctaa cggacaccaa
      661 ccagcgaacc agcagcgtcg cgtcgggcca agcgaagcag acggcacggc atctctgtcg
      721 ctgcctctgg acccctctcg agagttccgc tccaccgttg gacttgctcc gctgtcggca
      781 tccagaaatt gcgtggcgga gcggcagacg tgagccggca cggcaggcgg cctcctcctc
      841 ctctcacggc accggcagct acgggggatt cctttcccac cgctccttcg ctttcccttc
      901 ctcgcccgcc gtaataaata gacaccccct ccacaccctc tttccccaac ctcgtgttgt
      961 tcggagcgca cacacacaca accagaactc ccccaaatcc acccgtcggc acctccgctt
     1021 caaggtacgc cgctcgtcct cccccccccc ctctctacct tctcaagatc ggcgttccgg
     1081 tccatggtta gggcccggta gttctacttc tgttcatgtt tgtgttagat ccgtgtttgt
     1141 gttagatccg tgctactagc gttcgtacac ggatgcgacc tgtacgtcag acacgttctg
     1201 attgctaact tgccagtgtt tctctttggg gaatcctggg atggctctag ccgttccgca
     1261 gacgggatcg atttcatgat tttttttgtt tcgttgcata gggtttggtt tgcccttttc
     1321 ctttatttca atatatgccg tgcacttgtt tgtcgggtca tcttttcatg cttttttttg
     1381 tcttggttgt gatgatgtgg tctggttggg cggtcgttca agatcggagt agaattaatt
     1441 ctgtttcaaa ctacctggtg gatttattaa ttttggatct gtatgtgtgt gccatacata
     1501 ttcatagtta cgaattgaag atgatggatg gaaatatcga tctaggatag gtatacatgt
     1561 tgatgcgggt tttactgatg catatacaga gatgcttttt gttcgcttgg ttgtgatgat
     1621 gtggtgtggt tgggcggtcg ttcattcgtt caagatcgga gtagaatact gtttcaaact
     1681 acctggtgta tttattaatt ttggaactgt atgtgtgtgt catacatctt catagttacg
     1741 agtttaagat ggatggaaat atcgatctag gataggtata catgttgatg tgggttttac
     1801 tgatgcatat acatgatggc atatgcagca tctattcata tgctctaacc ttgagtacct
     1861 atctattata ataaacaagt atgttttata attattttga tcttgatata cttggatgat
     1921 ggcatatgca gcagctatat gtggattttt ttagccctgc cttcatacgc tatttatttg
     1981 cttggtactg tttcttttgt cgatgctcac cctgttgttt ggtgttactt ctgcagcctg
     2041 caggtagatc gctcgtcgac atggataaga agtactctat cggactcgct atcggaacta
     2101 actctgtggg atgggctgtg atcaccgatg agtacaaggt gccatctaag aagttcaagg
     2161 ttctcggaaa caccgatagg cactctatca agaaaaacct tatcggtgct ctcctcttcg
     2221 attctggtga aactgctgag gctaccagac tcaagagaac cgctagaaga aggtacacca
     2281 gaagaaagaa caggatctgc tacctccaag agatcttctc taacgagatg gctaaagtgg
     2341 atgattcatt cttccacagg ctcgaagagt cattcctcgt ggaagaagat aagaagcacg
     2401 agaggcaccc tatcttcgga aacatcgttg atgaggtggc ataccacgag aagtacccta
     2461 ctatctacca cctcagaaag aagctcgttg attctactga taaggctgat ctcaggctca
     2521 tctacctcgc tctcgctcac atgatcaagt tcagaggaca cttcctcatc gagggtgatc
     2581 tcaaccctga taactctgat gtggataagt tgttcatcca gctcgtgcag acctacaacc
     2641 agcttttcga agagaaccct atcaacgctt caggtgtgga tgctaaggct atcctctctg
     2701 ctaggctctc taagtcaaga aggcttgaga acctcattgc tcagctccct ggtgagaaga
     2761 agaacggact tttcggaaac ttgatcgctc tctctctcgg actcacccct aacttcaagt
     2821 ctaacttcga tctcgctgag gatgcaaagc tccagctctc aaaggatacc tacgatgatg
     2881 atctcgataa cctcctcgct cagatcggag atcagtacgc tgatttgttc ctcgctgcta
     2941 agaacctctc tgatgctatc ctcctcagtg atatcctcag agtgaacacc gagatcacca
     3001 aggctccact ctcagcttct atgatcaaga gatacgatga gcaccaccag gatctcacac
     3061 ttctcaaggc tcttgttaga cagcagctcc cagagaagta caaagagatt ttcttcgatc
     3121 agtctaagaa cggatacgct ggttacatcg atggtggtgc atctcaagaa gagttctaca
     3181 agttcatcaa gcctatcctc gagaagatgg atggaaccga ggaactcctc gtgaagctca
     3241 atagagagga tcttctcaga aagcagagga ccttcgataa cggatctatc cctcatcaga
     3301 tccacctcgg agagttgcac gctatcctta gaaggcaaga ggatttctac ccattcctca
     3361 aggataacag ggaaaagatt gagaagattc tcaccttcag aatcccttac tacgtgggac
     3421 ctctcgctag aggaaactca agattcgctt ggatgaccag aaagtctgag gaaaccatca
     3481 ccccttggaa cttcgaagag gtggtggata agggtgctag tgctcagtct ttcatcgaga
     3541 ggatgaccaa cttcgataag aaccttccaa acgagaaggt gctccctaag cactctttgc
     3601 tctacgagta cttcaccgtg tacaacgagt tgaccaaggt taagtacgtg accgagggaa
     3661 tgaggaagcc tgcttttttg tcaggtgagc aaaagaaggc tatcgttgat ctcttgttca
     3721 agaccaacag aaaggtgacc gtgaagcagc tcaaagagga ttacttcaag aaaatcgagt
     3781 gcttcgattc agttgagatt tctggtgttg aggataggtt caacgcatct ctcggaacct
     3841 accacgatct cctcaagatc attaaggata aggatttctt ggataacgag gaaaacgagg
     3901 atatcttgga ggatatcgtt cttaccctca ccctctttga agatagagag atgattgaag
     3961 aaaggctcaa gacctacgct catctcttcg atgataaggt gatgaagcag ttgaagagaa
     4021 gaagatacac tggttgggga aggctctcaa gaaagctcat taacggaatc agggataagc
     4081 agtctggaaa gacaatcctt gatttcctca agtctgatgg attcgctaac agaaacttca
     4141 tgcagctcat ccacgatgat tctctcacct ttaaagagga tatccagaag gctcaggttt
     4201 caggacaggg tgatagtctc catgagcata tcgctaacct cgctggatct cctgcaatca
     4261 agaagggaat cctccagact gtgaaggttg tggatgagtt ggtgaaggtg atgggaaggc
     4321 ataagcctga gaacatcgtg atcgaaatgg ctagagagaa ccagaccact cagaagggac
     4381 agaagaactc tagggaaagg atgaagagga tcgaggaagg tatcaaagag cttggatctc
     4441 agatcctcaa agagcaccct gttgagaaca ctcagctcca gaatgagaag ctctacctct
     4501 actacctcca gaacggaagg gatatgtatg tggatcaaga gttggatatc aacaggctct
     4561 ctgattacga tgttgatcat atcgtgccac agtcattctt gaaggatgat tctatcgata
     4621 acaaggtgct caccaggtct gataagaaca ggggtaagag tgataacgtg ccaagtgaag
     4681 aggttgtgaa gaaaatgaag aactattgga ggcagctcct caacgctaag ctcatcactc
     4741 agagaaagtt cgataacttg actaaggctg agaggggagg actctctgaa ttggataagg
     4801 caggattcat caagaggcag cttgtggaaa ccaggcagat cactaagcac gttgcacaga
     4861 tcctcgattc taggatgaac accaagtacg atgagaacga taagttgatc agggaagtga
     4921 aggttatcac cctcaagtca aagctcgtgt ctgatttcag aaaggatttc caattctaca
     4981 aggtgaggga aatcaacaac taccaccacg ctcacgatgc ttaccttaac gctgttgttg
     5041 gaaccgctct catcaagaag tatcctaagc tcgagtcaga gttcgtgtac ggtgattaca
     5101 aggtgtacga tgtgaggaag atgatcgcta agtctgagca agagatcgga aaggctaccg
     5161 ctaagtattt cttctactct aacatcatga atttcttcaa gaccgagatt accctcgcta
     5221 acggtgagat cagaaagagg ccactcatcg agacaaacgg tgaaacaggt gagatcgtgt
     5281 gggataaggg aagggatttc gctaccgtta gaaaggtgct ctctatgcca caggtgaaca
     5341 tcgttaagaa aaccgaggtg cagaccggtg gattctctaa agagtctatc ctccctaaga
     5401 ggaactctga taagctcatt gctaggaaga aggattggga ccctaagaaa tacggtggtt
     5461 tcgattctcc taccgtggct tactctgttc tcgttgtggc taaggttgag aagggaaaga
     5521 gtaagaagct caagtctgtt aaggaacttc tcggaatcac tatcatggaa aggtcatctt
     5581 tcgagaagaa cccaatcgat ttcctcgagg ctaagggata caaagaggtt aagaaggatc
     5641 tcatcatcaa gctcccaaag tactcactct tcgaactcga gaacggtaga aagaggatgc
     5701 tcgcttctgc tggtgagctt caaaagggaa acgagcttgc tctcccatct aagtacgtta
     5761 actttcttta cctcgcttct cactacgaga agttgaaggg atctccagaa gataacgagc
     5821 agaagcaact tttcgttgag cagcacaagc actacttgga tgagatcatc gagcagatct
     5881 ctgagttctc taaaagggtg atcctcgctg atgcaaacct cgataaggtg ttgtctgctt
     5941 acaacaagca cagagataag cctatcaggg aacaggcaga gaacatcatc catctcttca
     6001 cccttaccaa cctcggtgct cctgctgctt tcaagtactt cgatacaacc atcgatagga
     6061 agagatacac ctctaccaaa gaagtgctcg atgctaccct catccatcag tctatcactg
     6121 gactctacga gactaggatc gatctctcac agctcggtgg tgattcaagg gctgatccta
     6181 agaagaagag gaaggtttga ggatctaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     6241 aaaaaaaaaa aaaaaaggct tttgtcaccg gaggagacca cggtctcagt tttagagcta
     6301 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     6361 gtgcttttgt caccggagga gacgaccgtc tcagttttag agctagaaat agcaagttaa
     6421 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttgtcaccg
     6481 gatgtgtttt ccggtctgat gagtccgtga ggacgaaaca aaagcgagct caaactatca
     6541 gtgtttgaca ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat
     6601 cggatattta aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgcatgccaa
     6661 ccacagggtt cccctcggga tcaaagtact ttgatccaac ccctccgctg ctatagtgca
     6721 gtcggcttct gacgttcagt gcagccgtct tctgaaaacg acatgtcgca caagtcctaa
     6781 gttacgcgac aggctgccgc cctgcccttt tcctggcgtt ttcttgtcgc gtgttttagt
     6841 cgcataaagt agaatacttg cgactagaac cggagacatt acgccatgaa caagagcgcc
     6901 gccgctggcc tgctgggcta tgcccgcgtc agcaccgacg accaggactt gaccaaccaa
     6961 cgggccgaac tgcacgcggc cggctgcacc aagctgtttt ccgagaagat caccggcacc
     7021 aggcgcgacc gcccggagct ggccaggatg cttgaccacc tacgccctgg cgacgttgtg
     7081 acagtgacca ggctagaccg cctggcccgc agcacccgcg acctactgga cattgccgag
     7141 cgcatccagg aggccggcgc gggcctgcgt agcctggcag agccgtgggc cgacaccacc
     7201 acgccggccg gccgcatggt gttgaccgtg ttcgccggca ttgccgagtt cgagcgttcc
     7261 ctaatcatcg accgcacccg gagcgggcgc gaggccgcca aggcccgagg cgtgaagttt
     7321 ggcccccgcc ctaccctcac cccggcacag atcgcgcacg cccgcgagct gatcgaccag
     7381 gaaggccgca ccgtgaaaga ggcggctgca ctgcttggcg tgcatcgctc gaccctgtac
     7441 cgcgcacttg agcgcagcga ggaagtgacg cccaccgagg ccaggcggcg cggtgccttc
     7501 cgtgaggacg cattgaccga ggccgacgcc ctggcggccg ccgagaatga acgccaagag
     7561 gaacaagcat gaaaccgcac caggacggcc aggacgaacc gtttttcatt accgaagaga
     7621 tcgaggcgga gatgatcgcg gccgggtacg tgttcgagcc gcccgcgcac ggctcaaccg
     7681 tgcggctgca tgaaatcctg gccggtttgt ctgatgccaa gctggcggcc tggccggcca
     7741 gcttggccgc tgaagaaacc gagcgccgcc gtctaaaaag gtgatgtgta tttgagtaaa
     7801 acagcttgcg tcatgcggtc gctgcgtata tgatgcgatg agtaaataaa caaatacgca
     7861 aggggaacgc atgaaggtta tcgctgtact taaccagaaa ggcgggtcag gcaagacgac
     7921 catcgcaacc catctagccc gcgccctgca actcgccggg gccgatgttc tgttagtcga
     7981 ttccgatccc cagggcagtg cccgcgattg ggcggccgtg cgggaagatc aaccgctaac
     8041 cgttgtcggc atcgaccgcc cgacgattga ccgcgacgtg aaggccatcg gccggcgcga
     8101 cttcgtagtg atcgacggag cgccccaggc ggcggacttg gctgtgtccg cgatcaaggc
     8161 agccgacttc gtgctgattc cggtgcagcc aagcccttac gacatatggg ccaccgccga
     8221 cctggtggag ctggttaagc agcgcattga ggtcacggat ggaaggctac aagcggcctt
     8281 tgtcgtgtcg cgggcgatca aaggcacgcg catcggcggt gaggttgccg aggcgctggc
     8341 cgggtacgag ctgcccattc ttgagtcccg tatcacgcag cgcgtgagct acccaggcac
     8401 tgccgccgcc ggcacaaccg ttcttgaatc agaacccgag ggcgacgctg cccgcgaggt
     8461 ccaggcgctg gccgctgaaa ttaaatcaaa actcatttga gttaatgagg taaagagaaa
     8521 atgagcaaaa gcacaaacac gctaagtgcc ggccgtccga gcgcacgcag cagcaaggct
     8581 gcaacgttgg ccagcctggc agacacgcca gccatgaagc gggtcaactt tcagttgccg
     8641 gcggaggatc acaccaagct gaagatgtac gcggtacgcc aaggcaagac cattaccgag
     8701 ctgctatctg aatacatcgc gcagctacca gagtaaatga gcaaatgaat aaatgagtag
     8761 atgaatttta gcggctaaag gaggcggcat ggaaaatcaa gaacaaccag gcaccgacgc
     8821 cgtggaatgc cccatgtgtg gaggaacggg cggttggcca ggcgtaagcg gctgggttgc
     8881 ctgccggccc tgcaatggca ctggaacccc caagcccgag gaatcggcgt gagcggtcgc
     8941 aaaccatccg gcccggtaca aatcggcgcg gcgctgggtg atgacctggt ggagaagttg
     9001 aaggccgcgc aggccgccca gcggcaacgc atcgaggcag aagcacgccc cggtgaatcg
     9061 tggcaagcgg ccgctgatcg aatccgcaaa gaatcccggc aaccgccggc agccggtgcg
     9121 ccgtcgatta ggaagccgcc caagggcgac gagcaaccag attttttcgt tccgatgctc
     9181 tatgacgtgg gcacccgcga tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg
     9241 aagcgtgacc gacgagctgg cgaggtgatc cgctacgagc ttccagacgg gcacgtagag
     9301 gtttccgcag ggccggccgg catggccagt gtgtgggatt acgacctggt actgatggcg
     9361 gtttcccatc taaccgaatc catgaaccga taccgggaag ggaagggaga caagcccggc
     9421 cgcgtgttcc gtccacacgt tgcggacgta ctcaagttct gccggcgagc cgatggcgga
     9481 aagcagaaag acgacctggt agaaacctgc attcggttaa acaccacgca cgttgccatg
     9541 cagcgtacga agaaggccaa gaacggccgc ctggtgacgg tatccgaggg tgaagccttg
     9601 attagccgct acaagatcgt aaagagcgaa accgggcggc cggagtacat cgagatcgag
     9661 ctagctgatt ggatgtaccg cgagatcaca gaaggcaaga acccggacgt gctgacggtt
     9721 caccccgatt actttttgat cgatcccggc atcggccgtt ttctctaccg cctggcacgc
     9781 cgcgccgcag gcaaggcaga agccagatgg ttgttcaaga cgatctacga acgcagtggc
     9841 agcgccggag agttcaagaa gttctgtttc accgtgcgca agctgatcgg gtcaaatgac
     9901 ctgccggagt acgatttgaa ggaggaggcg gggcaggctg gcccgatcct agtcatgcgc
     9961 taccgcaacc tgatcgaggg cgaagcatcc gccggttcct aatgtacgga gcagatgcta
    10021 gggcaaattg ccctagcagg ggaaaaaggt cgaaaaggcc tctttcctgt ggatagcacg
    10081 tacattggga acccaaagcc gtacattggg aaccggaacc cgtacattgg gaacccaaag
    10141 ccgtacattg ggaaccggtc acacatgtaa gtgactgata taaaagagaa aaaaggcgat
    10201 ttttccgcct aaaactcttt aaaacttatt aaaactctta aaacccgcct ggcctgtgca
    10261 taactgtctg gccagcgcac agccgaagag ctgcaaaaag cgcctaccct tcggtcgctg
    10321 cgctccctac gccccgccgc ttcgcgtcgg cctatcgcgg ccgctggccg ctcaaaaatg
    10381 gctggcctac ggccaggcaa tctaccaggg cgcggacaag ccgcgccgtc gccactcgac
    10441 cgccggcgcc cacatcaagg caccctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
    10501 ctgacacatg cagctcccgg aaacggtcac agcttgtctg taagcggatg ccgggagcag
    10561 acaagcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
    10621 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
    10681 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
    10741 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
    10801 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
    10861 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
    10921 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
    10981 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
    11041 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
    11101 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
    11161 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
    11221 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
    11281 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
    11341 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
    11401 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
    11461 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
    11521 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
    11581 tcatgcattc taggtactaa aacaattcat ccagtaaaat ataatatttt attttctccc
    11641 aatcaggctt gatccccagt aagtcaaaaa atagctcgac atactgttct tccccgatat
    11701 cctccctgat cgaccggacg cagaaggcaa tgtcatacca cttgtccgcc ctgccgcttc
    11761 tcccaagatc aataaagcca cttactttgc catctttcac aaagatgttg ctgtctccca
    11821 ggtcgccgtg ggaaaagaca agttcctctt cgggcttttc cgtctttaaa aaatcataca
    11881 gctcgcgcgg atctttaaat ggagtgtctt cttcccagtt ttcgcaatcc acatcggcca
    11941 gatcgttatt cagtaagtaa tccaattcgg ctaagcggct gtctaagcta ttcgtatagg
    12001 gacaatccga tatgtcgatg gagtgaaaga gcctgatgca ctccgcatac agctcgataa
    12061 tcttttcagg gctttgttca tcttcatact cttccgagca aaggacgcca tcggcctcac
    12121 tcatgagcag attgctccag ccatcatgcc gttcaaagtg caggaccttt ggaacaggca
    12181 gctttccttc cagccatagc atcatgtcct tttcccgttc cacatcatag gtggtccctt
    12241 tataccggct gtccgtcatt tttaaatata ggttttcatt ttctcccacc agcttatata
    12301 ccttagcagg agacattcct tccgtatctt ttacgcagcg gtatttttcg atcagttttt
    12361 tcaattccgg tgatattctc attttagcca tttattattt ccttcctctt ttctacagta
    12421 tttaaagata ccccaagaag ctaattataa caagacgaac tccaattcac tgttccttgc
    12481 attctaaaac cttaaatacc agaaaacagc tttttcaaag ttgttttcaa agttggcgta
    12541 taacatagta tcgacggagc cgattttgaa accgcggtga tcacaggcag caacgctctg
    12601 tcatcgttac aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc
    12661 ttagttgccg ttcttccgaa tagcatcggt aacatgagca aagtctgccg ccttacaacg
    12721 gctctcccgc tgacgccgtc ccggactgat gggctgcctg tatcgagtgg tgattttgtg
    12781 ccgagctgcc ggtcggggag ctgttggctg gctggtggca ggatatattg tggtgtaaac
    12841 aaattgacgc ttagacaact taataacaca ttgcggacgt ttttaatgta ctgaattaac
    12901 gccgaattaa ttcgggggat ctggatttta gtactggatt ttggttttag gaattagaaa
    12961 ttttattgat agaagtattt tacaaataca aatacatact aagggtttct tatatgctca
    13021 acacatgagc gaaaccctat aggaacccta attcccttat ctgggaacta ctcacacatt
    13081 attatggaga aaatagagct tgtcgatcgt taattaatca cctccaagat cccggacgag
    13141 tgctggggcg tcggtttcca ctatcggcga gtacttctac acagccatcg gtccagacgg
    13201 ccgcgcttct gcgggcgatt tgtgtacgcc cgacagtccc ggctccggat cggacgattg
    13261 cgtcgcatcg accctgcgcc caagctgcat catcgaaatt gccgtcaacc aagctctgat
    13321 agagttggtc aagaccaatg cggagcatat acgcccggag ccgcggcgat cctgcaagct
    13381 ccggatgcct ccgctcgaag tagcgcgtct gctgctccat acaagccaac cacggcctcc
    13441 agaagaagat gttggcgacc tcgtattggg aatccccgaa catcgcctcg ctccagtcaa
    13501 tgaccgctgt tatgcggcca ttgtccgtca ggacattgtt ggagccgaaa tccgcgtgca
    13561 cgaggtgccg gacttcgggg cagtcctcgg cccaaagcat cagctcatcg agagcctgcg
    13621 cgacggacgc actgacggtg tcgtccatca cagtttgcca gtgatacaca tggggatcag
    13681 caatcgcgca tatgaaatca cgccatgtag tgtattgacc gattccttgc ggtccgaatg
    13741 ggccgaaccc gctcgtctgg ctaagatcgg ccgcagcgat cgcatccatg gcctccgcga
    13801 ccggctgcag aacagcgggc agttcggttt caggcaggtc ttgcaacgtg acaccctgtg
    13861 cacggcggga gatgcaatag gtcaggctct cgctgaattc cccaatgtca agcacttccg
    13921 gaatcgggag cgcggccgat gcaaagtgcc gataaacata acgatctttg tagaaaccat
    13981 cggcgcagct atttacccgc aggacatatc cacgccctcc tacatcgaag ctgaaagcac
    14041 gagattcttc gccctccgag agctgcatca ggtcggacac gctgtcgaac ttttcgatca
    14101 gaaacttctc gacagacgtc gcggtgagtt caggcttttt catcctagga gagatagatt
    14161 tgtagagaga gactggtgat ttcagcgtgt cctctccaaa tgaaatgaac ttccttatat
    14221 agaggaaggg tcttgcgaag gatagtggga ttgtgcgtca tcccttacgt cagtggagat
    14281 atcacatcaa tccacttgct ttgaagacgt ggttggaacg tcttcttttt ccacgatgct
    14341 cctcgtgggt gggggtccat ctttgggacc actgtcggca gaggcatctt gaacgatagc
    14401 ctttccttta tcgcaatgat ggcatttgta ggtgccacct tccttttcta ctgtcctttt
    14461 gatgaagtga cagatagctg ggcaatggaa tccgaggagg tttcccgata ttaccctttg
    14521 ttgaaaagtc tcaatagccc tttggtcttc tgagactgta tctttgatat tcttggagta
    14581 gacgagagtg tcgtgctcca ccatgttcac atcaatccac ttgctttgaa gacgtggttg
    14641 gaacgtcttc tttttccacg atgctcctcg tgggtggggg tccatctttg ggaccactgt
    14701 cggcagaggc atcttgaacg atagcctttc ctttatcgca atgatggcat ttgtaggtgc
    14761 caccttcctt ttctactgtc cttttgatga agtgacagat agctgggcaa tggaatccga
    14821 ggaggtttcc cgatattacc ctttgttgaa aagtctcaat agccctttgg tcttctgaga
    14881 ctgtatcttt gatattct
//