LOCUS       sequence-182756-       19065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  WDV replicon T-DNA for gene targeting in wheat scutella, H840A 
            single nickase (nTaCas9_H840A+gUbi1+donor).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 19065)
  AUTHORS   Cermak T, Curtin SJ, Gil-Humanes J, Cegan R, Kono TJY, Konecna E, 
            Belanto JJ, Starker CG, Mathre JW, Greenstein RL, Voytas DF
  TITLE     A multi-purpose toolkit to enable advanced genome engineering in 
            plants.
  JOURNAL   Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 
            10.1105/tpc.16.00922.
  PUBMED    28522548
REFERENCE   2  (bases 1 to 19065)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant Cell.
            2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922."
COMMENT     Sequence Label: pJG486
FEATURES             Location/Qualifiers
     source          1..19065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(291..1085)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    1510..1534
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             join(4061..6577,6578..6580,6581..8164)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the H840A mutation in the HNH catalytic domain"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8165..8212
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      8233..8940
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_RNA        9331..9406
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        9800..9875
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             10647..10700
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(10701..10703,10704..10706,10707..11420)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(10746..10767)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(11007..11026)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     11354..11375
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      11451..11703
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    14295..14319
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             15620..16249
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             16678..17751
                     /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      17817..18011
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(18247..18269)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    18355..18495
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     18410..18429
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(18510..18527)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(18681..19065,1..204))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(18761..18780)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
       61 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
      121 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
      181 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
      241 cagtggaacg aaaactcacg ttaagggatt ttggtcatgc attctaggta ctaaaacaat
      301 tcatccagta aaatataata ttttattttc tcccaatcag gcttgatccc cagtaagtca
      361 aaaaatagct cgacatactg ttcttccccg atatcctccc tgatcgaccg gacgcagaag
      421 gcaatgtcat accacttgtc cgccctgccg cttctcccaa gatcaataaa gccacttact
      481 ttgccatctt tcacaaagat gttgctgtct cccaggtcgc cgtgggaaaa gacaagttcc
      541 tcttcgggct tttccgtctt taaaaaatca tacagctcgc gcggatcttt aaatggagtg
      601 tcttcttccc agttttcgca atccacatcg gccagatcgt tattcagtaa gtaatccaat
      661 tcggctaagc ggctgtctaa gctattcgta tagggacaat ccgatatgtc gatggagtga
      721 aagagcctga tgcactccgc atacagctcg ataatctttt cagggctttg ttcatcttca
      781 tactcttccg agcaaaggac gccatcggcc tcactcatga gcagattgct ccagccatca
      841 tgccgttcaa agtgcaggac ctttggaaca ggcagctttc cttccagcca tagcatcatg
      901 tccttttccc gttccacatc ataggtggtc cctttatacc ggctgtccgt catttttaaa
      961 tataggtttt cattttctcc caccagctta tataccttag caggagacat tccttccgta
     1021 tcttttacgc agcggtattt ttcgatcagt tttttcaatt ccggtgatat tctcatttta
     1081 gccatttatt atttccttcc tcttttctac agtatttaaa gataccccaa gaagctaatt
     1141 ataacaagac gaactccaat tcactgttcc ttgcattcta aaaccttaaa taccagaaaa
     1201 cagctttttc aaagttgttt tcaaagttgg cgtataacat agtatcgacg gagccgattt
     1261 tgaaaccgcg gtgatcacag gcagcaacgc tctgtcatcg ttacaatcaa catgctaccc
     1321 tccgcgagat catccgtgtt tcaaacccgg cagcttagtt gccgttcttc cgaatagcat
     1381 cggtaacatg agcaaagtct gccgccttac aacggctctc ccgctgacgc cgtcccggac
     1441 tgatgggctg cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg
     1501 gctggctggt ggcaggatat attgtggtgt aaacaaattg acgcttagac aacttaataa
     1561 cacattgcgg acgtttttaa tgtagagctc gttcctgcgg ccgcttaatt aaggtagtga
     1621 acagaagtcc ggcaggtcct tagcgaaaaa acggggtgtg ccagaaaact ctatcctcta
     1681 ccctgcgtgg aggtgtgaat tctgcacact gcaaatgcaa tgtgtccaat gctttatata
     1741 gggcaggttt tggcgggaga acagggccct agtgttccca cggtagcgta gcgaatcgtg
     1801 tgggccctgt tcggtgtgcg gtcggggggc ctccacgcgg gttataatat taccccgcgt
     1861 ggtggccccc gacgcgcact cggcttttcg tgagtgcgcg gaggcttttg gaccacatct
     1921 tttctgatca ctttcgtgga agatgttgat ttatcacact tttgacgggg aaatctgtgc
     1981 catgccttag cttataagga agtgcgtggt agcccatctc gacaagtttg taccgatctg
     2041 caggcgcgcc tgcagtgcag cgtgacccgg tcgtgcccct ctctagagat aatgagcatt
     2101 gcatgtctaa gttataaaaa attaccacat attttttttg tcacacttgt ttgaagtgca
     2161 gtttatctat ctttatacat atatttaaac tttactctac gaataatata atctatagta
     2221 ctacaataat atcagtgttt tagagaatca tataaatgaa cagttagaca tggtctaaag
     2281 gacaattgag tattttgaca acaggactct acagttttat ctttttagtg tgcatgtgtt
     2341 ctcctttttt tttgcaaata gcttcaccta tataatactt catccatttt attagtacat
     2401 ccatttaggg tttagggtta atggttttta tagactaatt tttttagtac atctatttta
     2461 ttctatttta gcctctaaat taagaaaact aaaactctat tttagttttt ttatttaata
     2521 atttagatat aaaatagaat aaaataaagt gactaaaaat taaacaaata ccctttaaga
     2581 aattaaaaaa actaaggaaa catttttctt gtttcgagta gataatgcca gcctgttaaa
     2641 cgccgtcgac gagtctaacg gacaccaacc agcgaaccag cagcgtcgcg tcgggccaag
     2701 cgaagcagac ggcacggcat ctctgtcgct gcctctggac ccctctcgag agttccgctc
     2761 caccgttgga cttgctccgc tgtcggcatc cagaaattgc gtggcggagc ggcagacgtg
     2821 agccggcacg gcaggcggcc tcctcctcct ctcacggcac cggcagctac gggggattcc
     2881 tttcccaccg ctccttcgct ttcccttcct cgcccgccgt aataaataga caccccctcc
     2941 acaccctctt tccccaacct cgtgttgttc ggagcgcaca cacacacaac cagatctccc
     3001 ccaaatccac ccgtcggcac ctccgcttca aggtacgccg ctcgtcctcc cccccccccc
     3061 tctctacctt ctctagatcg gcgttccggt ccatggttag ggcccggtag ttctacttct
     3121 gttcatgttt gtgttagatc cgtgtttgtg ttagatccgt gctgctagcg ttcgtacacg
     3181 gatgcgacct gtacgtcaga cacgttctga ttgctaactt gccagtgttt ctctttgggg
     3241 aatcctggga tggctctagc cgttccgcag acgggatcga tttcatgatt ttttttgttt
     3301 cgttgcatag ggtttggttt gcccttttcc tttatttcaa tatatgccgt gcacttgttt
     3361 gtcgggtcat cttttcatgc ttttttttgt cttggttgtg atgatgtggt ctggttgggc
     3421 ggtcgttcta gatcggagta gaattaattc tgtttcaaac tacctggtgg atttattaat
     3481 tttggatctg tatgtgtgtg ccatacatat tcatagttac gaattgaaga tgatggatgg
     3541 aaatatcgat ctaggatagg tatacatgtt gatgcgggtt ttactgatgc atatacagag
     3601 atgctttttg ttcgcttggt tgtgatgatg tggtgtggtt gggcggtcgt tcattcgttc
     3661 tagatcggag tagaatactg tttcaaacta cctggtgtat ttattaattt tggaactgta
     3721 tgtgtgtgtc atacatcttc atagttacga gtttaagatg gatggaaata tcgatctagg
     3781 ataggtatac atgttgatgt gggttttact gatgcatata catgatggca tatgcagcat
     3841 ctattcatat gctctaacct tgagtaccta tctattataa taaacaagta tgttttataa
     3901 ttattttgat cttgatatac ttggatgatg gcatatgcag cagctatatg tggatttttt
     3961 tagccctgcc ttcatacgct atttatttgc ttggtactgt ttcttttgtc gatgctcacc
     4021 ctgttgtttg gtgttacttc ctgcagggcg atctattcga atggacaaga agtactcgat
     4081 cggcctcgac atcgggacga actcagttgg ctgggccgtg atcaccgacg agtacaaggt
     4141 gccctctaag aagttcaagg tcctggggaa caccgaccgc cattccatca agaagaacct
     4201 catcggcgct ctcctgttcg acagcgggga gaccgctgag gctacgaggc tcaagagaac
     4261 cgctaggcgc cggtacacga gaaggaagaa caggatctgc tacctccaag agattttctc
     4321 caacgagatg gccaaggttg acgattcatt cttccaccgc ctggaggagt ctttcctcgt
     4381 ggaggaggat aagaagcacg agcggcatcc catcttcggc aacatcgtgg acgaggttgc
     4441 ctaccacgag aagtacccta cgatctacca tctgcggaag aagctcgtgg actccaccga
     4501 taaggcggac ctcagactga tctacctcgc tctggcccac atgatcaagt tccgcggcca
     4561 tttcctgatc gagggggatc tcaacccaga caacagcgat gttgacaagc tgttcatcca
     4621 actcgtgcag acctacaacc aactcttcga ggagaacccg atcaacgcct ctggcgtgga
     4681 cgcgaaggct atcctgtccg cgaggctctc gaagtccagg aggctggaga acctgatcgc
     4741 tcagctccca ggcgagaaga agaacggcct gttcgggaac ctcatcgctc tcagcctggg
     4801 gctcaccccg aacttcaagt cgaacttcga tctcgctgag gacgccaagc tgcaactctc
     4861 caaggacacc tacgacgatg acctcgataa cctcctggcc cagatcggcg atcaatacgc
     4921 ggacctgttc ctcgctgcca agaacctgtc ggacgccatc ctcctgtcag atatcctccg
     4981 cgtgaacacc gagatcacga aggctccact ctctgcctcc atgatcaagc gctacgacga
     5041 gcaccatcag gatctgaccc tcctgaaggc gctggtccgc caacagctcc cggagaagta
     5101 caaggagatt ttcttcgatc agtcgaagaa cggctacgct gggtacatcg acggcggggc
     5161 ctcacaagag gagttctaca agttcatcaa gccaatcctg gagaagatgg acggcacgga
     5221 ggagctcctg gtgaagctca acagggagga cctcctgcgg aagcagagaa ccttcgataa
     5281 cggcagcatc ccccaccaaa tccatctcgg ggagctgcac gccatcctga gaaggcaaga
     5341 ggacttctac cctttcctca aggataaccg ggagaagatc gagaagatcc tgaccttcag
     5401 aatcccatac tacgtcggcc ctctcgcgcg ggggaactca agattcgctt ggatgacccg
     5461 caagtctgag gagaccatca cgccgtggaa cttcgaggag gtggtggaca agggcgctag
     5521 cgctcagtcg ttcatcgaga ggatgaccaa cttcgacaag aacctgccca acgagaaggt
     5581 gctccctaag cactcgctcc tgtacgagta cttcaccgtc tacaacgagc tcacgaaggt
     5641 gaagtacgtc accgagggca tgcgcaagcc agcgttcctg tccggggagc agaagaaggc
     5701 tatcgtggac ctcctgttca agaccaaccg gaaggtcacg gttaagcaac tcaaggagga
     5761 ctacttcaag aagatcgagt gcttcgattc ggtcgagatc agcggcgttg aggaccgctt
     5821 caacgccagc ctcgggacct accacgatct cctgaagatc atcaaggata aggacttcct
     5881 ggacaacgag gagaacgagg atatcctgga ggacatcgtg ctgaccctca cgctgttcga
     5941 ggacagggag atgatcgagg agcgcctgaa gacgtacgcc catctcttcg atgacaaggt
     6001 catgaagcaa ctcaagcgcc ggagatacac cggctggggg aggctgtccc gcaagctcat
     6061 caacggcatc cgggacaagc agtccgggaa gaccatcctc gacttcctga agagcgatgg
     6121 cttcgccaac aggaacttca tgcaactgat ccacgatgac agcctcacct tcaaggagga
     6181 tatccaaaag gctcaagtga gcggccaggg ggactcgctg cacgagcata tcgcgaacct
     6241 cgctggctcc cccgcgatca agaagggcat cctccagacc gtgaaggttg tggacgagct
     6301 cgtgaaggtc atgggccggc acaagcctga gaacatcgtc atcgagatgg ccagagagaa
     6361 ccaaaccacg cagaaggggc aaaagaactc tagggagcgc atgaagcgca tcgaggaggg
     6421 catcaaggag ctggggtccc aaatcctcaa ggagcaccca gtggagaaca cccaactgca
     6481 gaacgagaag ctctacctgt actacctcca gaacggcagg gatatgtacg tggaccaaga
     6541 gctggatatc aaccgcctca gcgattacga cgtcgatgct atcgttcccc agtctttcct
     6601 gaaggatgac tccatcgaca acaaggtcct caccaggtcg gacaagaacc gcggcaagtc
     6661 agataacgtt ccatctgagg aggtcgttaa gaagatgaag aactactgga ggcagctcct
     6721 gaacgccaag ctgatcacgc aaaggaagtt cgacaacctc accaaggctg agagaggcgg
     6781 gctctcagag ctggacaagg ccggcttcat caagcggcag ctggtcgaga ccagacaaat
     6841 cacgaagcac gttgcgcaaa tcctcgactc tcggatgaac acgaagtacg atgagaacga
     6901 caagctgatc agggaggtta aggtgatcac cctgaagtct aagctcgtct ccgacttcag
     6961 gaaggatttc cagttctaca aggttcgcga gatcaacaac taccaccatg cccatgacgc
     7021 ttacctcaac gctgtggtcg gcaccgctct gatcaagaag tacccaaagc tggagtccga
     7081 gttcgtgtac ggggactaca aggtttacga tgtgcgcaag atgatcgcca agtcggagca
     7141 agagatcggc aaggctaccg ccaagtactt cttctactca aacatcatga acttcttcaa
     7201 gaccgagatc acgctggcca acggcgagat ccggaagaga ccgctcatcg agaccaacgg
     7261 cgagacgggg gagatcgtgt gggacaaggg cagggatttc gcgaccgtcc gcaaggttct
     7321 ctccatgccc caggtgaaca tcgtcaagaa gaccgaggtc caaacgggcg ggttctcaaa
     7381 ggagtctatc ctgcctaagc ggaacagcga caagctcatc gccagaaaga aggactggga
     7441 cccaaagaag tacggcgggt tcgacagccc taccgtggcc tactcggtcc tggttgtggc
     7501 gaaggttgag aagggcaagt ccaagaagct caagagcgtg aaggagctcc tggggatcac
     7561 catcatggag aggtccagct tcgagaagaa cccaatcgac ttcctggagg ccaagggcta
     7621 caaggaggtg aagaaggacc tgatcatcaa gctcccgaag tactctctct tcgagctgga
     7681 gaacggcagg aagagaatgc tggcttccgc tggcgagctc cagaagggga acgagctcgc
     7741 gctgccaagc aagtacgtga acttcctcta cctggcttcc cactacgaga agctcaaggg
     7801 cagcccggag gacaacgagc aaaagcagct gttcgtcgag cagcacaagc attacctcga
     7861 cgagatcatc gagcaaatct ccgagttcag caagcgcgtg atcctcgccg acgcgaacct
     7921 ggataaggtc ctctccgcct acaacaagca ccgggacaag cccatcagag agcaagcgga
     7981 gaacatcatc catctcttca ccctgacgaa cctcggcgct cctgctgctt tcaagtactt
     8041 cgacaccacg atcgatcgga agagatacac ctccacgaag gaggtcctgg acgcgaccct
     8101 catccaccag tcgatcaccg gcctgtacga gacgaggatc gacctctcac aactcggcgg
     8161 ggataagaga cccgcagcaa ccaagaaggc agggcaagca aagaagaaga agtgactcga
     8221 gtccctagag tcctgcttta atgagatatg cgagacgcct atgatcgcat gatatttgct
     8281 ttcaattctg ttgtgcacgt tgtaaaaaac ctgagcatgt gtagctcaga tccttaccgc
     8341 cggtttcggt tcattctaat gaatatatca cccgttacta tcgtattttt atgaataata
     8401 ttctccgttc aatttactga ttgtacccta ctacttatat gtacaatatt aaaatgaaaa
     8461 caatatattg tgctgaatag gtttatagcg acatctatga tagagcgcca caataacaaa
     8521 caattgcgtt ttattattac aaatccaatt ttaaaaaaag cggcagaacc ggtcaaacct
     8581 aaaagactga ttacataaat cttattcaaa tttcaaaagt gccccagggg ctagtatcta
     8641 cgacacaccg agcggcgaac taataacgct cactgaaggg aactccggtt ccccgccggc
     8701 gcgcatgggt gagattcctt gaagttgagt attggccgtc cgctctaccg aaagttacgg
     8761 gcaccattca acccggtcca gcacggcggc cgggtaaccg acttgctgcc ccgagaatta
     8821 tgcagcattt ttttggtgta tgtgggcccc aaatgaagtg caggtcaaac cttgacagtg
     8881 acgacaaatc gttgggcggg tccagggcga attttgcgac aacatgtcga ggctcagcag
     8941 gaggacgacc aagcccgtta ttctgacagt tctggtgctc aacacattta tatttatcaa
     9001 ggagcacatt gttactcact gctaggaggg aatcgaacta ggaatattga tcagaggaac
     9061 tacgagagag ctgaagataa ctgccctcta gctctcactg atctgggtcg catagtgaga
     9121 tgcagcccac gtgagttcag caacggtcta gcgctgggct tttaggcccg catgatcggg
     9181 cttttgtcgg gtggtcgacg tgttcacgat tggggagagc aacgcagcag ttcctcttag
     9241 tttagtccca cctcgcctgt ccagcagagt tctgaccggt ttataaactc gcttgctgca
     9301 tcagacttgg agacggagtc gattcgtctc gttttagagc tagaaatagc aagttaaaat
     9361 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttccgggac
     9421 caagcccgtt attctgacag ttctggtgct caacacattt atatttatca aggagcacat
     9481 tgttactcac tgctaggagg gaatcgaact aggaatattg atcagaggaa ctacgagaga
     9541 gctgaagata actgccctct agctctcact gatctgggtc gcatagtgag atgcagccca
     9601 cgtgagttca gcaacggtct agcgctgggc ttttaggccc gcatgatcgg gcttttgtcg
     9661 ggtggtcgac gtgttcacga ttggggagag caacgcagca gttcctctta gtttagtccc
     9721 acctcgcctg tccagcagag ttctgaccgg tttataaact cgcttgctgc atcagacttg
     9781 ctggtgcaac tggtggcccg ttttagagct agaaatagca agttaaaata aggctagtcc
     9841 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttcgcgtagt cctcggtatg
     9901 gtgctactgg agctgctagt ggcaggccag caggtttatt tggggctgga cttccggaat
     9961 tagatcaaat gcagcaacag ttgagccaga atcccaacct tatgagggag ataatgaaca
    10021 tgccaatgat gcagagtctc atgaataacc ctgatctaat acgcaatatg attatgaata
    10081 atccacaaat gcgtgatatt attgatcgga atccagatct tgcccatgtc ctcaatgatc
    10141 ctagtgttct ccgccagacc cttgaagctg caagaaaccc tgaaattatg agggagatga
    10201 tgcggaacac agacagagca atgagcaaca tcgaagcttc ccctgaaggg tttaatatgc
    10261 tccggcgtat gtatgaaact gtacaggagc cttttcttaa tgcaacaaca atgggagggg
    10321 gtggggaagg caccccggcc tctaacccgt ttgcagctct tcttggaaat caggggccta
    10381 accaagccgg caatgctcca actaccggcc cagagtccac aacaggaacc cctgttccaa
    10441 atactaatcc acttccaaac ccctggagca acaatggtag gttctagtta tttagagttt
    10501 tttgtttgtt ttgttgttga atgttgataa ttacatgtgg tagtattttt attctcacag
    10561 ctgctgataa ttgcctgtga tactattata ttttcccagc tgggggtgcg caaggaacaa
    10621 cacggtcagg tcctgctgct agtccagagg gcagaggaag tcttctaaca tgcggtgacg
    10681 tggaggagaa tcccgggccc atggtgagca agggcgagga gctgttcacc ggggtggtgc
    10741 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
    10801 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
    10861 tgcccgtgcc ctggcccacc ctcgtgacca ccttcaccta cggcgtgcag tgcttcagcc
    10921 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
    10981 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
    11041 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
    11101 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
    11161 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
    11221 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
    11281 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
    11341 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actcacggca
    11401 tggacgagct gtacaagtaa agcggccggg taccgagctc gaatttcccc gatcgttcaa
    11461 acatttggca ataaagtttc ttaagattga atcctgttgc cggtcttgcg atgattatca
    11521 tataatttct gttgaattac gttaagcatg taataattaa catgtaatgc atgacgttat
    11581 ttatgagatg ggtttttatg attagagtcc cgcaattata catttaatac gcgatagaaa
    11641 acaaaatata gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct atgttactag
    11701 atcgcagggc tggtgcaact ggtggcccac cagggctggg ttcagcagat ttgagcagcc
    11761 tgctcggtgg tcttggtggg aatgcaagaa ctggtgctgc aggtggtcta ggagggttgg
    11821 gttcagcaga tttggggagt atgcttggtg gtccacctga tgctgctctt ttgagtcaga
    11881 tgctgcaaaa ccctgctatg atgcagatga tgcagaacat tatgtctgac ccacagtcaa
    11941 tgaaccaggt ccaatatttt tcaaaactag ttcttttatg atttttggag atgaccttgg
    12001 atcattctgt aacatttgct tgtcccacag ttgcttagca tgaacccaaa tgcacgtagc
    12061 ctgatggagt caaacactca gttgagggat atgttccaaa acccagaatt tcttcgccag
    12121 atggcatccc cagaggcttt gcaggtaaaa tctgttgtga tgcaagttaa caactgttct
    12181 cgtattttat tttctgataa aatttgtatt tgttctgcgc agcaattact ctcattccag
    12241 cagacactgt catcacagct tggccaaaat caacctagcc agtgagtaac tctttttttt
    12301 gcgagaaaaa agggaaaaag taacactcta attcaatagc atgattgtat cacccctttt
    12361 ttttatgaaa ttaaataaaa tagagattat gaagtgcagt tatgtttatc ttttgagggt
    12421 gcaattatgc gtttgctgag tcttttcttt tcagggctgg taacctaggg ggcaatggag
    12481 tgtacttcaa gtcacaccgg cgagtgtttg atcgccggcg gtacaaagtg gttaaaataa
    12541 tattttattt atctcatgtc attcgattac agaggctcgg ctacgagcaa agacaaacca
    12601 aatataacaa acaacaaccc ttacacaatg acatcggaaa acgaaataca acaccctgag
    12661 atattacatt tatagaaact gtacgccgtc cgcgctagga cagtcactgc gaagcagtga
    12721 cgtcttcgcc ggaggcgaac gagtagttga tgaacgtctc gccttcatac atgtagtgaa
    12781 caacagtgtt agagtacatg taatccgact gttcgggagt catatccttg agccaatctt
    12841 cgtctggatt aactaaaatg atgcaaggta ttccaccccg tatgaccttt cgcttaccat
    12901 attttggatt gaccgtgaag tcacgctgag ccccgacgaa gcacttccag ttgggtgtga
    12961 acttgaatgg aatgtcgtcg atgatattat acttggcgtt gacgtcatat gttgtgaaat
    13021 caactagact gttataataa ttgtgtgtcc ctagagacct tgcccaggaa gtctttcctg
    13081 ttctggttgg cccgcagatg tagatggact tatgcctccc cggtgactcc tggaataatc
    13141 gtccatccac tctaagtcag attgcgcttg atccgcagga gtggaagtac aaaggatata
    13201 ggattcgagg cttacggagt agagatgttc atttttccag ctttcaatgg tctcatggca
    13261 aatgagtgat tcggttggaa actcaggtgt gtaagtggca actgggtcag gaaatagatg
    13321 gcgtgccgtg tactcgaagt ctttgagacg gatagaccat tcaaacggaa aacgattgca
    13381 aaccatgctg aggaattcct cgcgagagga actagattca atgatctgtt tcatatccgc
    13441 atcacggtct ttacgacctg gagttgaaac agccacgaat gttccccact cagctgtgtt
    13501 tacatcggag tcaacctcct tcgtgatgta atcacgaact tggttgcagt ctttggcagc
    13561 ttgtatattt ggatggaata tggagaatgg agatgtatcc atacggaggt ttaaggcatt
    13621 gggattggtg atggaagcac gaagcttgtt ctgcacgaga acgtgcagat gtggtgatcc
    13681 atcttcgtgg agctctctaa cagcagcgat gtagaggggc tcatatttgt tcaagagagt
    13741 gcgaagtgaa tccaaggcgt actgtggctc aagggtacat tgaggatatg ttagaaagag
    13801 gtacttggaa tagacacgga acctgggtgc agatgaagag gccatggtag tgaacagaag
    13861 tccggcaggt ccttagcgaa aaaacggggt gtgccagaaa actctatcct ctaccctgcg
    13921 tggaggtgtg aattctgcac actgcaaatg caatgtgtcc aatgctttat atagggcagg
    13981 ttttggcggg agaacagggc cctagtgttc ccacggtagc gtagcgaatc gtgtgggccc
    14041 tgttcggtgt gcggtcgggg ggcctccacg cgggttataa tattaccccg cgtggtggcc
    14101 cccgacgcgc actcggcttt tcgtgagtgc gcggaggctt ttggaccaca tcttttctga
    14161 tcactttcgt ggaagatgtt gatttatcac acttttgacg gggaaatctg tgccatgcct
    14221 tagcttataa ggaagtgcgt ggtagcccat ctcggggccc tcgattcgac gttcctgttt
    14281 aaactatcag tgtttgacag gatatattgg cgggtaaacc taagagaaaa gagcgtttat
    14341 tagaataatc ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt
    14401 gcatgccaac cacagggttc ccctcgggat caaagtactt tgatccaacc cctccgctgc
    14461 tatagtgcag tcggcttctg acgttcagtg cagccgtctt ctgaaaacga catgtcgcac
    14521 aagtcctaag ttacgcgaca ggctgccgcc ctgccctttt cctggcgttt tcttgtcgcg
    14581 tgttttagtc gcataaagta gaatacttgc gactagaacc ggagacatta cgccatgaac
    14641 aagagcgccg ccgctggcct gctgggctat gcccgcgtca gcaccgacga ccaggacttg
    14701 accaaccaac gggccgaact gcacgcggcc ggctgcacca agctgttttc cgagaagatc
    14761 accggcacca ggcgcgaccg cccggagctg gccaggatgc ttgaccacct acgccctggc
    14821 gacgttgtga cagtgaccag gctagaccgc ctggcccgca gcacccgcga cctactggac
    14881 attgccgagc gcatccagga ggccggcgcg ggcctgcgta gcctggcaga gccgtgggcc
    14941 gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt tcgccggcat tgccgagttc
    15001 gagcgttccc taatcatcga ccgcacccgg agcgggcgcg aggccgccaa ggcccgaggc
    15061 gtgaagtttg gcccccgccc taccctcacc ccggcacaga tcgcgcacgc ccgcgagctg
    15121 atcgaccagg aaggccgcac cgtgaaagag gcggctgcac tgcttggcgt gcatcgctcg
    15181 accctgtacc gcgcacttga gcgcagcgag gaagtgacgc ccaccgaggc caggcggcgc
    15241 ggtgccttcc gtgaggacgc attgaccgag gccgacgccc tggcggccgc cgagaatgaa
    15301 cgccaagagg aacaagcatg aaaccgcacc aggacggcca ggacgaaccg tttttcatta
    15361 ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt gttcgagccg cccgcgcacg
    15421 gctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc tgatgccaag ctggcggcct
    15481 ggccggccag cttggccgct gaagaaaccg agcgccgccg tctaaaaagg tgatgtgtat
    15541 ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat gatgcgatga gtaaataaac
    15601 aaatacgcaa ggggaacgca tgaaggttat cgctgtactt aaccagaaag gcgggtcagg
    15661 caagacgacc atcgcaaccc atctagcccg cgccctgcaa ctcgccgggg ccgatgttct
    15721 gttagtcgat tccgatcccc agggcagtgc ccgcgattgg gcggccgtgc gggaagatca
    15781 accgctaacc gttgtcggca tcgaccgccc gacgattgac cgcgacgtga aggccatcgg
    15841 ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg gcggacttgg ctgtgtccgc
    15901 gatcaaggca gccgacttcg tgctgattcc ggtgcagcca agcccttacg acatatgggc
    15961 caccgccgac ctggtggagc tggttaagca gcgcattgag gtcacggatg gaaggctaca
    16021 agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc atcggcggtg aggttgccga
    16081 ggcgctggcc gggtacgagc tgcccattct tgagtcccgt atcacgcagc gcgtgagcta
    16141 cccaggcact gccgccgccg gcacaaccgt tcttgaatca gaacccgagg gcgacgctgc
    16201 ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa ctcatttgag ttaatgaggt
    16261 aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg gccgtccgag cgcacgcagc
    16321 agcaaggctg caacgttggc cagcctggca gacacgccag ccatgaagcg ggtcaacttt
    16381 cagttgccgg cggaggatca caccaagctg aagatgtacg cggtacgcca aggcaagacc
    16441 attaccgagc tgctatctga atacatcgcg cagctaccag agtaaatgag caaatgaata
    16501 aatgagtaga tgaattttag cggctaaagg aggcggcatg gaaaatcaag aacaaccagg
    16561 caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc ggttggccag gcgtaagcgg
    16621 ctgggttgcc tgccggccct gcaatggcac tggaaccccc aagcccgagg aatcggcgtg
    16681 agcggtcgca aaccatccgg cccggtacaa atcggcgcgg cgctgggtga tgacctggtg
    16741 gagaagttga aggccgcgca ggccgcccag cggcaacgca tcgaggcaga agcacgcccc
    16801 ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag aatcccggca accgccggca
    16861 gccggtgcgc cgtcgattag gaagccgccc aagggcgacg agcaaccaga ttttttcgtt
    16921 ccgatgctct atgacgtggg cacccgcgat agtcgcagca tcatggacgt ggccgttttc
    16981 cgtctgtcga agcgtgaccg acgagctggc gaggtgatcc gctacgagct tccagacggg
    17041 cacgtagagg tttccgcagg gccggccggc atggccagtg tgtgggatta cgacctggta
    17101 ctgatggcgg tttcccatct aaccgaatcc atgaaccgat accgggaagg gaagggagac
    17161 aagcccggcc gcgtgttccg tccacacgtt gcggacgtac tcaagttctg ccggcgagcc
    17221 gatggcggaa agcagaaaga cgacctggta gaaacctgca ttcggttaaa caccacgcac
    17281 gttgccatgc agcgtacgaa gaaggccaag aacggccgcc tggtgacggt atccgagggt
    17341 gaagccttga ttagccgcta caagatcgta aagagcgaaa ccgggcggcc ggagtacatc
    17401 gagatcgagc tagctgattg gatgtaccgc gagatcacag aaggcaagaa cccggacgtg
    17461 ctgacggttc accccgatta ctttttgatc gatcccggca tcggccgttt tctctaccgc
    17521 ctggcacgcc gcgccgcagg caaggcagaa gccagatggt tgttcaagac gatctacgaa
    17581 cgcagtggca gcgccggaga gttcaagaag ttctgtttca ccgtgcgcaa gctgatcggg
    17641 tcaaatgacc tgccggagta cgatttgaag gaggaggcgg ggcaggctgg cccgatccta
    17701 gtcatgcgct accgcaacct gatcgagggc gaagcatccg ccggttccta atgtacggag
    17761 cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc gaaaaggcct ctttcctgtg
    17821 gatagcacgt acattgggaa cccaaagccg tacattggga accggaaccc gtacattggg
    17881 aacccaaagc cgtacattgg gaaccggtca cacatgtaag tgactgatat aaaagagaaa
    17941 aaaggcgatt tttccgccta aaactcttta aaacttatta aaactcttaa aacccgcctg
    18001 gcctgtgcat aactgtctgg ccagcgcaca gccgaagagc tgcaaaaagc gcctaccctt
    18061 cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc ctatcgcggc cgctggccgc
    18121 tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc gcggacaagc cgcgccgtcg
    18181 ccactcgacc gccggcgccc acatcaaggc accctgcctc gcgcgtttcg gtgatgacgg
    18241 tgaaaacctc tgacacatgc agctcccgga aacggtcaca gcttgtctgt aagcggatgc
    18301 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
    18361 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
    18421 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
    18481 aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
    18541 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
    18601 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
    18661 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
    18721 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
    18781 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
    18841 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
    18901 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
    18961 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
    19021 ccactggcag cagccactgg taacaggatt agcagagcga ggtat
//
