LOCUS       sequence-182724-       18608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BeYDV replicon T-DNA for gene targeting in tobacco leaves, D10A 
            single nickase no gRNA control (nAtCas9_D10A+gRNActrl+donor).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18608)
  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 18608)
  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: pJG370
FEATURES             Location/Qualifiers
     source          1..18608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1629..2423)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    2848..2872
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        3813..4158
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     4101..4121
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             join(4192..4218,4219..4221,4222..8295)
                     /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             8308..8328
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      8339..8587
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     misc_RNA        9076..9151
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        9511..9586
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             11070..11861
                     /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="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(11121..11140)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     11731..11750
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      11888..12140
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    13838..13862
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             15163..15792
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             16221..17294
                     /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      17360..17554
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(17790..17812)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    17898..18038
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     17953..17972
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(18053..18070)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(18224..18608,1..204))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(18304..18323)
                     /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 cagtgctgat gaatccccta atgattttta tcaaaatcat taagttaagg tagatacaca
      301 tcttgtcata tgatcaaatg gtttcgccaa aaatcaataa tcagacaaca aaatgtgcga
      361 actcgatatt ttacacgact ctctttacca attctgcccc gaattacact taaaacgact
      421 caacagctta acgttggctt gccacgcctt acttgactgt aaaactctca ctcttaccga
      481 acttggccgt aacctgccaa ccaaagcgag aacaaaacat aacatcaaac gaatcgaccg
      541 attgttaggt aatcgtcacc tccacaaaga gcgactcgct gtataccgtt ggcatgctag
      601 ctttatctgt tcgggcaata cgatgcccat tgtacttgtt gactggtctg atatccgtga
      661 gcaaaaacgg cttatggtat tgcgagcttc agtcgcacta cacggtcgtt ctgttactct
      721 ttatgagaaa gcgttcccgc tttcagagca atgttcaaag aaagctcatg accaatttct
      781 agccgacctt gcgagcattc taccgagtaa caccacaccg ctcattgtca gtgatgctgg
      841 ctttaaagtg ccatggtata aatccgttga gaagctgggt tggtactggt taagtcgagt
      901 aagaggaaaa gtacaatatg cagacctagg agcggaaaac tggaaaccta tcagcaactt
      961 acatgatatg tcatctagtc actcaaagac tttaggctat aagaggctga ctaaaagcaa
     1021 tccaatctca tgccaaattc tattgtataa atctcgctct aaaggccgaa aaaatcagcg
     1081 ctcgacacgg actcattgtc accacccgtc acctaaaatc tactcagcgt cggcaaagga
     1141 gccatggatt ctagcaacta acttacctgt tgaaattcga acacccaaac aacttgttaa
     1201 tatctattcg aagcgaatgc agattgaaga aaccttccga gacttgaaaa gtcctgccta
     1261 cggactaggc ctacgccata gccgaacgag cagctcagag cgttttgata tcatgctgct
     1321 aatcgccctg atgcttcaac taacatgttg gcttgcgggc gttcatgctc agaaacaagg
     1381 ttgggacaag cacttccagg ctaacacagt cagaaatcga aacgtactct caacagttcg
     1441 cttaggcatg gaagttttgc ggcattctgg ctacacaata acaagggaag acttactcgt
     1501 ggctgcaacc ctactagctc aaaatttatt cacacatggt tacgctttgg ggaaattatg
     1561 aggggatctc tcagcgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgcat
     1621 tctaggtact aaaacaattc atccagtaaa atataatatt ttattttctc ccaatcaggc
     1681 ttgatcccca gtaagtcaaa aaatagctcg acatactgtt cttccccgat atcctccctg
     1741 atcgaccgga cgcagaaggc aatgtcatac cacttgtccg ccctgccgct tctcccaaga
     1801 tcaataaagc cacttacttt gccatctttc acaaagatgt tgctgtctcc caggtcgccg
     1861 tgggaaaaga caagttcctc ttcgggcttt tccgtcttta aaaaatcata cagctcgcgc
     1921 ggatctttaa atggagtgtc ttcttcccag ttttcgcaat ccacatcggc cagatcgtta
     1981 ttcagtaagt aatccaattc ggctaagcgg ctgtctaagc tattcgtata gggacaatcc
     2041 gatatgtcga tggagtgaaa gagcctgatg cactccgcat acagctcgat aatcttttca
     2101 gggctttgtt catcttcata ctcttccgag caaaggacgc catcggcctc actcatgagc
     2161 agattgctcc agccatcatg ccgttcaaag tgcaggacct ttggaacagg cagctttcct
     2221 tccagccata gcatcatgtc cttttcccgt tccacatcat aggtggtccc tttataccgg
     2281 ctgtccgtca tttttaaata taggttttca ttttctccca ccagcttata taccttagca
     2341 ggagacattc cttccgtatc ttttacgcag cggtattttt cgatcagttt tttcaattcc
     2401 ggtgatattc tcattttagc catttattat ttccttcctc ttttctacag tatttaaaga
     2461 taccccaaga agctaattat aacaagacga actccaattc actgttcctt gcattctaaa
     2521 accttaaata ccagaaaaca gctttttcaa agttgttttc aaagttggcg tataacatag
     2581 tatcgacgga gccgattttg aaaccgcggt gatcacaggc agcaacgctc tgtcatcgtt
     2641 acaatcaaca tgctaccctc cgcgagatca tccgtgtttc aaacccggca gcttagttgc
     2701 cgttcttccg aatagcatcg gtaacatgag caaagtctgc cgccttacaa cggctctccc
     2761 gctgacgccg tcccggactg atgggctgcc tgtatcgagt ggtgattttg tgccgagctg
     2821 ccggtcgggg agctgttggc tggctggtgg caggatatat tgtggtgtaa acaaattgac
     2881 gcttagacaa cttaataaca cattgcggac gtttttaatg tagagctcaa agtttaacgc
     2941 gttagcagaa ggcatgttgt tgtgactccg aggggttgcc tcaaactcta tcttataacc
     3001 ggcgtggagg catggaggca ggggtatttt ggtcatttta atagatagtg gaaaatgacg
     3061 tggaatttac ttaaagacga agtctttgcg acaagggggg gcccacgccg aatttaatat
     3121 taccggcgtg gccccccctt atcgcgagtg ctttagcacg agcggtccag atttaaagta
     3181 gaaaatttcc cgcccactag ggttaaaggt gttcacacta taaaagcata tacgatgtga
     3241 tggtatttga tggagcgtat attgtatcag gtatttccgt tggatacgaa ttattcgtac
     3301 gaccctcggt accgatcggc gcgccagatt tgccttttca atttcagaaa gaatgctaac
     3361 ccacagatgg ttagagaggc ttacgcagca ggtatcatca agacgatcta cccgagcaat
     3421 aatctccagg aaatcaaata ccttcccaag aaggttaaag atgcagtcaa aagattcagg
     3481 actaactgca tcaagaacac agagaaagat atatttctca agatcagaag tactattcca
     3541 gtatggacga ttcaaggctt gcttcacaaa ccaaggcaag taatagagat tggagtctct
     3601 aaaaaggtag ttcccactga atcaaaggcc atggagtcaa agattcaaat agaggaccta
     3661 acagaactcg ccgtaaagac tggcgaacag ttcatacaga gtctcttacg actcaatgac
     3721 aagaagaaaa tcttcgtcaa catggtggag cacgacacac ttgtctactc caaaaatatc
     3781 aaagatacag tctcagaaga ccaaagggca attgagactt ttcaacaaag ggtaatatcc
     3841 ggaaacctcc tcggattcca ttgcccagct atctgtcact ttattgtgaa gatagtggaa
     3901 aaggaaggtg gctcctacaa atgccatcat tgcgataaag gaaaggccat cgttgaagat
     3961 gcctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat cgtggaaaaa
     4021 gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgatatctc cactgacgta
     4081 agggatgacg cacaatccca ctatccttcg caagaccctt cctctatata aggaagttca
     4141 tttcatttgg agagaacacg ggggactcct gcaggtagat cgctcgtcga catggataag
     4201 aagtactcta tcggactcgc tatcggaact aactctgtgg gatgggctgt gatcaccgat
     4261 gagtacaagg tgccatctaa gaagttcaag gttctcggaa acaccgatag gcactctatc
     4321 aagaaaaacc ttatcggtgc tctcctcttc gattctggtg aaactgctga ggctaccaga
     4381 ctcaagagaa ccgctagaag aaggtacacc agaagaaaga acaggatctg ctacctccaa
     4441 gagatcttct ctaacgagat ggctaaagtg gatgattcat tcttccacag gctcgaagag
     4501 tcattcctcg tggaagaaga taagaagcac gagaggcacc ctatcttcgg aaacatcgtt
     4561 gatgaggtgg cataccacga gaagtaccct actatctacc acctcagaaa gaagctcgtt
     4621 gattctactg ataaggctga tctcaggctc atctacctcg ctctcgctca catgatcaag
     4681 ttcagaggac acttcctcat cgagggtgat ctcaaccctg ataactctga tgtggataag
     4741 ttgttcatcc agctcgtgca gacctacaac cagcttttcg aagagaaccc tatcaacgct
     4801 tcaggtgtgg atgctaaggc tatcctctct gctaggctct ctaagtcaag aaggcttgag
     4861 aacctcattg ctcagctccc tggtgagaag aagaacggac ttttcggaaa cttgatcgct
     4921 ctctctctcg gactcacccc taacttcaag tctaacttcg atctcgctga ggatgcaaag
     4981 ctccagctct caaaggatac ctacgatgat gatctcgata acctcctcgc tcagatcgga
     5041 gatcagtacg ctgatttgtt cctcgctgct aagaacctct ctgatgctat cctcctcagt
     5101 gatatcctca gagtgaacac cgagatcacc aaggctccac tctcagcttc tatgatcaag
     5161 agatacgatg agcaccacca ggatctcaca cttctcaagg ctcttgttag acagcagctc
     5221 ccagagaagt acaaagagat tttcttcgat cagtctaaga acggatacgc tggttacatc
     5281 gatggtggtg catctcaaga agagttctac aagttcatca agcctatcct cgagaagatg
     5341 gatggaaccg aggaactcct cgtgaagctc aatagagagg atcttctcag aaagcagagg
     5401 accttcgata acggatctat ccctcatcag atccacctcg gagagttgca cgctatcctt
     5461 agaaggcaag aggatttcta cccattcctc aaggataaca gggaaaagat tgagaagatt
     5521 ctcaccttca gaatccctta ctacgtggga cctctcgcta gaggaaactc aagattcgct
     5581 tggatgacca gaaagtctga ggaaaccatc accccttgga acttcgaaga ggtggtggat
     5641 aagggtgcta gtgctcagtc tttcatcgag aggatgacca acttcgataa gaaccttcca
     5701 aacgagaagg tgctccctaa gcactctttg ctctacgagt acttcaccgt gtacaacgag
     5761 ttgaccaagg ttaagtacgt gaccgaggga atgaggaagc ctgctttttt gtcaggtgag
     5821 caaaagaagg ctatcgttga tctcttgttc aagaccaaca gaaaggtgac cgtgaagcag
     5881 ctcaaagagg attacttcaa gaaaatcgag tgcttcgatt cagttgagat ttctggtgtt
     5941 gaggataggt tcaacgcatc tctcggaacc taccacgatc tcctcaagat cattaaggat
     6001 aaggatttct tggataacga ggaaaacgag gatatcttgg aggatatcgt tcttaccctc
     6061 accctctttg aagatagaga gatgattgaa gaaaggctca agacctacgc tcatctcttc
     6121 gatgataagg tgatgaagca gttgaagaga agaagataca ctggttgggg aaggctctca
     6181 agaaagctca ttaacggaat cagggataag cagtctggaa agacaatcct tgatttcctc
     6241 aagtctgatg gattcgctaa cagaaacttc atgcagctca tccacgatga ttctctcacc
     6301 tttaaagagg atatccagaa ggctcaggtt tcaggacagg gtgatagtct ccatgagcat
     6361 atcgctaacc tcgctggatc tcctgcaatc aagaagggaa tcctccagac tgtgaaggtt
     6421 gtggatgagt tggtgaaggt gatgggaagg cataagcctg agaacatcgt gatcgaaatg
     6481 gctagagaga accagaccac tcagaaggga cagaagaact ctagggaaag gatgaagagg
     6541 atcgaggaag gtatcaaaga gcttggatct cagatcctca aagagcaccc tgttgagaac
     6601 actcagctcc agaatgagaa gctctacctc tactacctcc agaacggaag ggatatgtat
     6661 gtggatcaag agttggatat caacaggctc tctgattacg atgttgatca tatcgtgcca
     6721 cagtcattct tgaaggatga ttctatcgat aacaaggtgc tcaccaggtc tgataagaac
     6781 aggggtaaga gtgataacgt gccaagtgaa gaggttgtga agaaaatgaa gaactattgg
     6841 aggcagctcc tcaacgctaa gctcatcact cagagaaagt tcgataactt gactaaggct
     6901 gagaggggag gactctctga attggataag gcaggattca tcaagaggca gcttgtggaa
     6961 accaggcaga tcactaagca cgttgcacag atcctcgatt ctaggatgaa caccaagtac
     7021 gatgagaacg ataagttgat cagggaagtg aaggttatca ccctcaagtc aaagctcgtg
     7081 tctgatttca gaaaggattt ccaattctac aaggtgaggg aaatcaacaa ctaccaccac
     7141 gctcacgatg cttaccttaa cgctgttgtt ggaaccgctc tcatcaagaa gtatcctaag
     7201 ctcgagtcag agttcgtgta cggtgattac aaggtgtacg atgtgaggaa gatgatcgct
     7261 aagtctgagc aagagatcgg aaaggctacc gctaagtatt tcttctactc taacatcatg
     7321 aatttcttca agaccgagat taccctcgct aacggtgaga tcagaaagag gccactcatc
     7381 gagacaaacg gtgaaacagg tgagatcgtg tgggataagg gaagggattt cgctaccgtt
     7441 agaaaggtgc tctctatgcc acaggtgaac atcgttaaga aaaccgaggt gcagaccggt
     7501 ggattctcta aagagtctat cctccctaag aggaactctg ataagctcat tgctaggaag
     7561 aaggattggg accctaagaa atacggtggt ttcgattctc ctaccgtggc ttactctgtt
     7621 ctcgttgtgg ctaaggttga gaagggaaag agtaagaagc tcaagtctgt taaggaactt
     7681 ctcggaatca ctatcatgga aaggtcatct ttcgagaaga acccaatcga tttcctcgag
     7741 gctaagggat acaaagaggt taagaaggat ctcatcatca agctcccaaa gtactcactc
     7801 ttcgaactcg agaacggtag aaagaggatg ctcgcttctg ctggtgagct tcaaaaggga
     7861 aacgagcttg ctctcccatc taagtacgtt aactttcttt acctcgcttc tcactacgag
     7921 aagttgaagg gatctccaga agataacgag cagaagcaac ttttcgttga gcagcacaag
     7981 cactacttgg atgagatcat cgagcagatc tctgagttct ctaaaagggt gatcctcgct
     8041 gatgcaaacc tcgataaggt gttgtctgct tacaacaagc acagagataa gcctatcagg
     8101 gaacaggcag agaacatcat ccatctcttc acccttacca acctcggtgc tcctgctgct
     8161 ttcaagtact tcgatacaac catcgatagg aagagataca cctctaccaa agaagtgctc
     8221 gatgctaccc tcatccatca gtctatcact ggactctacg agactaggat cgatctctca
     8281 cagctcggtg gtgattcaag ggctgatcct aagaagaaga ggaaggtttg actcgagata
     8341 tgaagatgaa gatgaaatat ttggtgtgtc aaataaaaag cttgtgtgct taagtttgtg
     8401 tttttttctt ggcttgttgt gttatgaatt tgtggctttt tctaatatta aatgaatgta
     8461 agatcacatt ataatgaata aacaaatgtt tctataatcc attgtgaatg ttttgttgga
     8521 tctcttctgc agcatataac tactgtatgt gctatggtat ggactatgga atatgattaa
     8581 agataaggag ctccggtgac ggacccatgg cttcgttgaa caacggaaac tcgacttgcc
     8641 ttccgcacaa tacatcattt cttcttagct ttttttcttc ttcttcgttc atacagtttt
     8701 tttttgttta tcagcttaca ttttcttgaa ccgtagcttt cgttttcttc tttttaactt
     8761 tccattcgga gtttttgtat cttgtttcat agtttgtccc aggattagaa tgattaggca
     8821 tcgaaccttc aagaatttga ttgaataaaa catcttcatt cttaagatat gaagataatc
     8881 ttcaaaaggc ccctgggaat ctgaaagaag agaagcaggc ccatttatat gggaaagaac
     8941 aatagtattt cttatatagg cccatttaag ttgaaaacaa tcttcaaaag tcccacatcg
     9001 cttagataag aaaacgaagc tgagtttata tacagctaga gtcgaagtag tgattgagag
     9061 ccttatagga ggaaagtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
     9121 tcaacttgaa aaagtggcac cgagtcggtg ctttttttcc cggcgccatg gatgttgttg
     9181 ttaccagaaa gtaaataaat gttcaatctc tgatgttctc aagtaagtga gttttattgg
     9241 gaataatatt aacttatgtt cttcttgcat ttgatttctt tgccgctctc ttcttctatc
     9301 ttaaatctgt gtatactatt tcactattgg gctttttatt agtctataat gggactcaaa
     9361 ataaggcttt ggcccacatc aaaaagataa gtcacaaatc aaaactaaat tcagagtctt
     9421 ttctcccaca tcggtcactg tactcatttt gtgtttgttt atatattaca cgaaccgatc
     9481 tttggtacgg agacggagtc gattcgtctc gttttagagc tagaaatagc aagttaaaat
     9541 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttcgcgcgt
     9601 agtcctcggt acagtcttac ttccatgatt tctttaacta tgccggaatc catcgcagcg
     9661 taatgctcta caccacgccg aacacctggg tggacgatat caccgtggtg acgcatgtcg
     9721 cgcaagactg taaccacgcg tctgttgact ggcaggtggt ggccaatggt gatgtcagcg
     9781 ttgaactgcg tgatgcggat caacaggtgg ttgcaactgg acaaggcact agcgggactt
     9841 tgcaagtggt gaatccgcac ctctggcaac cgggtgaagg ttatctctat gaactgtgcg
     9901 tcacagccaa aagccagaca gagtgtgata tctacccgct tcgcgtcggc atccggtcag
     9961 tggcagtgaa gggcgaacag ttcctgatta accacaaacc gttctacttt actggctttg
    10021 gtcgtcatga agatgcggac ttgcgtggca aaggattcga taacgtgctg atggtgcacg
    10081 accacgcatt aatggactgg attggggcca actcctaccg tacctcgcat tacccttacg
    10141 ctgaagagat gctcgactgg gcagatgaac atggcatcgt ggtgattgat gaaactgctg
    10201 ctgtcggctt taacctctct ttaggcattg gtttcgaagc gggcaacaag ccgaaagaac
    10261 tgtacagcga agaggcagtc aacggggaaa ctcagcaagc gcacttacag gcgattaaag
    10321 agctgatagc gcgtgacaaa aaccacccaa gcgtggtgat gtggagtatt gccaacgaac
    10381 cggatacccg tccgcaaggt gcacgggaat atttcgcgcc actggcggaa gcaacgcgta
    10441 aactcgaccc gacgcgtccg atcacctgcg tcaatgtaat gttctgcgac gctcacaccg
    10501 ataccatcag cgatctcttt gatgtgctgt gcctgaaccg ttattacgga tggtatgtcc
    10561 aaagcggcga tttggaaacg gcagagaagg tactggaaaa agaacttctg gcctggcagg
    10621 agaaactgca tcagccgatt atcatcaccg aatacggcgt ggatacgtta gccgggctgc
    10681 actcaatgta caccgacatg tggagtgaag agtatcagtg tgcatggctg gatatgtatc
    10741 accgcgtctt tgatcgcgtc agcgccgtcg tcggtgaaca ggtatggaat ttcgccgatt
    10801 ttgcgacctc gcaaggcata ttgcgcgttg gcggtaacaa gaaagggatc ttcactcgcg
    10861 accgcaaacc gaagtcggcg gcttttctgc tgcaaaaacg ctggactggc atgaacttcg
    10921 gtgaaaaacc gcagcaggga ggcaaacaac gcagggaggc aaacaatgat atcacaactc
    10981 tcctgacgcg tcatcgtcgg ctacagcctc gggaattgct acctagctcg agcaagatcc
    11041 aaggagatat aacaatggct tcctcctgga ttgaacaaga tggattgcac gcaggttctc
    11101 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
    11161 ctgatgccgc cgtgttccgg ctgtcagcgc agggtagacc ggttcttttt gtcaagaccg
    11221 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
    11281 cgacgggcgt accttgcgct gctgtgctcg acgttgtcac tgaagcggga agggactggc
    11341 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
    11401 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
    11461 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
    11521 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
    11581 ccaggctcaa ggcgagaatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
    11641 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
    11701 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
    11761 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
    11821 agcgcatcgc cttctatcgc cttcttgacg agttcttctg ataaccgcgg agagctcgaa
    11881 tttccccgat cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg
    11941 tcttgcgatg attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat
    12001 gtaatgcatg acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat
    12061 ttaatacgcg atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt
    12121 gtcatctatg ttactagatc ggagtgtact tcaagtcaca ccggcgagtg tttgatcgcc
    12181 ggcggtaccg agtgtacttc aagtcagtgg gaaatcaata aaatgattat tttatgaata
    12241 tatttcattg tgcaagtaga tagaaattac atatgttaca taacacacga aataaacaaa
    12301 aaaagacaat ccaaaaacaa acaccccaaa aaaaataatc actttagata aactcgtatg
    12361 aggagaggca cgttcagtga ctcgacgatt cccgagcaaa aaaagtctcc ccgtcacaca
    12421 tgtagtgggt gacgcaatta tctttaaagt aatccttctg ttgacttgtc attgataaca
    12481 tccagtcttc gtcaggattg caaagaatta tagaagggat cccacctttt attttcttct
    12541 tttttccata tttagggttg acagtgaaat cagactggca acctattaat tgcttccaca
    12601 atgggacgaa cttgaagggg atgtcgtcga tgatattata ggtggcgtgt tcatcgtagt
    12661 tggtgaaatc gatggtaccg ttccaatagt tgtgtcgtcc gagacttcta gcccaggtgg
    12721 tctttccggt acgagttggt ccgcagatgt agaggctggg gtgtcggatt ccattccttc
    12781 cattgtcctt gttaaatcgg ccatccattc aaggtcagat tgagcttgtt ggtatgagac
    12841 aggatgtatg taagtataag cgtctatgct tacatggtat agatgggttt ccctccagga
    12901 gtgtagatct tcgtggcagc gaagatctga ttctgtgaag ggcgacacat acggttcagg
    12961 ttgtggaggg aataatttgt tggctgaata ttccagccat tgaagctttg ttgcccattc
    13021 atgagggaat tcttccttga tcatgtcaag atattcctcc ttagacgttg cagtctggat
    13081 aatagttctc catcgtgcgt cagatttgcg aggagaaacc ttatgatctc ggaaatctcc
    13141 tctggtttta atatctccgt cctttgatat gtaatcaagg acttgtttag agtttctagc
    13201 tggctggata ttagggtgat ttccttcaaa atcgaaaaaa gaaggatccc taatacaagg
    13261 ttttttatca agctggagaa gagcatgata gtgggtagtg ccatcttgat gaagctcaga
    13321 agcaacacca aggaagaaaa taagaaaagg tgtgagtttc tcccagagaa actggaataa
    13381 atcatctctt tgagatgagc acttgggata ggtaaggaaa acatatttag attggagtct
    13441 gaagttctta ctagcagaag gcatgttgtt gtgactccga ggggttgcct caaactctat
    13501 cttataaccg gcgtggaggc atggaggcag gggtattttg gtcattttaa tagatagtgg
    13561 aaaatgacgt ggaatttact taaagacgaa gtctttgcga caaggggggg cccacgccga
    13621 atttaatatt accggcgtgg ccccccctta tcgcgagtgc tttagcacga gcggtccaga
    13681 tttaaagtag aaaatttccc gcccactagg gttaaaggtg ttcacactat aaaagcatat
    13741 acgatgtgat ggtatttgat ggagcgtata ttgtatcagg tatttccgtt ggatacgaat
    13801 tattcgtacg accctcatag tttaaactat cagtgtttga caggatatat tggcgggtaa
    13861 acctaagaga aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt
    13921 tatccgttcg tccatttgta tgtgcatgcc aaccacaggg ttcccctcgg gatcaaagta
    13981 ctttgatcca acccctccgc tgctatagtg cagtcggctt ctgacgttca gtgcagccgt
    14041 cttctgaaaa cgacatgtcg cacaagtcct aagttacgcg acaggctgcc gccctgccct
    14101 tttcctggcg ttttcttgtc gcgtgtttta gtcgcataaa gtagaatact tgcgactaga
    14161 accggagaca ttacgccatg aacaagagcg ccgccgctgg cctgctgggc tatgcccgcg
    14221 tcagcaccga cgaccaggac ttgaccaacc aacgggccga actgcacgcg gccggctgca
    14281 ccaagctgtt ttccgagaag atcaccggca ccaggcgcga ccgcccggag ctggccagga
    14341 tgcttgacca cctacgccct ggcgacgttg tgacagtgac caggctagac cgcctggccc
    14401 gcagcacccg cgacctactg gacattgccg agcgcatcca ggaggccggc gcgggcctgc
    14461 gtagcctggc agagccgtgg gccgacacca ccacgccggc cggccgcatg gtgttgaccg
    14521 tgttcgccgg cattgccgag ttcgagcgtt ccctaatcat cgaccgcacc cggagcgggc
    14581 gcgaggccgc caaggcccga ggcgtgaagt ttggcccccg ccctaccctc accccggcac
    14641 agatcgcgca cgcccgcgag ctgatcgacc aggaaggccg caccgtgaaa gaggcggctg
    14701 cactgcttgg cgtgcatcgc tcgaccctgt accgcgcact tgagcgcagc gaggaagtga
    14761 cgcccaccga ggccaggcgg cgcggtgcct tccgtgagga cgcattgacc gaggccgacg
    14821 ccctggcggc cgccgagaat gaacgccaag aggaacaagc atgaaaccgc accaggacgg
    14881 ccaggacgaa ccgtttttca ttaccgaaga gatcgaggcg gagatgatcg cggccgggta
    14941 cgtgttcgag ccgcccgcgc acggctcaac cgtgcggctg catgaaatcc tggccggttt
    15001 gtctgatgcc aagctggcgg cctggccggc cagcttggcc gctgaagaaa ccgagcgccg
    15061 ccgtctaaaa aggtgatgtg tatttgagta aaacagcttg cgtcatgcgg tcgctgcgta
    15121 tatgatgcga tgagtaaata aacaaatacg caaggggaac gcatgaaggt tatcgctgta
    15181 cttaaccaga aaggcgggtc aggcaagacg accatcgcaa cccatctagc ccgcgccctg
    15241 caactcgccg gggccgatgt tctgttagtc gattccgatc cccagggcag tgcccgcgat
    15301 tgggcggccg tgcgggaaga tcaaccgcta accgttgtcg gcatcgaccg cccgacgatt
    15361 gaccgcgacg tgaaggccat cggccggcgc gacttcgtag tgatcgacgg agcgccccag
    15421 gcggcggact tggctgtgtc cgcgatcaag gcagccgact tcgtgctgat tccggtgcag
    15481 ccaagccctt acgacatatg ggccaccgcc gacctggtgg agctggttaa gcagcgcatt
    15541 gaggtcacgg atggaaggct acaagcggcc tttgtcgtgt cgcgggcgat caaaggcacg
    15601 cgcatcggcg gtgaggttgc cgaggcgctg gccgggtacg agctgcccat tcttgagtcc
    15661 cgtatcacgc agcgcgtgag ctacccaggc actgccgccg ccggcacaac cgttcttgaa
    15721 tcagaacccg agggcgacgc tgcccgcgag gtccaggcgc tggccgctga aattaaatca
    15781 aaactcattt gagttaatga ggtaaagaga aaatgagcaa aagcacaaac acgctaagtg
    15841 ccggccgtcc gagcgcacgc agcagcaagg ctgcaacgtt ggccagcctg gcagacacgc
    15901 cagccatgaa gcgggtcaac tttcagttgc cggcggagga tcacaccaag ctgaagatgt
    15961 acgcggtacg ccaaggcaag accattaccg agctgctatc tgaatacatc gcgcagctac
    16021 cagagtaaat gagcaaatga ataaatgagt agatgaattt tagcggctaa aggaggcggc
    16081 atggaaaatc aagaacaacc aggcaccgac gccgtggaat gccccatgtg tggaggaacg
    16141 ggcggttggc caggcgtaag cggctgggtt gcctgccggc cctgcaatgg cactggaacc
    16201 cccaagcccg aggaatcggc gtgagcggtc gcaaaccatc cggcccggta caaatcggcg
    16261 cggcgctggg tgatgacctg gtggagaagt tgaaggccgc gcaggccgcc cagcggcaac
    16321 gcatcgaggc agaagcacgc cccggtgaat cgtggcaagc ggccgctgat cgaatccgca
    16381 aagaatcccg gcaaccgccg gcagccggtg cgccgtcgat taggaagccg cccaagggcg
    16441 acgagcaacc agattttttc gttccgatgc tctatgacgt gggcacccgc gatagtcgca
    16501 gcatcatgga cgtggccgtt ttccgtctgt cgaagcgtga ccgacgagct ggcgaggtga
    16561 tccgctacga gcttccagac gggcacgtag aggtttccgc agggccggcc ggcatggcca
    16621 gtgtgtggga ttacgacctg gtactgatgg cggtttccca tctaaccgaa tccatgaacc
    16681 gataccggga agggaaggga gacaagcccg gccgcgtgtt ccgtccacac gttgcggacg
    16741 tactcaagtt ctgccggcga gccgatggcg gaaagcagaa agacgacctg gtagaaacct
    16801 gcattcggtt aaacaccacg cacgttgcca tgcagcgtac gaagaaggcc aagaacggcc
    16861 gcctggtgac ggtatccgag ggtgaagcct tgattagccg ctacaagatc gtaaagagcg
    16921 aaaccgggcg gccggagtac atcgagatcg agctagctga ttggatgtac cgcgagatca
    16981 cagaaggcaa gaacccggac gtgctgacgg ttcaccccga ttactttttg atcgatcccg
    17041 gcatcggccg ttttctctac cgcctggcac gccgcgccgc aggcaaggca gaagccagat
    17101 ggttgttcaa gacgatctac gaacgcagtg gcagcgccgg agagttcaag aagttctgtt
    17161 tcaccgtgcg caagctgatc gggtcaaatg acctgccgga gtacgatttg aaggaggagg
    17221 cggggcaggc tggcccgatc ctagtcatgc gctaccgcaa cctgatcgag ggcgaagcat
    17281 ccgccggttc ctaatgtacg gagcagatgc tagggcaaat tgccctagca ggggaaaaag
    17341 gtcgaaaagg cctctttcct gtggatagca cgtacattgg gaacccaaag ccgtacattg
    17401 ggaaccggaa cccgtacatt gggaacccaa agccgtacat tgggaaccgg tcacacatgt
    17461 aagtgactga tataaaagag aaaaaaggcg atttttccgc ctaaaactct ttaaaactta
    17521 ttaaaactct taaaacccgc ctggcctgtg cataactgtc tggccagcgc acagccgaag
    17581 agctgcaaaa agcgcctacc cttcggtcgc tgcgctccct acgccccgcc gcttcgcgtc
    17641 ggcctatcgc ggccgctggc cgctcaaaaa tggctggcct acggccaggc aatctaccag
    17701 ggcgcggaca agccgcgccg tcgccactcg accgccggcg cccacatcaa ggcaccctgc
    17761 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggaaacggtc
    17821 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
    17881 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
    17941 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
    18001 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
    18061 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
    18121 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
    18181 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
    18241 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
    18301 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
    18361 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
    18421 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
    18481 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
    18541 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
    18601 cgaggtat
//
