LOCUS       sequence-182459-       20401 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  WDV replicon T-DNA for gene targeting in wheat scutella, H840A 
            double nickase (nTaCas9_H840A+gUbi8+gUbi1+donor).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 20401)
  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 20401)
  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: pJG488
FEATURES             Location/Qualifiers
     source          1..20401
                     /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    2848..2872
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             join(5398..7914,7915..7917,7918..9501)
                     /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             9502..9549
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      9570..10277
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_RNA        10667..10742
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        11136..11211
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             11983..12036
                     /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(12037..12039,12040..12042,12043..12756)
                     /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(12082..12103)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(12343..12362)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     12690..12711
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      12787..13039
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    15631..15655
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             16956..17585
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             18014..19087
                     /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      19153..19347
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(19583..19605)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    19691..19831
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     19746..19765
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(19846..19863)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(20017..20401,1..204))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(20097..20116)
                     /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 gctgagagat
     1141 cccctcataa tttccccaaa gcgtaaccat gtgtgaataa attttgagct agtagggttg
     1201 cagccacgag taagtcttcc cttgttattg tgtagccaga atgccgcaaa acttccatgc
     1261 ctaagcgaac tgttgagagt acgtttcgat ttctgactgt gttagcctgg aagtgcttgt
     1321 cccaaccttg tttctgagca tgaacgcccg caagccaaca tgttagttga agcatcaggg
     1381 cgattagcag catgatatca aaacgctctg agctgctcgt tcggctatgg cgtaggccta
     1441 gtccgtaggc aggacttttc aagtctcgga aggtttcttc aatctgcatt cgcttcgaat
     1501 agatattaac aagttgtttg ggtgttcgaa tttcaacagg taagttagtt gctagaaccc
     1561 atggctcctt tgccgacgct gagtagattt taggtgacgg gtggtgacaa tgagtccgtg
     1621 tcgagcgctg attttttcgg cctttagagc gagatttata caatagaatt tggcatgaga
     1681 ttggattgct tttagtcagc ctcttatagc ctaaagtctt tgagtgacta gatgacatat
     1741 catgtaagtt gctgataggt ttccagtttt ccgctcctag gtctgcatat tgtacttttc
     1801 ctcttactcg acttaaccag taccaaccca gcttctcaac ggatttatac catggcactt
     1861 taaagccagc atcactgaca atgagcggtg tggtgttact cggtagaatg ctcgcaaggt
     1921 cggctagaaa ttggtcatga gctttctttg aacattgctc tgaaagcggg aacgctttct
     1981 cataaagagt aacagaacga ccgtgtagtg cgactgaagc tcgcaatacc ataagtcgtt
     2041 tttgctcacg aatatcagac cagtcaacaa gtacaatggg catcgtattg cccgaacaga
     2101 taaagctagc atgccaacgg tatacagcga gtcgctcttt gtggaggtga cgattaccta
     2161 acaatcggtc gattcgtttg atgttatgtt ttgttctcgc tttggttggc aggttacggc
     2221 caagttcggt aagagtgaga gttttacagt caagtaatgc gtggcaagcc aacgttaagc
     2281 tgttgagtcg ttttaagtgt aattcggggc agaattggta aagagagtcg tgtaaaatat
     2341 cgagttcgca catcttgttg tctgattatt gatttttcgc gaaaccattt gatcatatga
     2401 caagatgtgt atccacctta acttaatgat ttttaccaaa atcattaggg gattcatcag
     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 tagagctcgt tcctgcggcc
     2941 gcttaattaa ggtagtgaac agaagtccgg caggtcctta gcgaaaaaac ggggtgtgcc
     3001 agaaaactct atcctctacc ctgcgtggag gtgtgaattc tgcacactgc aaatgcaatg
     3061 tgtccaatgc tttatatagg gcaggttttg gcgggagaac agggccctag tgttcccacg
     3121 gtagcgtagc gaatcgtgtg ggccctgttc ggtgtgcggt cggggggcct ccacgcgggt
     3181 tataatatta ccccgcgtgg tggcccccga cgcgcactcg gcttttcgtg agtgcgcgga
     3241 ggcttttgga ccacatcttt tctgatcact ttcgtggaag atgttgattt atcacacttt
     3301 tgacggggaa atctgtgcca tgccttagct tataaggaag tgcgtggtag cccatctcga
     3361 caagtttgta ccgatctgca ggcgcgcctg cagtgcagcg tgacccggtc gtgcccctct
     3421 ctagagataa tgagcattgc atgtctaagt tataaaaaat taccacatat tttttttgtc
     3481 acacttgttt gaagtgcagt ttatctatct ttatacatat atttaaactt tactctacga
     3541 ataatataat ctatagtact acaataatat cagtgtttta gagaatcata taaatgaaca
     3601 gttagacatg gtctaaagga caattgagta ttttgacaac aggactctac agttttatct
     3661 ttttagtgtg catgtgttct cctttttttt tgcaaatagc ttcacctata taatacttca
     3721 tccattttat tagtacatcc atttagggtt tagggttaat ggtttttata gactaatttt
     3781 tttagtacat ctattttatt ctattttagc ctctaaatta agaaaactaa aactctattt
     3841 tagttttttt atttaataat ttagatataa aatagaataa aataaagtga ctaaaaatta
     3901 aacaaatacc ctttaagaaa ttaaaaaaac taaggaaaca tttttcttgt ttcgagtaga
     3961 taatgccagc ctgttaaacg ccgtcgacga gtctaacgga caccaaccag cgaaccagca
     4021 gcgtcgcgtc gggccaagcg aagcagacgg cacggcatct ctgtcgctgc ctctggaccc
     4081 ctctcgagag ttccgctcca ccgttggact tgctccgctg tcggcatcca gaaattgcgt
     4141 ggcggagcgg cagacgtgag ccggcacggc aggcggcctc ctcctcctct cacggcaccg
     4201 gcagctacgg gggattcctt tcccaccgct ccttcgcttt cccttcctcg cccgccgtaa
     4261 taaatagaca ccccctccac accctctttc cccaacctcg tgttgttcgg agcgcacaca
     4321 cacacaacca gatctccccc aaatccaccc gtcggcacct ccgcttcaag gtacgccgct
     4381 cgtcctcccc ccccccctct ctaccttctc tagatcggcg ttccggtcca tggttagggc
     4441 ccggtagttc tacttctgtt catgtttgtg ttagatccgt gtttgtgtta gatccgtgct
     4501 gctagcgttc gtacacggat gcgacctgta cgtcagacac gttctgattg ctaacttgcc
     4561 agtgtttctc tttggggaat cctgggatgg ctctagccgt tccgcagacg ggatcgattt
     4621 catgattttt tttgtttcgt tgcatagggt ttggtttgcc cttttccttt atttcaatat
     4681 atgccgtgca cttgtttgtc gggtcatctt ttcatgcttt tttttgtctt ggttgtgatg
     4741 atgtggtctg gttgggcggt cgttctagat cggagtagaa ttaattctgt ttcaaactac
     4801 ctggtggatt tattaatttt ggatctgtat gtgtgtgcca tacatattca tagttacgaa
     4861 ttgaagatga tggatggaaa tatcgatcta ggataggtat acatgttgat gcgggtttta
     4921 ctgatgcata tacagagatg ctttttgttc gcttggttgt gatgatgtgg tgtggttggg
     4981 cggtcgttca ttcgttctag atcggagtag aatactgttt caaactacct ggtgtattta
     5041 ttaattttgg aactgtatgt gtgtgtcata catcttcata gttacgagtt taagatggat
     5101 ggaaatatcg atctaggata ggtatacatg ttgatgtggg ttttactgat gcatatacat
     5161 gatggcatat gcagcatcta ttcatatgct ctaaccttga gtacctatct attataataa
     5221 acaagtatgt tttataatta ttttgatctt gatatacttg gatgatggca tatgcagcag
     5281 ctatatgtgg atttttttag ccctgccttc atacgctatt tatttgcttg gtactgtttc
     5341 ttttgtcgat gctcaccctg ttgtttggtg ttacttcctg cagggcgatc tattcgaatg
     5401 gacaagaagt actcgatcgg cctcgacatc gggacgaact cagttggctg ggccgtgatc
     5461 accgacgagt acaaggtgcc ctctaagaag ttcaaggtcc tggggaacac cgaccgccat
     5521 tccatcaaga agaacctcat cggcgctctc ctgttcgaca gcggggagac cgctgaggct
     5581 acgaggctca agagaaccgc taggcgccgg tacacgagaa ggaagaacag gatctgctac
     5641 ctccaagaga ttttctccaa cgagatggcc aaggttgacg attcattctt ccaccgcctg
     5701 gaggagtctt tcctcgtgga ggaggataag aagcacgagc ggcatcccat cttcggcaac
     5761 atcgtggacg aggttgccta ccacgagaag taccctacga tctaccatct gcggaagaag
     5821 ctcgtggact ccaccgataa ggcggacctc agactgatct acctcgctct ggcccacatg
     5881 atcaagttcc gcggccattt cctgatcgag ggggatctca acccagacaa cagcgatgtt
     5941 gacaagctgt tcatccaact cgtgcagacc tacaaccaac tcttcgagga gaacccgatc
     6001 aacgcctctg gcgtggacgc gaaggctatc ctgtccgcga ggctctcgaa gtccaggagg
     6061 ctggagaacc tgatcgctca gctcccaggc gagaagaaga acggcctgtt cgggaacctc
     6121 atcgctctca gcctggggct caccccgaac ttcaagtcga acttcgatct cgctgaggac
     6181 gccaagctgc aactctccaa ggacacctac gacgatgacc tcgataacct cctggcccag
     6241 atcggcgatc aatacgcgga cctgttcctc gctgccaaga acctgtcgga cgccatcctc
     6301 ctgtcagata tcctccgcgt gaacaccgag atcacgaagg ctccactctc tgcctccatg
     6361 atcaagcgct acgacgagca ccatcaggat ctgaccctcc tgaaggcgct ggtccgccaa
     6421 cagctcccgg agaagtacaa ggagattttc ttcgatcagt cgaagaacgg ctacgctggg
     6481 tacatcgacg gcggggcctc acaagaggag ttctacaagt tcatcaagcc aatcctggag
     6541 aagatggacg gcacggagga gctcctggtg aagctcaaca gggaggacct cctgcggaag
     6601 cagagaacct tcgataacgg cagcatcccc caccaaatcc atctcgggga gctgcacgcc
     6661 atcctgagaa ggcaagagga cttctaccct ttcctcaagg ataaccggga gaagatcgag
     6721 aagatcctga ccttcagaat cccatactac gtcggccctc tcgcgcgggg gaactcaaga
     6781 ttcgcttgga tgacccgcaa gtctgaggag accatcacgc cgtggaactt cgaggaggtg
     6841 gtggacaagg gcgctagcgc tcagtcgttc atcgagagga tgaccaactt cgacaagaac
     6901 ctgcccaacg agaaggtgct ccctaagcac tcgctcctgt acgagtactt caccgtctac
     6961 aacgagctca cgaaggtgaa gtacgtcacc gagggcatgc gcaagccagc gttcctgtcc
     7021 ggggagcaga agaaggctat cgtggacctc ctgttcaaga ccaaccggaa ggtcacggtt
     7081 aagcaactca aggaggacta cttcaagaag atcgagtgct tcgattcggt cgagatcagc
     7141 ggcgttgagg accgcttcaa cgccagcctc gggacctacc acgatctcct gaagatcatc
     7201 aaggataagg acttcctgga caacgaggag aacgaggata tcctggagga catcgtgctg
     7261 accctcacgc tgttcgagga cagggagatg atcgaggagc gcctgaagac gtacgcccat
     7321 ctcttcgatg acaaggtcat gaagcaactc aagcgccgga gatacaccgg ctgggggagg
     7381 ctgtcccgca agctcatcaa cggcatccgg gacaagcagt ccgggaagac catcctcgac
     7441 ttcctgaaga gcgatggctt cgccaacagg aacttcatgc aactgatcca cgatgacagc
     7501 ctcaccttca aggaggatat ccaaaaggct caagtgagcg gccaggggga ctcgctgcac
     7561 gagcatatcg cgaacctcgc tggctccccc gcgatcaaga agggcatcct ccagaccgtg
     7621 aaggttgtgg acgagctcgt gaaggtcatg ggccggcaca agcctgagaa catcgtcatc
     7681 gagatggcca gagagaacca aaccacgcag aaggggcaaa agaactctag ggagcgcatg
     7741 aagcgcatcg aggagggcat caaggagctg gggtcccaaa tcctcaagga gcacccagtg
     7801 gagaacaccc aactgcagaa cgagaagctc tacctgtact acctccagaa cggcagggat
     7861 atgtacgtgg accaagagct ggatatcaac cgcctcagcg attacgacgt cgatgctatc
     7921 gttccccagt ctttcctgaa ggatgactcc atcgacaaca aggtcctcac caggtcggac
     7981 aagaaccgcg gcaagtcaga taacgttcca tctgaggagg tcgttaagaa gatgaagaac
     8041 tactggaggc agctcctgaa cgccaagctg atcacgcaaa ggaagttcga caacctcacc
     8101 aaggctgaga gaggcgggct ctcagagctg gacaaggccg gcttcatcaa gcggcagctg
     8161 gtcgagacca gacaaatcac gaagcacgtt gcgcaaatcc tcgactctcg gatgaacacg
     8221 aagtacgatg agaacgacaa gctgatcagg gaggttaagg tgatcaccct gaagtctaag
     8281 ctcgtctccg acttcaggaa ggatttccag ttctacaagg ttcgcgagat caacaactac
     8341 caccatgccc atgacgctta cctcaacgct gtggtcggca ccgctctgat caagaagtac
     8401 ccaaagctgg agtccgagtt cgtgtacggg gactacaagg tttacgatgt gcgcaagatg
     8461 atcgccaagt cggagcaaga gatcggcaag gctaccgcca agtacttctt ctactcaaac
     8521 atcatgaact tcttcaagac cgagatcacg ctggccaacg gcgagatccg gaagagaccg
     8581 ctcatcgaga ccaacggcga gacgggggag atcgtgtggg acaagggcag ggatttcgcg
     8641 accgtccgca aggttctctc catgccccag gtgaacatcg tcaagaagac cgaggtccaa
     8701 acgggcgggt tctcaaagga gtctatcctg cctaagcgga acagcgacaa gctcatcgcc
     8761 agaaagaagg actgggaccc aaagaagtac ggcgggttcg acagccctac cgtggcctac
     8821 tcggtcctgg ttgtggcgaa ggttgagaag ggcaagtcca agaagctcaa gagcgtgaag
     8881 gagctcctgg ggatcaccat catggagagg tccagcttcg agaagaaccc aatcgacttc
     8941 ctggaggcca agggctacaa ggaggtgaag aaggacctga tcatcaagct cccgaagtac
     9001 tctctcttcg agctggagaa cggcaggaag agaatgctgg cttccgctgg cgagctccag
     9061 aaggggaacg agctcgcgct gccaagcaag tacgtgaact tcctctacct ggcttcccac
     9121 tacgagaagc tcaagggcag cccggaggac aacgagcaaa agcagctgtt cgtcgagcag
     9181 cacaagcatt acctcgacga gatcatcgag caaatctccg agttcagcaa gcgcgtgatc
     9241 ctcgccgacg cgaacctgga taaggtcctc tccgcctaca acaagcaccg ggacaagccc
     9301 atcagagagc aagcggagaa catcatccat ctcttcaccc tgacgaacct cggcgctcct
     9361 gctgctttca agtacttcga caccacgatc gatcggaaga gatacacctc cacgaaggag
     9421 gtcctggacg cgaccctcat ccaccagtcg atcaccggcc tgtacgagac gaggatcgac
     9481 ctctcacaac tcggcgggga taagagaccc gcagcaacca agaaggcagg gcaagcaaag
     9541 aagaagaagt gactcgagtc cctagagtcc tgctttaatg agatatgcga gacgcctatg
     9601 atcgcatgat atttgctttc aattctgttg tgcacgttgt aaaaaacctg agcatgtgta
     9661 gctcagatcc ttaccgccgg tttcggttca ttctaatgaa tatatcaccc gttactatcg
     9721 tatttttatg aataatattc tccgttcaat ttactgattg taccctacta cttatatgta
     9781 caatattaaa atgaaaacaa tatattgtgc tgaataggtt tatagcgaca tctatgatag
     9841 agcgccacaa taacaaacaa ttgcgtttta ttattacaaa tccaatttta aaaaaagcgg
     9901 cagaaccggt caaacctaaa agactgatta cataaatctt attcaaattt caaaagtgcc
     9961 ccaggggcta gtatctacga cacaccgagc ggcgaactaa taacgctcac tgaagggaac
    10021 tccggttccc cgccggcgcg catgggtgag attccttgaa gttgagtatt ggccgtccgc
    10081 tctaccgaaa gttacgggca ccattcaacc cggtccagca cggcggccgg gtaaccgact
    10141 tgctgccccg agaattatgc agcatttttt tggtgtatgt gggccccaaa tgaagtgcag
    10201 gtcaaacctt gacagtgacg acaaatcgtt gggcgggtcc agggcgaatt ttgcgacaac
    10261 atgtcgaggc tcagcaggag gacgaccaag cccgttattc tgacagttct ggtgctcaac
    10321 acatttatat ttatcaagga gcacattgtt actcactgct aggagggaat cgaactagga
    10381 atattgatca gaggaactac gagagagctg aagataactg ccctctagct ctcactgatc
    10441 tgggtcgcat agtgagatgc agcccacgtg agttcagcaa cggtctagcg ctgggctttt
    10501 aggcccgcat gatcgggctt ttgtcgggtg gtcgacgtgt tcacgattgg ggagagcaac
    10561 gcagcagttc ctcttagttt agtcccacct cgcctgtcca gcagagttct gaccggttta
    10621 taaactcgct tgctgcatca gacttgccag ccctgggact agcagcgttt tagagctaga
    10681 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
    10741 gctttttttc cgggaccaag cccgttattc tgacagttct ggtgctcaac acatttatat
    10801 ttatcaagga gcacattgtt actcactgct aggagggaat cgaactagga atattgatca
    10861 gaggaactac gagagagctg aagataactg ccctctagct ctcactgatc tgggtcgcat
    10921 agtgagatgc agcccacgtg agttcagcaa cggtctagcg ctgggctttt aggcccgcat
    10981 gatcgggctt ttgtcgggtg gtcgacgtgt tcacgattgg ggagagcaac gcagcagttc
    11041 ctcttagttt agtcccacct cgcctgtcca gcagagttct gaccggttta taaactcgct
    11101 tgctgcatca gacttgctgg tgcaactggt ggcccgtttt agagctagaa atagcaagtt
    11161 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttcg
    11221 cgtagtcctc ggtatggtgc tactggagct gctagtggca ggccagcagg tttatttggg
    11281 gctggacttc cggaattaga tcaaatgcag caacagttga gccagaatcc caaccttatg
    11341 agggagataa tgaacatgcc aatgatgcag agtctcatga ataaccctga tctaatacgc
    11401 aatatgatta tgaataatcc acaaatgcgt gatattattg atcggaatcc agatcttgcc
    11461 catgtcctca atgatcctag tgttctccgc cagacccttg aagctgcaag aaaccctgaa
    11521 attatgaggg agatgatgcg gaacacagac agagcaatga gcaacatcga agcttcccct
    11581 gaagggttta atatgctccg gcgtatgtat gaaactgtac aggagccttt tcttaatgca
    11641 acaacaatgg gagggggtgg ggaaggcacc ccggcctcta acccgtttgc agctcttctt
    11701 ggaaatcagg ggcctaacca agccggcaat gctccaacta ccggcccaga gtccacaaca
    11761 ggaacccctg ttccaaatac taatccactt ccaaacccct ggagcaacaa tggtaggttc
    11821 tagttattta gagttttttg tttgttttgt tgttgaatgt tgataattac atgtggtagt
    11881 atttttattc tcacagctgc tgataattgc ctgtgatact attatatttt cccagctggg
    11941 ggtgcgcaag gaacaacacg gtcaggtcct gctgctagtc cagagggcag aggaagtctt
    12001 ctaacatgcg gtgacgtgga ggagaatccc gggcccatgg tgagcaaggg cgaggagctg
    12061 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
    12121 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
    12181 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cacctacggc
    12241 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
    12301 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
    12361 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
    12421 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
    12481 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
    12541 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
    12601 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
    12661 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
    12721 gggatcactc acggcatgga cgagctgtac aagtaaagcg gccgggtacc gagctcgaat
    12781 ttccccgatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc tgttgccggt
    12841 cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat aattaacatg
    12901 taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca attatacatt
    12961 taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcgcggtg
    13021 tcatctatgt tactagatcg cagggctggt gcaactggtg gcccaccagg gctgggttca
    13081 gcagatttga gcagcctgct cggtggtctt ggtgggaatg caagaactgg tgctgcaggt
    13141 ggtctaggag ggttgggttc agcagatttg gggagtatgc ttggtggtcc acctgatgct
    13201 gctcttttga gtcagatgct gcaaaaccct gctatgatgc agatgatgca gaacattatg
    13261 tctgacccac agtcaatgaa ccaggtccaa tatttttcaa aactagttct tttatgattt
    13321 ttggagatga ccttggatca ttctgtaaca tttgcttgtc ccacagttgc ttagcatgaa
    13381 cccaaatgca cgtagcctga tggagtcaaa cactcagttg agggatatgt tccaaaaccc
    13441 agaatttctt cgccagatgg catccccaga ggctttgcag gtaaaatctg ttgtgatgca
    13501 agttaacaac tgttctcgta ttttattttc tgataaaatt tgtatttgtt ctgcgcagca
    13561 attactctca ttccagcaga cactgtcatc acagcttggc caaaatcaac ctagccagtg
    13621 agtaactctt ttttttgcga gaaaaaaggg aaaaagtaac actctaattc aatagcatga
    13681 ttgtatcacc cctttttttt atgaaattaa ataaaataga gattatgaag tgcagttatg
    13741 tttatctttt gagggtgcaa ttatgcgttt gctgagtctt ttcttttcag ggctggtaac
    13801 ctagggggca atggagtgta cttcaagtca caccggcgag tgtttgatcg ccggcggtac
    13861 aaagtggtta aaataatatt ttatttatct catgtcattc gattacagag gctcggctac
    13921 gagcaaagac aaaccaaata taacaaacaa caacccttac acaatgacat cggaaaacga
    13981 aatacaacac cctgagatat tacatttata gaaactgtac gccgtccgcg ctaggacagt
    14041 cactgcgaag cagtgacgtc ttcgccggag gcgaacgagt agttgatgaa cgtctcgcct
    14101 tcatacatgt agtgaacaac agtgttagag tacatgtaat ccgactgttc gggagtcata
    14161 tccttgagcc aatcttcgtc tggattaact aaaatgatgc aaggtattcc accccgtatg
    14221 acctttcgct taccatattt tggattgacc gtgaagtcac gctgagcccc gacgaagcac
    14281 ttccagttgg gtgtgaactt gaatggaatg tcgtcgatga tattatactt ggcgttgacg
    14341 tcatatgttg tgaaatcaac tagactgtta taataattgt gtgtccctag agaccttgcc
    14401 caggaagtct ttcctgttct ggttggcccg cagatgtaga tggacttatg cctccccggt
    14461 gactcctgga ataatcgtcc atccactcta agtcagattg cgcttgatcc gcaggagtgg
    14521 aagtacaaag gatataggat tcgaggctta cggagtagag atgttcattt ttccagcttt
    14581 caatggtctc atggcaaatg agtgattcgg ttggaaactc aggtgtgtaa gtggcaactg
    14641 ggtcaggaaa tagatggcgt gccgtgtact cgaagtcttt gagacggata gaccattcaa
    14701 acggaaaacg attgcaaacc atgctgagga attcctcgcg agaggaacta gattcaatga
    14761 tctgtttcat atccgcatca cggtctttac gacctggagt tgaaacagcc acgaatgttc
    14821 cccactcagc tgtgtttaca tcggagtcaa cctccttcgt gatgtaatca cgaacttggt
    14881 tgcagtcttt ggcagcttgt atatttggat ggaatatgga gaatggagat gtatccatac
    14941 ggaggtttaa ggcattggga ttggtgatgg aagcacgaag cttgttctgc acgagaacgt
    15001 gcagatgtgg tgatccatct tcgtggagct ctctaacagc agcgatgtag aggggctcat
    15061 atttgttcaa gagagtgcga agtgaatcca aggcgtactg tggctcaagg gtacattgag
    15121 gatatgttag aaagaggtac ttggaataga cacggaacct gggtgcagat gaagaggcca
    15181 tggtagtgaa cagaagtccg gcaggtcctt agcgaaaaaa cggggtgtgc cagaaaactc
    15241 tatcctctac cctgcgtgga ggtgtgaatt ctgcacactg caaatgcaat gtgtccaatg
    15301 ctttatatag ggcaggtttt ggcgggagaa cagggcccta gtgttcccac ggtagcgtag
    15361 cgaatcgtgt gggccctgtt cggtgtgcgg tcggggggcc tccacgcggg ttataatatt
    15421 accccgcgtg gtggcccccg acgcgcactc ggcttttcgt gagtgcgcgg aggcttttgg
    15481 accacatctt ttctgatcac tttcgtggaa gatgttgatt tatcacactt ttgacgggga
    15541 aatctgtgcc atgccttagc ttataaggaa gtgcgtggta gcccatctcg gggccctcga
    15601 ttcgacgttc ctgtttaaac tatcagtgtt tgacaggata tattggcggg taaacctaag
    15661 agaaaagagc gtttattaga ataatcggat atttaaaagg gcgtgaaaag gtttatccgt
    15721 tcgtccattt gtatgtgcat gccaaccaca gggttcccct cgggatcaaa gtactttgat
    15781 ccaacccctc cgctgctata gtgcagtcgg cttctgacgt tcagtgcagc cgtcttctga
    15841 aaacgacatg tcgcacaagt cctaagttac gcgacaggct gccgccctgc ccttttcctg
    15901 gcgttttctt gtcgcgtgtt ttagtcgcat aaagtagaat acttgcgact agaaccggag
    15961 acattacgcc atgaacaaga gcgccgccgc tggcctgctg ggctatgccc gcgtcagcac
    16021 cgacgaccag gacttgacca accaacgggc cgaactgcac gcggccggct gcaccaagct
    16081 gttttccgag aagatcaccg gcaccaggcg cgaccgcccg gagctggcca ggatgcttga
    16141 ccacctacgc cctggcgacg ttgtgacagt gaccaggcta gaccgcctgg cccgcagcac
    16201 ccgcgaccta ctggacattg ccgagcgcat ccaggaggcc ggcgcgggcc tgcgtagcct
    16261 ggcagagccg tgggccgaca ccaccacgcc ggccggccgc atggtgttga ccgtgttcgc
    16321 cggcattgcc gagttcgagc gttccctaat catcgaccgc acccggagcg ggcgcgaggc
    16381 cgccaaggcc cgaggcgtga agtttggccc ccgccctacc ctcaccccgg cacagatcgc
    16441 gcacgcccgc gagctgatcg accaggaagg ccgcaccgtg aaagaggcgg ctgcactgct
    16501 tggcgtgcat cgctcgaccc tgtaccgcgc acttgagcgc agcgaggaag tgacgcccac
    16561 cgaggccagg cggcgcggtg ccttccgtga ggacgcattg accgaggccg acgccctggc
    16621 ggccgccgag aatgaacgcc aagaggaaca agcatgaaac cgcaccagga cggccaggac
    16681 gaaccgtttt tcattaccga agagatcgag gcggagatga tcgcggccgg gtacgtgttc
    16741 gagccgcccg cgcacggctc aaccgtgcgg ctgcatgaaa tcctggccgg tttgtctgat
    16801 gccaagctgg cggcctggcc ggccagcttg gccgctgaag aaaccgagcg ccgccgtcta
    16861 aaaaggtgat gtgtatttga gtaaaacagc ttgcgtcatg cggtcgctgc gtatatgatg
    16921 cgatgagtaa ataaacaaat acgcaagggg aacgcatgaa ggttatcgct gtacttaacc
    16981 agaaaggcgg gtcaggcaag acgaccatcg caacccatct agcccgcgcc ctgcaactcg
    17041 ccggggccga tgttctgtta gtcgattccg atccccaggg cagtgcccgc gattgggcgg
    17101 ccgtgcggga agatcaaccg ctaaccgttg tcggcatcga ccgcccgacg attgaccgcg
    17161 acgtgaaggc catcggccgg cgcgacttcg tagtgatcga cggagcgccc caggcggcgg
    17221 acttggctgt gtccgcgatc aaggcagccg acttcgtgct gattccggtg cagccaagcc
    17281 cttacgacat atgggccacc gccgacctgg tggagctggt taagcagcgc attgaggtca
    17341 cggatggaag gctacaagcg gcctttgtcg tgtcgcgggc gatcaaaggc acgcgcatcg
    17401 gcggtgaggt tgccgaggcg ctggccgggt acgagctgcc cattcttgag tcccgtatca
    17461 cgcagcgcgt gagctaccca ggcactgccg ccgccggcac aaccgttctt gaatcagaac
    17521 ccgagggcga cgctgcccgc gaggtccagg cgctggccgc tgaaattaaa tcaaaactca
    17581 tttgagttaa tgaggtaaag agaaaatgag caaaagcaca aacacgctaa gtgccggccg
    17641 tccgagcgca cgcagcagca aggctgcaac gttggccagc ctggcagaca cgccagccat
    17701 gaagcgggtc aactttcagt tgccggcgga ggatcacacc aagctgaaga tgtacgcggt
    17761 acgccaaggc aagaccatta ccgagctgct atctgaatac atcgcgcagc taccagagta
    17821 aatgagcaaa tgaataaatg agtagatgaa ttttagcggc taaaggaggc ggcatggaaa
    17881 atcaagaaca accaggcacc gacgccgtgg aatgccccat gtgtggagga acgggcggtt
    17941 ggccaggcgt aagcggctgg gttgcctgcc ggccctgcaa tggcactgga acccccaagc
    18001 ccgaggaatc ggcgtgagcg gtcgcaaacc atccggcccg gtacaaatcg gcgcggcgct
    18061 gggtgatgac ctggtggaga agttgaaggc cgcgcaggcc gcccagcggc aacgcatcga
    18121 ggcagaagca cgccccggtg aatcgtggca agcggccgct gatcgaatcc gcaaagaatc
    18181 ccggcaaccg ccggcagccg gtgcgccgtc gattaggaag ccgcccaagg gcgacgagca
    18241 accagatttt ttcgttccga tgctctatga cgtgggcacc cgcgatagtc gcagcatcat
    18301 ggacgtggcc gttttccgtc tgtcgaagcg tgaccgacga gctggcgagg tgatccgcta
    18361 cgagcttcca gacgggcacg tagaggtttc cgcagggccg gccggcatgg ccagtgtgtg
    18421 ggattacgac ctggtactga tggcggtttc ccatctaacc gaatccatga accgataccg
    18481 ggaagggaag ggagacaagc ccggccgcgt gttccgtcca cacgttgcgg acgtactcaa
    18541 gttctgccgg cgagccgatg gcggaaagca gaaagacgac ctggtagaaa cctgcattcg
    18601 gttaaacacc acgcacgttg ccatgcagcg tacgaagaag gccaagaacg gccgcctggt
    18661 gacggtatcc gagggtgaag ccttgattag ccgctacaag atcgtaaaga gcgaaaccgg
    18721 gcggccggag tacatcgaga tcgagctagc tgattggatg taccgcgaga tcacagaagg
    18781 caagaacccg gacgtgctga cggttcaccc cgattacttt ttgatcgatc ccggcatcgg
    18841 ccgttttctc taccgcctgg cacgccgcgc cgcaggcaag gcagaagcca gatggttgtt
    18901 caagacgatc tacgaacgca gtggcagcgc cggagagttc aagaagttct gtttcaccgt
    18961 gcgcaagctg atcgggtcaa atgacctgcc ggagtacgat ttgaaggagg aggcggggca
    19021 ggctggcccg atcctagtca tgcgctaccg caacctgatc gagggcgaag catccgccgg
    19081 ttcctaatgt acggagcaga tgctagggca aattgcccta gcaggggaaa aaggtcgaaa
    19141 aggcctcttt cctgtggata gcacgtacat tgggaaccca aagccgtaca ttgggaaccg
    19201 gaacccgtac attgggaacc caaagccgta cattgggaac cggtcacaca tgtaagtgac
    19261 tgatataaaa gagaaaaaag gcgatttttc cgcctaaaac tctttaaaac ttattaaaac
    19321 tcttaaaacc cgcctggcct gtgcataact gtctggccag cgcacagccg aagagctgca
    19381 aaaagcgcct acccttcggt cgctgcgctc cctacgcccc gccgcttcgc gtcggcctat
    19441 cgcggccgct ggccgctcaa aaatggctgg cctacggcca ggcaatctac cagggcgcgg
    19501 acaagccgcg ccgtcgccac tcgaccgccg gcgcccacat caaggcaccc tgcctcgcgc
    19561 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggaaacg gtcacagctt
    19621 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
    19681 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa
    19741 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
    19801 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
    19861 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
    19921 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
    19981 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
    20041 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
    20101 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
    20161 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
    20221 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
    20281 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
    20341 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
    20401 t
//
