LOCUS       sequence-182478-       17270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BeYDV replicon T-DNA for gene targeting in tobacco leaves, nuclease 
            (AtCas9+gNt_R2+donor).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17270)
  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 17270)
  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: pJG380
FEATURES             Location/Qualifiers
     source          1..17270
                     /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"
     promoter        2475..2820
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2763..2783
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             2854..6957
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2854..6957
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6970..6990
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      7001..7249
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     misc_RNA        7738..7813
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        8173..8248
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             9732..10523
                     /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(9783..9802)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     10393..10412
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      10550..10802
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    12500..12524
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             13825..14454
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             14883..15956
                     /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      16022..16216
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(16452..16474)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    16560..16700
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     16615..16634
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(16715..16732)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(16886..17270,1..204))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(16966..16985)
                     /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 aaagtttaac gcgttagcag aaggcatgtt
     1621 gttgtgactc cgaggggttg cctcaaactc tatcttataa ccggcgtgga ggcatggagg
     1681 caggggtatt ttggtcattt taatagatag tggaaaatga cgtggaattt acttaaagac
     1741 gaagtctttg cgacaagggg gggcccacgc cgaatttaat attaccggcg tggccccccc
     1801 ttatcgcgag tgctttagca cgagcggtcc agatttaaag tagaaaattt cccgcccact
     1861 agggttaaag gtgttcacac tataaaagca tatacgatgt gatggtattt gatggagcgt
     1921 atattgtatc aggtatttcc gttggatacg aattattcgt acgaccctcg gtaccgatcg
     1981 gcgcgccaga tttgcctttt caatttcaga aagaatgcta acccacagat ggttagagag
     2041 gcttacgcag caggtatcat caagacgatc tacccgagca ataatctcca ggaaatcaaa
     2101 taccttccca agaaggttaa agatgcagtc aaaagattca ggactaactg catcaagaac
     2161 acagagaaag atatatttct caagatcaga agtactattc cagtatggac gattcaaggc
     2221 ttgcttcaca aaccaaggca agtaatagag attggagtct ctaaaaaggt agttcccact
     2281 gaatcaaagg ccatggagtc aaagattcaa atagaggacc taacagaact cgccgtaaag
     2341 actggcgaac agttcataca gagtctctta cgactcaatg acaagaagaa aatcttcgtc
     2401 aacatggtgg agcacgacac acttgtctac tccaaaaata tcaaagatac agtctcagaa
     2461 gaccaaaggg caattgagac ttttcaacaa agggtaatat ccggaaacct cctcggattc
     2521 cattgcccag ctatctgtca ctttattgtg aagatagtgg aaaaggaagg tggctcctac
     2581 aaatgccatc attgcgataa aggaaaggcc atcgttgaag atgcctctgc cgacagtggt
     2641 cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt tccaaccacg
     2701 tcttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga cgcacaatcc
     2761 cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt ggagagaaca
     2821 cgggggactc ctgcaggtag atcgctcgtc gacatggata agaagtactc tatcggactc
     2881 gatatcggaa ctaactctgt gggatgggct gtgatcaccg atgagtacaa ggtgccatct
     2941 aagaagttca aggttctcgg aaacaccgat aggcactcta tcaagaaaaa ccttatcggt
     3001 gctctcctct tcgattctgg tgaaactgct gaggctacca gactcaagag aaccgctaga
     3061 agaaggtaca ccagaagaaa gaacaggatc tgctacctcc aagagatctt ctctaacgag
     3121 atggctaaag tggatgattc attcttccac aggctcgaag agtcattcct cgtggaagaa
     3181 gataagaagc acgagaggca ccctatcttc ggaaacatcg ttgatgaggt ggcataccac
     3241 gagaagtacc ctactatcta ccacctcaga aagaagctcg ttgattctac tgataaggct
     3301 gatctcaggc tcatctacct cgctctcgct cacatgatca agttcagagg acacttcctc
     3361 atcgagggtg atctcaaccc tgataactct gatgtggata agttgttcat ccagctcgtg
     3421 cagacctaca accagctttt cgaagagaac cctatcaacg cttcaggtgt ggatgctaag
     3481 gctatcctct ctgctaggct ctctaagtca agaaggcttg agaacctcat tgctcagctc
     3541 cctggtgaga agaagaacgg acttttcgga aacttgatcg ctctctctct cggactcacc
     3601 cctaacttca agtctaactt cgatctcgct gaggatgcaa agctccagct ctcaaaggat
     3661 acctacgatg atgatctcga taacctcctc gctcagatcg gagatcagta cgctgatttg
     3721 ttcctcgctg ctaagaacct ctctgatgct atcctcctca gtgatatcct cagagtgaac
     3781 accgagatca ccaaggctcc actctcagct tctatgatca agagatacga tgagcaccac
     3841 caggatctca cacttctcaa ggctcttgtt agacagcagc tcccagagaa gtacaaagag
     3901 attttcttcg atcagtctaa gaacggatac gctggttaca tcgatggtgg tgcatctcaa
     3961 gaagagttct acaagttcat caagcctatc ctcgagaaga tggatggaac cgaggaactc
     4021 ctcgtgaagc tcaatagaga ggatcttctc agaaagcaga ggaccttcga taacggatct
     4081 atccctcatc agatccacct cggagagttg cacgctatcc ttagaaggca agaggatttc
     4141 tacccattcc tcaaggataa cagggaaaag attgagaaga ttctcacctt cagaatccct
     4201 tactacgtgg gacctctcgc tagaggaaac tcaagattcg cttggatgac cagaaagtct
     4261 gaggaaacca tcaccccttg gaacttcgaa gaggtggtgg ataagggtgc tagtgctcag
     4321 tctttcatcg agaggatgac caacttcgat aagaaccttc caaacgagaa ggtgctccct
     4381 aagcactctt tgctctacga gtacttcacc gtgtacaacg agttgaccaa ggttaagtac
     4441 gtgaccgagg gaatgaggaa gcctgctttt ttgtcaggtg agcaaaagaa ggctatcgtt
     4501 gatctcttgt tcaagaccaa cagaaaggtg accgtgaagc agctcaaaga ggattacttc
     4561 aagaaaatcg agtgcttcga ttcagttgag atttctggtg ttgaggatag gttcaacgca
     4621 tctctcggaa cctaccacga tctcctcaag atcattaagg ataaggattt cttggataac
     4681 gaggaaaacg aggatatctt ggaggatatc gttcttaccc tcaccctctt tgaagataga
     4741 gagatgattg aagaaaggct caagacctac gctcatctct tcgatgataa ggtgatgaag
     4801 cagttgaaga gaagaagata cactggttgg ggaaggctct caagaaagct cattaacgga
     4861 atcagggata agcagtctgg aaagacaatc cttgatttcc tcaagtctga tggattcgct
     4921 aacagaaact tcatgcagct catccacgat gattctctca cctttaaaga ggatatccag
     4981 aaggctcagg tttcaggaca gggtgatagt ctccatgagc atatcgctaa cctcgctgga
     5041 tctcctgcaa tcaagaaggg aatcctccag actgtgaagg ttgtggatga gttggtgaag
     5101 gtgatgggaa ggcataagcc tgagaacatc gtgatcgaaa tggctagaga gaaccagacc
     5161 actcagaagg gacagaagaa ctctagggaa aggatgaaga ggatcgagga aggtatcaaa
     5221 gagcttggat ctcagatcct caaagagcac cctgttgaga acactcagct ccagaatgag
     5281 aagctctacc tctactacct ccagaacgga agggatatgt atgtggatca agagttggat
     5341 atcaacaggc tctctgatta cgatgttgat catatcgtgc cacagtcatt cttgaaggat
     5401 gattctatcg ataacaaggt gctcaccagg tctgataaga acaggggtaa gagtgataac
     5461 gtgccaagtg aagaggttgt gaagaaaatg aagaactatt ggaggcagct cctcaacgct
     5521 aagctcatca ctcagagaaa gttcgataac ttgactaagg ctgagagggg aggactctct
     5581 gaattggata aggcaggatt catcaagagg cagcttgtgg aaaccaggca gatcactaag
     5641 cacgttgcac agatcctcga ttctaggatg aacaccaagt acgatgagaa cgataagttg
     5701 atcagggaag tgaaggttat caccctcaag tcaaagctcg tgtctgattt cagaaaggat
     5761 ttccaattct acaaggtgag ggaaatcaac aactaccacc acgctcacga tgcttacctt
     5821 aacgctgttg ttggaaccgc tctcatcaag aagtatccta agctcgagtc agagttcgtg
     5881 tacggtgatt acaaggtgta cgatgtgagg aagatgatcg ctaagtctga gcaagagatc
     5941 ggaaaggcta ccgctaagta tttcttctac tctaacatca tgaatttctt caagaccgag
     6001 attaccctcg ctaacggtga gatcagaaag aggccactca tcgagacaaa cggtgaaaca
     6061 ggtgagatcg tgtgggataa gggaagggat ttcgctaccg ttagaaaggt gctctctatg
     6121 ccacaggtga acatcgttaa gaaaaccgag gtgcagaccg gtggattctc taaagagtct
     6181 atcctcccta agaggaactc tgataagctc attgctagga agaaggattg ggaccctaag
     6241 aaatacggtg gtttcgattc tcctaccgtg gcttactctg ttctcgttgt ggctaaggtt
     6301 gagaagggaa agagtaagaa gctcaagtct gttaaggaac ttctcggaat cactatcatg
     6361 gaaaggtcat ctttcgagaa gaacccaatc gatttcctcg aggctaaggg atacaaagag
     6421 gttaagaagg atctcatcat caagctccca aagtactcac tcttcgaact cgagaacggt
     6481 agaaagagga tgctcgcttc tgctggtgag cttcaaaagg gaaacgagct tgctctccca
     6541 tctaagtacg ttaactttct ttacctcgct tctcactacg agaagttgaa gggatctcca
     6601 gaagataacg agcagaagca acttttcgtt gagcagcaca agcactactt ggatgagatc
     6661 atcgagcaga tctctgagtt ctctaaaagg gtgatcctcg ctgatgcaaa cctcgataag
     6721 gtgttgtctg cttacaacaa gcacagagat aagcctatca gggaacaggc agagaacatc
     6781 atccatctct tcacccttac caacctcggt gctcctgctg ctttcaagta cttcgataca
     6841 accatcgata ggaagagata cacctctacc aaagaagtgc tcgatgctac cctcatccat
     6901 cagtctatca ctggactcta cgagactagg atcgatctct cacagctcgg tggtgattca
     6961 agggctgatc ctaagaagaa gaggaaggtt tgactcgaga tatgaagatg aagatgaaat
     7021 atttggtgtg tcaaataaaa agcttgtgtg cttaagtttg tgtttttttc ttggcttgtt
     7081 gtgttatgaa tttgtggctt tttctaatat taaatgaatg taagatcaca ttataatgaa
     7141 taaacaaatg tttctataat ccattgtgaa tgttttgttg gatctcttct gcagcatata
     7201 actactgtat gtgctatggt atggactatg gaatatgatt aaagataagg agctccggtg
     7261 acggacccat ggcttcgttg aacaacggaa actcgacttg ccttccgcac aatacatcat
     7321 ttcttcttag ctttttttct tcttcttcgt tcatacagtt tttttttgtt tatcagctta
     7381 cattttcttg aaccgtagct ttcgttttct tctttttaac tttccattcg gagtttttgt
     7441 atcttgtttc atagtttgtc ccaggattag aatgattagg catcgaacct tcaagaattt
     7501 gattgaataa aacatcttca ttcttaagat atgaagataa tcttcaaaag gcccctggga
     7561 atctgaaaga agagaagcag gcccatttat atgggaaaga acaatagtat ttcttatata
     7621 ggcccattta agttgaaaac aatcttcaaa agtcccacat cgcttagata agaaaacgaa
     7681 gctgagttta tatacagcta gagtcgaagt agtgattgcg tcccgggtcg ctaccttgtt
     7741 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
     7801 accgagtcgg tgcttttttt cccggcgcca tggatgttgt tgttaccaga aagtaaataa
     7861 atgttcaatc tctgatgttc tcaagtaagt gagttttatt gggaataata ttaacttatg
     7921 ttcttcttgc atttgatttc tttgccgctc tcttcttcta tcttaaatct gtgtatacta
     7981 tttcactatt gggcttttta ttagtctata atgggactca aaataaggct ttggcccaca
     8041 tcaaaaagat aagtcacaaa tcaaaactaa attcagagtc ttttctccca catcggtcac
     8101 tgtactcatt ttgtgtttgt ttatatatta cacgaaccga tctttggtac ggagacggag
     8161 tcgattcgtc tcgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
     8221 acttgaaaaa gtggcaccga gtcggtgctt tttttcgcgc gtagtcctcg gtacagtctt
     8281 acttccatga tttctttaac tatgccggaa tccatcgcag cgtaatgctc tacaccacgc
     8341 cgaacacctg ggtggacgat atcaccgtgg tgacgcatgt cgcgcaagac tgtaaccacg
     8401 cgtctgttga ctggcaggtg gtggccaatg gtgatgtcag cgttgaactg cgtgatgcgg
     8461 atcaacaggt ggttgcaact ggacaaggca ctagcgggac tttgcaagtg gtgaatccgc
     8521 acctctggca accgggtgaa ggttatctct atgaactgtg cgtcacagcc aaaagccaga
     8581 cagagtgtga tatctacccg cttcgcgtcg gcatccggtc agtggcagtg aagggcgaac
     8641 agttcctgat taaccacaaa ccgttctact ttactggctt tggtcgtcat gaagatgcgg
     8701 acttgcgtgg caaaggattc gataacgtgc tgatggtgca cgaccacgca ttaatggact
     8761 ggattggggc caactcctac cgtacctcgc attaccctta cgctgaagag atgctcgact
     8821 gggcagatga acatggcatc gtggtgattg atgaaactgc tgctgtcggc tttaacctct
     8881 ctttaggcat tggtttcgaa gcgggcaaca agccgaaaga actgtacagc gaagaggcag
     8941 tcaacgggga aactcagcaa gcgcacttac aggcgattaa agagctgata gcgcgtgaca
     9001 aaaaccaccc aagcgtggtg atgtggagta ttgccaacga accggatacc cgtccgcaag
     9061 gtgcacggga atatttcgcg ccactggcgg aagcaacgcg taaactcgac ccgacgcgtc
     9121 cgatcacctg cgtcaatgta atgttctgcg acgctcacac cgataccatc agcgatctct
     9181 ttgatgtgct gtgcctgaac cgttattacg gatggtatgt ccaaagcggc gatttggaaa
     9241 cggcagagaa ggtactggaa aaagaacttc tggcctggca ggagaaactg catcagccga
     9301 ttatcatcac cgaatacggc gtggatacgt tagccgggct gcactcaatg tacaccgaca
     9361 tgtggagtga agagtatcag tgtgcatggc tggatatgta tcaccgcgtc tttgatcgcg
     9421 tcagcgccgt cgtcggtgaa caggtatgga atttcgccga ttttgcgacc tcgcaaggca
     9481 tattgcgcgt tggcggtaac aagaaaggga tcttcactcg cgaccgcaaa ccgaagtcgg
     9541 cggcttttct gctgcaaaaa cgctggactg gcatgaactt cggtgaaaaa ccgcagcagg
     9601 gaggcaaaca acgcagggag gcaaacaatg atatcacaac tctcctgacg cgtcatcgtc
     9661 ggctacagcc tcgggaattg ctacctagct cgagcaagat ccaaggagat ataacaatgg
     9721 cttcctcctg gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     9781 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     9841 ggctgtcagc gcagggtaga ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     9901 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gtaccttgcg
     9961 ctgctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
    10021 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
    10081 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
    10141 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
    10201 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagaa
    10261 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
    10321 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
    10381 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
    10441 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
    10501 gccttcttga cgagttcttc tgataaccgc ggagagctcg aatttccccg atcgttcaaa
    10561 catttggcaa taaagtttct taagattgaa tcctgttgcc ggtcttgcga tgattatcat
    10621 ataatttctg ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt
    10681 tatgagatgg gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa
    10741 caaaatatag cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga
    10801 tcggagtgta cttcaagtca caccggcgag tgtttgatcg ccggcggtac cgagtgtact
    10861 tcaagtcagt gggaaatcaa taaaatgatt attttatgaa tatatttcat tgtgcaagta
    10921 gatagaaatt acatatgtta cataacacac gaaataaaca aaaaaagaca atccaaaaac
    10981 aaacacccca aaaaaaataa tcactttaga taaactcgta tgaggagagg cacgttcagt
    11041 gactcgacga ttcccgagca aaaaaagtct ccccgtcaca catgtagtgg gtgacgcaat
    11101 tatctttaaa gtaatccttc tgttgacttg tcattgataa catccagtct tcgtcaggat
    11161 tgcaaagaat tatagaaggg atcccacctt ttattttctt cttttttcca tatttagggt
    11221 tgacagtgaa atcagactgg caacctatta attgcttcca caatgggacg aacttgaagg
    11281 ggatgtcgtc gatgatatta taggtggcgt gttcatcgta gttggtgaaa tcgatggtac
    11341 cgttccaata gttgtgtcgt ccgagacttc tagcccaggt ggtctttccg gtacgagttg
    11401 gtccgcagat gtagaggctg gggtgtcgga ttccattcct tccattgtcc ttgttaaatc
    11461 ggccatccat tcaaggtcag attgagcttg ttggtatgag acaggatgta tgtaagtata
    11521 agcgtctatg cttacatggt atagatgggt ttccctccag gagtgtagat cttcgtggca
    11581 gcgaagatct gattctgtga agggcgacac atacggttca ggttgtggag ggaataattt
    11641 gttggctgaa tattccagcc attgaagctt tgttgcccat tcatgaggga attcttcctt
    11701 gatcatgtca agatattcct ccttagacgt tgcagtctgg ataatagttc tccatcgtgc
    11761 gtcagatttg cgaggagaaa ccttatgatc tcggaaatct cctctggttt taatatctcc
    11821 gtcctttgat atgtaatcaa ggacttgttt agagtttcta gctggctgga tattagggtg
    11881 atttccttca aaatcgaaaa aagaaggatc cctaatacaa ggttttttat caagctggag
    11941 aagagcatga tagtgggtag tgccatcttg atgaagctca gaagcaacac caaggaagaa
    12001 aataagaaaa ggtgtgagtt tctcccagag aaactggaat aaatcatctc tttgagatga
    12061 gcacttggga taggtaagga aaacatattt agattggagt ctgaagttct tactagcaga
    12121 aggcatgttg ttgtgactcc gaggggttgc ctcaaactct atcttataac cggcgtggag
    12181 gcatggaggc aggggtattt tggtcatttt aatagatagt ggaaaatgac gtggaattta
    12241 cttaaagacg aagtctttgc gacaaggggg ggcccacgcc gaatttaata ttaccggcgt
    12301 ggccccccct tatcgcgagt gctttagcac gagcggtcca gatttaaagt agaaaatttc
    12361 ccgcccacta gggttaaagg tgttcacact ataaaagcat atacgatgtg atggtatttg
    12421 atggagcgta tattgtatca ggtatttccg ttggatacga attattcgta cgaccctcat
    12481 agtttaaact atcagtgttt gacaggatat attggcgggt aaacctaaga gaaaagagcg
    12541 tttattagaa taatcggata tttaaaaggg cgtgaaaagg tttatccgtt cgtccatttg
    12601 tatgtgcatg ccaaccacag ggttcccctc gggatcaaag tactttgatc caacccctcc
    12661 gctgctatag tgcagtcggc ttctgacgtt cagtgcagcc gtcttctgaa aacgacatgt
    12721 cgcacaagtc ctaagttacg cgacaggctg ccgccctgcc cttttcctgg cgttttcttg
    12781 tcgcgtgttt tagtcgcata aagtagaata cttgcgacta gaaccggaga cattacgcca
    12841 tgaacaagag cgccgccgct ggcctgctgg gctatgcccg cgtcagcacc gacgaccagg
    12901 acttgaccaa ccaacgggcc gaactgcacg cggccggctg caccaagctg ttttccgaga
    12961 agatcaccgg caccaggcgc gaccgcccgg agctggccag gatgcttgac cacctacgcc
    13021 ctggcgacgt tgtgacagtg accaggctag accgcctggc ccgcagcacc cgcgacctac
    13081 tggacattgc cgagcgcatc caggaggccg gcgcgggcct gcgtagcctg gcagagccgt
    13141 gggccgacac caccacgccg gccggccgca tggtgttgac cgtgttcgcc ggcattgccg
    13201 agttcgagcg ttccctaatc atcgaccgca cccggagcgg gcgcgaggcc gccaaggccc
    13261 gaggcgtgaa gtttggcccc cgccctaccc tcaccccggc acagatcgcg cacgcccgcg
    13321 agctgatcga ccaggaaggc cgcaccgtga aagaggcggc tgcactgctt ggcgtgcatc
    13381 gctcgaccct gtaccgcgca cttgagcgca gcgaggaagt gacgcccacc gaggccaggc
    13441 ggcgcggtgc cttccgtgag gacgcattga ccgaggccga cgccctggcg gccgccgaga
    13501 atgaacgcca agaggaacaa gcatgaaacc gcaccaggac ggccaggacg aaccgttttt
    13561 cattaccgaa gagatcgagg cggagatgat cgcggccggg tacgtgttcg agccgcccgc
    13621 gcacggctca accgtgcggc tgcatgaaat cctggccggt ttgtctgatg ccaagctggc
    13681 ggcctggccg gccagcttgg ccgctgaaga aaccgagcgc cgccgtctaa aaaggtgatg
    13741 tgtatttgag taaaacagct tgcgtcatgc ggtcgctgcg tatatgatgc gatgagtaaa
    13801 taaacaaata cgcaagggga acgcatgaag gttatcgctg tacttaacca gaaaggcggg
    13861 tcaggcaaga cgaccatcgc aacccatcta gcccgcgccc tgcaactcgc cggggccgat
    13921 gttctgttag tcgattccga tccccagggc agtgcccgcg attgggcggc cgtgcgggaa
    13981 gatcaaccgc taaccgttgt cggcatcgac cgcccgacga ttgaccgcga cgtgaaggcc
    14041 atcggccggc gcgacttcgt agtgatcgac ggagcgcccc aggcggcgga cttggctgtg
    14101 tccgcgatca aggcagccga cttcgtgctg attccggtgc agccaagccc ttacgacata
    14161 tgggccaccg ccgacctggt ggagctggtt aagcagcgca ttgaggtcac ggatggaagg
    14221 ctacaagcgg cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt
    14281 gccgaggcgc tggccgggta cgagctgccc attcttgagt cccgtatcac gcagcgcgtg
    14341 agctacccag gcactgccgc cgccggcaca accgttcttg aatcagaacc cgagggcgac
    14401 gctgcccgcg aggtccaggc gctggccgct gaaattaaat caaaactcat ttgagttaat
    14461 gaggtaaaga gaaaatgagc aaaagcacaa acacgctaag tgccggccgt ccgagcgcac
    14521 gcagcagcaa ggctgcaacg ttggccagcc tggcagacac gccagccatg aagcgggtca
    14581 actttcagtt gccggcggag gatcacacca agctgaagat gtacgcggta cgccaaggca
    14641 agaccattac cgagctgcta tctgaataca tcgcgcagct accagagtaa atgagcaaat
    14701 gaataaatga gtagatgaat tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa
    14761 ccaggcaccg acgccgtgga atgccccatg tgtggaggaa cgggcggttg gccaggcgta
    14821 agcggctggg ttgcctgccg gccctgcaat ggcactggaa cccccaagcc cgaggaatcg
    14881 gcgtgagcgg tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc
    14941 tggtggagaa gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac
    15001 gccccggtga atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc
    15061 cggcagccgg tgcgccgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt
    15121 tcgttccgat gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg
    15181 ttttccgtct gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag
    15241 acgggcacgt agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc
    15301 tggtactgat ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg
    15361 gagacaagcc cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc
    15421 gagccgatgg cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca
    15481 cgcacgttgc catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg
    15541 agggtgaagc cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt
    15601 acatcgagat cgagctagct gattggatgt accgcgagat cacagaaggc aagaacccgg
    15661 acgtgctgac ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct
    15721 accgcctggc acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct
    15781 acgaacgcag tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga
    15841 tcgggtcaaa tgacctgccg gagtacgatt tgaaggagga ggcggggcag gctggcccga
    15901 tcctagtcat gcgctaccgc aacctgatcg agggcgaagc atccgccggt tcctaatgta
    15961 cggagcagat gctagggcaa attgccctag caggggaaaa aggtcgaaaa ggcctctttc
    16021 ctgtggatag cacgtacatt gggaacccaa agccgtacat tgggaaccgg aacccgtaca
    16081 ttgggaaccc aaagccgtac attgggaacc ggtcacacat gtaagtgact gatataaaag
    16141 agaaaaaagg cgatttttcc gcctaaaact ctttaaaact tattaaaact cttaaaaccc
    16201 gcctggcctg tgcataactg tctggccagc gcacagccga agagctgcaa aaagcgccta
    16261 cccttcggtc gctgcgctcc ctacgccccg ccgcttcgcg tcggcctatc gcggccgctg
    16321 gccgctcaaa aatggctggc ctacggccag gcaatctacc agggcgcgga caagccgcgc
    16381 cgtcgccact cgaccgccgg cgcccacatc aaggcaccct gcctcgcgcg tttcggtgat
    16441 gacggtgaaa acctctgaca catgcagctc ccggaaacgg tcacagcttg tctgtaagcg
    16501 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
    16561 gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat
    16621 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
    16681 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
    16741 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
    16801 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
    16861 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
    16921 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
    16981 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
    17041 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
    17101 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
    17161 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
    17221 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
//
