LOCUS       sequence-151858-       15385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  recipient plasmid [multiplexing, nickase]; pPcUbi:Cas9 D10A, BASTA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15385)
  AUTHORS   Ordon J, Gantner J, Kemna J, Schwalgun L, Reschke M, Streubel J, 
            Boch J, Stuttmann J
  TITLE     Generation of chromosomal deletions in dicotyledonous plants 
            employing a user-friendly genome editing toolkit.
  JOURNAL   Plant J. 2016 Aug 31. doi: 10.1111/tpj.13319.
  PUBMED    27579989
REFERENCE   2  (bases 1 to 15385)
  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 J. 
            2016 Aug 31. doi: 10.1111/tpj.13319."
COMMENT     Sequence Label: pDGE79
FEATURES             Location/Qualifiers
     source          1..15385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    302..326
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1627..2256
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2690..3757
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      3823..4017
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4253..4275)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4361..4501
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4416..4435
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4516..4533)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4687..5275)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4767..4786)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5519..6310)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    6838..6862
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(6905..7157)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(7177..7728)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(7771..7950)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      complement(8044..8751)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     CDS             complement(8762..8782)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(8795..12868,12869..12871,12872..12898))
                     /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             complement(8795..12898)
                     /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="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             complement(12905..12925)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        13936..14038
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             14039..14698
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(14105..14124)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             15040..15345
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     15224..15243
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
ORIGIN
        1 ccatgctaga gtccgcaaaa atcaccagtc tctctctaca aatctatctc tctctatttt
       61 tctccagaat aatgtgtgag tagttcccag ataagggaat tagggttctt atagggtttc
      121 gctcatgtgt tgagcatata agaaaccctt agtatgtatt tgtatttgta aaatacttct
      181 atcaataaaa tttctaattc ctaaaaccaa aatccagtga cctgcaggca tgcgacgtcg
      241 agcttagctt gagcttggat cagattgtcg tttcccgcct tcagtttaaa ctatcagtgt
      301 ttgacaggat atattggcgg gtaaacctaa gagaaaagag cgtttattag aataatcgga
      361 tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac
      421 agggttcccc tcgggatcaa agtactttga tccaacccct ccgctgctat agtgcagtcg
      481 gcttctgacg ttcagtgcag ccgtcttctg aaaacgacat gtcgcacaag tcctaagtta
      541 cgcgacaggc tgccgccctg cccttttcct ggcgttttct tgtcgcgtgt tttagtcgca
      601 taaagtagaa tacttgcgac tagaaccgga gacattacgc catgaacaag agcgccgccg
      661 ctggcctgct gggctatgcc cgcgtcagca ccgacgacca ggacttgacc aaccaacggg
      721 ccgaactgca cgcggccggc tgcaccaagc tgttttccga gaagatcacc ggcaccaggc
      781 gcgaccgccc ggagctggcc aggatgcttg accacctacg ccctggcgac gttgtgacag
      841 tgaccaggct agaccgcctg gcccgcagca cccgcgacct actggacatt gccgagcgca
      901 tccaggaggc cggcgcgggc ctgcgtagcc tggcagagcc gtgggccgac accaccacgc
      961 cggccggccg catggtgttg accgtgttcg ccggcattgc cgagttcgag cgttccctaa
     1021 tcatcgaccg cacccggagc gggcgcgagg ccgccaaggc ccgaggcgtg aagtttggcc
     1081 cccgccctac cctcaccccg gcacagatcg cgcacgcccg cgagctgatc gaccaggaag
     1141 gccgcaccgt gaaagaggcg gctgcactgc ttggcgtgca tcgctcgacc ctgtaccgcg
     1201 cacttgagcg cagcgaggaa gtgacgccca ccgaggccag gcggcgcggt gccttccgtg
     1261 aggacgcatt gaccgaggcc gacgccctgg cggccgccga gaatgaacgc caagaggaac
     1321 aagcatgaaa ccgcaccagg acggccagga cgaaccgttt ttcattaccg aagagatcga
     1381 ggcggagatg atcgcggccg ggtacgtgtt cgagccgccc gcgcacgtct caaccgtgcg
     1441 gctgcatgaa atcctggccg gtttgtctga tgccaagctg gcggcctggc cggccagctt
     1501 ggccgctgaa gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag
     1561 cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta aataaacaaa tacgcaaggg
     1621 gaacgcatga aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc
     1681 gcaacccatc tagcccgcgc cctgcaactc gccggggccg atgttctgtt agtcgattcc
     1741 gatccccagg gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt
     1801 gtcggcatcg accgcccgac gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc
     1861 gtagtgatcg acggagcgcc ccaggcggcg gacttggctg tgtccgcgat caaggcagcc
     1921 gacttcgtgc tgattccggt gcagccaagc ccttacgaca tatgggccac cgccgacctg
     1981 gtggagctgg ttaagcagcg cattgaggtc acggatggaa ggctacaagc ggcctttgtc
     2041 gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg
     2101 tacgagctgc ccattcttga gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc
     2161 gccgccggca caaccgttct tgaatcagaa cccgagggcg acgctgcccg cgaggtccag
     2221 gcgctggccg ctgaaattaa atcaaaactc atttgagtta atgaggtaaa gagaaaatga
     2281 gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa
     2341 cgttggccag cctggcagac acgccagcca tgaagcgggt caactttcag ttgccggcgg
     2401 aggatcacac caagctgaag atgtacgcgg tacgccaagg caagaccatt accgagctgc
     2461 tatctgaata catcgcgcag ctaccagagt aaatgagcaa atgaataaat gagtagatga
     2521 attttagcgg ctaaaggagg cggcatggaa aatcaagaac aaccaggcac cgacgccgtg
     2581 gaatgcccca tgtgtggagg aacgggcggt tggccaggcg taagcggctg ggttgtctgc
     2641 cggccctgca atggcactgg aacccccaag cccgaggaat cggcgtgacg gtcgcaaacc
     2701 atccggcccg gtacaaatcg gcgcggcgct gggtgatgac ctggtggaga agttgaaggc
     2761 cgcgcaggcc gcccagcggc aacgcatcga ggcagaagca cgccccggtg aatcgtggca
     2821 agcggccgct gatcgaatcc gcaaagaatc ccggcaaccg ccggcagccg gtgcgccgtc
     2881 gattaggaag ccgcccaagg gcgacgagca accagatttt ttcgttccga tgctctatga
     2941 cgtgggcacc cgcgatagtc gcagcatcat ggacgtggcc gttttccgtc tgtcgaagcg
     3001 tgaccgacga gctggcgagg tgatccgcta cgagcttcca gacgggcacg tagaggtttc
     3061 cgcagggccg gccggcatgg ccagtgtgtg ggattacgac ctggtactga tggcggtttc
     3121 ccatctaacc gaatccatga accgataccg ggaagggaag ggagacaagc ccggccgcgt
     3181 gttccgtcca cacgttgcgg acgtactcaa gttctgccgg cgagccgatg gcggaaagca
     3241 gaaagacgac ctggtagaaa cctgcattcg gttaaacacc acgcacgttg ccatgcagcg
     3301 tacgaagaag gccaagaacg gccgcctggt gacggtatcc gagggtgaag ccttgattag
     3361 ccgctacaag atcgtaaaga gcgaaaccgg gcggccggag tacatcgaga tcgagctagc
     3421 tgattggatg taccgcgaga tcacagaagg caagaacccg gacgtgctga cggttcaccc
     3481 cgattacttt ttgatcgatc ccggcatcgg ccgttttctc taccgcctgg cacgccgcgc
     3541 cgcaggcaag gcagaagcca gatggttgtt caagacgatc tacgaacgca gtggcagcgc
     3601 cggagagttc aagaagttct gtttcaccgt gcgcaagctg atcgggtcaa atgacctgcc
     3661 ggagtacgat ttgaaggagg aggcggggca ggctggcccg atcctagtca tgcgctaccg
     3721 caacctgatc gagggcgaag catccgccgg ttcctaatgt acggagcaga tgctagggca
     3781 aattgcccta gcaggggaaa aaggtcgaaa aggtgtcttt cctgtggata gcacgtacat
     3841 tgggaaccca aagccgtaca ttgggaaccg gaacccgtac attgggaacc caaagccgta
     3901 cattgggaac cggtcacaca tgtaagtgac tgatataaaa gagaaaaaag gcgatttttc
     3961 cgcctaaaac tctttaaaac ttattaaaac tcttaaaacc cgcctggcct gtgcataact
     4021 gtctggccag cgcacagccg aagagctgca aaaagcgcct acccttcggt cgctgcgctc
     4081 cctacgcccc gccgcttcgc gtcggcctat cgcggccgct ggccgctcaa aaatggctgg
     4141 cctacggcca ggcaatctac cagggcgcgg acaagccgcg ccgtcgccac tcgaccgccg
     4201 gcgcccacat caaggcaccc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
     4261 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     4321 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac
     4381 gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag
     4441 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     4501 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4561 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4621 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4681 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4741 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4801 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4861 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4921 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4981 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5041 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5101 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     5161 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5221 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5281 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5341 tttggtcatg catgatatat ctcccaattt gtgtagggct tattatgcac gcttaaaaat
     5401 aataaaagca gacttgacct gatagtttgg ctgtgagcaa ttatgtgctt agtgcatcta
     5461 atcgcttgag ttaacgccgg cgaagcggcg tcggcttgaa cgaatttcta gctagacatt
     5521 atttgccgac taccttggtg atctcgcctt tcacgtagtg gacaaattct tccaactgat
     5581 ctgcgcgcga ggccaagcga tcttcttctt gtccaagata agcctgtcta gcttcaagta
     5641 tgacgggctg atactgggcc ggcaggcgct ccattgccca gtcggcagcg acatccttcg
     5701 gcgcgatttt gccggttact gcgctgtacc aaatgcggga caacgtaagc actacatttc
     5761 gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt taaggtttca tttagcgcct
     5821 caaatagatc ctgttcagga accggatcaa agagttcctc cgccgctgga cctaccaagg
     5881 caacgctatg ttctcttgct tttgtcagca agatagccag atcaatgtcg atcgtggctg
     5941 gctcgaagat acctgcaaga atgtcattgc gctgccattc tccaaattgc agttcgcgct
     6001 tagctggata acgccacgga atgatgtcgt cgtgcacaac aatggtgact tctacagcgc
     6061 ggagaatctc gctctctcca ggggaagccg aagtttccaa aaggtcgttg atcaaagctc
     6121 gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag caaatcaata tcactgtgtg
     6181 gcttcaggcc gccatccact gcggagccgt acaaatgtac ggccagcaac gtcggttcga
     6241 gatggcgctc gatgacgcca actacctctg atagttgagt cgatacttcg gcgatcaccg
     6301 cttcccccat gatgtttaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc
     6361 tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag
     6421 gcatagactg taccccaaaa aaacatgtca taacaagaag ccatgaaaac cgccactgcg
     6481 ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgacgg cagttacgct
     6541 acttgcatta cagcttacga accgaacgag gcttatgtcc actgggttcg tgcccgaatt
     6601 gatcacaggc agcaacgctc tgtcatcgtt acaatcaaca tgctaccctc cgcgagatca
     6661 tccgtgtttc aaacccggca gcttagttgc cgttcttccg aatagcatcg gtaacatgag
     6721 caaagtctgc cgccttacaa cggctctccc gctgacgccg tcccggactg atgggctgcc
     6781 tgtatcgagt ggtgattttg tgccgagctg ccggtcgggg agctgttggc tggctggtgg
     6841 caggatatat tgtggtgtaa acaaattgac gcttagacaa cttaataaca cattgcggaa
     6901 ttccgatcta gtaacataga tgacaccgcg cgcgataatt tatcctagtt tgcgcgctat
     6961 attttgtttt ctatcgcgta ttaaatgtat aattgcggga ctctaatcat aaaaacccat
     7021 ctcataaata acgtcatgca ttacatgtta attattacat gcttaacgta attcaacaga
     7081 aattatatga taatcatcgc aagaccggca acaggattca atcttaagaa actttattgc
     7141 caaatgtttg aacgatctgc ttgactctag gggtcatcag atttcggtga cgggcaggac
     7201 cggacggggc ggcaccggca ggctgaagtc cagctgccag aaacccacgt catgccagtt
     7261 cccgtgcttg aagccggccg cccgcagcat gccgcggggg gcatatccga gcgcctcgtg
     7321 catgcgcacg ctcgggtcgt tgggcagccc gatgacagcg accacgctct tgaagccctg
     7381 tgcctccagg gacttcagca ggtgggtgta gagcgtggag cccagtcccg tccgctggtg
     7441 gcggggggag acgtacacgg tcgactcggc cgtccagtcg taggcgttgc gtgccttcca
     7501 ggggcccgcg taggcgatgc cggcgacctc gccgtccacc tcggcgacga gccagggata
     7561 gcgctcccgc agacggacga ggtcgtccgt ccactcctgc ggttcctgcg gctcggtacg
     7621 gaagttgacc gtgcttgtct cgatgtagtg gttgacgatg gtgcagaccg ccggcatgtc
     7681 cgcctcggtg gcacggcgga tgtcggccgg gcgtcgttct gggctcatgg tagatccccc
     7741 tcgatcgagt tgagagtgaa tatgagactc taattggata ccgaggggaa tttatggaac
     7801 gtcagtggag catttttgac aagaaatatt tgctagctga tagtgacctt aggcgacttt
     7861 tgaacgcgca ataatggttt ctgacgtatg tgcttagctc attaaactcc agaaacccgc
     7921 ggctgagtgg ctccttcaac gttgcggttc tgtcagttcc aaacgtaaaa cggcttgtcc
     7981 cgcgtcatcg gcgggggtca taacgtgact cccttaattc tccgctcatg attagtagcg
     8041 gtcctgctga gcctcgacat gttgtcgcaa aattcgccct ggacccgccc aacgatttgt
     8101 cgtcactgtc aaggtttgac ctgcacttca tttggggccc acatacacca aaaaaatgct
     8161 gcataattct cggggcagca agtcggttac ccggccgccg tgctggaccg ggttgaatgg
     8221 tgcccgtaac tttcggtaga gcggacggcc aatactcaac ttcaaggaat ctcacccatg
     8281 cgcgccggcg gggaaccgga gttcccttca gtgagcgtta ttagttcgcc gctcggtgtg
     8341 tcgtagatac tagcccctgg ggcacttttg aaatttgaat aagatttatg taatcagtct
     8401 tttaggtttg accggttctg ccgctttttt taaaattgga tttgtaataa taaaacgcaa
     8461 ttgtttgtta ttgtggcgct ctatcataga tgtcgctata aacctattca gcacaatata
     8521 ttgttttcat tttaatattg tacatataag tagtagggta caatcagtaa attgaacgga
     8581 gaatattatt cataaaaata cgatagtaac gggtgatata ttcattagaa tgaaccgaaa
     8641 ccggcggtaa ggatctgagc tacacatgct caggtttttt acaacgtgca caacagaatt
     8701 gaaagcaaat atcatgcgat cataggcttc tcgcatatct cattaaagca ggacaagctc
     8761 acaccttcct cttcttcttg gggtcagccc tgctgtctcc accgagctga gagaggtcga
     8821 ttcttgtttc atagagcccc gtaattgact gatgaatcag tgtggcgtcc aggacctcct
     8881 ttgtagaggt gtaccgcttt ctgtctatgg tggtgtcgaa gtacttgaag gctgcaggcg
     8941 cgcccaagtt ggtcagagta aacaagtgga taatgttttc tgcctgctcc ctgatgggct
     9001 tatccctgtg cttattgtaa gcagaaagca ccttatcgag gttagcgtcg gcgaggatca
     9061 ctcttttgga gaattcgctt atttgctcga tgatctcatc aaggtagtgt ttgtgttgtt
     9121 ccacgaacag ctgcttctgc tcattatctt cgggagatcc tttgagcttt tcatagtggc
     9181 tggccagata caagaaatta acgtatttag agggcagtgc cagctcgtta cctttctgca
     9241 gctcgcccgc actagcgagc attcgtttcc ggccgttttc aagctcaaag agagagtact
     9301 tgggaagctt aatgatgagg tcttttttga cctctttata tcctttcgcc tcgagaaagt
     9361 cgatggggtt tttttcgaag cttgatcgct ccatgattgt gatgcccagc agttccttga
     9421 cgcttttgag ttttttagac ttccctttct ccactttggc cacaaccagt acactgtaag
     9481 cgactgtagg agaatcgaat ccgccgtatt tcttggggtc ccaatctttt ttgcgtgcga
     9541 tcagcttgtc gctgttcctt ttcgggagga tactttcctt ggagaagcct ccggtctgta
     9601 cttcggtctt tttaacgatg ttcacctgcg gcatggacag gaccttccgg actgtcgcga
     9661 aatccctacc cttgtcccac acgatttctc ctgtttctcc gtttgtttcg ataagtggtc
     9721 gcttccgaat ctctccattg gccagtgtaa tctcggtctt gaaaaaattc ataatattgc
     9781 tgtaaaagaa gtacttagcg gtggccttgc ctatttcctg ctcagacttt gcgatcattt
     9841 tcctaacatc gtacacttta tagtctccgt aaacaaattc agattcaagc ttgggatatt
     9901 ttttgataag tgcagtgcct accactgcat tcaggtaggc atcatgcgca tggtggtaat
     9961 tgttgatctc tctcacctta taaaactgaa agtcctttct gaaatctgaa accagcttag
    10021 acttcagagt aataactttc acctctcgaa tcagtttgtc attttcatcg tacttggtgt
    10081 tcatgcgtga atcgagaatt tgggccacgt gcttggtgat ctggcgtgtc tcaacaagct
    10141 gccttttgat gaagccggct ttatccaact cagacaggcc acctcgttca gccttagtca
    10201 gattatcgaa cttccgttgt gtgatcagtt tggcgttcag cagctgccgc caataatttt
    10261 tcattttctt gacaacttct tctgagggga cgttatcact cttccctcta tttttatcgg
    10321 atcttgtcaa cactttatta tcaatagaat catctttgag aaaagactgg ggcacgatat
    10381 gatccacgtc gtagtcggag agccgattga tgtccagttc ctgatccacg tacatgtccc
    10441 tgccgttctg caggtagtac aggtagagct tctcattctg aagctgggtg ttttcaactg
    10501 ggtgttcctt aaggatttgg gaccccagtt cttttatacc ctcttcaatc ctcttcatcc
    10561 tttccctact gttcttctgt cccttctggg tagtttggtt ctctcgggcc atctcgataa
    10621 cgatattctc gggcttatgc cttcccatta ctttgacgag ttcatccacg accttaacgg
    10681 tctgcagtat tccctttttg atagctgggc tacctgcaag attagcgatg tgctcgtgga
    10741 gactgtcccc ctggccagaa acttgtgctt tctggatgtc ctccttaaag gtgagagagt
    10801 catcatggat caactgcatg aagttccggt tggcaaatcc atcggactta agaaaatcca
    10861 ggattgtctt tccactctgc ttgtctcgga tcccattgat cagttttctt gacagccgcc
    10921 cccatcctgt atatcggcgc ctcttgagct gtttcatgac tttgtcgtcg aagagatgag
    10981 cgtaagtttt caagcgttct tcaatcatct ccctatcttc aaacaacgta agggtgagga
    11041 caatgtcctc aagaatgtcc tcgttctcct cattgtccag gaagtccttg tctttaatga
    11101 ttttcaggag atcgtgatac gttcccaggg atgcgttgaa gcgatcctcc actccgctga
    11161 tttcaacaga gtcgaaacat tcaatctttt tgaaataatc ttctttgagc tgtttcacgg
    11221 taactttccg gttcgtcttg aagaggaggt ccacgatagc tttcttctgc tctccagaca
    11281 ggaatgctgg ctttctcatc ccttctgtga cgtatttgac cttggtgagc tcgttataaa
    11341 ctgtgaagta ctcgtacagc agagagtgtt taggaagcac cttttcgtta ggcagatttt
    11401 tatcaaagtt agtcatcctt tcgatgaagg actgggcaga ggccccctta tccacgactt
    11461 cctcgaagtt ccagggagtg atagtctctt ctgatttgcg agtcatccac gcgaatctgg
    11521 aatttccccg ggcgaggggg cctacatagt agggtatccg aaatgtgagg attttctcaa
    11581 tcttttccct gttatctttc aaaaaggggt agaaatcctc ttgccgcctg aggatagcgt
    11641 gcagttcgcc caggtgaatc tggtggggga tgcttccatt gtcgaaagtg cgctgtttgc
    11701 gcaacagatc ttctctgtta agctttacca gcagctcctc ggtgccgtcc attttttcca
    11761 agatgggctt aataaatttg taaaattcct cctggcttgc tccgccgtca atgtatccgg
    11821 cgtagccatt tttagactga tcgaagaaaa tttccttgta cttctcaggc agttgctgtc
    11881 tgacaagggc cttcagcaaa gtcaagtctt ggtggtgctc atcatagcgc ttgatcatac
    11941 tagcgctcag cggagctttg gtgatctccg tgttcactcg cagaatatca ctcagcagaa
    12001 tggcgtctga caggttcttt gccgccaaaa aaaggtctgc gtactggtcg ccgatctggg
    12061 ccagcagatt gtcgagatca tcatcgtagg tgtctttgct cagttgaagc ttggcatctt
    12121 cggccaggtc gaagttagat ttaaagttgg gggtcagccc gagtgacagg gcgataagat
    12181 taccaaacag gccgttcttc ttctccccag ggagctgtgc gatgaggttt tcgagccgcc
    12241 gggatttgga cagcctagcg ctcaggattg ctttggcgtc aactccggat gcgttgatcg
    12301 ggttctcttc gaaaagctga ttgtaagtct gaaccagttg gataaagagt ttgtcgacat
    12361 cgctgttgtc tgggttcagg tccccctcga tgaggaagtg tccccgaaat ttgatcatat
    12421 gcgccagcgc gagatagatc aaccgcaagt cagccttatc agtactgtct acaagcttct
    12481 tcctcagatg atatatggtt gggtactttt catggtacgc cacctcgtcc acgatattgc
    12541 caaagattgg gtggcgctcg tgctttttat cctcctccac caaaaaggac tcctccagcc
    12601 tatggaagaa agagtcatcc accttagcca tctcattact aaagatctcc tgcaggtagc
    12661 agatccgatt ctttctgcgg gtatatctgc gccgtgctgt tcttttgagc cgcgtggctt
    12721 cggccgtctc cccggagtcg aacaggaggg cgccaatgag gttcttcttt atgctgtggc
    12781 gatcggtatt gcccagaact ttgaattttt tgctcggcac cttgtactcg tccgtaatga
    12841 cggcccagcc gacgctgttt gtgccgatag cgagcccaat ggagtacttc ttgtccatgc
    12901 caccaacctt ccgcttcttc ttaggagcca ttgctgcaca tacataacat atcaagatca
    12961 gaacacacat atacacacac aaatacaatc aagtcaacaa ctccaaaaag tccagatcta
    13021 catatataca tacgtaaata acaaaatcat gtaaataatc acaatcatgt aatccagatc
    13081 tatgcacata tatatataca caattaataa aaaaaatgat ataacagatc tatatctatg
    13141 tatgtaacaa cacaatcaga tgagagaagt gatgttttca gatctgtata catacaaaca
    13201 caaacagatg aacaattgat acgtagatcc atatgtatac gtacaattag ctacacgatt
    13261 aaatgaaaaa aatcaacgat ttcggattgg tacacacaaa cgcaacaata tgaagaaatt
    13321 catatctgat tagatataaa cataaccacg tgtagataca cagtcaaatc aacaaattta
    13381 tagcttctaa acggatgaga tgaacaagat aaagatattc acataaggca tacataagat
    13441 aagcagatta acaaactagc aataatacat acctaattaa aacaaggaat aacagagaga
    13501 gagagagaga gagagattta ccttgaaaat gaagaggaga agagaggatt tcttaaaatt
    13561 gggggtagag aaagaaagat gatgaattgt gagaaaggag agatagaagg gggggttgta
    13621 tatataggct gtagaagatt atttttgtgt ttgaggcggt gaaggaagag gggatctgac
    13681 tatgacacgt ttgcggttac gtatttcgat aggagtcttt caacgcttaa cgccgttact
    13741 ctatatgacc gtttgggccg taacggggcc gtttgttaac gctgatgttg attcttttct
    13801 ttctttcttt cttccttttt taaagaagca attgtacaat cgttgctagc tgtcaaacgg
    13861 ataattcgga tacggatatg cctatattca tatccgtaat ctccaagctt gaattcttgc
    13921 agagacctga gacgttgatc ggcacgtaag aggttccaac tttcaccata atgaaataag
    13981 atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat
    14041 ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca
    14101 ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat
    14161 tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca
    14221 cattcttgcc cgcctgatga atgctcatcc ggaattccgt atggcaatga aagacggtga
    14281 gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac
    14341 gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc
    14401 gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa
    14461 tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc
    14521 caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga
    14581 caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtctgtgatg gcttccatgt
    14641 cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatc
    14701 gcgtggatcc ggcttactaa aagccagata acagtatgcg tatttgcgcg ctgatttttg
    14761 cggtataaga atatatactg atatgtatac ccgaagtatg tcaaaaagag gtatgctatg
    14821 aagcagcgta ttacagtgac agttgacagc gacagctatc agttgctcaa ggcatatatg
    14881 atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa tgaagcccgt cgtctgcgtg
    14941 ccgaacgctg gaaagcggaa aatcaggaag ggatggctga ggtcgcccgg tttattgaaa
    15001 tgaacggctc ttttgctgac gagaacaggg gctggtgaaa tgcagtttaa ggtttacacc
    15061 tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat tattgacacg
    15121 cccgggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga taaagtctcc
    15181 cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat gaccaccgat
    15241 atggccagtg tgccggtttc cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa
    15301 aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc aggctccctt
    15361 atacacagcc agtcggtctc agctt
//
