LOCUS       sequence-151852-       15432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  recipient plasmid [multiplexing, nickase]; p35S:Cas9 D10A, nptII.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15432)
  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 15432)
  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: pDGE76
FEATURES             Location/Qualifiers
     source          1..15432
                     /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(7547..8341)
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7658..7677)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     8268..8287
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(8362..8545)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(8398..8577)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      complement(8670..8922)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(8938..8958)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(8971..13044,13045..13047,13048..13074))
                     /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(8971..13074)
                     /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(13081..13101)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(13198..13543)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(13235..13255)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        13983..14085
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             14086..14745
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(14152..14171)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             15087..15392
                     /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     15271..15290
                     /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 aacgatcggg gatcatccgg gtctgtggcg ggaactccac gaaaatatcc
     7201 gaacgcagca agatatcgcg gtgcatctcg gtcttgcctg ggcagtcgcc gccgacgccg
     7261 ttgatgtgga cgccgggccc gatcatattg tcgctcagga tcgtggcgtt gtgcttgtcg
     7321 gccgttgctg tcgtaatgat atcggcacct tcgaccgcct gttccgcaga gatcccgtgg
     7381 gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa
     7441 acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc
     7501 aggttgggcg tcgcttggtc ggtcatttcg aaccccagag tcccgctcag aagaactcgt
     7561 caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga
     7621 ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta
     7681 tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc
     7741 cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcatcgc
     7801 cgtcgggcat gcgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct
     7861 cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga
     7921 tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc
     7981 gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat
     8041 cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga
     8101 gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct
     8161 gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg
     8221 ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc
     8281 cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca
     8341 tgcgaaacga tccagatccg gtgcagatta tttggattga gagtgaatat gagactctaa
     8401 ttggataccg aggggaattt atggaacgtc agtggagcat ttttgacaag aaatatttgc
     8461 tagctgatag tgaccttagg cgacttttga acgcgcaata atggtttctg acgtatgtgc
     8521 ttagctcatt aaactccaga aacccgcggc tgagtggctc cttcaacgtt gcggttctgt
     8581 cagttccaaa cgtaaaacgg cttgtcccgc gtcatcggcg ggggtcataa cgtgactccc
     8641 ttaattctcc gctcatgatt agtagcgtcg atctagtaac atagatgaca ccgcgcgcga
     8701 taatttatcc tagtttgcgc gctatatttt gttttctatc gcgtattaaa tgtataattg
     8761 cgggactcta atcataaaaa cccatctcat aaataacgtc atgcattaca tgttaattat
     8821 tacatgctta acgtaattca acagaaatta tatgataatc atcgcaagac cggcaacagg
     8881 attcaatctt aagaaacttt attgccaaat gtttgaacga tctgcttgac aagctcacac
     8941 cttcctcttc ttcttggggt cagccctgct gtctccaccg agctgagaga ggtcgattct
     9001 tgtttcatag agccccgtaa ttgactgatg aatcagtgtg gcgtccagga cctcctttgt
     9061 agaggtgtac cgctttctgt ctatggtggt gtcgaagtac ttgaaggctg caggcgcgcc
     9121 caagttggtc agagtaaaca agtggataat gttttctgcc tgctccctga tgggcttatc
     9181 cctgtgctta ttgtaagcag aaagcacctt atcgaggtta gcgtcggcga ggatcactct
     9241 tttggagaat tcgcttattt gctcgatgat ctcatcaagg tagtgtttgt gttgttccac
     9301 gaacagctgc ttctgctcat tatcttcggg agatcctttg agcttttcat agtggctggc
     9361 cagatacaag aaattaacgt atttagaggg cagtgccagc tcgttacctt tctgcagctc
     9421 gcccgcacta gcgagcattc gtttccggcc gttttcaagc tcaaagagag agtacttggg
     9481 aagcttaatg atgaggtctt ttttgacctc tttatatcct ttcgcctcga gaaagtcgat
     9541 ggggtttttt tcgaagcttg atcgctccat gattgtgatg cccagcagtt ccttgacgct
     9601 tttgagtttt ttagacttcc ctttctccac tttggccaca accagtacac tgtaagcgac
     9661 tgtaggagaa tcgaatccgc cgtatttctt ggggtcccaa tcttttttgc gtgcgatcag
     9721 cttgtcgctg ttccttttcg ggaggatact ttccttggag aagcctccgg tctgtacttc
     9781 ggtcttttta acgatgttca cctgcggcat ggacaggacc ttccggactg tcgcgaaatc
     9841 cctacccttg tcccacacga tttctcctgt ttctccgttt gtttcgataa gtggtcgctt
     9901 ccgaatctct ccattggcca gtgtaatctc ggtcttgaaa aaattcataa tattgctgta
     9961 aaagaagtac ttagcggtgg ccttgcctat ttcctgctca gactttgcga tcattttcct
    10021 aacatcgtac actttatagt ctccgtaaac aaattcagat tcaagcttgg gatatttttt
    10081 gataagtgca gtgcctacca ctgcattcag gtaggcatca tgcgcatggt ggtaattgtt
    10141 gatctctctc accttataaa actgaaagtc ctttctgaaa tctgaaacca gcttagactt
    10201 cagagtaata actttcacct ctcgaatcag tttgtcattt tcatcgtact tggtgttcat
    10261 gcgtgaatcg agaatttggg ccacgtgctt ggtgatctgg cgtgtctcaa caagctgcct
    10321 tttgatgaag ccggctttat ccaactcaga caggccacct cgttcagcct tagtcagatt
    10381 atcgaacttc cgttgtgtga tcagtttggc gttcagcagc tgccgccaat aatttttcat
    10441 tttcttgaca acttcttctg aggggacgtt atcactcttc cctctatttt tatcggatct
    10501 tgtcaacact ttattatcaa tagaatcatc tttgagaaaa gactggggca cgatatgatc
    10561 cacgtcgtag tcggagagcc gattgatgtc cagttcctga tccacgtaca tgtccctgcc
    10621 gttctgcagg tagtacaggt agagcttctc attctgaagc tgggtgtttt caactgggtg
    10681 ttccttaagg atttgggacc ccagttcttt tataccctct tcaatcctct tcatcctttc
    10741 cctactgttc ttctgtccct tctgggtagt ttggttctct cgggccatct cgataacgat
    10801 attctcgggc ttatgccttc ccattacttt gacgagttca tccacgacct taacggtctg
    10861 cagtattccc tttttgatag ctgggctacc tgcaagatta gcgatgtgct cgtggagact
    10921 gtccccctgg ccagaaactt gtgctttctg gatgtcctcc ttaaaggtga gagagtcatc
    10981 atggatcaac tgcatgaagt tccggttggc aaatccatcg gacttaagaa aatccaggat
    11041 tgtctttcca ctctgcttgt ctcggatccc attgatcagt tttcttgaca gccgccccca
    11101 tcctgtatat cggcgcctct tgagctgttt catgactttg tcgtcgaaga gatgagcgta
    11161 agttttcaag cgttcttcaa tcatctccct atcttcaaac aacgtaaggg tgaggacaat
    11221 gtcctcaaga atgtcctcgt tctcctcatt gtccaggaag tccttgtctt taatgatttt
    11281 caggagatcg tgatacgttc ccagggatgc gttgaagcga tcctccactc cgctgatttc
    11341 aacagagtcg aaacattcaa tctttttgaa ataatcttct ttgagctgtt tcacggtaac
    11401 tttccggttc gtcttgaaga ggaggtccac gatagctttc ttctgctctc cagacaggaa
    11461 tgctggcttt ctcatccctt ctgtgacgta tttgaccttg gtgagctcgt tataaactgt
    11521 gaagtactcg tacagcagag agtgtttagg aagcaccttt tcgttaggca gatttttatc
    11581 aaagttagtc atcctttcga tgaaggactg ggcagaggcc cccttatcca cgacttcctc
    11641 gaagttccag ggagtgatag tctcttctga tttgcgagtc atccacgcga atctggaatt
    11701 tccccgggcg agggggccta catagtaggg tatccgaaat gtgaggattt tctcaatctt
    11761 ttccctgtta tctttcaaaa aggggtagaa atcctcttgc cgcctgagga tagcgtgcag
    11821 ttcgcccagg tgaatctggt gggggatgct tccattgtcg aaagtgcgct gtttgcgcaa
    11881 cagatcttct ctgttaagct ttaccagcag ctcctcggtg ccgtccattt tttccaagat
    11941 gggcttaata aatttgtaaa attcctcctg gcttgctccg ccgtcaatgt atccggcgta
    12001 gccattttta gactgatcga agaaaatttc cttgtacttc tcaggcagtt gctgtctgac
    12061 aagggccttc agcaaagtca agtcttggtg gtgctcatca tagcgcttga tcatactagc
    12121 gctcagcgga gctttggtga tctccgtgtt cactcgcaga atatcactca gcagaatggc
    12181 gtctgacagg ttctttgccg ccaaaaaaag gtctgcgtac tggtcgccga tctgggccag
    12241 cagattgtcg agatcatcat cgtaggtgtc tttgctcagt tgaagcttgg catcttcggc
    12301 caggtcgaag ttagatttaa agttgggggt cagcccgagt gacagggcga taagattacc
    12361 aaacaggccg ttcttcttct ccccagggag ctgtgcgatg aggttttcga gccgccggga
    12421 tttggacagc ctagcgctca ggattgcttt ggcgtcaact ccggatgcgt tgatcgggtt
    12481 ctcttcgaaa agctgattgt aagtctgaac cagttggata aagagtttgt cgacatcgct
    12541 gttgtctggg ttcaggtccc cctcgatgag gaagtgtccc cgaaatttga tcatatgcgc
    12601 cagcgcgaga tagatcaacc gcaagtcagc cttatcagta ctgtctacaa gcttcttcct
    12661 cagatgatat atggttgggt acttttcatg gtacgccacc tcgtccacga tattgccaaa
    12721 gattgggtgg cgctcgtgct ttttatcctc ctccaccaaa aaggactcct ccagcctatg
    12781 gaagaaagag tcatccacct tagccatctc attactaaag atctcctgca ggtagcagat
    12841 ccgattcttt ctgcgggtat atctgcgccg tgctgttctt ttgagccgcg tggcttcggc
    12901 cgtctccccg gagtcgaaca ggagggcgcc aatgaggttc ttctttatgc tgtggcgatc
    12961 ggtattgccc agaactttga attttttgct cggcaccttg tactcgtccg taatgacggc
    13021 ccagccgacg ctgtttgtgc cgatagcgag cccaatggag tacttcttgt ccatgccacc
    13081 aaccttccgc ttcttcttag gagccattgt atcgataatt gtaaatgtaa ttgtaatgtt
    13141 gtttgttgtt tgttgttgtt ggtaattgtt gtaaaaatga gctcttatac tcgagcgtgt
    13201 cctctccaaa tgaaatgaac ttccttatat agaggaaggg tcttgcgaag gatagtggga
    13261 ttgtgcgtca tcccttacgt cagtggagat gtcacatcaa tccacttgct ttgtagacgt
    13321 ggttggaacc tcttcttttt ccacgatgct cctcgtgggt gggggtccat ctttgggacc
    13381 actgtcggca gagagatctt gaatgatagc ctttccttta tcgcaatgat ggcatttgta
    13441 ggagccacct tccttttcta ctgtcctttc gatgaagtga cagatagctg ggcaatggaa
    13501 tccgaggagg tttcccgaaa ttatcctttg ttgaaaagtc tcaatagccc tttgatcttc
    13561 tgagactgta tctttgacat ttttggagta gaccagagtg tcgtgctcca ccatgttatc
    13621 acatcaatcc acttgctttg tagacgtggt tggaacctct tctttttcca cgatgctcct
    13681 cgtgggtggg ggtccatctt tgggaccact gtcggcagag agatcttgaa tgatagcctt
    13741 tcctttatcg caatgatggc atttgtagga gccaccttcc ttttctactg tcctttcgat
    13801 gaagtgacag atagctgggc aatggaatcc gaggaggttt cccgaaatta tcctttgttg
    13861 aaaagtctca atagcccttt gatcttctga gactgtatct ttgacatttt tggagtagac
    13921 cagagtgtcg tgctccacca tgttgacctc caagcttgaa ttcttgcaga gacctgagac
    13981 gttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc
    14041 gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
    14101 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
    14161 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
    14221 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
    14281 ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg
    14341 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
    14401 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
    14461 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
    14521 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
    14581 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
    14641 ccgctggcga ttcaggttca tcatgccgtc tgtgatggct tccatgtcgg cagaatgctt
    14701 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaatcgcg tggatccggc
    14761 ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata
    14821 tatactgata tgtatacccg aagtatgtca aaaagaggta tgctatgaag cagcgtatta
    14881 cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc
    14941 cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa
    15001 agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt
    15061 tgctgacgag aacaggggct ggtgaaatgc agtttaaggt ttacacctat aaaagagaga
    15121 gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga
    15181 tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc
    15241 cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc
    15301 cggtttccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa
    15361 acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt
    15421 cggtctcagc tt
//
