LOCUS       sequence-151856-       16012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  recipient plasmid [multiplexing, nickase]; pPcUbi:Cas9 D10A, nptII.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16012)
  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 16012)
  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: pDGE78
FEATURES             Location/Qualifiers
     source          1..16012
                     /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(8671..9378)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     CDS             complement(9389..9409)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(9422..13495,13496..13498,13499..13525))
                     /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(9422..13525)
                     /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(13532..13552)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        14563..14665
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             14666..15325
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(14732..14751)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             15667..15972
                     /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     15851..15870
                     /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 agtagcggtc ctgctgagcc tcgacatgtt gtcgcaaaat
     8701 tcgccctgga cccgcccaac gatttgtcgt cactgtcaag gtttgacctg cacttcattt
     8761 ggggcccaca tacaccaaaa aaatgctgca taattctcgg ggcagcaagt cggttacccg
     8821 gccgccgtgc tggaccgggt tgaatggtgc ccgtaacttt cggtagagcg gacggccaat
     8881 actcaacttc aaggaatctc acccatgcgc gccggcgggg aaccggagtt cccttcagtg
     8941 agcgttatta gttcgccgct cggtgtgtcg tagatactag cccctggggc acttttgaaa
     9001 tttgaataag atttatgtaa tcagtctttt aggtttgacc ggttctgccg ctttttttaa
     9061 aattggattt gtaataataa aacgcaattg tttgttattg tggcgctcta tcatagatgt
     9121 cgctataaac ctattcagca caatatattg ttttcatttt aatattgtac atataagtag
     9181 tagggtacaa tcagtaaatt gaacggagaa tattattcat aaaaatacga tagtaacggg
     9241 tgatatattc attagaatga accgaaaccg gcggtaagga tctgagctac acatgctcag
     9301 gttttttaca acgtgcacaa cagaattgaa agcaaatatc atgcgatcat aggcttctcg
     9361 catatctcat taaagcagga caagctcaca ccttcctctt cttcttgggg tcagccctgc
     9421 tgtctccacc gagctgagag aggtcgattc ttgtttcata gagccccgta attgactgat
     9481 gaatcagtgt ggcgtccagg acctcctttg tagaggtgta ccgctttctg tctatggtgg
     9541 tgtcgaagta cttgaaggct gcaggcgcgc ccaagttggt cagagtaaac aagtggataa
     9601 tgttttctgc ctgctccctg atgggcttat ccctgtgctt attgtaagca gaaagcacct
     9661 tatcgaggtt agcgtcggcg aggatcactc ttttggagaa ttcgcttatt tgctcgatga
     9721 tctcatcaag gtagtgtttg tgttgttcca cgaacagctg cttctgctca ttatcttcgg
     9781 gagatccttt gagcttttca tagtggctgg ccagatacaa gaaattaacg tatttagagg
     9841 gcagtgccag ctcgttacct ttctgcagct cgcccgcact agcgagcatt cgtttccggc
     9901 cgttttcaag ctcaaagaga gagtacttgg gaagcttaat gatgaggtct tttttgacct
     9961 ctttatatcc tttcgcctcg agaaagtcga tggggttttt ttcgaagctt gatcgctcca
    10021 tgattgtgat gcccagcagt tccttgacgc ttttgagttt tttagacttc cctttctcca
    10081 ctttggccac aaccagtaca ctgtaagcga ctgtaggaga atcgaatccg ccgtatttct
    10141 tggggtccca atcttttttg cgtgcgatca gcttgtcgct gttccttttc gggaggatac
    10201 tttccttgga gaagcctccg gtctgtactt cggtcttttt aacgatgttc acctgcggca
    10261 tggacaggac cttccggact gtcgcgaaat ccctaccctt gtcccacacg atttctcctg
    10321 tttctccgtt tgtttcgata agtggtcgct tccgaatctc tccattggcc agtgtaatct
    10381 cggtcttgaa aaaattcata atattgctgt aaaagaagta cttagcggtg gccttgccta
    10441 tttcctgctc agactttgcg atcattttcc taacatcgta cactttatag tctccgtaaa
    10501 caaattcaga ttcaagcttg ggatattttt tgataagtgc agtgcctacc actgcattca
    10561 ggtaggcatc atgcgcatgg tggtaattgt tgatctctct caccttataa aactgaaagt
    10621 cctttctgaa atctgaaacc agcttagact tcagagtaat aactttcacc tctcgaatca
    10681 gtttgtcatt ttcatcgtac ttggtgttca tgcgtgaatc gagaatttgg gccacgtgct
    10741 tggtgatctg gcgtgtctca acaagctgcc ttttgatgaa gccggcttta tccaactcag
    10801 acaggccacc tcgttcagcc ttagtcagat tatcgaactt ccgttgtgtg atcagtttgg
    10861 cgttcagcag ctgccgccaa taatttttca ttttcttgac aacttcttct gaggggacgt
    10921 tatcactctt ccctctattt ttatcggatc ttgtcaacac tttattatca atagaatcat
    10981 ctttgagaaa agactggggc acgatatgat ccacgtcgta gtcggagagc cgattgatgt
    11041 ccagttcctg atccacgtac atgtccctgc cgttctgcag gtagtacagg tagagcttct
    11101 cattctgaag ctgggtgttt tcaactgggt gttccttaag gatttgggac cccagttctt
    11161 ttataccctc ttcaatcctc ttcatccttt ccctactgtt cttctgtccc ttctgggtag
    11221 tttggttctc tcgggccatc tcgataacga tattctcggg cttatgcctt cccattactt
    11281 tgacgagttc atccacgacc ttaacggtct gcagtattcc ctttttgata gctgggctac
    11341 ctgcaagatt agcgatgtgc tcgtggagac tgtccccctg gccagaaact tgtgctttct
    11401 ggatgtcctc cttaaaggtg agagagtcat catggatcaa ctgcatgaag ttccggttgg
    11461 caaatccatc ggacttaaga aaatccagga ttgtctttcc actctgcttg tctcggatcc
    11521 cattgatcag ttttcttgac agccgccccc atcctgtata tcggcgcctc ttgagctgtt
    11581 tcatgacttt gtcgtcgaag agatgagcgt aagttttcaa gcgttcttca atcatctccc
    11641 tatcttcaaa caacgtaagg gtgaggacaa tgtcctcaag aatgtcctcg ttctcctcat
    11701 tgtccaggaa gtccttgtct ttaatgattt tcaggagatc gtgatacgtt cccagggatg
    11761 cgttgaagcg atcctccact ccgctgattt caacagagtc gaaacattca atctttttga
    11821 aataatcttc tttgagctgt ttcacggtaa ctttccggtt cgtcttgaag aggaggtcca
    11881 cgatagcttt cttctgctct ccagacagga atgctggctt tctcatccct tctgtgacgt
    11941 atttgacctt ggtgagctcg ttataaactg tgaagtactc gtacagcaga gagtgtttag
    12001 gaagcacctt ttcgttaggc agatttttat caaagttagt catcctttcg atgaaggact
    12061 gggcagaggc ccccttatcc acgacttcct cgaagttcca gggagtgata gtctcttctg
    12121 atttgcgagt catccacgcg aatctggaat ttccccgggc gagggggcct acatagtagg
    12181 gtatccgaaa tgtgaggatt ttctcaatct tttccctgtt atctttcaaa aaggggtaga
    12241 aatcctcttg ccgcctgagg atagcgtgca gttcgcccag gtgaatctgg tgggggatgc
    12301 ttccattgtc gaaagtgcgc tgtttgcgca acagatcttc tctgttaagc tttaccagca
    12361 gctcctcggt gccgtccatt ttttccaaga tgggcttaat aaatttgtaa aattcctcct
    12421 ggcttgctcc gccgtcaatg tatccggcgt agccattttt agactgatcg aagaaaattt
    12481 ccttgtactt ctcaggcagt tgctgtctga caagggcctt cagcaaagtc aagtcttggt
    12541 ggtgctcatc atagcgcttg atcatactag cgctcagcgg agctttggtg atctccgtgt
    12601 tcactcgcag aatatcactc agcagaatgg cgtctgacag gttctttgcc gccaaaaaaa
    12661 ggtctgcgta ctggtcgccg atctgggcca gcagattgtc gagatcatca tcgtaggtgt
    12721 ctttgctcag ttgaagcttg gcatcttcgg ccaggtcgaa gttagattta aagttggggg
    12781 tcagcccgag tgacagggcg ataagattac caaacaggcc gttcttcttc tccccaggga
    12841 gctgtgcgat gaggttttcg agccgccggg atttggacag cctagcgctc aggattgctt
    12901 tggcgtcaac tccggatgcg ttgatcgggt tctcttcgaa aagctgattg taagtctgaa
    12961 ccagttggat aaagagtttg tcgacatcgc tgttgtctgg gttcaggtcc ccctcgatga
    13021 ggaagtgtcc ccgaaatttg atcatatgcg ccagcgcgag atagatcaac cgcaagtcag
    13081 ccttatcagt actgtctaca agcttcttcc tcagatgata tatggttggg tacttttcat
    13141 ggtacgccac ctcgtccacg atattgccaa agattgggtg gcgctcgtgc tttttatcct
    13201 cctccaccaa aaaggactcc tccagcctat ggaagaaaga gtcatccacc ttagccatct
    13261 cattactaaa gatctcctgc aggtagcaga tccgattctt tctgcgggta tatctgcgcc
    13321 gtgctgttct tttgagccgc gtggcttcgg ccgtctcccc ggagtcgaac aggagggcgc
    13381 caatgaggtt cttctttatg ctgtggcgat cggtattgcc cagaactttg aattttttgc
    13441 tcggcacctt gtactcgtcc gtaatgacgg cccagccgac gctgtttgtg ccgatagcga
    13501 gcccaatgga gtacttcttg tccatgccac caaccttccg cttcttctta ggagccattg
    13561 ctgcacatac ataacatatc aagatcagaa cacacatata cacacacaaa tacaatcaag
    13621 tcaacaactc caaaaagtcc agatctacat atatacatac gtaaataaca aaatcatgta
    13681 aataatcaca atcatgtaat ccagatctat gcacatatat atatacacaa ttaataaaaa
    13741 aaatgatata acagatctat atctatgtat gtaacaacac aatcagatga gagaagtgat
    13801 gttttcagat ctgtatacat acaaacacaa acagatgaac aattgatacg tagatccata
    13861 tgtatacgta caattagcta cacgattaaa tgaaaaaaat caacgatttc ggattggtac
    13921 acacaaacgc aacaatatga agaaattcat atctgattag atataaacat aaccacgtgt
    13981 agatacacag tcaaatcaac aaatttatag cttctaaacg gatgagatga acaagataaa
    14041 gatattcaca taaggcatac ataagataag cagattaaca aactagcaat aatacatacc
    14101 taattaaaac aaggaataac agagagagag agagagagag agatttacct tgaaaatgaa
    14161 gaggagaaga gaggatttct taaaattggg ggtagagaaa gaaagatgat gaattgtgag
    14221 aaaggagaga tagaaggggg ggttgtatat ataggctgta gaagattatt tttgtgtttg
    14281 aggcggtgaa ggaagagggg atctgactat gacacgtttg cggttacgta tttcgatagg
    14341 agtctttcaa cgcttaacgc cgttactcta tatgaccgtt tgggccgtaa cggggccgtt
    14401 tgttaacgct gatgttgatt cttttctttc tttctttctt ccttttttaa agaagcaatt
    14461 gtacaatcgt tgctagctgt caaacggata attcggatac ggatatgcct atattcatat
    14521 ccgtaatctc caagcttgaa ttcttgcaga gacctgagac gttgatcggc acgtaagagg
    14581 ttccaacttt caccataatg aaataagatc actaccgggc gtattttttg agttatcgag
    14641 attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga
    14701 tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac
    14761 ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa
    14821 gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga
    14881 attccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta
    14941 caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga
    15001 tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc
    15061 ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag
    15121 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac
    15181 catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca
    15241 tcatgccgtc tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg
    15301 cgatgagtgg cagggcgggg cgtaatcgcg tggatccggc ttactaaaag ccagataaca
    15361 gtatgcgtat ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg
    15421 aagtatgtca aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac
    15481 agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca
    15541 tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga
    15601 tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct
    15661 ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg
    15721 gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt
    15781 gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat
    15841 gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtttccgt tatcggggaa
    15901 gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc
    15961 tggggaatat aaatgtcagg ctcccttata cacagccagt cggtctcagc tt
//
