LOCUS       sequence-153536-       15531 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  recipient plasmid [single nuclease]; pPcUbi:Cas9, BASTA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15531)
  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 15531)
  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: pDGE65
FEATURES             Location/Qualifiers
     source          1..15531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    266..290
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(333..585)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(605..1156)
                     /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(1198..1377)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      complement(1471..2178)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     CDS             complement(2189..2209)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2222..6325)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(2222..6325)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(2222..6325)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     promoter        7421..7523
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             7524..8183
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(7590..7609)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             8529..8834
                     /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     8713..8732
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_RNA        8872..8947
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    9261..9285
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             10586..11215
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             11649..12716
                     /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      12782..12976
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(13212..13234)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    13320..13460
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     13375..13394
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(13475..13492)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13646..14234)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13726..13745)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(14478..15269)
                     /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"
ORIGIN
        1 ttatgtccac tgggttcgtg cccgaattga tcacaggcag caacgctctg tcatcgttac
       61 aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc ttagttgccg
      121 ttcttccgaa tagcatcggt aacatgagca aagtctgccg ccttacaacg gctctcccgc
      181 tgacgccgtc ccggactgat gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc
      241 ggtcggggag ctgttggctg gctggtggca ggatatattg tggtgtaaac aaattgacgc
      301 ttagacaact taataacaca ttgcggaatt ccgatctagt aacatagatg acaccgcgcg
      361 cgataattta tcctagtttg cgcgctatat tttgttttct atcgcgtatt aaatgtataa
      421 ttgcgggact ctaatcataa aaacccatct cataaataac gtcatgcatt acatgttaat
      481 tattacatgc ttaacgtaat tcaacagaaa ttatatgata atcatcgcaa gaccggcaac
      541 aggattcaat cttaagaaac tttattgcca aatgtttgaa cgatctgctt gactctaggg
      601 gtcatcagat ttcggtgacg ggcaggaccg gacggggcgg caccggcagg ctgaagtcca
      661 gctgccagaa acccacgtca tgccagttcc cgtgcttgaa gccggccgcc cgcagcatgc
      721 cgcggggggc atatccgagc gcctcgtgca tgcgcacgct cgggtcgttg ggcagcccga
      781 tgacagcgac cacgctcttg aagccctgtg cctccaggga cttcagcagg tgggtgtaga
      841 gcgtggagcc cagtcccgtc cgctggtggc ggggggagac gtacacggtc gactcggccg
      901 tccagtcgta ggcgttgcgt gccttccagg gacccgcgta ggcgatgccg gcgacctcgc
      961 cgtccacctc ggcgacgagc cagggatagc gctcccgcag acggacgagg tcgtccgtcc
     1021 actcctgcgg ttcctgcggc tcggtacgga agttgaccgt gcttgtctcg atgtagtggt
     1081 tgacgatggt gcagaccgcc ggcatgtccg cctcggtggc acggcggatg tcggccgggc
     1141 gtcgttctgg gctcatggta gatcccctcg atcgagttga gagtgaatat gagactctaa
     1201 ttggataccg aggggaattt atggaacgtc agtggagcat ttttgacaag aaatatttgc
     1261 tagctgatag tgaccttagg cgacttttga acgcgcaata atggtttctg acgtatgtgc
     1321 ttagctcatt aaactccaga aacccgcggc tcagtggctc cttcaacgtt gcggttctgt
     1381 cagttccaaa cgtaaaacgg cttgtcccgc gtcatcggcg ggggtcataa cgtgactccc
     1441 ttaattctcc gctcatgatt agtagcggtc ctgctgagcc tcgacatgtt gtcgcaaaat
     1501 tcgccctgga cccgcccaac gatttgtcgt cactgtcaag gtttgacctg cacttcattt
     1561 ggggcccaca tacaccaaaa aaatgctgca taattctcgg ggcagcaagt cggttacccg
     1621 gccgccgtgc tggaccgggt tgaatggtgc ccgtaacttt cggtagagcg gacggccaat
     1681 actcaacttc aaggaatctc acccatgcgc gccggcgggg aaccggagtt cccttcagtg
     1741 agcgttatta gttcgccgct cggtgtgtcg tagatactag cccctggggc acttttgaaa
     1801 tttgaataag atttatgtaa tcagtctttt aggtttgacc ggttctgccg ctttttttaa
     1861 aattggattt gtaataataa aacgcaattg tttgttattg tggcgctcta tcatagatgt
     1921 cgctataaac ctattcagca caatatattg ttttcatttt aatattgtac atataagtag
     1981 tagggtacaa tcagtaaatt gaacggagaa tattattcat aaaaatacga tagtaacggg
     2041 tgatatattc attagaatga accgaaaccg gcggtaagga tctgagctac acatgctcag
     2101 gttttttaca acgtgcacaa cagaattgaa agcaaatatc atgcgatcat aggcttctcg
     2161 catatctcat taaagcagga caagctcaca ccttcctctt cttcttgggg tcagccctgc
     2221 tgtctccacc gagctgagag aggtcgattc ttgtttcata gagccccgta attgactgat
     2281 gaatcagtgt ggcgtccagg acctcctttg tagaggtgta ccgctttctg tctatggtgg
     2341 tgtcgaagta cttgaaggct gcaggcgcgc ccaagttggt cagagtaaac aagtggataa
     2401 tgttttctgc ctgctccctg atgggcttat ccctgtgctt attgtaagca gaaagcacct
     2461 tatcgaggtt agcgtcggcg aggatcactc ttttggagaa ttcgcttatt tgctcgatga
     2521 tctcatcaag gtagtgtttg tgttgttcca cgaacagctg cttctgctca ttatcttcgg
     2581 gagatccttt gagcttttca tagtggctgg ccagatacaa gaaattaacg tatttagagg
     2641 gcagtgccag ctcgttacct ttctgcagct cgcccgcact agcgagcatt cgtttccggc
     2701 cgttttcaag ctcaaagaga gagtacttgg gaagcttaat gatgaggtct tttttgacct
     2761 ctttatatcc tttcgcctcg agaaagtcga tggggttttt ttcgaagctt gatcgctcca
     2821 tgattgtgat gcccagcagt tccttgacgc ttttgagttt tttagacttc cctttctcca
     2881 ctttggccac aaccagtaca ctgtaagcga ctgtaggaga atcgaatccg ccgtatttct
     2941 tggggtccca atcttttttg cgtgcgatca gcttgtcgct gttccttttc gggaggatac
     3001 tttccttgga gaagcctccg gtctgtactt cggtcttttt aacgatgttc acctgcggca
     3061 tggacaggac cttccggact gtcgcgaaat ccctaccctt gtcccacacg atttctcctg
     3121 tttctccgtt tgtttcgata agtggtcgct tccgaatctc tccattggcc agtgtaatct
     3181 cggtcttgaa aaaattcata atattgctgt aaaagaagta cttagcggtg gccttgccta
     3241 tttcctgctc agactttgcg atcattttcc taacatcgta cactttatag tctccgtaaa
     3301 caaattcaga ttcaagcttg ggatattttt tgataagtgc agtgcctacc actgcattca
     3361 ggtaggcatc atgcgcatgg tggtaattgt tgatctctct caccttataa aactgaaagt
     3421 cctttctgaa atctgaaacc agcttagact tcagagtaat aactttcacc tctcgaatca
     3481 gtttgtcatt ttcatcgtac ttggtgttca tgcgtgaatc gagaatttgg gccacgtgct
     3541 tggtgatctg gcgtgtctca acaagctgcc ttttgatgaa gccggcttta tccaactcag
     3601 acaggccacc tcgttcagcc ttagtcagat tatcgaactt ccgttgtgtg atcagtttgg
     3661 cgttcagcag ctgccgccaa taatttttca ttttcttgac aacttcttct gaggggacgt
     3721 tatcactctt ccctctattt ttatcggatc ttgtcaacac tttattatca atagaatcat
     3781 ctttgagaaa agactggggc acgatatgat ccacgtcgta gtcggagagc cgattgatgt
     3841 ccagttcctg atccacgtac atgtccctgc cgttctgcag gtagtacagg tagagcttct
     3901 cattctgaag ctgggtgttt tcaactgggt gttccttaag gatttgggac cccagttctt
     3961 ttataccctc ttcaatcctc ttcatccttt ccctactgtt cttctgtccc ttctgggtag
     4021 tttggttctc tcgggccatc tcgataacga tattctcggg cttatgcctt cccattactt
     4081 tgacgagttc atccacgacc ttaacggtct gcagtattcc ctttttgata gctgggctac
     4141 ctgcaagatt agcgatgtgc tcgtggagac tgtccccctg gccagaaact tgtgctttct
     4201 ggatgtcctc cttaaaggtg agagagtcat catggatcaa ctgcatgaag ttccggttgg
     4261 caaatccatc ggacttaaga aaatccagga ttgtctttcc actctgcttg tctcggatcc
     4321 cattgatcag ttttcttgac agccgccccc atcctgtata tcggcgcctc ttgagctgtt
     4381 tcatgacttt gtcgtcgaag agatgagcgt aagttttcaa gcgttcttca atcatctccc
     4441 tatcttcaaa caacgtaagg gtgaggacaa tgtcctcaag aatgtcctcg ttctcctcat
     4501 tgtccaggaa gtccttgtct ttaatgattt tcaggagatc gtgatacgtt cccagggatg
     4561 cgttgaagcg atcctccact ccgctgattt caacagagtc gaaacattca atctttttga
     4621 aataatcttc tttgagctgt ttcacggtaa ctttccggtt cgtcttgaag aggaggtcca
     4681 cgatagcttt cttctgctct ccagacagga atgctggctt tctcatccct tctgtgacgt
     4741 atttgacctt ggtgagctcg ttataaactg tgaagtactc gtacagcaga gagtgtttag
     4801 gaagcacctt ttcgttaggc agatttttat caaagttagt catcctttcg atgaaggact
     4861 gggcagaggc ccccttatcc acgacttcct cgaagttcca gggagtgata gtctcttctg
     4921 atttgcgagt catccacgcg aatctggaat ttccccgggc gagggggcct acatagtagg
     4981 gtatccgaaa tgtgaggatt ttctcaatct tttccctgtt atctttcaaa aaggggtaga
     5041 aatcctcttg ccgcctgagg atagcgtgca gttcgcccag gtgaatctgg tgggggatgc
     5101 ttccattgtc gaaagtgcgc tgtttgcgca acagatcttc tctgttaagc tttaccagca
     5161 gctcctcggt gccgtccatt ttttccaaga tgggcttaat aaatttgtaa aattcctcct
     5221 ggcttgctcc gccgtcaatg tatccggcgt agccattttt agactgatcg aagaaaattt
     5281 ccttgtactt ctcaggcagt tgctgtctga caagggcctt cagcaaagtc aagtcttggt
     5341 ggtgctcatc atagcgcttg atcatactag cgctcagcgg agctttggtg atctccgtgt
     5401 tcactcgcag aatatcactc agcagaatgg cgtctgacag gttctttgcc gccaaaaaaa
     5461 ggtctgcgta ctggtcgccg atctgggcca gcagattgtc gagatcatca tcgtaggtgt
     5521 ctttgctcag ttgaagcttg gcatcttcgg ccaggtcgaa gttagattta aagttggggg
     5581 tcagcccgag tgacagggcg ataagattac caaacaggcc gttcttcttc tccccaggga
     5641 gctgtgcgat gaggttttcg agccgccggg atttggacag cctagcgctc aggattgctt
     5701 tggcgtcaac tccggatgcg ttgatcgggt tctcttcgaa aagctgattg taagtctgaa
     5761 ccagttggat aaagagtttg tcgacatcgc tgttgtctgg gttcaggtcc ccctcgatga
     5821 ggaagtgtcc ccgaaatttg atcatatgcg ccagcgcgag atagatcaac cgcaagtcag
     5881 ccttatcagt actgtctaca agcttcttcc tcagatgata tatggttggg tacttttcat
     5941 ggtacgccac ctcgtccacg atattgccaa agattgggtg gcgctcgtgc tttttatcct
     6001 cctccaccaa aaaggactcc tccagcctat ggaagaaaga gtcatccacc ttagccatct
     6061 cattactaaa gatctcctgc aggtagcaga tccgattctt tctgcgggta tatctgcgcc
     6121 gtgctgttct tttgagccgc gtggcttcgg ccgtctcccc ggagtcgaac aggagggcgc
     6181 caatgaggtt cttctttatg ctgtggcgat cggtattgcc cagaactttg aattttttgc
     6241 tcggcacctt gtactcgtcc gtaatgacgg cccagccgac gctgtttgtg ccgatatcga
     6301 gcccaatgga gtacttcttg tccattgctg cacatacata acatatcaag atcagaacac
     6361 acatatacac acacaaatac aatcaagtca acaactccaa aaagtccaga tctacatata
     6421 tacatacgta aataacaaaa tcatgtaaat aatcacaatc atgtaatcca gatctatgca
     6481 catatatata tacacaatta ataaaaaaaa tgatataaca gatctatatc tatgtatgta
     6541 acaacacaat cagatgagag aagtgatgtt ttcagatctg tatacataca aacacaaaca
     6601 gatgaacaat tgatacgtag atccatatgt atacgtacaa ttagctacac gattaaatga
     6661 aaaaaatcaa cgatttcgga ttggtacaca caaacgcaac aatatgaaga aattcatatc
     6721 tgattagata taaacataac cacgtgtaga tacacagtca aatcaacaaa tttatagctt
     6781 ctaaacggat gagatgaaca agataaagat attcacataa ggcatacata agataagcag
     6841 attaacaaac tagcaataat acatacctaa ttaaaacaag gaataacaga gagagagaga
     6901 gagagagaga tttaccttga aaatgaagag gagaagagag gatttcttaa aattgggggt
     6961 agagaaagaa agatgatgaa ttgtgagaaa ggagagatag aagggggggt tgtatatata
     7021 ggctgtagaa gattattttt gtgtttgagg cggtgaagga agaggggatc tgactatgac
     7081 acgtttgcgg ttacgtattt cgataggagt ctttcaacgc ttaacgccgt tactctatat
     7141 gaccgtttgg gccgtaacgg ggccgtttgt taacgctgat gttgattctt ttctttcttt
     7201 ctttcttcct tttttaaaga agcaattgta caatcgttgc tagctgtcaa acggataatt
     7261 cggatacgga tatgcctata ttcatatccg taatctccaa gcttgaattc ttgctaagtt
     7321 gaaaacaatc ttcaaaagtc ccacatcgct tagataagaa aacgaagctg agtttatata
     7381 cagctagagt cgaagtagtg attgaagtct tctgagacgt tgatcggcac gtaagaggtt
     7441 ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat
     7501 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
     7561 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
     7621 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     7681 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     7741 tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
     7801 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
     7861 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
     7921 atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt
     7981 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     8041 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     8101 atgccgtctg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
     8161 atgagtggca ggggggggcg taaacgctcg cgtggatccg gcttactaaa agccagataa
     8221 cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa tatatactga tatgtatacc
     8281 cgaagtatgt caaaaagagg tatgctatga agcagcgtat tacagtgaca gttgacagcg
     8341 acagctatca gttgctcaag gcatatatga tgtcaatatc tccggtctgg taagcacaac
     8401 catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg
     8461 gatggctgag gtcgcccggt ttattgaaat gaacggctct tttgctgacg agaacagggg
     8521 ctggtgaaat gcagtttaag gtttacacct ataaaagaga gagccgttat cgtctgtttg
     8581 tggatgtaca gagtgatatt attgacacgc ccgggcgacg gatggtgatc cccctggcca
     8641 gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg catatcgggg
     8701 atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtttcc gttatcgggg
     8761 aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt aacctgatgt
     8821 tctggggaat ataaatgtca ggctccctta tacacagcca gtcgaagact tgttttagag
     8881 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
     8941 tcggtgcttt tttttgcttc catgctagag tccgcaaaaa tcaccagtct ctctctacaa
     9001 atctatctct ctctattttt ctccagaata atgtgtgagt agttcccaga taagggaatt
     9061 agggttctta tagggtttcg ctcatgtgtt gagcatataa gaaaccctta gtatgtattt
     9121 gtatttgtaa aatacttcta tcaataaaat ttctaattcc taaaaccaaa atccagtgac
     9181 ctgcaggcat gcgacgtcga gcttagcttg agcttggatc agattgtcgt ttcccgcctt
     9241 cagtttaaac tatcagtgtt tgacaggata tattggcggg taaacctaag agaaaagagc
     9301 gtttattaga ataatcggat atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt
     9361 gtatgtgcat gccaaccaca gggttcccct cgggatcaaa gtactttgat ccaacccctc
     9421 cgctgctata gtgcagtcgg cttctgacgt tcagtgcagc cgtgttctga aaacgacatg
     9481 tcgcacaagt cctaagttac gcgacaggct gccgccctgc ccttttcctg gcgttttctt
     9541 gtcgcgtgtt ttagtcgcat aaagtagaat acttgcgact agaaccggag acattacgcc
     9601 atgaacaaga gcgccgccgc tggcctgctg ggctatgccc gcgtcagcac cgacgaccag
     9661 gacttgacca accaacgggc cgaactgcac gcggccggct gcaccaagct gttttccgag
     9721 aagatcaccg gcaccaggcg cgaccgcccg gagctggcca ggatgcttga ccacctacgc
     9781 cctggcgacg ttgtgacagt gaccaggcta gaccgcctgg cccgcagcac ccgcgaccta
     9841 ctggacattg ccgagcgcat ccaggaggcc ggcgcgggcc tgcgtagcct ggcagagccg
     9901 tgggccgaca ccaccacgcc ggccggccgc atggtgttga ccgtgttcgc cggcattgcc
     9961 gagttcgagc gttccctaat catcgaccgc acccggagcg ggcgcgaggc cgccaaggcc
    10021 cgaggcgtga agtttggccc ccgccctacc ctcaccccgg cacagatcgc gcacgcccgc
    10081 gagctgatcg accaggaagg ccgcaccgtg aaagaggcgg ctgcactgct tggcgtgcat
    10141 cgctcgaccc tgtaccgcgc acttgagcgc agcgaggaag tgacgcccac cgaggccagg
    10201 cggcgcggtg ccttccgtga ggacgcattg accgaggccg acgccctggc ggccgccgag
    10261 aatgaacgcc aagaggaaca agcatgaaac cgcaccagga cggccaggac gaaccgtttt
    10321 tcattaccga agagatcgag gcggagatga tcgcggccgg gtacgtgttc gagccgcccg
    10381 cgcacgtctc aaccgtgcgg ctgcatgaaa tcctggccgg tttgtctgat gccaagctgg
    10441 cggcctggcc ggccagcttg gccgctgaag aaaccgagcg ccgccgtcta aaaaggtgat
    10501 gtgtatttga gtaaaacagc ttgcgtcatg cggtcgctgc gtatatgatg cgatgagtaa
    10561 ataaacaaat acgcaagggg aacgcatgaa ggttatcgct gtacttaacc agaaaggcgg
    10621 gtcaggcaag acgaccatcg caacccatct agcccgcgcc ctgcaactcg ccggggccga
    10681 tgttctgtta gtcgattccg atccccaggg cagtgcccgc gattgggcgg ccgtgcggga
    10741 agatcaaccg ctaaccgttg tcggcatcga ccgcccgacg attgaccgcg acgtgaaggc
    10801 catcggccgg cgcgacttcg tagtgatcga cggagcgccc caggcggcgg acttggctgt
    10861 gtccgcgatc aaggcagccg acttcgtgct gattccggtg cagccaagcc cttacgacat
    10921 atgggccacc gccgacctgg tggagctggt taagcagcgc attgaggtca cggatggaag
    10981 gctacaagcg gcctttgtcg tgtcgcgggc gatcaaaggc acgcgcatcg gcggtgaggt
    11041 tgccgaggcg ctggccgggt acgagctgcc cattcttgag tcccgtatca cgcagcgcgt
    11101 gagctaccca ggcactgccg ccgccggcac aaccgttctt gaatcagaac ccgagggcga
    11161 cgctgcccgc gaggtccagg cgctggccgc tgaaattaaa tcaaaactca tttgagttaa
    11221 tgaggtaaag agaaaatgag caaaagcaca aacacgctaa gtgccggccg tccgagcgca
    11281 cgcagcagca aggctgcaac gttggccagc ctggcagaca cgccagccat gaagcgggtc
    11341 aactttcagt tgccggcgga ggatcacacc aagctgaaga tgtacgcggt acgccaaggc
    11401 aagaccatta ccgagctgct atctgaatac atcgcgcagc taccagagta aatgagcaaa
    11461 tgaataaatg agtagatgaa ttttagcggc taaaggaggc ggcatggaaa atcaagaaca
    11521 accaggcacc gacgccgtgg aatgccccat gtgtggagga acgggcggtt ggccaggcgt
    11581 aagcggctgg gttgtctgcc ggccctgcaa tggcactgga acccccaagc ccgaggaatc
    11641 ggcgtgacgg tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc
    11701 tggtggagaa gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac
    11761 gccccggtga atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc
    11821 cggcagccgg tgcgccgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt
    11881 tcgttccgat gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg
    11941 ttttccgtct gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag
    12001 acgggcacgt agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc
    12061 tggtactgat ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg
    12121 gagacaagcc cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc
    12181 gagccgatgg cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca
    12241 cgcacgttgc catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg
    12301 agggtgaagc cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt
    12361 acatcgagat cgagctagct gattggatgt accgcgagat cacagaaggc aagaacccgg
    12421 acgtgctgac ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct
    12481 accgcctggc acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct
    12541 acgaacgcag tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga
    12601 tcgggtcaaa tgacctgccg gagtacgatt tgaaggagga ggcggggcag gctggcccga
    12661 tcctagtcat gcgctaccgc aacctgatcg agggcgaagc atccgccggt tcctaatgta
    12721 cggagcagat gctagggcaa attgccctag caggggaaaa aggtcgaaaa ggtgtctttc
    12781 ctgtggatag cacgtacatt gggaacccaa agccgtacat tgggaaccgg aacccgtaca
    12841 ttgggaaccc aaagccgtac attgggaacc ggtcacacat gtaagtgact gatataaaag
    12901 agaaaaaagg cgatttttcc gcctaaaact ctttaaaact tattaaaact cttaaaaccc
    12961 gcctggcctg tgcataactg tctggccagc gcacagccga agagctgcaa aaagcgccta
    13021 cccttcggtc gctgcgctcc ctacgccccg ccgcttcgcg tcggcctatc gcggccgctg
    13081 gccgctcaaa aatggctggc ctacggccag gcaatctacc agggcgcgga caagccgcgc
    13141 cgtcgccact cgaccgccgg cgcccacatc aaggcaccct gcctcgcgcg tttcggtgat
    13201 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
    13261 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
    13321 gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat
    13381 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
    13441 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
    13501 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
    13561 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
    13621 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
    13681 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
    13741 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
    13801 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
    13861 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
    13921 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
    13981 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
    14041 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
    14101 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
    14161 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
    14221 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
    14281 aaaactcacg ttaagggatt ttggtcatgc atgatatatc tcccaatttg tgtagggctt
    14341 attatgcacg cttaaaaata ataaaagcag acttgacctg atagtttggc tgtgagcaat
    14401 tatgtgctta gtgcatctaa tcgcttgagt taacgccggc gaagcggcgt cggcttgaac
    14461 gaatttctag ctagacatta tttgccgact accttggtga tctcgccttt cacgtagtgg
    14521 acaaattctt ccaactgatc tgcgcgcgag gccaagcgat cttcttcttg tccaagataa
    14581 gcctgtctag cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag
    14641 tcggcagcga catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac
    14701 aacgtaagca ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt
    14761 aaggtttcat ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc
    14821 gccgctggac ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga
    14881 tcaatgtcga tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct
    14941 ccaaattgca gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca
    15001 atggtgactt ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa
    15061 aggtcgttga tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc
    15121 aaatcaatat cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg
    15181 gccagcaacg tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc
    15241 gatacttcgg cgatcaccgc ttcccccatg atgtttaact ttgttttagg gcgactgccc
    15301 tgctgcgtaa catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct
    15361 tgctgcttgg atgcccgagg catagactgt accccaaaaa aacatgtcat aacaagaagc
    15421 catgaaaacc gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt
    15481 gcgtgacggc agttacgcta cttgcattac agcttacgaa ccgaacgagg c
//
