LOCUS       sequence-148612-       15532 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 15532)
  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 15532)
  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..15532
                     /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(1199..1378)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      complement(1472..2179)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     CDS             complement(2190..2210)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2223..6326)
                     /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(2223..6326)
                     /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(2223..6326)
                     /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        7422..7524
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             7525..8184
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(7591..7610)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             8530..8835
                     /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     8714..8733
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_RNA        8873..8948
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    9262..9286
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             10587..11216
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             11650..12717
                     /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      12783..12977
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(13213..13235)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    13321..13461
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     13376..13395
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(13476..13493)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13647..14235)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13727..13746)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(14479..15270)
                     /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 ggcccgcgta 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 gatccccctc gatcgagttg agagtgaata tgagactcta
     1201 attggatacc gaggggaatt tatggaacgt cagtggagca tttttgacaa gaaatatttg
     1261 ctagctgata gtgaccttag gcgacttttg aacgcgcaat aatggtttct gacgtatgtg
     1321 cttagctcat taaactccag aaacccgcgg ctgagtggct ccttcaacgt tgcggttctg
     1381 tcagttccaa acgtaaaacg gcttgtcccg cgtcatcggc gggggtcata acgtgactcc
     1441 cttaattctc cgctcatgat tagtagcggt cctgctgagc ctcgacatgt tgtcgcaaaa
     1501 ttcgccctgg acccgcccaa cgatttgtcg tcactgtcaa ggtttgacct gcacttcatt
     1561 tggggcccac atacaccaaa aaaatgctgc ataattctcg gggcagcaag tcggttaccc
     1621 ggccgccgtg ctggaccggg ttgaatggtg cccgtaactt tcggtagagc ggacggccaa
     1681 tactcaactt caaggaatct cacccatgcg cgccggcggg gaaccggagt tcccttcagt
     1741 gagcgttatt agttcgccgc tcggtgtgtc gtagatacta gcccctgggg cacttttgaa
     1801 atttgaataa gatttatgta atcagtcttt taggtttgac cggttctgcc gcttttttta
     1861 aaattggatt tgtaataata aaacgcaatt gtttgttatt gtggcgctct atcatagatg
     1921 tcgctataaa cctattcagc acaatatatt gttttcattt taatattgta catataagta
     1981 gtagggtaca atcagtaaat tgaacggaga atattattca taaaaatacg atagtaacgg
     2041 gtgatatatt cattagaatg aaccgaaacc ggcggtaagg atctgagcta cacatgctca
     2101 ggttttttac aacgtgcaca acagaattga aagcaaatat catgcgatca taggcttctc
     2161 gcatatctca ttaaagcagg acaagctcac accttcctct tcttcttggg gtcagccctg
     2221 ctgtctccac cgagctgaga gaggtcgatt cttgtttcat agagccccgt aattgactga
     2281 tgaatcagtg tggcgtccag gacctccttt gtagaggtgt accgctttct gtctatggtg
     2341 gtgtcgaagt acttgaaggc tgcaggcgcg cccaagttgg tcagagtaaa caagtggata
     2401 atgttttctg cctgctccct gatgggctta tccctgtgct tattgtaagc agaaagcacc
     2461 ttatcgaggt tagcgtcggc gaggatcact cttttggaga attcgcttat ttgctcgatg
     2521 atctcatcaa ggtagtgttt gtgttgttcc acgaacagct gcttctgctc attatcttcg
     2581 ggagatcctt tgagcttttc atagtggctg gccagataca agaaattaac gtatttagag
     2641 ggcagtgcca gctcgttacc tttctgcagc tcgcccgcac tagcgagcat tcgtttccgg
     2701 ccgttttcaa gctcaaagag agagtacttg ggaagcttaa tgatgaggtc ttttttgacc
     2761 tctttatatc ctttcgcctc gagaaagtcg atggggtttt tttcgaagct tgatcgctcc
     2821 atgattgtga tgcccagcag ttccttgacg cttttgagtt ttttagactt ccctttctcc
     2881 actttggcca caaccagtac actgtaagcg actgtaggag aatcgaatcc gccgtatttc
     2941 ttggggtccc aatctttttt gcgtgcgatc agcttgtcgc tgttcctttt cgggaggata
     3001 ctttccttgg agaagcctcc ggtctgtact tcggtctttt taacgatgtt cacctgcggc
     3061 atggacagga ccttccggac tgtcgcgaaa tccctaccct tgtcccacac gatttctcct
     3121 gtttctccgt ttgtttcgat aagtggtcgc ttccgaatct ctccattggc cagtgtaatc
     3181 tcggtcttga aaaaattcat aatattgctg taaaagaagt acttagcggt ggccttgcct
     3241 atttcctgct cagactttgc gatcattttc ctaacatcgt acactttata gtctccgtaa
     3301 acaaattcag attcaagctt gggatatttt ttgataagtg cagtgcctac cactgcattc
     3361 aggtaggcat catgcgcatg gtggtaattg ttgatctctc tcaccttata aaactgaaag
     3421 tcctttctga aatctgaaac cagcttagac ttcagagtaa taactttcac ctctcgaatc
     3481 agtttgtcat tttcatcgta cttggtgttc atgcgtgaat cgagaatttg ggccacgtgc
     3541 ttggtgatct ggcgtgtctc aacaagctgc cttttgatga agccggcttt atccaactca
     3601 gacaggccac ctcgttcagc cttagtcaga ttatcgaact tccgttgtgt gatcagtttg
     3661 gcgttcagca gctgccgcca ataatttttc attttcttga caacttcttc tgaggggacg
     3721 ttatcactct tccctctatt tttatcggat cttgtcaaca ctttattatc aatagaatca
     3781 tctttgagaa aagactgggg cacgatatga tccacgtcgt agtcggagag ccgattgatg
     3841 tccagttcct gatccacgta catgtccctg ccgttctgca ggtagtacag gtagagcttc
     3901 tcattctgaa gctgggtgtt ttcaactggg tgttccttaa ggatttggga ccccagttct
     3961 tttataccct cttcaatcct cttcatcctt tccctactgt tcttctgtcc cttctgggta
     4021 gtttggttct ctcgggccat ctcgataacg atattctcgg gcttatgcct tcccattact
     4081 ttgacgagtt catccacgac cttaacggtc tgcagtattc cctttttgat agctgggcta
     4141 cctgcaagat tagcgatgtg ctcgtggaga ctgtccccct ggccagaaac ttgtgctttc
     4201 tggatgtcct ccttaaaggt gagagagtca tcatggatca actgcatgaa gttccggttg
     4261 gcaaatccat cggacttaag aaaatccagg attgtctttc cactctgctt gtctcggatc
     4321 ccattgatca gttttcttga cagccgcccc catcctgtat atcggcgcct cttgagctgt
     4381 ttcatgactt tgtcgtcgaa gagatgagcg taagttttca agcgttcttc aatcatctcc
     4441 ctatcttcaa acaacgtaag ggtgaggaca atgtcctcaa gaatgtcctc gttctcctca
     4501 ttgtccagga agtccttgtc tttaatgatt ttcaggagat cgtgatacgt tcccagggat
     4561 gcgttgaagc gatcctccac tccgctgatt tcaacagagt cgaaacattc aatctttttg
     4621 aaataatctt ctttgagctg tttcacggta actttccggt tcgtcttgaa gaggaggtcc
     4681 acgatagctt tcttctgctc tccagacagg aatgctggct ttctcatccc ttctgtgacg
     4741 tatttgacct tggtgagctc gttataaact gtgaagtact cgtacagcag agagtgttta
     4801 ggaagcacct tttcgttagg cagattttta tcaaagttag tcatcctttc gatgaaggac
     4861 tgggcagagg cccccttatc cacgacttcc tcgaagttcc agggagtgat agtctcttct
     4921 gatttgcgag tcatccacgc gaatctggaa tttccccggg cgagggggcc tacatagtag
     4981 ggtatccgaa atgtgaggat tttctcaatc ttttccctgt tatctttcaa aaaggggtag
     5041 aaatcctctt gccgcctgag gatagcgtgc agttcgccca ggtgaatctg gtgggggatg
     5101 cttccattgt cgaaagtgcg ctgtttgcgc aacagatctt ctctgttaag ctttaccagc
     5161 agctcctcgg tgccgtccat tttttccaag atgggcttaa taaatttgta aaattcctcc
     5221 tggcttgctc cgccgtcaat gtatccggcg tagccatttt tagactgatc gaagaaaatt
     5281 tccttgtact tctcaggcag ttgctgtctg acaagggcct tcagcaaagt caagtcttgg
     5341 tggtgctcat catagcgctt gatcatacta gcgctcagcg gagctttggt gatctccgtg
     5401 ttcactcgca gaatatcact cagcagaatg gcgtctgaca ggttctttgc cgccaaaaaa
     5461 aggtctgcgt actggtcgcc gatctgggcc agcagattgt cgagatcatc atcgtaggtg
     5521 tctttgctca gttgaagctt ggcatcttcg gccaggtcga agttagattt aaagttgggg
     5581 gtcagcccga gtgacagggc gataagatta ccaaacaggc cgttcttctt ctccccaggg
     5641 agctgtgcga tgaggttttc gagccgccgg gatttggaca gcctagcgct caggattgct
     5701 ttggcgtcaa ctccggatgc gttgatcggg ttctcttcga aaagctgatt gtaagtctga
     5761 accagttgga taaagagttt gtcgacatcg ctgttgtctg ggttcaggtc cccctcgatg
     5821 aggaagtgtc cccgaaattt gatcatatgc gccagcgcga gatagatcaa ccgcaagtca
     5881 gccttatcag tactgtctac aagcttcttc ctcagatgat atatggttgg gtacttttca
     5941 tggtacgcca cctcgtccac gatattgcca aagattgggt ggcgctcgtg ctttttatcc
     6001 tcctccacca aaaaggactc ctccagccta tggaagaaag agtcatccac cttagccatc
     6061 tcattactaa agatctcctg caggtagcag atccgattct ttctgcgggt atatctgcgc
     6121 cgtgctgttc ttttgagccg cgtggcttcg gccgtctccc cggagtcgaa caggagggcg
     6181 ccaatgaggt tcttctttat gctgtggcga tcggtattgc ccagaacttt gaattttttg
     6241 ctcggcacct tgtactcgtc cgtaatgacg gcccagccga cgctgtttgt gccgatatcg
     6301 agcccaatgg agtacttctt gtccattgct gcacatacat aacatatcaa gatcagaaca
     6361 cacatataca cacacaaata caatcaagtc aacaactcca aaaagtccag atctacatat
     6421 atacatacgt aaataacaaa atcatgtaaa taatcacaat catgtaatcc agatctatgc
     6481 acatatatat atacacaatt aataaaaaaa atgatataac agatctatat ctatgtatgt
     6541 aacaacacaa tcagatgaga gaagtgatgt tttcagatct gtatacatac aaacacaaac
     6601 agatgaacaa ttgatacgta gatccatatg tatacgtaca attagctaca cgattaaatg
     6661 aaaaaaatca acgatttcgg attggtacac acaaacgcaa caatatgaag aaattcatat
     6721 ctgattagat ataaacataa ccacgtgtag atacacagtc aaatcaacaa atttatagct
     6781 tctaaacgga tgagatgaac aagataaaga tattcacata aggcatacat aagataagca
     6841 gattaacaaa ctagcaataa tacataccta attaaaacaa ggaataacag agagagagag
     6901 agagagagag atttaccttg aaaatgaaga ggagaagaga ggatttctta aaattggggg
     6961 tagagaaaga aagatgatga attgtgagaa aggagagata gaaggggggg ttgtatatat
     7021 aggctgtaga agattatttt tgtgtttgag gcggtgaagg aagaggggat ctgactatga
     7081 cacgtttgcg gttacgtatt tcgataggag tctttcaacg cttaacgccg ttactctata
     7141 tgaccgtttg ggccgtaacg gggccgtttg ttaacgctga tgttgattct tttctttctt
     7201 tctttcttcc ttttttaaag aagcaattgt acaatcgttg ctagctgtca aacggataat
     7261 tcggatacgg atatgcctat attcatatcc gtaatctcca agcttgaatt cttgctaagt
     7321 tgaaaacaat cttcaaaagt cccacatcgc ttagataaga aaacgaagct gagtttatat
     7381 acagctagag tcgaagtagt gattgaagtc ttctgagacg ttgatcggca cgtaagaggt
     7441 tccaactttc accataatga aataagatca ctaccgggcg tattttttga gttatcgaga
     7501 ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat
     7561 atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc
     7621 tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag
     7681 cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa
     7741 ttccgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac
     7801 accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat
     7861 ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc
     7921 tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt
     7981 ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc
     8041 atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat
     8101 catgccgtct gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc
     8161 gatgagtggc aggggggggc gtaaacgctc gcgtggatcc ggcttactaa aagccagata
     8221 acagtatgcg tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac
     8281 ccgaagtatg tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc
     8341 gacagctatc agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa
     8401 ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag
     8461 ggatggctga ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg
     8521 gctggtgaaa tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt
     8581 gtggatgtac agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc
     8641 agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg
     8701 gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtttc cgttatcggg
     8761 gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg
     8821 ttctggggaa tataaatgtc aggctccctt atacacagcc agtcgaagac ttgttttaga
     8881 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
     8941 gtcggtgctt ttttttgctt ccatgctaga gtccgcaaaa atcaccagtc tctctctaca
     9001 aatctatctc tctctatttt tctccagaat aatgtgtgag tagttcccag ataagggaat
     9061 tagggttctt atagggtttc gctcatgtgt tgagcatata agaaaccctt agtatgtatt
     9121 tgtatttgta aaatacttct atcaataaaa tttctaattc ctaaaaccaa aatccagtga
     9181 cctgcaggca tgcgacgtcg agcttagctt gagcttggat cagattgtcg tttcccgcct
     9241 tcagtttaaa ctatcagtgt ttgacaggat atattggcgg gtaaacctaa gagaaaagag
     9301 cgtttattag aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt
     9361 tgtatgtgca tgccaaccac agggttcccc tcgggatcaa agtactttga tccaacccct
     9421 ccgctgctat agtgcagtcg gcttctgacg ttcagtgcag ccgtgttctg aaaacgacat
     9481 gtcgcacaag tcctaagtta cgcgacaggc tgccgccctg cccttttcct ggcgttttct
     9541 tgtcgcgtgt tttagtcgca taaagtagaa tacttgcgac tagaaccgga gacattacgc
     9601 catgaacaag agcgccgccg ctggcctgct gggctatgcc cgcgtcagca ccgacgacca
     9661 ggacttgacc aaccaacggg ccgaactgca cgcggccggc tgcaccaagc tgttttccga
     9721 gaagatcacc ggcaccaggc gcgaccgccc ggagctggcc aggatgcttg accacctacg
     9781 ccctggcgac gttgtgacag tgaccaggct agaccgcctg gcccgcagca cccgcgacct
     9841 actggacatt gccgagcgca tccaggaggc cggcgcgggc ctgcgtagcc tggcagagcc
     9901 gtgggccgac accaccacgc cggccggccg catggtgttg accgtgttcg ccggcattgc
     9961 cgagttcgag cgttccctaa tcatcgaccg cacccggagc gggcgcgagg ccgccaaggc
    10021 ccgaggcgtg aagtttggcc cccgccctac cctcaccccg gcacagatcg cgcacgcccg
    10081 cgagctgatc gaccaggaag gccgcaccgt gaaagaggcg gctgcactgc ttggcgtgca
    10141 tcgctcgacc ctgtaccgcg cacttgagcg cagcgaggaa gtgacgccca ccgaggccag
    10201 gcggcgcggt gccttccgtg aggacgcatt gaccgaggcc gacgccctgg cggccgccga
    10261 gaatgaacgc caagaggaac aagcatgaaa ccgcaccagg acggccagga cgaaccgttt
    10321 ttcattaccg aagagatcga ggcggagatg atcgcggccg ggtacgtgtt cgagccgccc
    10381 gcgcacgtct caaccgtgcg gctgcatgaa atcctggccg gtttgtctga tgccaagctg
    10441 gcggcctggc cggccagctt ggccgctgaa gaaaccgagc gccgccgtct aaaaaggtga
    10501 tgtgtatttg agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta
    10561 aataaacaaa tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg
    10621 ggtcaggcaa gacgaccatc gcaacccatc tagcccgcgc cctgcaactc gccggggccg
    10681 atgttctgtt agtcgattcc gatccccagg gcagtgcccg cgattgggcg gccgtgcggg
    10741 aagatcaacc gctaaccgtt gtcggcatcg accgcccgac gattgaccgc gacgtgaagg
    10801 ccatcggccg gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg gacttggctg
    10861 tgtccgcgat caaggcagcc gacttcgtgc tgattccggt gcagccaagc ccttacgaca
    10921 tatgggccac cgccgacctg gtggagctgg ttaagcagcg cattgaggtc acggatggaa
    10981 ggctacaagc ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg
    11041 ttgccgaggc gctggccggg tacgagctgc ccattcttga gtcccgtatc acgcagcgcg
    11101 tgagctaccc aggcactgcc gccgccggca caaccgttct tgaatcagaa cccgagggcg
    11161 acgctgcccg cgaggtccag gcgctggccg ctgaaattaa atcaaaactc atttgagtta
    11221 atgaggtaaa gagaaaatga gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc
    11281 acgcagcagc aaggctgcaa cgttggccag cctggcagac acgccagcca tgaagcgggt
    11341 caactttcag ttgccggcgg aggatcacac caagctgaag atgtacgcgg tacgccaagg
    11401 caagaccatt accgagctgc tatctgaata catcgcgcag ctaccagagt aaatgagcaa
    11461 atgaataaat gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac
    11521 aaccaggcac cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggccaggcg
    11581 taagcggctg ggttgtctgc cggccctgca atggcactgg aacccccaag cccgaggaat
    11641 cggcgtgacg gtcgcaaacc atccggcccg gtacaaatcg gcgcggcgct gggtgatgac
    11701 ctggtggaga agttgaaggc cgcgcaggcc gcccagcggc aacgcatcga ggcagaagca
    11761 cgccccggtg aatcgtggca agcggccgct gatcgaatcc gcaaagaatc ccggcaaccg
    11821 ccggcagccg gtgcgccgtc gattaggaag ccgcccaagg gcgacgagca accagatttt
    11881 ttcgttccga tgctctatga cgtgggcacc cgcgatagtc gcagcatcat ggacgtggcc
    11941 gttttccgtc tgtcgaagcg tgaccgacga gctggcgagg tgatccgcta cgagcttcca
    12001 gacgggcacg tagaggtttc cgcagggccg gccggcatgg ccagtgtgtg ggattacgac
    12061 ctggtactga tggcggtttc ccatctaacc gaatccatga accgataccg ggaagggaag
    12121 ggagacaagc ccggccgcgt gttccgtcca cacgttgcgg acgtactcaa gttctgccgg
    12181 cgagccgatg gcggaaagca gaaagacgac ctggtagaaa cctgcattcg gttaaacacc
    12241 acgcacgttg ccatgcagcg tacgaagaag gccaagaacg gccgcctggt gacggtatcc
    12301 gagggtgaag ccttgattag ccgctacaag atcgtaaaga gcgaaaccgg gcggccggag
    12361 tacatcgaga tcgagctagc tgattggatg taccgcgaga tcacagaagg caagaacccg
    12421 gacgtgctga cggttcaccc cgattacttt ttgatcgatc ccggcatcgg ccgttttctc
    12481 taccgcctgg cacgccgcgc cgcaggcaag gcagaagcca gatggttgtt caagacgatc
    12541 tacgaacgca gtggcagcgc cggagagttc aagaagttct gtttcaccgt gcgcaagctg
    12601 atcgggtcaa atgacctgcc ggagtacgat ttgaaggagg aggcggggca ggctggcccg
    12661 atcctagtca tgcgctaccg caacctgatc gagggcgaag catccgccgg ttcctaatgt
    12721 acggagcaga tgctagggca aattgcccta gcaggggaaa aaggtcgaaa aggtgtcttt
    12781 cctgtggata gcacgtacat tgggaaccca aagccgtaca ttgggaaccg gaacccgtac
    12841 attgggaacc caaagccgta cattgggaac cggtcacaca tgtaagtgac tgatataaaa
    12901 gagaaaaaag gcgatttttc cgcctaaaac tctttaaaac ttattaaaac tcttaaaacc
    12961 cgcctggcct gtgcataact gtctggccag cgcacagccg aagagctgca aaaagcgcct
    13021 acccttcggt cgctgcgctc cctacgcccc gccgcttcgc gtcggcctat cgcggccgct
    13081 ggccgctcaa aaatggctgg cctacggcca ggcaatctac cagggcgcgg acaagccgcg
    13141 ccgtcgccac tcgaccgccg gcgcccacat caaggcaccc tgcctcgcgc gtttcggtga
    13201 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
    13261 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
    13321 cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca
    13381 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
    13441 aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
    13501 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
    13561 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
    13621 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
    13681 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
    13741 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
    13801 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
    13861 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
    13921 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
    13981 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
    14041 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
    14101 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
    14161 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
    14221 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
    14281 gaaaactcac gttaagggat tttggtcatg catgatatat ctcccaattt gtgtagggct
    14341 tattatgcac gcttaaaaat aataaaagca gacttgacct gatagtttgg ctgtgagcaa
    14401 ttatgtgctt agtgcatcta atcgcttgag ttaacgccgg cgaagcggcg tcggcttgaa
    14461 cgaatttcta gctagacatt atttgccgac taccttggtg atctcgcctt tcacgtagtg
    14521 gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt gtccaagata
    14581 agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca
    14641 gtcggcagcg acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga
    14701 caacgtaagc actacatttc gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt
    14761 taaggtttca tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc
    14821 cgccgctgga cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag
    14881 atcaatgtcg atcgtggctg gctcgaagat acctgcaaga atgtcattgc gctgccattc
    14941 tccaaattgc agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac
    15001 aatggtgact tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa
    15061 aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag
    15121 caaatcaata tcactgtgtg gcttcaggcc gccatccact gcggagccgt acaaatgtac
    15181 ggccagcaac gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt
    15241 cgatacttcg gcgatcaccg cttcccccat gatgtttaac tttgttttag ggcgactgcc
    15301 ctgctgcgta acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc
    15361 ttgctgcttg gatgcccgag gcatagactg taccccaaaa aaacatgtca taacaagaag
    15421 ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt
    15481 tgcgtgacgg cagttacgct acttgcatta cagcttacga accgaacgag gc
//
