LOCUS       sequence-148608-       15352 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  recipient plasmid [multiplexing]; pPcUbi:Cas9, BASTA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15352)
  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 15352)
  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: pDGE4
FEATURES             Location/Qualifiers
     source          1..15352
                     /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        7331..7433
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             7434..8093
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(7500..7519)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             8435..8740
                     /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     8619..8638
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_feature    9082..9106
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             10407..11036
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             11470..12537
                     /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      12603..12797
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(13033..13055)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    13141..13281
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     13196..13215
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(13296..13313)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13467..14055)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13547..13566)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(14299..15090)
                     /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 cttgcagaga
     7321 cctgagacgt tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac
     7381 taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct aaaatggaga
     7441 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
     7501 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
     7561 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
     7621 ttgcccgcct gatgaatgct catccggaat tccgtatggc aatgaaagac ggtgagctgg
     7681 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
     7741 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
     7801 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
     7861 ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
     7921 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
     7981 tgctgatgcc gctggcgatt caggttcatc atgccgtctg tgatggcttc catgtcggca
     8041 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taatcgcgtg
     8101 gatccggctt actaaaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta
     8161 taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca
     8221 gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc
     8281 aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa
     8341 cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac
     8401 ggctcttttg ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa
     8461 aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg
     8521 gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga
     8581 actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc
     8641 cagtgtgccg gtttccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga
     8701 catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca
     8761 cagccagtcg gtctcagctt ccatgctaga gtccgcaaaa atcaccagtc tctctctaca
     8821 aatctatctc tctctatttt tctccagaat aatgtgtgag tagttcccag ataagggaat
     8881 tagggttctt atagggtttc gctcatgtgt tgagcatata agaaaccctt agtatgtatt
     8941 tgtatttgta aaatacttct atcaataaaa tttctaattc ctaaaaccaa aatccagtga
     9001 cctgcaggca tgcgacgtcg agcttagctt gagcttggat cagattgtcg tttcccgcct
     9061 tcagtttaaa ctatcagtgt ttgacaggat atattggcgg gtaaacctaa gagaaaagag
     9121 cgtttattag aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt
     9181 tgtatgtgca tgccaaccac agggttcccc tcgggatcaa agtactttga tccaacccct
     9241 ccgctgctat agtgcagtcg gcttctgacg ttcagtgcag ccgtgttctg aaaacgacat
     9301 gtcgcacaag tcctaagtta cgcgacaggc tgccgccctg cccttttcct ggcgttttct
     9361 tgtcgcgtgt tttagtcgca taaagtagaa tacttgcgac tagaaccgga gacattacgc
     9421 catgaacaag agcgccgccg ctggcctgct gggctatgcc cgcgtcagca ccgacgacca
     9481 ggacttgacc aaccaacggg ccgaactgca cgcggccggc tgcaccaagc tgttttccga
     9541 gaagatcacc ggcaccaggc gcgaccgccc ggagctggcc aggatgcttg accacctacg
     9601 ccctggcgac gttgtgacag tgaccaggct agaccgcctg gcccgcagca cccgcgacct
     9661 actggacatt gccgagcgca tccaggaggc cggcgcgggc ctgcgtagcc tggcagagcc
     9721 gtgggccgac accaccacgc cggccggccg catggtgttg accgtgttcg ccggcattgc
     9781 cgagttcgag cgttccctaa tcatcgaccg cacccggagc gggcgcgagg ccgccaaggc
     9841 ccgaggcgtg aagtttggcc cccgccctac cctcaccccg gcacagatcg cgcacgcccg
     9901 cgagctgatc gaccaggaag gccgcaccgt gaaagaggcg gctgcactgc ttggcgtgca
     9961 tcgctcgacc ctgtaccgcg cacttgagcg cagcgaggaa gtgacgccca ccgaggccag
    10021 gcggcgcggt gccttccgtg aggacgcatt gaccgaggcc gacgccctgg cggccgccga
    10081 gaatgaacgc caagaggaac aagcatgaaa ccgcaccagg acggccagga cgaaccgttt
    10141 ttcattaccg aagagatcga ggcggagatg atcgcggccg ggtacgtgtt cgagccgccc
    10201 gcgcacgtct caaccgtgcg gctgcatgaa atcctggccg gtttgtctga tgccaagctg
    10261 gcggcctggc cggccagctt ggccgctgaa gaaaccgagc gccgccgtct aaaaaggtga
    10321 tgtgtatttg agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta
    10381 aataaacaaa tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg
    10441 ggtcaggcaa gacgaccatc gcaacccatc tagcccgcgc cctgcaactc gccggggccg
    10501 atgttctgtt agtcgattcc gatccccagg gcagtgcccg cgattgggcg gccgtgcggg
    10561 aagatcaacc gctaaccgtt gtcggcatcg accgcccgac gattgaccgc gacgtgaagg
    10621 ccatcggccg gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg gacttggctg
    10681 tgtccgcgat caaggcagcc gacttcgtgc tgattccggt gcagccaagc ccttacgaca
    10741 tatgggccac cgccgacctg gtggagctgg ttaagcagcg cattgaggtc acggatggaa
    10801 ggctacaagc ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg
    10861 ttgccgaggc gctggccggg tacgagctgc ccattcttga gtcccgtatc acgcagcgcg
    10921 tgagctaccc aggcactgcc gccgccggca caaccgttct tgaatcagaa cccgagggcg
    10981 acgctgcccg cgaggtccag gcgctggccg ctgaaattaa atcaaaactc atttgagtta
    11041 atgaggtaaa gagaaaatga gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc
    11101 acgcagcagc aaggctgcaa cgttggccag cctggcagac acgccagcca tgaagcgggt
    11161 caactttcag ttgccggcgg aggatcacac caagctgaag atgtacgcgg tacgccaagg
    11221 caagaccatt accgagctgc tatctgaata catcgcgcag ctaccagagt aaatgagcaa
    11281 atgaataaat gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac
    11341 aaccaggcac cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggccaggcg
    11401 taagcggctg ggttgtctgc cggccctgca atggcactgg aacccccaag cccgaggaat
    11461 cggcgtgacg gtcgcaaacc atccggcccg gtacaaatcg gcgcggcgct gggtgatgac
    11521 ctggtggaga agttgaaggc cgcgcaggcc gcccagcggc aacgcatcga ggcagaagca
    11581 cgccccggtg aatcgtggca agcggccgct gatcgaatcc gcaaagaatc ccggcaaccg
    11641 ccggcagccg gtgcgccgtc gattaggaag ccgcccaagg gcgacgagca accagatttt
    11701 ttcgttccga tgctctatga cgtgggcacc cgcgatagtc gcagcatcat ggacgtggcc
    11761 gttttccgtc tgtcgaagcg tgaccgacga gctggcgagg tgatccgcta cgagcttcca
    11821 gacgggcacg tagaggtttc cgcagggccg gccggcatgg ccagtgtgtg ggattacgac
    11881 ctggtactga tggcggtttc ccatctaacc gaatccatga accgataccg ggaagggaag
    11941 ggagacaagc ccggccgcgt gttccgtcca cacgttgcgg acgtactcaa gttctgccgg
    12001 cgagccgatg gcggaaagca gaaagacgac ctggtagaaa cctgcattcg gttaaacacc
    12061 acgcacgttg ccatgcagcg tacgaagaag gccaagaacg gccgcctggt gacggtatcc
    12121 gagggtgaag ccttgattag ccgctacaag atcgtaaaga gcgaaaccgg gcggccggag
    12181 tacatcgaga tcgagctagc tgattggatg taccgcgaga tcacagaagg caagaacccg
    12241 gacgtgctga cggttcaccc cgattacttt ttgatcgatc ccggcatcgg ccgttttctc
    12301 taccgcctgg cacgccgcgc cgcaggcaag gcagaagcca gatggttgtt caagacgatc
    12361 tacgaacgca gtggcagcgc cggagagttc aagaagttct gtttcaccgt gcgcaagctg
    12421 atcgggtcaa atgacctgcc ggagtacgat ttgaaggagg aggcggggca ggctggcccg
    12481 atcctagtca tgcgctaccg caacctgatc gagggcgaag catccgccgg ttcctaatgt
    12541 acggagcaga tgctagggca aattgcccta gcaggggaaa aaggtcgaaa aggtgtcttt
    12601 cctgtggata gcacgtacat tgggaaccca aagccgtaca ttgggaaccg gaacccgtac
    12661 attgggaacc caaagccgta cattgggaac cggtcacaca tgtaagtgac tgatataaaa
    12721 gagaaaaaag gcgatttttc cgcctaaaac tctttaaaac ttattaaaac tcttaaaacc
    12781 cgcctggcct gtgcataact gtctggccag cgcacagccg aagagctgca aaaagcgcct
    12841 acccttcggt cgctgcgctc cctacgcccc gccgcttcgc gtcggcctat cgcggccgct
    12901 ggccgctcaa aaatggctgg cctacggcca ggcaatctac cagggcgcgg acaagccgcg
    12961 ccgtcgccac tcgaccgccg gcgcccacat caaggcaccc tgcctcgcgc gtttcggtga
    13021 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
    13081 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
    13141 cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca
    13201 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
    13261 aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
    13321 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
    13381 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
    13441 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
    13501 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
    13561 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
    13621 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
    13681 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
    13741 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
    13801 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
    13861 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
    13921 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
    13981 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
    14041 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
    14101 gaaaactcac gttaagggat tttggtcatg catgatatat ctcccaattt gtgtagggct
    14161 tattatgcac gcttaaaaat aataaaagca gacttgacct gatagtttgg ctgtgagcaa
    14221 ttatgtgctt agtgcatcta atcgcttgag ttaacgccgg cgaagcggcg tcggcttgaa
    14281 cgaatttcta gctagacatt atttgccgac taccttggtg atctcgcctt tcacgtagtg
    14341 gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt gtccaagata
    14401 agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca
    14461 gtcggcagcg acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga
    14521 caacgtaagc actacatttc gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt
    14581 taaggtttca tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc
    14641 cgccgctgga cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag
    14701 atcaatgtcg atcgtggctg gctcgaagat acctgcaaga atgtcattgc gctgccattc
    14761 tccaaattgc agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac
    14821 aatggtgact tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa
    14881 aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag
    14941 caaatcaata tcactgtgtg gcttcaggcc gccatccact gcggagccgt acaaatgtac
    15001 ggccagcaac gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt
    15061 cgatacttcg gcgatcaccg cttcccccat gatgtttaac tttgttttag ggcgactgcc
    15121 ctgctgcgta acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc
    15181 ttgctgcttg gatgcccgag gcatagactg taccccaaaa aaacatgtca taacaagaag
    15241 ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt
    15301 tgcgtgacgg cagttacgct acttgcatta cagcttacga accgaacgag gc
//
