LOCUS       sequence-311489-       16055 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module for the constitutive expression of the nptII, Cas9 and DsRed 
            genes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16055)
  AUTHORS   Vazquez-Vilar M, Garcia-Carpintero V, Selma S, Bernabe-Orts JM, 
            Sanchez-Vicente J, Salazar-Sarasua B, Ressa A, de Paola C, Ajenjo M,
            Quintela JC, Fernandez-Del-Carmen A, Granell A, Orzaez D
  TITLE     The GB4.0 Platform, an All-In-One Tool for CRISPR/Cas-Based 
            Multiplex Genome Engineering in Plants.
  JOURNAL   Front Plant Sci. 2021 Jul 1;12:689937. doi: 
            10.3389/fpls.2021.689937. eCollection 2021.
  PUBMED    34276739
REFERENCE   2  (bases 1 to 16055)
  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: "Front Plant
            Sci."; date: "2021-07-1"; volume: "12:689937. doi"; pages: " 
            10.3389/fpls.2021.689937. eCollection 2021"
COMMENT     Sequence Label: pDGB3 omega1 Tnos:nptII:Pnos - P35S:Cas9:Tnos ...
FEATURES             Location/Qualifiers
     source          1..16055
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(28..280)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(320..1114)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(431..450)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1041..1060
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1148..1327)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        2029..2373
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             2446..6549
                     /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             2446..6549
                     /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             2446..6549
                     /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             6562..6582
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6609..6861
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(6896..6914)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7705..8049
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             8122..8799
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(8302..8322)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     8714..8737
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    9375..9399
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             10700..11329
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             11763..12830
                     /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      12896..13090
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(13326..13348)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    13434..13574
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     13489..13508
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(13589..13606)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13760..14348)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13840..13859)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(14436..15227)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    16013..16037
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggagagcgat cagcttgcat gccggtcgat ctagtaacat agatgacacc gcgcgcgata
       61 atttatccta gtttgcgcgc tatattttgt tttctatcgc gtattaaatg tataattgcg
      121 ggactctaat cataaaaacc catctcataa ataacgtcat gcattacatg ttaattatta
      181 catgcttaac gtaattcaac agaaattata tgataatcat ggcaagaccg gcaacaggat
      241 tcaatcttaa gaaactttat tgccaaatgt ttgaacgatc tgcttgactc tagctagagt
      301 ccgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag gctatgcgct
      361 gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa
      421 gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca
      481 gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc
      541 aggcgtcgcc gtgggtcacg acgagatcct cgccgtcggg catccgcgcc ttgagcctgg
      601 cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa
      661 gaccggcttc catccgagta cgtgctcgct cgatgcggtg tttcgcttgg tggtcgaatg
      721 ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt
      781 tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca
      841 gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg
      901 tggccagcca cgatagccgc gctgcctcgt cttggagttc attcagggca ccggacaggt
      961 cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag
     1021 agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg
     1081 gagaacctgc gtgcaatcca tcttgttcaa tcatgcctcg atcgagttga gagtgaatat
     1141 gagactctaa ttggataccg aggggaattt atggaacgtc agtggagcat ttttgacaag
     1201 aaatatttgc tagctgatag tgaccttagg cgacttttga acgcgcaata atggtttctg
     1261 acgtatgtgc ttagctcatt aaactccaga aacccgcggc tgagtggctc cttcaacgtt
     1321 gcggttctgt cagttccaaa cgtaaaacgg cttgtcccgc gtcatcggcg ggggtcataa
     1381 cgtgactccc ttaattctca tgtatctccg tcaggagact agagccaagc tgatctcctt
     1441 tgccccggag atcaccatgg acgactttct ctatctctac gatctaggaa gaaagttcga
     1501 cggagaaggt gacgatacca tgttcaccac cgataatgag aagattagcc tcttcaattt
     1561 cagaaagaat gctgacccac agatggttag agaggcctac gcggcaggtc tgatcaagac
     1621 gatctacccg agtaataatc tccaggagat caaatacctt cccaagaagg ttaaagatgc
     1681 agtcaaaaga ttcaggacta actgcatcaa gaacacagag aaagatatat ttctcaagat
     1741 cagaagtact attccagtat ggacgattca aggcttgctt cataaaccaa ggcaagtaat
     1801 agagattgga gtctctaaga aagtagttcc tactgaatca aaggccatgg agtcaaaaat
     1861 tcagatcgag gatctaacag aactcgccgt gaagactggc gaacagttca tacagagtct
     1921 tttacgactc aatgacaaga agaaaatctt cgtcaacatg gtggagcacg acactctcgt
     1981 ctactccaag aatatcaaag atacagtctc agaagaccaa agggctattg agacttttca
     2041 acaaagggta atatcgggaa acctcctcgg attccattgc ccagctatct gtcacttcat
     2101 caaaaggaca gtagaaaagg aaggtggcac ctacaaatgc catcattgcg ataaaggaaa
     2161 ggctatcgtt caagatgcct ctgccgacag tggtcccaaa gatggacccc cacccacgag
     2221 gagcatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg attgatgtga
     2281 tatctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag acccttcctc
     2341 tatataagga agttcatttc atttggagag gactccggta tttttacaac aataccacaa
     2401 caaaacaaac aacaaacaac attacaattt actattctag tcgaaatgga caagaagtac
     2461 tccattgggc tcgatatcgg cacaaacagc gtcggctggg ccgtcattac ggacgagtac
     2521 aaggtgccga gcaaaaaatt caaagttctg ggcaataccg atcgccacag cataaagaag
     2581 aacctcattg gcgccctcct gttcgactcc ggggaaacgg ccgaagccac gcggctcaaa
     2641 agaacagcac ggcgcagata tacccgcaga aagaatcgga tctgctacct gcaggagatc
     2701 tttagtaatg agatggctaa ggtggatgac tctttcttcc ataggctgga ggagtccttt
     2761 ttggtggagg aggataaaaa gcacgagcgc cacccaatct ttggcaatat cgtggacgag
     2821 gtggcgtacc atgaaaagta cccaaccata tatcatctga ggaagaagct tgtagacagt
     2881 actgataagg ctgacttgcg gttgatctat ctcgcgctgg cgcatatgat caaatttcgg
     2941 ggacacttcc tcatcgaggg ggacctgaac ccagacaaca gcgatgtcga caaactcttt
     3001 atccaactgg ttcagactta caatcagctt ttcgaagaga acccgatcaa cgcatccgga
     3061 gttgacgcca aagcaatcct gagcgctagg ctgtccaaat cccggcggct cgaaaacctc
     3121 atcgcacagc tccctgggga gaagaagaac ggcctgtttg gtaatcttat cgccctgtca
     3181 ctcgggctga cccccaactt taaatctaac ttcgacctgg ccgaagatgc caagcttcaa
     3241 ctgagcaaag acacctacga tgatgatctc gacaatctgc tggcccagat cggcgaccag
     3301 tacgcagacc tttttttggc ggcaaagaac ctgtcagacg ccattctgct gagtgatatt
     3361 ctgcgagtga acacggagat caccaaagct ccgctgagcg ctagtatgat caagcgctat
     3421 gatgagcacc accaagactt gactttgctg aaggcccttg tcagacagca actgcctgag
     3481 aagtacaagg aaattttctt cgatcagtct aaaaatggct acgccggata cattgacggc
     3541 ggagcaagcc aggaggaatt ttacaaattt attaagccca tcttggaaaa aatggacggc
     3601 accgaggagc tgctggtaaa gcttaacaga gaagatctgt tgcgcaaaca gcgcactttc
     3661 gacaatggaa gcatccccca ccagattcac ctgggcgaac tgcacgctat cctcaggcgg
     3721 caagaggatt tctacccctt tttgaaagat aacagggaaa agattgagaa aatcctcaca
     3781 tttcggatac cctactatgt aggccccctc gcccggggaa attccagatt cgcgtggatg
     3841 actcgcaaat cagaagagac tatcactccc tggaacttcg aggaagtcgt ggataagggg
     3901 gcctctgccc agtccttcat cgaaaggatg actaactttg ataaaaatct gcctaacgaa
     3961 aaggtgcttc ctaaacactc tctgctgtac gagtacttca cagtttataa cgagctcacc
     4021 aaggtcaaat acgtcacaga agggatgaga aagccagcat tcctgtctgg agagcagaag
     4081 aaagctatcg tggacctcct cttcaagacg aaccggaaag ttaccgtgaa acagctcaaa
     4141 gaagattatt tcaaaaagat tgaatgtttc gactctgttg aaatcagcgg agtggaggat
     4201 cgcttcaacg catccctggg aacgtatcac gatctcctga aaatcattaa agacaaggac
     4261 ttcctggaca atgaggagaa cgaggacatt cttgaggaca ttgtcctcac ccttacgttg
     4321 tttgaagata gggagatgat tgaagaacgc ttgaaaactt acgctcatct cttcgacgac
     4381 aaagtcatga aacagctcaa gaggcgccga tatacaggat gggggcggct gtcaagaaaa
     4441 ctgatcaatg ggatccgaga caagcagagt ggaaagacaa tcctggattt tcttaagtcc
     4501 gatggatttg ccaaccggaa cttcatgcag ttgatccatg atgactctct cacctttaag
     4561 gaggacatcc agaaagcaca agtttctggc cagggggaca gtctccacga gcacatcgct
     4621 aatcttgcag gtagcccagc tatcaaaaag ggaatactgc agaccgttaa ggtcgtggat
     4681 gaactcgtca aagtaatggg aaggcataag cccgagaata tcgttatcga gatggcccga
     4741 gagaaccaaa ctacccagaa gggacagaag aacagtaggg aaaggatgaa gaggattgaa
     4801 gagggtataa aagaactggg gtcccaaatc cttaaggaac acccagttga aaacacccag
     4861 cttcagaatg agaagctcta cctgtactac ctgcagaacg gcagggacat gtacgtggat
     4921 caggaactgg acatcaatcg gctctccgac tacgacgtgg atcatatcgt gccccagtct
     4981 tttctcaaag atgattctat tgataataaa gtgttgacaa gatccgataa aaatagaggg
     5041 aagagtgata acgtcccctc agaagaagtt gtcaagaaaa tgaaaaatta ttggcggcag
     5101 ctgctgaacg ccaaactgat cacacaacgg aagttcgata atctgactaa ggctgaacga
     5161 ggtggcctgt ctgagttgga taaagccggc ttcatcaaaa ggcagcttgt tgagacacgc
     5221 cagatcacca agcacgtggc ccaaattctc gattcacgca tgaacaccaa gtacgatgaa
     5281 aatgacaaac tgattcgaga ggtgaaagtt attactctga agtctaagct ggtttcagat
     5341 ttcagaaagg actttcagtt ttataaggtg agagagatca acaattacca ccatgcgcat
     5401 gatgcctacc tgaatgcagt ggtaggcact gcacttatca aaaaatatcc caagcttgaa
     5461 tctgaatttg tttacggaga ctataaagtg tacgatgtta ggaaaatgat cgcaaagtct
     5521 gagcaggaaa taggcaaggc caccgctaag tacttctttt acagcaatat tatgaatttt
     5581 ttcaagaccg agattacact ggccaatgga gagattcgga agcgaccact tatcgaaaca
     5641 aacggagaaa caggagaaat cgtgtgggac aagggtaggg atttcgcgac agtccggaag
     5701 gtcctgtcca tgccgcaggt gaacatcgtt aaaaagaccg aagtacagac cggaggcttc
     5761 tccaaggaaa gtatcctccc gaaaaggaac agcgacaagc tgatcgcacg caaaaaagat
     5821 tgggacccca agaaatacgg cggattcgat tctcctacag tcgcttacag tgtactggtt
     5881 gtggccaaag tggagaaagg gaagtctaaa aaactcaaaa gcgtcaagga actgctgggc
     5941 atcacaatca tggagcgatc aagcttcgaa aaaaacccca tcgactttct cgaggcgaaa
     6001 ggatataaag aggtcaaaaa agacctcatc attaagcttc ccaagtactc tctctttgag
     6061 cttgaaaacg gccggaaacg aatgctcgct agtgcgggcg agctgcagaa aggtaacgag
     6121 ctggcactgc cctctaaata cgttaatttc ttgtatctgg ccagccacta tgaaaagctc
     6181 aaaggatctc ccgaagataa tgagcagaag cagctgttcg tggaacaaca caaacactac
     6241 cttgatgaga tcatcgagca aataagcgaa ttctccaaaa gagtgatcct cgccgacgct
     6301 aacctcgata aggtgctttc tgcttacaat aagcacaggg ataagcccat cagggagcag
     6361 gcagaaaaca ttatccactt gtttactctg accaacttgg gcgcgcctgc agccttcaag
     6421 tacttcgaca ccaccataga cagaaagcgg tacacctcta caaaggaggt cctggacgcc
     6481 acactgattc atcagtcaat tacggggctc tatgaaacaa gaatcgacct ctctcagctc
     6541 ggtggagaca gcagggctga ccccaagaag aagaggaagg tgtgagcttg gaatggatct
     6601 tcgatcccga tcgttcaaac atttggcaat aaagtttctt aagattgaat cctgttgccg
     6661 gtcttgcgac gattatcata taatttctgt tgaattacgt taagcatgta ataattaaca
     6721 tgtaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg caattataca
     6781 tttaatacgc gatagaaaac aaaatatagc gcgcaaacta ggataaatta tcgcgcdcgg
     6841 tgtcatctat gttactagat cgggaattgc caagctaatt cttgaagacg aaagggcctc
     6901 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     6961 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     7021 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataatg ggaccgactc
     7081 gcgctgtcag gagactagag ccaagctgat ctcctttgcc ccggagatca ccatggacga
     7141 ctttctctat ctctacgatc taggaagaaa gttcgacgga gaaggtgacg ataccatgtt
     7201 caccaccgat aatgagaaga ttagcctctt caatttcaga aagaatgctg acccacagat
     7261 ggttagagag gcctacgcgg caggtctgat caagacgatc tacccgagta ataatctcca
     7321 ggagatcaaa taccttccca agaaggttaa agatgcagtc aaaagattca ggactaactg
     7381 catcaagaac acagagaaag atatatttct caagatcaga agtactattc cagtatggac
     7441 gattcaaggc ttgcttcata aaccaaggca agtaatagag attggagtct ctaagaaagt
     7501 agttcctact gaatcaaagg ccatggagtc aaaaattcag atcgaggatc taacagaact
     7561 cgccgtgaag actggcgaac agttcataca gagtctttta cgactcaatg acaagaagaa
     7621 aatcttcgtc aacatggtgg agcacgacac tctcgtctac tccaagaata tcaaagatac
     7681 agtctcagaa gaccaaaggg ctattgagac ttttcaacaa agggtaatat cgggaaacct
     7741 cctcggattc cattgcccag ctatctgtca cttcatcaaa aggacagtag aaaaggaagg
     7801 tggcacctac aaatgccatc attgcgataa aggaaaggct atcgttcaag atgcctctgc
     7861 cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt
     7921 tccaaccacg tcttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga
     7981 cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt
     8041 ggagaggact ccggtatttt tacaacaata ccacaacaaa acaaacaaca aacaacatta
     8101 caatttacta ttctagtcga aatggcctcc tccgagaacg tcatcaccga gttcatgcgc
     8161 ttcaaggtgc gcatggaggg caccgtgaac ggccacgagt tcgagatcga gggcgagggc
     8221 gagggccgcc cctacgaggg ccacaacacc gtgaagctga aggtgaccaa gggcggcccc
     8281 ctgcccttcg cctgggacat cctgtccccc cagttccagt acggctccaa ggtgtacgtg
     8341 aagcaccccg ccgacatccc cgactacaag aagctgtcct tccccgaggg cttcaagtgg
     8401 gagcgcgtga tgaacttcga ggacggcggc gtggcgaccg tgacccagga ctcctccctg
     8461 caggacggct gcttcatcta caaggtgaag ttcatcggcg tgaacttccc ctccgacggc
     8521 cccgtgatgc agaagaagac gatgggctgg gaggcctcca ccgagcgcct gtacccccgc
     8581 gacggcgtgc tgaagggcga gacacacaag gccctgaagc tgaaggacgg cggccactac
     8641 ctggtggagt tcaagtccat ctacatggcc aagaagcccg tgcagctgcc cggctactac
     8701 tacgtggacg ccaagctgga catcacctcc cacaacgagg actacaccat cgtggagcag
     8761 tacgagcgca ccgagggccg ccaccacctg ttcctgtagg cttcggccat gctagagtcc
     8821 gcaaaaatca ccagtctctc tctacaaatc tatctctctc tatttttctc cagaataatg
     8881 tgtgagtagt tcccagataa gggaattagg gttcttatag ggtttcgctc atgtgttgag
     8941 catataagaa acccttagta tgtatttgta tttgtaaaat acttctatca ataaaatttc
     9001 taattcctaa aaccaaaatc cagtgacctc gctgtcagga gtctcaatgg taactttact
     9061 ctttatttaa ccatacattt ttttttattt ttttcacttt gttcttcatc cactattgtt
     9121 ctttgttcat cttgaacaaa agctccctcc ttctttgttc ttcatccacc attgttcttc
     9181 atcaatcatt tcgctgtcag gagtctcaat ggtaacttta ctctttattt aaccatacat
     9241 ttttttttat ttttttcact ttgttcttca tccactattg ttctttgttc atcttgaaca
     9301 aaagctccct ccttctttgt tcttcatcca ccattgttct tcatcaatca tttcgctgtc
     9361 atgagaccgg atcctgacag gatatattgg cgggtaaacc taagagaaaa gagcgtttat
     9421 tagaataatc ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt
     9481 gcatgccaac cacagggttc ccctcgggat caaagtactt tgatccaacc cctccgctgc
     9541 tatagtgcag tcggcttctg acgttcagtg cagccgtcat ctgaaaacga catgtcgcac
     9601 aagtcctaag ttacgcgaca ggctgccgcc ctgccctttt cctggcgttt tcttgtcgcg
     9661 tgttttagtc gcataaagta gaatacttgc gactagaacc ggagacatta cgccatgaac
     9721 aagagcgccg ccgctggcct gctgggctat gcccgcgtca gcaccgacga ccaggacttg
     9781 accaaccaac gggccgaact gcacgcggcc ggctgcacca agctgttttc cgagaagatc
     9841 accggcacca ggcgcgaccg cccggagctg gccaggatgc ttgaccacct acgccctggc
     9901 gacgttgtga cagtgaccag gctagaccgc ctggcccgca gcacccgcga cctactggac
     9961 attgccgagc gcatccagga ggccggcgcg ggcctgcgta gcctggcaga gccgtgggcc
    10021 gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt tcgccggcat tgccgagttc
    10081 gagcgttccc taatcatcga ccgcacccgg agcgggcgcg aggccgccaa ggcccgaggc
    10141 gtgaagtttg gcccccgccc taccctcacc ccggcacaga tcgcgcacgc ccgcgagctg
    10201 atcgaccagg aaggccgcac cgtgaaagag gcggctgcac tgcttggcgt gcatcgctcg
    10261 accctgtacc gcgcacttga gcgcagcgag gaagtgacgc ccaccgaggc caggcggcgc
    10321 ggtgccttcc gtgaggacgc attgaccgag gccgacgccc tggcggccgc cgagaatgaa
    10381 cgccaagagg aacaagcatg aaaccgcacc aggacggcca ggacgaaccg tttttcatta
    10441 ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt gttcgagccg cccgcgcacc
    10501 tctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc tgatgccaag ctggcggcct
    10561 ggccggccag cttggccgct gaagaaaccg agcgccgccg tctaaaaagg tgatgtgtat
    10621 ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat gatccgatga gtaaataaac
    10681 aaatacgcaa ggggaacgca tgaaggttat cgctgtactt aaccagaaag gcgggtcagg
    10741 caagacgacc atcggaaccc atctagcccg cgccctgcaa ctcgccgggg ccgatgttct
    10801 gttagtcgat tccgatcccc agggcagtgc ccgcgattgg gcggccgtgc gggaagatca
    10861 accgctaacc gttgtcggca tcgaccgccc gacgattgac cgcgacgtga aggccatcgg
    10921 ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg gcggacttgg ctgtgtccgc
    10981 gatcaaggca gccgacttcg tgctgattcc ggtgcagcca agcccttacg acatatgggc
    11041 caccgccgac ctggtggagc tggttaagca gcgcattgag gtcacggatg gaaggctaca
    11101 agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc atcggcggtg aggttgccga
    11161 ggcgctggcc gggtacgagc tgcccattct tgagtcccgt atcacgcagc gcgtgagcta
    11221 cccaggcact gccgccgccg gcacaaccgt tcttgaatca gaacccgagg gcgacgctgc
    11281 ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa ctcatttgag ttaatgaggt
    11341 aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg gccgtccgag cgcacgcagc
    11401 agcaaggctg caacgttggc cagcctggca gacacgccag ccatgaagcg ggtcaacttt
    11461 cagttgccgg cggaggatca caccaagctg aagatgtacg cggtacgcca aggcaagacc
    11521 attaccgagc tgctatctga atagatcgcg cagctaccag agtaaatgag caaatgaata
    11581 aatgagtaga tgaattttag cggctaaagg aggcggcatg gaaaatcaag aacaaccagg
    11641 caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc ggttggccag gcgtaagcgg
    11701 ctgggttgtc tgccggccct gcaatggcac tggaaccccc aagcccgagg aatcggcgtg
    11761 acggtcgcaa accatccggc ccggtacaaa tcggcgcggc gctgggtgat gacctggtgg
    11821 agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat cgaggcagaa gcacgccccg
    11881 gtgaatcgtg gcaagcggcc gctgatcgaa tccgcaaaga atcccggcaa ccgccggcag
    11941 ccggtgcgcc gtcgattagg aagccgccca agggcgacga gcaaccagat tttttcgttc
    12001 cgatgctcta tgacgtgggc acccgcgata gtcgcagcat catggacgtg gccgttttcc
    12061 gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg ctacgagctt ccagacgggc
    12121 acgtagaggt ttccgcaggg ccggccggca tggccagtgt gtgggattac gacctggtac
    12181 tgatggcggt ttcccatcta accgaatcca tgaaccgata ccgggaaggg aagggagaca
    12241 agcccggccg cgtgttccgt ccacacgttg cggacgtact caagttctgc cggcgagccg
    12301 atggcggaaa gcagaaagac gacctggtag aaacctgcat tcggttaaac accacgcacg
    12361 ttgccatgca gcgtacgaag aaggccaaga acggccgcct ggtgacggta tccgagggtg
    12421 aagccttgat tagccgctac aagatcgtaa agagcgaaac cgggcggccg gagtacatcg
    12481 agatcgagct agctgattgg atgtaccgcg agatcacaga aggcaagaac ccggacgtgc
    12541 tgacggttca ccccgattac tttttgatcg atcccggcat cggccgtttt ctctaccgcc
    12601 tggcacgccg cgccgcaggc aaggcagaag ccagatggtt gttcaagacg atctacgaac
    12661 gcagtggcag cgccggagag ttcaagaagt tctgtttcac cgtgcgcaag ctgatcgggt
    12721 caaatgacct gccggagtac gatttgaagg aggaggcggg gcaggctggc ccgatcctag
    12781 tcatgcgcta ccgcaacctg atcgagggcg aagcatccgc cggttcctaa tgtacggagc
    12841 agatgctagg gcaaattgcc ctagcagggg aaaaaggtcg aaaaggactc tttcctgtgg
    12901 atagcacgta cattgggaac ccaaagccgt acattgggaa ccggaacccg tacattggga
    12961 acccaaagcc gtacattggg aaccggtcac acatgtaagt gactgatata aaagagaaaa
    13021 aaggcgattt ttccgcctaa aactctttaa aacttattaa aactcttaaa acccgcctgg
    13081 cctgtgcata actgtctggc cagcgcacag ccgaagagct gcaaaaagcg cctacccttc
    13141 ggtcgctgcg ctccctacgc cccgccgctt cgcgtcggcc tatcgcggcc gctggccgct
    13201 caaaaatggc tggcctacgg ccaggcaatc taccagggcg cggacaagcc gcgccgtcgc
    13261 cactcgaccg ccggcgccca catcaaggca ccctgcctcg cgcgtttcgg tgatgacggt
    13321 gaaaacctct gacacatgca gctcccggtg acggtcacag cttgtctgta agcggatgcc
    13381 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
    13441 atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc
    13501 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
    13561 aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
    13621 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
    13681 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
    13741 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
    13801 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
    13861 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
    13921 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
    13981 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
    14041 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
    14101 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
    14161 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
    14221 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
    14281 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
    14341 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
    14401 cacgttaagg gattttggtc atgcattcta ggtgattatt tgccgactac cttggtgatc
    14461 tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc caagcgatct
    14521 tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata ctgggccggc
    14581 aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg
    14641 ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcaccagc ccagtcgggc
    14701 ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc
    14761 ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt
    14821 gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc tgcaagaatg
    14881 tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg
    14941 atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg
    15001 gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt
    15061 acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc atccactgcg
    15121 gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat gacgccaact
    15181 acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcataat gtttaacttt
    15241 gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tccataacat caaacatcga
    15301 cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac cccaaaaaaa
    15361 cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg ttcggtcaag
    15421 gttctggacc agttgcgtga gcgcatacgc tacttgcatt acagcttacg aaccgaacag
    15481 gcttatgtcc actgggttcg tgccttcatc cgtttccacg gtgtgcgtca cccggcaacc
    15541 ttgggtagca gcgaagtcga ggcatttctg tcctggctgg aacagaactt attatttcct
    15601 tcctcttttc tacagtattt aaagataccc caagaagcta attataacaa gacgaactcc
    15661 aattcactgt tccttgcatt ctaaaacctt aaataccaga aaacagcttt ttcaaagttg
    15721 ttttcaaagt tggcgtataa catagtatcg acggagccga ttttgaaacc gcggtgatca
    15781 caggcagcaa cgctctgtca tcgttacaat caacatgcta ccctccgcga gatcatccgt
    15841 gtttcaaacc cggcagctta gttgccgttc ttccgaatag catcggtaac atgagcaaag
    15901 tctgccgcct tacaacggct ctcccgctga cgccgtcccg gactgatggg ctgcctgtat
    15961 cgagtggtga ttttgtgccg agctgccggt cggggagctg ttggctggct ggtggcagga
    16021 tatattgtgg tgtaaacata acggatccgg tctca
//
