LOCUS       sequence-244377-       12267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcriptional unit of (human codon optimized) inactivated Cas9 
            fused to the VP64 Transcriptional Activator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12267)
  AUTHORS   Vazquez-Vilar M, Bernabe-Orts JM, Fernandez-Del-Carmen A, Ziarsolo 
            P, Blanca J, Granell A, Orzaez D
  TITLE     A modular toolbox for gRNA-Cas9 genome engineering in plants based 
            on the GoldenBraid standard.
  JOURNAL   Plant Methods. 2016 Feb 1;12:10. doi: 10.1186/s13007-016-0101-2. 
            eCollection 2016.
  PUBMED    26839579
REFERENCE   2  (bases 1 to 12267)
  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 
            Methods."; date: "2016-02-1"; volume: "12:10. doi"; pages: " 
            10.1186/s13007-016-0101-2. eCollection 2016"
COMMENT     Sequence Label: pEGB 35s:dCas:VP64:tNos (GB1189)
FEATURES             Location/Qualifiers
     source          1..12267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        616..960
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     904..924
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             1037..1057
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1058..1081,1082..1084,1085..3568,3569..3574,3575..3640,
                     3641..3643,3644..5158)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(1058..1081,1082..1084,1085..3571,3572..3574,
                     3575..5158)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1058..5158
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5177..5197
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5201..5221
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5225..5245
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5246..5395
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     terminator      5422..5674
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(5709..5727)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5911..5935
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             7236..7865
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             8294..9367
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      9433..9627
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(9863..9885)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    9971..10111
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     10026..10045
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(10126..10143)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10297..10885)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10377..10396)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(10973..11788)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(10973..11788)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     11696..11715
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    12221..12245
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggagactaga gccaagctga tctcctttgc cccggagatc accatggacg actttctcta
       61 tctctacgat ctaggaagaa agttcgacgg agaaggtgac gataccatgt tcaccaccga
      121 taatgagaag attagcctct tcaatttcag aaagaatgct gacccacaga tggttagaga
      181 ggcctacgcg gcaggtctga tcaagacgat ctacccgagt aataatctcc aggagatcaa
      241 ataccttccc aagaaggtta aagatgcagt caaaagattc aggactaact gcatcaagaa
      301 cacagagaaa gatatatttc tcaagatcag aagtactatt ccagtatgga cgattcaagg
      361 cttgcttcat aaaccaaggc aagtaataga gattggagtc tctaagaaag tagttcctac
      421 tgaatcaaag gccatggagt caaaaattca gatcgaggat ctaacagaac tcgccgtgaa
      481 gactggcgaa cagttcatac agagtctttt acgactcaat gacaagaaga aaatcttcgt
      541 caacatggtg gagcacgaca ctctcgtcta ctccaagaat atcaaagata cagtctcaga
      601 agaccaaagg gctattgaga cttttcaaca aagggtaata tcgggaaacc tcctcggatt
      661 ccattgccca gctatctgtc acttcatcaa aaggacagta gaaaaggaag gtggcaccta
      721 caaatgccat cattgcgata aaggaaaggc tatcgttcaa gatgcccctg ccgacagtgg
      781 tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg ttccaaccac
      841 gtcttcaaag caagtggatt gatgtgatat ctccactgac gtaagggatg acgcacaatc
      901 ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagaggac
      961 tccggtattt ttacaacaat taccacaaca aaacaaacaa caaacaacat tacaatttac
     1021 tattctagtc gaaatgccca agaagaagag gaaggtggac aagaagtact ccattgggct
     1081 cgctatcggc acaaacagcg tcggctgggc cgtcattacg gacgagtaca aggtgccgag
     1141 caaaaaattc aaagttctgg gcaataccga tcgccacagc ataaagaaga acctcattgg
     1201 cgccctcctg ttcgactccg gggaaacggc cgaagccacg cggctcaaaa gaacagcacg
     1261 gcgcagatat acccgcagaa agaatcggat ctgctacctg caggagatct ttagtaatga
     1321 gatggctaag gtggatgact ctttcttcca taggctggag gagtcctttt tggtggagga
     1381 ggataaaaag cacgagcgcc acccaatctt tggcaatatc gtggacgagg tggcgtacca
     1441 tgaaaagtac ccaaccatat atcatctgag gaagaagctt gtagacagta ctgataaggc
     1501 tgacttgcgg ttgatctatc tcgcgctggc gcatatgatc aaatttcggg gacacttcct
     1561 catcgagggg gacctgaacc cagacaacag cgatgtcgac aaactcttta tccaactggt
     1621 tcagacttac aatcagcttt tcgaagagaa cccgatcaac gcatccggag ttgacgccaa
     1681 agcaatcctg agcgctaggc tgtccaaatc ccggcggctc gaaaacctca tcgcacagct
     1741 ccctggggag aagaagaacg gcctgtttgg taatcttatc gccctgtcac tcgggctgac
     1801 ccccaacttt aaatctaact tcgacctggc cgaagatgcc aagcttcaac tgagcaaaga
     1861 cacctacgat gatgatctcg acaatctgct ggcccagatc ggcgaccagt acgcagacct
     1921 ttttttggcg gcaaagaacc tgtcagacgc cattctgctg agtgatattc tgcgagtgaa
     1981 cacggagatc accaaagctc cgctgagcgc tagtatgatc aagcgctatg atgagcacca
     2041 ccaagacttg actttgctga aggcccttgt cagacagcaa ctgcctgaga agtacaagga
     2101 aattttcttc gatcagtcta aaaatggcta cgccggatac attgacggcg gagcaagcca
     2161 ggaggaattt tacaaattta ttaagcccat cttggaaaaa atggacggca ccgaggagct
     2221 gctggtaaag cttaacagag aagatctgtt gcgcaaacag cgcactttcg acaatggaag
     2281 catcccccac cagattcacc tgggcgaact gcacgctatc ctcaggcggc aagaggattt
     2341 ctaccccttt ttgaaagata acagggaaaa gattgagaaa atcctcacat ttcggatacc
     2401 ctactatgta ggccccctcg cccggggaaa ttccagattc gcgtggatga ctcgcaaatc
     2461 agaagagact atcactccct ggaacttcga ggaagtcgtg gataaggggg cctctgccca
     2521 gtccttcatc gaaaggatga ctaactttga taaaaatctg cctaacgaaa aggtgcttcc
     2581 taaacactct ctgctgtacg agtacttcac agtttataac gagctcacca aggtcaaata
     2641 cgtcacagaa gggatgagaa agccagcatt cctgtctgga gagcagaaga aagctatcgt
     2701 ggacctcctc ttcaagacga accggaaagt taccgtgaaa cagctcaaag aagattattt
     2761 caaaaagatt gaatgtttcg actctgttga aatcagcgga gtggaggatc gcttcaacgc
     2821 atccctggga acgtatcacg atctcctgaa aatcattaaa gacaaggact tcctggacaa
     2881 tgaggagaac gaggacattc ttgaggacat tgtcctcacc cttacgttgt ttgaagatag
     2941 ggagatgatt gaagaacgct tgaaaactta cgctcatctc ttcgacgaca aagtcatgaa
     3001 acagctcaag aggcgccgat atacaggatg ggggcggctg tcaagaaaac tgatcaatgg
     3061 gatccgagac aagcagagtg gaaagacaat cctggatttt cttaagtccg atggatttgc
     3121 caaccggaac ttcatgcagt tgatccatga tgactctctc acctttaagg aggacatcca
     3181 gaaagcacaa gtttctggcc agggggacag tctccacgag cacatcgcta atcttgcagg
     3241 tagcccagct atcaaaaagg gaatactgca gaccgttaag gtcgtggatg aactcgtcaa
     3301 agtaatggga aggcataagc ccgagaatat cgttatcgag atggcccgag agaaccaaac
     3361 tacccagaag ggacagaaga acagtaggga aaggatgaag aggattgaag agggtataaa
     3421 agaactgggg tcccaaatcc ttaaggaaca cccagttgaa aacacccagc ttcagaatga
     3481 gaagctctac ctgtactacc tgcagaacgg cagggacatg tacgtggatc aggaactgga
     3541 catcaatcgg ctctccgact acgacgtgga tgccatcgtg ccccagtctt ttctcaaaga
     3601 tgattctatt gataataaag tgttgacaag atccgataaa aatagaggga agagtgataa
     3661 cgtcccctca gaagaagttg tcaagaaaat gaaaaattat tggcggcagc tgctgaacgc
     3721 caaactgatc acacaacgga agttcgataa tctgactaag gctgaacgag gtggcctgtc
     3781 tgagttggat aaagccggct tcatcaaaag gcagcttgtt gagacacgcc agatcaccaa
     3841 gcacgtggcc caaattctcg attcacgcat gaacaccaag tacgatgaaa atgacaaact
     3901 gattcgagag gtgaaagtta ttactctgaa gtctaagctg gtttcagatt tcagaaagga
     3961 ctttcagttt tataaggtga gagagatcaa caattaccac catgcgcatg atgcctacct
     4021 gaatgcagtg gtaggcactg cacttatcaa aaaatatccc aagcttgaat ctgaatttgt
     4081 ttacggagac tataaagtgt acgatgttag gaaaatgatc gcaaagtctg agcaggaaat
     4141 aggcaaggcc accgctaagt acttctttta cagcaatatt atgaattttt tcaagaccga
     4201 gattacactg gccaatggag agattcggaa gcgaccactt atcgaaacaa acggagaaac
     4261 aggagaaatc gtgtgggaca agggtaggga tttcgcgaca gtccggaagg tcctgtccat
     4321 gccgcaggtg aacatcgtta aaaagaccga agtacagacc ggaggcttct ccaaggaaag
     4381 tatcctcccg aaaaggaaca gcgacaagct gatcgcacgc aaaaaagatt gggaccccaa
     4441 gaaatacggc ggattcgatt ctcctacagt cgcttacagt gtactggttg tggccaaagt
     4501 ggagaaaggg aagtctaaaa aactcaaaag cgtcaaggaa ctgctgggca tcacaatcat
     4561 ggagcgatca agcttcgaaa aaaaccccat cgactttctc gaggcgaaag gatataaaga
     4621 ggtcaaaaaa gacctcatca ttaagcttcc caagtactct ctctttgagc ttgaaaacgg
     4681 ccggaaacga atgctcgcta gtgcgggcga gctgcagaaa ggtaacgagc tggcactgcc
     4741 ctctaaatac gttaatttct tgtatctggc cagccactat gaaaagctca aaggatctcc
     4801 cgaagataat gagcagaagc agctgttcgt ggaacaacac aaacactacc ttgatgagat
     4861 catcgagcaa ataagcgaat tctccaaaag agtgatcctc gccgacgcta acctcgataa
     4921 ggtgctttct gcttacaata agcacaggga taagcccatc agggagcagg cagaaaacat
     4981 tatccacttg tttactctga ccaacttggg cgcgcctgca gccttcaagt acttcgacac
     5041 caccatagac agaaagcggt acacctctac aaaggaggtc ctggacgcca cactgattca
     5101 tcagtcaatt acggggctct atgaaacaag aatcgacctc tctcagctcg gtggagacag
     5161 cagggctgat tcggacccaa aaaagaagcg taaggtcgat cccaagaaga agagaaaggt
     5221 agatcctaag aagaagagaa aggtagacgc attggatgac ttcgacttag acatgcttgg
     5281 gtcagatgct ttggacgact ttgacctcga catgttgggt tctgacgcgc tagatgactt
     5341 cgacttggac atgttgggct ccgacgcttt ggacgacttt gatctggaca tgttatgagc
     5401 ttggaatgga tcttcgatcc cgatcgttca aacatttggc aataaagttt cttaagattg
     5461 aatcctgttg ccggtcttgc gacgattatc atataatttc tgttgaatta cgttaagcat
     5521 gtaataatta acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc
     5581 ccgcaattat acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa
     5641 ttatcgcgcg cggtgtcatc tatgttacta gatcgggaat tgccaagcta attcttgaag
     5701 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     5761 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5821 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5881 atgggaccga ctcgcgcttg agacgaagct tgacaggata tattggcggg taaacctaag
     5941 agaaaagagc gtttattaga ataatcggat atttaaaagg gcgtgaaaag gtttatccgt
     6001 tcgtccattt gtatgtgcat gccaaccaca gggttcccct cgggatcaaa gtactttgat
     6061 ccaacccctc cgctgctata gtgcagtcgg cttctgacgt tcagtgcagc cgtcatctga
     6121 aaacgacatg tcgcacaagt cctaagttac gcgacaggct gccgccctgc ccttttcctg
     6181 gcgttttctt gtcgcgtgtt ttagtcgcat aaagtagaat acttgcgact agaaccggag
     6241 acattacgcc atgaacaaga gcgccgccgc tggcctgctg ggctatgccc gcgtcagcac
     6301 cgacgaccag gacttgacca accaacgggc cgaactgcac gcggccggct gcaccaagct
     6361 gttttccgag aagatcaccg gcaccaggcg cgaccgcccg gagctggcca ggatgcttga
     6421 ccacctacgc cctggcgacg ttgtgacagt gaccaggcta gaccgcctgg cccgcagcac
     6481 ccgcgaccta ctggacattg ccgagcgcat ccaggaggcc ggcgcgggcc tgcgtagcct
     6541 ggcagagccg tgggccgaca ccaccacgcc ggccggccgc atggtgttga ccgtgttcgc
     6601 cggcattgcc gagttcgagc gttccctaat catcgaccgc acccggagcg ggcgcgaggc
     6661 cgccaaggcc cgaggcgtga agtttggccc ccgccctacc ctcaccccgg cacagatcgc
     6721 gcacgcccgc gagctgatcg accaggaagg ccgcaccgtg aaagaggcgg ctgcactgct
     6781 tggcgtgcat cgctcgaccc tgtaccgcgc acttgagcgc agcgaggaag tgacgcccac
     6841 cgaggccagg cggcgcggtg ccttccgtga ggacgcattg accgaggccg acgccctggc
     6901 ggccgccgag aatgaacgcc aagaggaaca agcatgaaac cgcaccagga cggccaggac
     6961 gaaccgtttt tcattaccga agagatcgag gcggagatga tcgcggccgg gtacgtgttc
     7021 gagccgcccg cgcacctctc aaccgtgcgg ctgcatgaaa tcctggccgg tttgtctgat
     7081 gccaagctgg cggcctggcc ggccagcttg gccgctgaag aaaccgagcg ccgccgtcta
     7141 aaaaggtgat gtgtatttga gtaaaacagc ttgcgtcatg cggtcgctgc gtatatgatg
     7201 cgatgagtaa ataaacaaat acgcaagggg aacgcatgaa ggttatcgct gtacttaacc
     7261 agaaaggcgg gtcaggcaag acgaccatcg caacccatct agcccgcgcc ctgcaactcg
     7321 ccggggccga tgttctgtta gtcgattccg atccccaggg cagtgcccgc gattgggcgg
     7381 ccgtgcggga agatcaaccg ctaaccgttg tcggcatcga ccgcccgacg attgaccgcg
     7441 acgtgaaggc catcggccgg cgcgacttcg tagtgatcga cggagcgccc caggcggcgg
     7501 acttggctgt gtccgcgatc aaggcagccg acttcgtgct gattccggtg cagccaagcc
     7561 cttacgacat atgggccacc gccgacctgg tggagctggt taagcagcgc attgaggtca
     7621 cggatggaag gctacaagcg gcctttgtcg tgtcgcgggc gatcaaaggc acgcgcatcg
     7681 gcggtgaggt tgccgaggcg ctggccgggt acgagctgcc cattcttgag tcccgtatca
     7741 cgcagcgcgt gagctaccca ggcactgccg ccgccggcac aaccgttctt gaatcagaac
     7801 ccgagggcga cgctgcccgc gaggtccagg cgctggccgc tgaaattaaa tcaaaactca
     7861 tttgagttaa tgaggtaaag agaaaatgag caaaagcaca aacacgctaa gtgccggccg
     7921 tccgagcgca cgcagcagca aggctgcaac gttggccagc ctggcagaca cgccagccat
     7981 gaagcgggtc aactttcagt tgccggcgga ggatcacacc aagctgaaga tgtacgcggt
     8041 acgccaaggc aagaccatta ccgagctgct atctgaatac atcgcgcagc taccagagta
     8101 aatgagcaaa tgaataaatg agtagatgaa ttttagcggc taaaggaggc ggcatggaaa
     8161 atcaagaaca accaggcacc gacgccgtgg aatgccccat gtgtggagga acgggcggtt
     8221 ggccaggcgt aagcggctgg gttgcctgcc ggccctgcaa tggcactgga acccccaagc
     8281 ccgaggaatc ggcgtgagcg gtcgcaaacc atccggcccg gtacaaatcg gcgcggcgct
     8341 gggtgatgac ctggtggaga agttgaaggc cgcgcaggcc gcccagcggc aacgcatcga
     8401 ggcagaagca cgccccggtg aatcgtggca agcggccgct gatcgaatcc gcaaagaatc
     8461 ccggcaaccg ccggcagccg gtgcgccgtc gattaggaag ccgcccaagg gcgacgagca
     8521 accagatttt ttcgttccga tgctctatga cgtgggcacc cgcgatagtc gcagcatcat
     8581 ggacgtggcc gttttccgtc tgtcgaagcg tgaccgacga gctggcgagg tgatccgcta
     8641 cgagcttcca gacgggcacg tagaggtttc cgcagggccg gccggcatgg ccagtgtgtg
     8701 ggattacgac ctggtactga tggcggtttc ccatctaacc gaatccatga accgataccg
     8761 ggaagggaag ggagacaagc ccggccgcgt gttccgtcca cacgttgcgg acgtactcaa
     8821 gttctgccgg cgagccgatg gcggaaagca gaaagacgac ctggtagaaa cctgcattcg
     8881 gttaaacacc acgcacgttg ccatgcagcg tacgaagaag gccaagaacg gccgcctggt
     8941 gacggtatcc gagggtgaag ccttgattag ccgctacaag atcgtaaaga gcgaaaccgg
     9001 gcggccggag tacatcgaga tcgagctagc tgattggatg taccgcgaga tcacagaagg
     9061 caagaacccg gacgtgctga cggttcaccc cgattacttt ttgatcgatc ccggcatcgg
     9121 ccgttttctc taccgcctgg cacgccgcgc cgcaggcaag gcagaagcca gatggttgtt
     9181 caagacgatc tacgaacgca gtggcagcgc cggagagttc aagaagttct gtttcaccgt
     9241 gcgcaagctg atcgggtcaa atgacctgcc ggagtacgat ttgaaggagg aggcggggca
     9301 ggctggcccg atcctagtca tgcgctaccg caacctgatc gagggcgaag catccgccgg
     9361 ttcctaatgt acggagcaga tgctagggca aattgcccta gcaggggaaa aaggtcgaaa
     9421 aggactcttt cctgtggata gcacgtacat tgggaaccca aagccgtaca ttgggaaccg
     9481 gaacccgtac attgggaacc caaagccgta cattgggaac cggtcacaca tgtaagtgac
     9541 tgatataaaa gagaaaaaag gcgatttttc cgcctaaaac tctttaaaac ttattaaaac
     9601 tcttaaaacc cgcctggcct gtgcataact gtctggccag cgcacagccg aagagctgca
     9661 aaaagcgcct acccttcggt cgctgcgctc cctacgcccc gccgcttcgc gtcggcctat
     9721 cgcggccgct ggccgctcaa aaatggctgg cctacggcca ggcaatctac cagggcgcgg
     9781 acaagccgcg ccgtcgccac tcgaccgccg gcgcccacat caaggcaccc tgcctcgcgc
     9841 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggtgacg gtcacagctt
     9901 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     9961 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa
    10021 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
    10081 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
    10141 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
    10201 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
    10261 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
    10321 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
    10381 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
    10441 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
    10501 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
    10561 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
    10621 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
    10681 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
    10741 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
    10801 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
    10861 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
    10921 tcagtggaac gaaaactcac gttaagggat tttggtcatg cattctaggt gattagaaaa
    10981 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
    11041 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
    11101 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
    11161 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
    11221 gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac
    11281 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
    11341 cgagtcgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
    11401 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
    11461 atacctggaa tgctgttttc cctgggatcg cagtggtgag taaccatgca tcatcaggag
    11521 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
    11581 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
    11641 gcgcatcggg cttcccatac aatcggtaga ttgtcgcacc tgattgcccg acattatcgc
    11701 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggccttgagc
    11761 aagacgtttc ccgttgaata tggctcataa cagaacttat tatttccttc ctcttttcta
    11821 cagtatttaa agatacccca agaagctaat tataacaaga cgaactccaa ttcactgttc
    11881 cttgcattct aaaaccttaa ataccagaaa acagcttttt caaagttgtt ttcaaagttg
    11941 gcgtataaca tagtatcgac ggagccgatt ttgaaaccgc ggtgatcaca ggcagcaacg
    12001 ctctgtcatc gttacaatca acatgctacc ctccgcgaga tcatccgtgt ttcaaacccg
    12061 gcagcttagt tgccgttctt ccgaatagca tcggtaacat gagcaaagtc tgccgcctta
    12121 caacggctct cccgctgacg ccgtcccgga ctgatgggct gcctgtatcg agtggtgatt
    12181 ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg tggcaggata tattgtggtg
    12241 taaacataac aagcttcgtc tcagtca
//
