LOCUS       sequence-177256-       12111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcriptional unit of (human codon optimized) inactivated Cas9 
            fused to the BRD Transcriptional Repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12111)
  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 12111)
  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:BRD:tNos (GB1172)
FEATURES             Location/Qualifiers
     source          1..12111
                     /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             5174..5194
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5266..5518
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(5553..5571)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5755..5779
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             7080..7709
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             8138..9211
                     /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      9277..9471
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(9707..9729)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    9815..9955
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     9870..9889
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(9970..9987)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10141..10729)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10221..10240)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(10817..11632)
                     /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(10817..11632)
                     /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     11540..11559
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    12065..12089
                     /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 tcgcccaaga agaagaggaa ggtgggtaat tctaagactc ttagactttt
     5221 tggtgttaat atggaatgtt aggcttggaa tggatcttcg atcccgatcg ttcaaacatt
     5281 tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgacgat tatcatataa
     5341 tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg
     5401 agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa
     5461 atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt actagatcgg
     5521 gaattgccaa gctaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     5581 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     5641 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5701 ataaccctga taaatgcttc aataatggga ccgactcgcg cttgagacga agcttgacag
     5761 gatatattgg cgggtaaacc taagagaaaa gagcgtttat tagaataatc ggatatttaa
     5821 aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt gcatgccaac cacagggttc
     5881 ccctcgggat caaagtactt tgatccaacc cctccgctgc tatagtgcag tcggcttctg
     5941 acgttcagtg cagccgtcat ctgaaaacga catgtcgcac aagtcctaag ttacgcgaca
     6001 ggctgccgcc ctgccctttt cctggcgttt tcttgtcgcg tgttttagtc gcataaagta
     6061 gaatacttgc gactagaacc ggagacatta cgccatgaac aagagcgccg ccgctggcct
     6121 gctgggctat gcccgcgtca gcaccgacga ccaggacttg accaaccaac gggccgaact
     6181 gcacgcggcc ggctgcacca agctgttttc cgagaagatc accggcacca ggcgcgaccg
     6241 cccggagctg gccaggatgc ttgaccacct acgccctggc gacgttgtga cagtgaccag
     6301 gctagaccgc ctggcccgca gcacccgcga cctactggac attgccgagc gcatccagga
     6361 ggccggcgcg ggcctgcgta gcctggcaga gccgtgggcc gacaccacca cgccggccgg
     6421 ccgcatggtg ttgaccgtgt tcgccggcat tgccgagttc gagcgttccc taatcatcga
     6481 ccgcacccgg agcgggcgcg aggccgccaa ggcccgaggc gtgaagtttg gcccccgccc
     6541 taccctcacc ccggcacaga tcgcgcacgc ccgcgagctg atcgaccagg aaggccgcac
     6601 cgtgaaagag gcggctgcac tgcttggcgt gcatcgctcg accctgtacc gcgcacttga
     6661 gcgcagcgag gaagtgacgc ccaccgaggc caggcggcgc ggtgccttcc gtgaggacgc
     6721 attgaccgag gccgacgccc tggcggccgc cgagaatgaa cgccaagagg aacaagcatg
     6781 aaaccgcacc aggacggcca ggacgaaccg tttttcatta ccgaagagat cgaggcggag
     6841 atgatcgcgg ccgggtacgt gttcgagccg cccgcgcacc tctcaaccgt gcggctgcat
     6901 gaaatcctgg ccggtttgtc tgatgccaag ctggcggcct ggccggccag cttggccgct
     6961 gaagaaaccg agcgccgccg tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt
     7021 catgcggtcg ctgcgtatat gatgcgatga gtaaataaac aaatacgcaa ggggaacgca
     7081 tgaaggttat cgctgtactt aaccagaaag gcgggtcagg caagacgacc atcgcaaccc
     7141 atctagcccg cgccctgcaa ctcgccgggg ccgatgttct gttagtcgat tccgatcccc
     7201 agggcagtgc ccgcgattgg gcggccgtgc gggaagatca accgctaacc gttgtcggca
     7261 tcgaccgccc gacgattgac cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga
     7321 tcgacggagc gccccaggcg gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg
     7381 tgctgattcc ggtgcagcca agcccttacg acatatgggc caccgccgac ctggtggagc
     7441 tggttaagca gcgcattgag gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc
     7501 gggcgatcaa aggcacgcgc atcggcggtg aggttgccga ggcgctggcc gggtacgagc
     7561 tgcccattct tgagtcccgt atcacgcagc gcgtgagcta cccaggcact gccgccgccg
     7621 gcacaaccgt tcttgaatca gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg
     7681 ccgctgaaat taaatcaaaa ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag
     7741 cacaaacacg ctaagtgccg gccgtccgag cgcacgcagc agcaaggctg caacgttggc
     7801 cagcctggca gacacgccag ccatgaagcg ggtcaacttt cagttgccgg cggaggatca
     7861 caccaagctg aagatgtacg cggtacgcca aggcaagacc attaccgagc tgctatctga
     7921 atacatcgcg cagctaccag agtaaatgag caaatgaata aatgagtaga tgaattttag
     7981 cggctaaagg aggcggcatg gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc
     8041 ccatgtgtgg aggaacgggc ggttggccag gcgtaagcgg ctgggttgcc tgccggccct
     8101 gcaatggcac tggaaccccc aagcccgagg aatcggcgtg agcggtcgca aaccatccgg
     8161 cccggtacaa atcggcgcgg cgctgggtga tgacctggtg gagaagttga aggccgcgca
     8221 ggccgcccag cggcaacgca tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc
     8281 cgctgatcga atccgcaaag aatcccggca accgccggca gccggtgcgc cgtcgattag
     8341 gaagccgccc aagggcgacg agcaaccaga ttttttcgtt ccgatgctct atgacgtggg
     8401 cacccgcgat agtcgcagca tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg
     8461 acgagctggc gaggtgatcc gctacgagct tccagacggg cacgtagagg tttccgcagg
     8521 gccggccggc atggccagtg tgtgggatta cgacctggta ctgatggcgg tttcccatct
     8581 aaccgaatcc atgaaccgat accgggaagg gaagggagac aagcccggcc gcgtgttccg
     8641 tccacacgtt gcggacgtac tcaagttctg ccggcgagcc gatggcggaa agcagaaaga
     8701 cgacctggta gaaacctgca ttcggttaaa caccacgcac gttgccatgc agcgtacgaa
     8761 gaaggccaag aacggccgcc tggtgacggt atccgagggt gaagccttga ttagccgcta
     8821 caagatcgta aagagcgaaa ccgggcggcc ggagtacatc gagatcgagc tagctgattg
     8881 gatgtaccgc gagatcacag aaggcaagaa cccggacgtg ctgacggttc accccgatta
     8941 ctttttgatc gatcccggca tcggccgttt tctctaccgc ctggcacgcc gcgccgcagg
     9001 caaggcagaa gccagatggt tgttcaagac gatctacgaa cgcagtggca gcgccggaga
     9061 gttcaagaag ttctgtttca ccgtgcgcaa gctgatcggg tcaaatgacc tgccggagta
     9121 cgatttgaag gaggaggcgg ggcaggctgg cccgatccta gtcatgcgct accgcaacct
     9181 gatcgagggc gaagcatccg ccggttccta atgtacggag cagatgctag ggcaaattgc
     9241 cctagcaggg gaaaaaggtc gaaaaggact ctttcctgtg gatagcacgt acattgggaa
     9301 cccaaagccg tacattggga accggaaccc gtacattggg aacccaaagc cgtacattgg
     9361 gaaccggtca cacatgtaag tgactgatat aaaagagaaa aaaggcgatt tttccgccta
     9421 aaactcttta aaacttatta aaactcttaa aacccgcctg gcctgtgcat aactgtctgg
     9481 ccagcgcaca gccgaagagc tgcaaaaagc gcctaccctt cggtcgctgc gctccctacg
     9541 ccccgccgct tcgcgtcggc ctatcgcggc cgctggccgc tcaaaaatgg ctggcctacg
     9601 gccaggcaat ctaccagggc gcggacaagc cgcgccgtcg ccactcgacc gccggcgccc
     9661 acatcaaggc accctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     9721 agctcccggt gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     9781 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
     9841 atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     9901 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc
     9961 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
    10021 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
    10081 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
    10141 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
    10201 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
    10261 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
    10321 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
    10381 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
    10441 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
    10501 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
    10561 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
    10621 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
    10681 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
    10741 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
    10801 catgcattct aggtgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca
    10861 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
    10921 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
    10981 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
    11041 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
    11101 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
    11161 tattcattcg tgattgcgcc tgagcgagtc gaaatacgcg atcgctgtta aaaggacaat
    11221 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
    11281 cacctgaatc aggatattct tctaatacct ggaatgctgt tttccctggg atcgcagtgg
    11341 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
    11401 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
    11461 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcgg tagattgtcg
    11521 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
    11581 tggaatttaa tcgcggcctt gagcaagacg tttcccgttg aatatggctc ataacagaac
    11641 ttattatttc cttcctcttt tctacagtat ttaaagatac cccaagaagc taattataac
    11701 aagacgaact ccaattcact gttccttgca ttctaaaacc ttaaatacca gaaaacagct
    11761 ttttcaaagt tgttttcaaa gttggcgtat aacatagtat cgacggagcc gattttgaaa
    11821 ccgcggtgat cacaggcagc aacgctctgt catcgttaca atcaacatgc taccctccgc
    11881 gagatcatcc gtgtttcaaa cccggcagct tagttgccgt tcttccgaat agcatcggta
    11941 acatgagcaa agtctgccgc cttacaacgg ctctcccgct gacgccgtcc cggactgatg
    12001 ggctgcctgt atcgagtggt gattttgtgc cgagctgccg gtcggggagc tgttggctgg
    12061 ctggtggcag gatatattgt ggtgtaaaca taacaagctt cgtctcagtc a
//
