LOCUS       sequence-244379-       12189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcriptional unit of (human codon optimized) inactivated Cas9 
            fused to the EDLL Transcriptional Activator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12189)
  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 12189)
  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:EDLL:tNos (GB1190)
FEATURES             Location/Qualifiers
     source          1..12189
                     /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"
     terminator      5344..5596
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(5631..5649)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5833..5857
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             7158..7787
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             8216..9289
                     /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      9355..9549
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(9785..9807)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    9893..10033
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     9948..9967
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(10048..10065)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10219..10807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10299..10318)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(10895..11710)
                     /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(10895..11710)
                     /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     11618..11637
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    12143..12167
                     /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 tcggatccta aaaagaagcg aaaggtagac ccgaagaaaa agaggaaggt
     5221 agatccaaag aagaagcgca aggtagaagt gtttgaattt gaatatctgg atgataaagt
     5281 gctggaagaa ctgctggata gcgaagaacg caaacgctga gcttggaatg gatcttcgat
     5341 cccgatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt tgccggtctt
     5401 gcgacgatta tcatataatt tctgttgaat tacgttaagc atgtaataat taacatgtaa
     5461 tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt atacatttaa
     5521 tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg cgcggtgtca
     5581 tctatgttac tagatcggga attgccaagc taattcttga agacgaaagg gcctcgtgat
     5641 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     5701 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5761 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatgggacc gactcgcgct
     5821 tgagacgaag cttgacagga tatattggcg ggtaaaccta agagaaaaga gcgtttatta
     5881 gaataatcgg atatttaaaa gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc
     5941 atgccaacca cagggttccc ctcgggatca aagtactttg atccaacccc tccgctgcta
     6001 tagtgcagtc ggcttctgac gttcagtgca gccgtcatct gaaaacgaca tgtcgcacaa
     6061 gtcctaagtt acgcgacagg ctgccgccct gcccttttcc tggcgttttc ttgtcgcgtg
     6121 ttttagtcgc ataaagtaga atacttgcga ctagaaccgg agacattacg ccatgaacaa
     6181 gagcgccgcc gctggcctgc tgggctatgc ccgcgtcagc accgacgacc aggacttgac
     6241 caaccaacgg gccgaactgc acgcggccgg ctgcaccaag ctgttttccg agaagatcac
     6301 cggcaccagg cgcgaccgcc cggagctggc caggatgctt gaccacctac gccctggcga
     6361 cgttgtgaca gtgaccaggc tagaccgcct ggcccgcagc acccgcgacc tactggacat
     6421 tgccgagcgc atccaggagg ccggcgcggg cctgcgtagc ctggcagagc cgtgggccga
     6481 caccaccacg ccggccggcc gcatggtgtt gaccgtgttc gccggcattg ccgagttcga
     6541 gcgttcccta atcatcgacc gcacccggag cgggcgcgag gccgccaagg cccgaggcgt
     6601 gaagtttggc ccccgcccta ccctcacccc ggcacagatc gcgcacgccc gcgagctgat
     6661 cgaccaggaa ggccgcaccg tgaaagaggc ggctgcactg cttggcgtgc atcgctcgac
     6721 cctgtaccgc gcacttgagc gcagcgagga agtgacgccc accgaggcca ggcggcgcgg
     6781 tgccttccgt gaggacgcat tgaccgaggc cgacgccctg gcggccgccg agaatgaacg
     6841 ccaagaggaa caagcatgaa accgcaccag gacggccagg acgaaccgtt tttcattacc
     6901 gaagagatcg aggcggagat gatcgcggcc gggtacgtgt tcgagccgcc cgcgcacctc
     6961 tcaaccgtgc ggctgcatga aatcctggcc ggtttgtctg atgccaagct ggcggcctgg
     7021 ccggccagct tggccgctga agaaaccgag cgccgccgtc taaaaaggtg atgtgtattt
     7081 gagtaaaaca gcttgcgtca tgcggtcgct gcgtatatga tgcgatgagt aaataaacaa
     7141 atacgcaagg ggaacgcatg aaggttatcg ctgtacttaa ccagaaaggc gggtcaggca
     7201 agacgaccat cgcaacccat ctagcccgcg ccctgcaact cgccggggcc gatgttctgt
     7261 tagtcgattc cgatccccag ggcagtgccc gcgattgggc ggccgtgcgg gaagatcaac
     7321 cgctaaccgt tgtcggcatc gaccgcccga cgattgaccg cgacgtgaag gccatcggcc
     7381 ggcgcgactt cgtagtgatc gacggagcgc cccaggcggc ggacttggct gtgtccgcga
     7441 tcaaggcagc cgacttcgtg ctgattccgg tgcagccaag cccttacgac atatgggcca
     7501 ccgccgacct ggtggagctg gttaagcagc gcattgaggt cacggatgga aggctacaag
     7561 cggcctttgt cgtgtcgcgg gcgatcaaag gcacgcgcat cggcggtgag gttgccgagg
     7621 cgctggccgg gtacgagctg cccattcttg agtcccgtat cacgcagcgc gtgagctacc
     7681 caggcactgc cgccgccggc acaaccgttc ttgaatcaga acccgagggc gacgctgccc
     7741 gcgaggtcca ggcgctggcc gctgaaatta aatcaaaact catttgagtt aatgaggtaa
     7801 agagaaaatg agcaaaagca caaacacgct aagtgccggc cgtccgagcg cacgcagcag
     7861 caaggctgca acgttggcca gcctggcaga cacgccagcc atgaagcggg tcaactttca
     7921 gttgccggcg gaggatcaca ccaagctgaa gatgtacgcg gtacgccaag gcaagaccat
     7981 taccgagctg ctatctgaat acatcgcgca gctaccagag taaatgagca aatgaataaa
     8041 tgagtagatg aattttagcg gctaaaggag gcggcatgga aaatcaagaa caaccaggca
     8101 ccgacgccgt ggaatgcccc atgtgtggag gaacgggcgg ttggccaggc gtaagcggct
     8161 gggttgcctg ccggccctgc aatggcactg gaacccccaa gcccgaggaa tcggcgtgag
     8221 cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg ctgggtgatg acctggtgga
     8281 gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg
     8341 tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc
     8401 cggtgcgccg tcgattagga agccgcccaa gggcgacgag caaccagatt ttttcgttcc
     8461 gatgctctat gacgtgggca cccgcgatag tcgcagcatc atggacgtgg ccgttttccg
     8521 tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc tacgagcttc cagacgggca
     8581 cgtagaggtt tccgcagggc cggccggcat ggccagtgtg tgggattacg acctggtact
     8641 gatggcggtt tcccatctaa ccgaatccat gaaccgatac cgggaaggga agggagacaa
     8701 gcccggccgc gtgttccgtc cacacgttgc ggacgtactc aagttctgcc ggcgagccga
     8761 tggcggaaag cagaaagacg acctggtaga aacctgcatt cggttaaaca ccacgcacgt
     8821 tgccatgcag cgtacgaaga aggccaagaa cggccgcctg gtgacggtat ccgagggtga
     8881 agccttgatt agccgctaca agatcgtaaa gagcgaaacc gggcggccgg agtacatcga
     8941 gatcgagcta gctgattgga tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct
     9001 gacggttcac cccgattact ttttgatcga tcccggcatc ggccgttttc tctaccgcct
     9061 ggcacgccgc gccgcaggca aggcagaagc cagatggttg ttcaagacga tctacgaacg
     9121 cagtggcagc gccggagagt tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc
     9181 aaatgacctg ccggagtacg atttgaagga ggaggcgggg caggctggcc cgatcctagt
     9241 catgcgctac cgcaacctga tcgagggcga agcatccgcc ggttcctaat gtacggagca
     9301 gatgctaggg caaattgccc tagcagggga aaaaggtcga aaaggactct ttcctgtgga
     9361 tagcacgtac attgggaacc caaagccgta cattgggaac cggaacccgt acattgggaa
     9421 cccaaagccg tacattggga accggtcaca catgtaagtg actgatataa aagagaaaaa
     9481 aggcgatttt tccgcctaaa actctttaaa acttattaaa actcttaaaa cccgcctggc
     9541 ctgtgcataa ctgtctggcc agcgcacagc cgaagagctg caaaaagcgc ctacccttcg
     9601 gtcgctgcgc tccctacgcc ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc
     9661 aaaaatggct ggcctacggc caggcaatct accagggcgc ggacaagccg cgccgtcgcc
     9721 actcgaccgc cggcgcccac atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg
     9781 aaaacctctg acacatgcag ctcccggtga cggtcacagc ttgtctgtaa gcggatgccg
     9841 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
     9901 tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca
     9961 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
    10021 ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
    10081 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
    10141 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
    10201 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
    10261 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
    10321 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
    10381 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
    10441 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
    10501 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
    10561 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
    10621 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
    10681 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
    10741 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
    10801 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
    10861 acgttaaggg attttggtca tgcattctag gtgattagaa aaactcatcg agcatcaaat
    10921 gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa agccgtttct
    10981 gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt
    11041 ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa
    11101 ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt
    11161 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac
    11221 tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagtcga aatacgcgat
    11281 cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca
    11341 gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt
    11401 tccctgggat cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga
    11461 tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat
    11521 cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat
    11581 acaatcggta gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat
    11641 ataaatcagc atccatgttg gaatttaatc gcggccttga gcaagacgtt tcccgttgaa
    11701 tatggctcat aacagaactt attatttcct tcctcttttc tacagtattt aaagataccc
    11761 caagaagcta attataacaa gacgaactcc aattcactgt tccttgcatt ctaaaacctt
    11821 aaataccaga aaacagcttt ttcaaagttg ttttcaaagt tggcgtataa catagtatcg
    11881 acggagccga ttttgaaacc gcggtgatca caggcagcaa cgctctgtca tcgttacaat
    11941 caacatgcta ccctccgcga gatcatccgt gtttcaaacc cggcagctta gttgccgttc
    12001 ttccgaatag catcggtaac atgagcaaag tctgccgcct tacaacggct ctcccgctga
    12061 cgccgtcccg gactgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt
    12121 cggggagctg ttggctggct ggtggcagga tatattgtgg tgtaaacata acaagcttcg
    12181 tctcagtca
//
