LOCUS       sequence-321998-       13263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TU for the constitutive expression of dCas9 fused to VPR 
            (VP64-p65-Rta) activation domains.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13263)
  AUTHORS   Selma S, Bernabe-Orts JM, Vazquez-Vilar M, Diego-Martin B, Ajenjo M,
            Garcia-Carpintero V, Granell A, Orzaez D
  TITLE     Strong gene activation in plants with genome-wide specificity using 
            a new orthogonal CRISPR/Cas9-based programmable transcriptional 
            activator.
  JOURNAL   Plant Biotechnol J. 2019 Sep;17(9):1703-1705. doi: 
            10.1111/pbi.13138. Epub 2019 May 23.
  PUBMED    31034138
REFERENCE   2  (bases 1 to 13263)
  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"; doi: "10.1111/pbi.13138";
            journalName: "Plant Biotechnol J"; date: "2019-09"; volume: "17"; 
            issue: "9"; pages: "1703-1705"
COMMENT     Sequence Label: pDGB3 alpha2 P35S:dCas9:VPR:Tnos (GB1826)
FEATURES             Location/Qualifiers
     source          1..13263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        637..981
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     925..945
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             1058..1078
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1079..1102,1103..1105,1106..3589,3590..3595,3596..3661,
                     3662..3664,3665..5179)
                     /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(1079..1102,1103..1105,1106..3592,3593..3595,
                     3596..5179)
                     /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             1079..5179
                     /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             5198..5218
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5222..5242
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5246..5266
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5267..5416
                     /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"
     CDS             5417..5437
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5441..5461
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5462..5818
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             5843..6412
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     terminator      6439..6691
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(6726..6744)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6928..6952
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             8253..8882
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             9311..10384
                     /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      10450..10644
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(10880..10902)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    10988..11128
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     11043..11062
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(11143..11160)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11314..11902)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11394..11413)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11990..12805)
                     /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(11990..12805)
                     /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     12713..12732
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    13238..13262
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 taacaagctt cgtctcagtc aggagactag agccaagctg atctcctttg ccccggagat
       61 caccatggac gactttctct atctctacga tctaggaaga aagttcgacg gagaaggtga
      121 cgataccatg ttcaccaccg ataatgagaa gattagcctc ttcaatttca gaaagaatgc
      181 tgacccacag atggttagag aggcctacgc ggcaggtctg atcaagacga tctacccgag
      241 taataatctc caggagatca aataccttcc caagaaggtt aaagatgcag tcaaaagatt
      301 caggactaac tgcatcaaga acacagagaa agatatattt ctcaagatca gaagtactat
      361 tccagtatgg acgattcaag gcttgcttca taaaccaagg caagtaatag agattggagt
      421 ctctaagaaa gtagttccta ctgaatcaaa ggccatggag tcaaaaattc agatcgagga
      481 tctaacagaa ctcgccgtga agactggcga acagttcata cagagtcttt tacgactcaa
      541 tgacaagaag aaaatcttcg tcaacatggt ggagcacgac actctcgtct actccaagaa
      601 tatcaaagat acagtctcag aagaccaaag ggctattgag acttttcaac aaagggtaat
      661 atcgggaaac ctcctcggat tccattgccc agctatctgt cacttcatca aaaggacagt
      721 agaaaaggaa ggtggcacct acaaatgcca tcattgcgat aaaggaaagg ctatcgttca
      781 agatgcccct gccgacagtg gtcccaaaga tggaccccca cccacgagga gcatcgtgga
      841 aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgata tctccactga
      901 cgtaagggat gacgcacaat cccactatcc ttcgcaagac ccttcctcta tataaggaag
      961 ttcatttcat ttggagagga ctccggtatt tttacaacaa ttaccacaac aaaacaaaca
     1021 acaaacaaca ttacaattta ctattctagt cgaaatgccc aagaagaaga ggaaggtgga
     1081 caagaagtac tccattgggc tcgctatcgg cacaaacagc gtcggctggg ccgtcattac
     1141 ggacgagtac aaggtgccga gcaaaaaatt caaagttctg ggcaataccg atcgccacag
     1201 cataaagaag aacctcattg gcgccctcct gttcgactcc ggggaaacgg ccgaagccac
     1261 gcggctcaaa agaacagcac ggcgcagata tacccgcaga aagaatcgga tctgctacct
     1321 gcaggagatc tttagtaatg agatggctaa ggtggatgac tctttcttcc ataggctgga
     1381 ggagtccttt ttggtggagg aggataaaaa gcacgagcgc cacccaatct ttggcaatat
     1441 cgtggacgag gtggcgtacc atgaaaagta cccaaccata tatcatctga ggaagaagct
     1501 tgtagacagt actgataagg ctgacttgcg gttgatctat ctcgcgctgg cgcatatgat
     1561 caaatttcgg ggacacttcc tcatcgaggg ggacctgaac ccagacaaca gcgatgtcga
     1621 caaactcttt atccaactgg ttcagactta caatcagctt ttcgaagaga acccgatcaa
     1681 cgcatccgga gttgacgcca aagcaatcct gagcgctagg ctgtccaaat cccggcggct
     1741 cgaaaacctc atcgcacagc tccctgggga gaagaagaac ggcctgtttg gtaatcttat
     1801 cgccctgtca ctcgggctga cccccaactt taaatctaac ttcgacctgg ccgaagatgc
     1861 caagcttcaa ctgagcaaag acacctacga tgatgatctc gacaatctgc tggcccagat
     1921 cggcgaccag tacgcagacc tttttttggc ggcaaagaac ctgtcagacg ccattctgct
     1981 gagtgatatt ctgcgagtga acacggagat caccaaagct ccgctgagcg ctagtatgat
     2041 caagcgctat gatgagcacc accaagactt gactttgctg aaggcccttg tcagacagca
     2101 actgcctgag aagtacaagg aaattttctt cgatcagtct aaaaatggct acgccggata
     2161 cattgacggc ggagcaagcc aggaggaatt ttacaaattt attaagccca tcttggaaaa
     2221 aatggacggc accgaggagc tgctggtaaa gcttaacaga gaagatctgt tgcgcaaaca
     2281 gcgcactttc gacaatggaa gcatccccca ccagattcac ctgggcgaac tgcacgctat
     2341 cctcaggcgg caagaggatt tctacccctt tttgaaagat aacagggaaa agattgagaa
     2401 aatcctcaca tttcggatac cctactatgt aggccccctc gcccggggaa attccagatt
     2461 cgcgtggatg actcgcaaat cagaagagac tatcactccc tggaacttcg aggaagtcgt
     2521 ggataagggg gcctctgccc agtccttcat cgaaaggatg actaactttg ataaaaatct
     2581 gcctaacgaa aaggtgcttc ctaaacactc tctgctgtac gagtacttca cagtttataa
     2641 cgagctcacc aaggtcaaat acgtcacaga agggatgaga aagccagcat tcctgtctgg
     2701 agagcagaag aaagctatcg tggacctcct cttcaagacg aaccggaaag ttaccgtgaa
     2761 acagctcaaa gaagattatt tcaaaaagat tgaatgtttc gactctgttg aaatcagcgg
     2821 agtggaggat cgcttcaacg catccctggg aacgtatcac gatctcctga aaatcattaa
     2881 agacaaggac ttcctggaca atgaggagaa cgaggacatt cttgaggaca ttgtcctcac
     2941 ccttacgttg tttgaagata gggagatgat tgaagaacgc ttgaaaactt acgctcatct
     3001 cttcgacgac aaagtcatga aacagctcaa gaggcgccga tatacaggat gggggcggct
     3061 gtcaagaaaa ctgatcaatg ggatccgaga caagcagagt ggaaagacaa tcctggattt
     3121 tcttaagtcc gatggatttg ccaaccggaa cttcatgcag ttgatccatg atgactctct
     3181 cacctttaag gaggacatcc agaaagcaca agtttctggc cagggggaca gtctccacga
     3241 gcacatcgct aatcttgcag gtagcccagc tatcaaaaag ggaatactgc agaccgttaa
     3301 ggtcgtggat gaactcgtca aagtaatggg aaggcataag cccgagaata tcgttatcga
     3361 gatggcccga gagaaccaaa ctacccagaa gggacagaag aacagtaggg aaaggatgaa
     3421 gaggattgaa gagggtataa aagaactggg gtcccaaatc cttaaggaac acccagttga
     3481 aaacacccag cttcagaatg agaagctcta cctgtactac ctgcagaacg gcagggacat
     3541 gtacgtggat caggaactgg acatcaatcg gctctccgac tacgacgtgg atgccatcgt
     3601 gccccagtct tttctcaaag atgattctat tgataataaa gtgttgacaa gatccgataa
     3661 aaatagaggg aagagtgata acgtcccctc agaagaagtt gtcaagaaaa tgaaaaatta
     3721 ttggcggcag ctgctgaacg ccaaactgat cacacaacgg aagttcgata atctgactaa
     3781 ggctgaacga ggtggcctgt ctgagttgga taaagccggc ttcatcaaaa ggcagcttgt
     3841 tgagacacgc cagatcacca agcacgtggc ccaaattctc gattcacgca tgaacaccaa
     3901 gtacgatgaa aatgacaaac tgattcgaga ggtgaaagtt attactctga agtctaagct
     3961 ggtttcagat ttcagaaagg actttcagtt ttataaggtg agagagatca acaattacca
     4021 ccatgcgcat gatgcctacc tgaatgcagt ggtaggcact gcacttatca aaaaatatcc
     4081 caagcttgaa tctgaatttg tttacggaga ctataaagtg tacgatgtta ggaaaatgat
     4141 cgcaaagtct gagcaggaaa taggcaaggc caccgctaag tacttctttt acagcaatat
     4201 tatgaatttt ttcaagaccg agattacact ggccaatgga gagattcgga agcgaccact
     4261 tatcgaaaca aacggagaaa caggagaaat cgtgtgggac aagggtaggg atttcgcgac
     4321 agtccggaag gtcctgtcca tgccgcaggt gaacatcgtt aaaaagaccg aagtacagac
     4381 cggaggcttc tccaaggaaa gtatcctccc gaaaaggaac agcgacaagc tgatcgcacg
     4441 caaaaaagat tgggacccca agaaatacgg cggattcgat tctcctacag tcgcttacag
     4501 tgtactggtt gtggccaaag tggagaaagg gaagtctaaa aaactcaaaa gcgtcaagga
     4561 actgctgggc atcacaatca tggagcgatc aagcttcgaa aaaaacccca tcgactttct
     4621 cgaggcgaaa ggatataaag aggtcaaaaa agacctcatc attaagcttc ccaagtactc
     4681 tctctttgag cttgaaaacg gccggaaacg aatgctcgct agtgcgggcg agctgcagaa
     4741 aggtaacgag ctggcactgc cctctaaata cgttaatttc ttgtatctgg ccagccacta
     4801 tgaaaagctc aaaggatctc ccgaagataa tgagcagaag cagctgttcg tggaacaaca
     4861 caaacactac cttgatgaga tcatcgagca aataagcgaa ttctccaaaa gagtgatcct
     4921 cgccgacgct aacctcgata aggtgctttc tgcttacaat aagcacaggg ataagcccat
     4981 cagggagcag gcagaaaaca ttatccactt gtttactctg accaacttgg gcgcgcctgc
     5041 agccttcaag tacttcgaca ccaccataga cagaaagcgg tacacctcta caaaggaggt
     5101 cctggacgcc acactgattc atcagtcaat tacggggctc tatgaaacaa gaatcgacct
     5161 ctctcagctc ggtggagaca gcagggctga ttcggaccca aaaaagaagc gtaaggtcga
     5221 tcccaagaag aagagaaagg tagatcctaa gaagaagaga aaggtagacg cattggatga
     5281 cttcgactta gacatgcttg ggtcagatgc tttggacgac tttgacctcg acatgttggg
     5341 ttctgacgcg ctagatgact tcgacttgga catgttgggc tccgacgctt tggacgactt
     5401 tgatctggac atgttaccga aaaagaaacg caaagttgat cctaagaaga agagaaaggt
     5461 acctacacag gccggcgagg gcacactgtc tgaagctctg ctgcagctgc agttcgacga
     5521 cgaggatctg ggagccctgc tgggaaacag caccgatcct gccgtgttca ccgacctggc
     5581 cagcgtggac aacagcgagt tccagcagct gctgaaccag ggcatccctg tggcccctca
     5641 caccaccgag cccatgctga tggaataccc cgaggccatc acccggctcg tgacaggcgc
     5701 tcagaggcct cctgatccag ctcctgcccc tctgggagca ccaggcctgc ctaatggact
     5761 gctgtctggc gacgaggact tcagctctat cgccgatatg gatttctcag ccttgctgat
     5821 taactctaga agttccggat ctcgggattc cagggaaggg atgtttttgc cgaagcctga
     5881 ggccggctcc gctattagtg acgtgtttga gggccgcgag gtgtgccagc caaaacgaat
     5941 ccggccattt catcctccag gaagtccatg ggccaaccgc ccactccccg ccagcctcgc
     6001 accaacacca accggtccag tacatgagcc agtcgggtca ctgaccccgg caccagtccc
     6061 tcagccactg gatccagcgc ccgcagtgac tcccgaggcc agtcacctgt tggaggatcc
     6121 cgatgaagaa acgagccagg ctgtcaaagc ccttcgggag atggccgata ctgtgattcc
     6181 ccagaaggaa gaggctgcaa tctgtggcca aatggacctt tcccatccgc ccccaagggg
     6241 ccatctggat gagctgacaa ccacacttga gtccatgacc gaggatctga acctggactc
     6301 acccctgacc ccggaattga acgagattct ggataccttc ctgaacgacg agtgcctctt
     6361 gcatgccatg catatcagca caggactgtc catcttcgac acatctctgt tttgagcttg
     6421 gaatggatct tcgatcccga tcgttcaaac atttggcaat aaagtttctt aagattgaat
     6481 cctgttgccg gtcttgcgac gattatcata taatttctgt tgaattacgt taagcatgta
     6541 ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg
     6601 caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta ggataaatta
     6661 tcgcgcgcgg tgtcatctat gttactagat cgggaattgc caagctaatt cttgaagacg
     6721 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     6781 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     6841 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataatg
     6901 ggaccgactc gcgcttgaga cgaagcttga caggatatat tggcgggtaa acctaagaga
     6961 aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt tatccgttcg
     7021 tccatttgta tgtgcatgcc aaccacaggg ttcccctcgg gatcaaagta ctttgatcca
     7081 acccctccgc tgctatagtg cagtcggctt ctgacgttca gtgcagccgt catctgaaaa
     7141 cgacatgtcg cacaagtcct aagttacgcg acaggctgcc gccctgccct tttcctggcg
     7201 ttttcttgtc gcgtgtttta gtcgcataaa gtagaatact tgcgactaga accggagaca
     7261 ttacgccatg aacaagagcg ccgccgctgg cctgctgggc tatgcccgcg tcagcaccga
     7321 cgaccaggac ttgaccaacc aacgggccga actgcacgcg gccggctgca ccaagctgtt
     7381 ttccgagaag atcaccggca ccaggcgcga ccgcccggag ctggccagga tgcttgacca
     7441 cctacgccct ggcgacgttg tgacagtgac caggctagac cgcctggccc gcagcacccg
     7501 cgacctactg gacattgccg agcgcatcca ggaggccggc gcgggcctgc gtagcctggc
     7561 agagccgtgg gccgacacca ccacgccggc cggccgcatg gtgttgaccg tgttcgccgg
     7621 cattgccgag ttcgagcgtt ccctaatcat cgaccgcacc cggagcgggc gcgaggccgc
     7681 caaggcccga ggcgtgaagt ttggcccccg ccctaccctc accccggcac agatcgcgca
     7741 cgcccgcgag ctgatcgacc aggaaggccg caccgtgaaa gaggcggctg cactgcttgg
     7801 cgtgcatcgc tcgaccctgt accgcgcact tgagcgcagc gaggaagtga cgcccaccga
     7861 ggccaggcgg cgcggtgcct tccgtgagga cgcattgacc gaggccgacg ccctggcggc
     7921 cgccgagaat gaacgccaag aggaacaagc atgaaaccgc accaggacgg ccaggacgaa
     7981 ccgtttttca ttaccgaaga gatcgaggcg gagatgatcg cggccgggta cgtgttcgag
     8041 ccgcccgcgc acctctcaac cgtgcggctg catgaaatcc tggccggttt gtctgatgcc
     8101 aagctggcgg cctggccggc cagcttggcc gctgaagaaa ccgagcgccg ccgtctaaaa
     8161 aggtgatgtg tatttgagta aaacagcttg cgtcatgcgg tcgctgcgta tatgatgcga
     8221 tgagtaaata aacaaatacg caaggggaac gcatgaaggt tatcgctgta cttaaccaga
     8281 aaggcgggtc aggcaagacg accatcgcaa cccatctagc ccgcgccctg caactcgccg
     8341 gggccgatgt tctgttagtc gattccgatc cccagggcag tgcccgcgat tgggcggccg
     8401 tgcgggaaga tcaaccgcta accgttgtcg gcatcgaccg cccgacgatt gaccgcgacg
     8461 tgaaggccat cggccggcgc gacttcgtag tgatcgacgg agcgccccag gcggcggact
     8521 tggctgtgtc cgcgatcaag gcagccgact tcgtgctgat tccggtgcag ccaagccctt
     8581 acgacatatg ggccaccgcc gacctggtgg agctggttaa gcagcgcatt gaggtcacgg
     8641 atggaaggct acaagcggcc tttgtcgtgt cgcgggcgat caaaggcacg cgcatcggcg
     8701 gtgaggttgc cgaggcgctg gccgggtacg agctgcccat tcttgagtcc cgtatcacgc
     8761 agcgcgtgag ctacccaggc actgccgccg ccggcacaac cgttcttgaa tcagaacccg
     8821 agggcgacgc tgcccgcgag gtccaggcgc tggccgctga aattaaatca aaactcattt
     8881 gagttaatga ggtaaagaga aaatgagcaa aagcacaaac acgctaagtg ccggccgtcc
     8941 gagcgcacgc agcagcaagg ctgcaacgtt ggccagcctg gcagacacgc cagccatgaa
     9001 gcgggtcaac tttcagttgc cggcggagga tcacaccaag ctgaagatgt acgcggtacg
     9061 ccaaggcaag accattaccg agctgctatc tgaatacatc gcgcagctac cagagtaaat
     9121 gagcaaatga ataaatgagt agatgaattt tagcggctaa aggaggcggc atggaaaatc
     9181 aagaacaacc aggcaccgac gccgtggaat gccccatgtg tggaggaacg ggcggttggc
     9241 caggcgtaag cggctgggtt gcctgccggc cctgcaatgg cactggaacc cccaagcccg
     9301 aggaatcggc gtgagcggtc gcaaaccatc cggcccggta caaatcggcg cggcgctggg
     9361 tgatgacctg gtggagaagt tgaaggccgc gcaggccgcc cagcggcaac gcatcgaggc
     9421 agaagcacgc cccggtgaat cgtggcaagc ggccgctgat cgaatccgca aagaatcccg
     9481 gcaaccgccg gcagccggtg cgccgtcgat taggaagccg cccaagggcg acgagcaacc
     9541 agattttttc gttccgatgc tctatgacgt gggcacccgc gatagtcgca gcatcatgga
     9601 cgtggccgtt ttccgtctgt cgaagcgtga ccgacgagct ggcgaggtga tccgctacga
     9661 gcttccagac gggcacgtag aggtttccgc agggccggcc ggcatggcca gtgtgtggga
     9721 ttacgacctg gtactgatgg cggtttccca tctaaccgaa tccatgaacc gataccggga
     9781 agggaaggga gacaagcccg gccgcgtgtt ccgtccacac gttgcggacg tactcaagtt
     9841 ctgccggcga gccgatggcg gaaagcagaa agacgacctg gtagaaacct gcattcggtt
     9901 aaacaccacg cacgttgcca tgcagcgtac gaagaaggcc aagaacggcc gcctggtgac
     9961 ggtatccgag ggtgaagcct tgattagccg ctacaagatc gtaaagagcg aaaccgggcg
    10021 gccggagtac atcgagatcg agctagctga ttggatgtac cgcgagatca cagaaggcaa
    10081 gaacccggac gtgctgacgg ttcaccccga ttactttttg atcgatcccg gcatcggccg
    10141 ttttctctac cgcctggcac gccgcgccgc aggcaaggca gaagccagat ggttgttcaa
    10201 gacgatctac gaacgcagtg gcagcgccgg agagttcaag aagttctgtt tcaccgtgcg
    10261 caagctgatc gggtcaaatg acctgccgga gtacgatttg aaggaggagg cggggcaggc
    10321 tggcccgatc ctagtcatgc gctaccgcaa cctgatcgag ggcgaagcat ccgccggttc
    10381 ctaatgtacg gagcagatgc tagggcaaat tgccctagca ggggaaaaag gtcgaaaagg
    10441 actctttcct gtggatagca cgtacattgg gaacccaaag ccgtacattg ggaaccggaa
    10501 cccgtacatt gggaacccaa agccgtacat tgggaaccgg tcacacatgt aagtgactga
    10561 tataaaagag aaaaaaggcg atttttccgc ctaaaactct ttaaaactta ttaaaactct
    10621 taaaacccgc ctggcctgtg cataactgtc tggccagcgc acagccgaag agctgcaaaa
    10681 agcgcctacc cttcggtcgc tgcgctccct acgccccgcc gcttcgcgtc ggcctatcgc
    10741 ggccgctggc cgctcaaaaa tggctggcct acggccaggc aatctaccag ggcgcggaca
    10801 agccgcgccg tcgccactcg accgccggcg cccacatcaa ggcaccctgc ctcgcgcgtt
    10861 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggtgacggtc acagcttgtc
    10921 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
    10981 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
    11041 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
    11101 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
    11161 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
    11221 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
    11281 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
    11341 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
    11401 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
    11461 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
    11521 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
    11581 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
    11641 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
    11701 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
    11761 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
    11821 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
    11881 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
    11941 gtggaacgaa aactcacgtt aagggatttt ggtcatgcat tctaggtgat tagaaaaact
    12001 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
    12061 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
    12121 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
    12181 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
    12241 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
    12301 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
    12361 gtcgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
    12421 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
    12481 cctggaatgc tgttttccct gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
    12541 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
    12601 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
    12661 catcgggctt cccatacaat cggtagattg tcgcacctga ttgcccgaca ttatcgcgag
    12721 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc cttgagcaag
    12781 acgtttcccg ttgaatatgg ctcataacag aacttattat ttccttcctc ttttctacag
    12841 tatttaaaga taccccaaga agctaattat aacaagacga actccaattc actgttcctt
    12901 gcattctaaa accttaaata ccagaaaaca gctttttcaa agttgttttc aaagttggcg
    12961 tataacatag tatcgacgga gccgattttg aaaccgcggt gatcacaggc agcaacgctc
    13021 tgtcatcgtt acaatcaaca tgctaccctc cgcgagatca tccgtgtttc aaacccggca
    13081 gcttagttgc cgttcttccg aatagcatcg gtaacatgag caaagtctgc cgccttacaa
    13141 cggctctccc gctgacgccg tcccggactg atgggctgcc tgtatcgagt ggtgattttg
    13201 tgccgagctg ccggtcgggg agctgttggc tggctggtgg caggatatat tgtggtgtaa
    13261 aca
//
