LOCUS       sequence-146044-       12511 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Transcriptional unit for (human codon optimized) Cas9 plant 
            expression driven by the 35S promoter. Specially conceived to be 
            linked with the TU of the kanamycin resistance gene GB1181..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12511)
  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 12511)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pEGB2Ω2_SF-35s:hCas9:tNos (GB1103)
FEATURES             Location/Qualifiers
     source          1..12511
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        778..1122
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1066..1086
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             1196..5299
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             1196..5299
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             1196..5299
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5312..5332
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5359..5611
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(5646..5664)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             7158..7787
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             8221..9288
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      9354..9548
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(9784..9806)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    9892..10032
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     9947..9966
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(10047..10064)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10218..10806)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10298..10317)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(10894..11685)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    12471..12493
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
ORIGIN
        1 ggagtctcaa tggtaacttt actctttatt taaccataca ttttttttta tttttttcac
       61 tttgttcttc atccactatt gttctttgtt catcttgaac aaaagctccc tccttctttg
      121 ttcttcatcc accattgttc ttcatcaatc atttcgctgt caggagacta gagccaagct
      181 gatctccttt gccccggaga tcaccatgga cgactttctc tatctctacg atctaggaag
      241 aaagttcgac ggagaaggtg acgataccat gttcaccacc gataatgaga agattagcct
      301 cttcaatttc agaaagaatg ctgacccaca gatggttaga gaggcctacg cggcaggtct
      361 gatcaagacg atctacccga gtaataatct ccaggagatc aaataccttc ccaagaaggt
      421 taaagatgca gtcaaaagat tcaggactaa ctgcatcaag aacacagaga aagatatatt
      481 tctcaagatc agaagtacta ttccagtatg gacgattcaa ggcttgcttc ataaaccaag
      541 gcaagtaata gagattggag tctctaagaa agtagttcct actgaatcaa aggccatgga
      601 gtcaaaaatt cagatcgagg atctaacaga actcgccgtg aagactggcg aacagttcat
      661 acagagtctt ttacgactca atgacaagaa gaaaatcttc gtcaacatgg tggagcacga
      721 cactctcgtc tactccaaga atatcaaaga tacagtctca gaagaccaaa gggctattga
      781 gacttttcaa caaagggtaa tatcgggaaa cctcctcgga ttccattgcc cagctatctg
      841 tcacttcatc aaaaggacag tagaaaagga aggtggcacc tacaaatgcc atcattgcga
      901 taaaggaaag gctatcgttc aagatgcccc tgccgacagt ggtcccaaag atggaccccc
      961 acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga
     1021 ttgatgtgat atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga
     1081 cccttcctct atataaggaa gttcatttca tttggagagg actccggtat ttttacaaca
     1141 attaccacaa caaaacaaac aacaaacaac attacaattt actattctag tcgaaatgga
     1201 caagaagtac tccattgggc tcgatatcgg cacaaacagc gtcggctggg ccgtcattac
     1261 ggacgagtac aaggtgccga gcaaaaaatt caaagttctg ggcaataccg atcgccacag
     1321 cataaagaag aacctcattg gcgccctcct gttcgactcc ggggaaacgg ccgaagccac
     1381 gcggctcaaa agaacagcac ggcgcagata tacccgcaga aagaatcgga tctgctacct
     1441 gcaggagatc tttagtaatg agatggctaa ggtggatgac tctttcttcc ataggctgga
     1501 ggagtccttt ttggtggagg aggataaaaa gcacgagcgc cacccaatct ttggcaatat
     1561 cgtggacgag gtggcgtacc atgaaaagta cccaaccata tatcatctga ggaagaagct
     1621 tgtagacagt actgataagg ctgacttgcg gttgatctat ctcgcgctgg cgcatatgat
     1681 caaatttcgg ggacacttcc tcatcgaggg ggacctgaac ccagacaaca gcgatgtcga
     1741 caaactcttt atccaactgg ttcagactta caatcagctt ttcgaagaga acccgatcaa
     1801 cgcatccgga gttgacgcca aagcaatcct gagcgctagg ctgtccaaat cccggcggct
     1861 cgaaaacctc atcgcacagc tccctgggga gaagaagaac ggcctgtttg gtaatcttat
     1921 cgccctgtca ctcgggctga cccccaactt taaatctaac ttcgacctgg ccgaagatgc
     1981 caagcttcaa ctgagcaaag acacctacga tgatgatctc gacaatctgc tggcccagat
     2041 cggcgaccag tacgcagacc tttttttggc ggcaaagaac ctgtcagacg ccattctgct
     2101 gagtgatatt ctgcgagtga acacggagat caccaaagct ccgctgagcg ctagtatgat
     2161 caagcgctat gatgagcacc accaagactt gactttgctg aaggcccttg tcagacagca
     2221 actgcctgag aagtacaagg aaattttctt cgatcagtct aaaaatggct acgccggata
     2281 cattgacggc ggagcaagcc aggaggaatt ttacaaattt attaagccca tcttggaaaa
     2341 aatggacggc accgaggagc tgctggtaaa gcttaacaga gaagatctgt tgcgcaaaca
     2401 gcgcactttc gacaatggaa gcatccccca ccagattcac ctgggcgaac tgcacgctat
     2461 cctcaggcgg caagaggatt tctacccctt tttgaaagat aacagggaaa agattgagaa
     2521 aatcctcaca tttcggatac cctactatgt aggccccctc gcccggggaa attccagatt
     2581 cgcgtggatg actcgcaaat cagaagagac tatcactccc tggaacttcg aggaagtcgt
     2641 ggataagggg gcctctgccc agtccttcat cgaaaggatg actaactttg ataaaaatct
     2701 gcctaacgaa aaggtgcttc ctaaacactc tctgctgtac gagtacttca cagtttataa
     2761 cgagctcacc aaggtcaaat acgtcacaga agggatgaga aagccagcat tcctgtctgg
     2821 agagcagaag aaagctatcg tggacctcct cttcaagacg aaccggaaag ttaccgtgaa
     2881 acagctcaaa gaagattatt tcaaaaagat tgaatgtttc gactctgttg aaatcagcgg
     2941 agtggaggat cgcttcaacg catccctggg aacgtatcac gatctcctga aaatcattaa
     3001 agacaaggac ttcctggaca atgaggagaa cgaggacatt cttgaggaca ttgtcctcac
     3061 ccttacgttg tttgaagata gggagatgat tgaagaacgc ttgaaaactt acgctcatct
     3121 cttcgacgac aaagtcatga aacagctcaa gaggcgccga tatacaggat gggggcggct
     3181 gtcaagaaaa ctgatcaatg ggatccgaga caagcagagt ggaaagacaa tcctggattt
     3241 tcttaagtcc gatggatttg ccaaccggaa cttcatgcag ttgatccatg atgactctct
     3301 cacctttaag gaggacatcc agaaagcaca agtttctggc cagggggaca gtctccacga
     3361 gcacatcgct aatcttgcag gtagcccagc tatcaaaaag ggaatactgc agaccgttaa
     3421 ggtcgtggat gaactcgtca aagtaatggg aaggcataag cccgagaata tcgttatcga
     3481 gatggcccga gagaaccaaa ctacccagaa gggacagaag aacagtaggg aaaggatgaa
     3541 gaggattgaa gagggtataa aagaactggg gtcccaaatc cttaaggaac acccagttga
     3601 aaacacccag cttcagaatg agaagctcta cctgtactac ctgcagaacg gcagggacat
     3661 gtacgtggat caggaactgg acatcaatcg gctctccgac tacgacgtgg atcatatcgt
     3721 gccccagtct tttctcaaag atgattctat tgataataaa gtgttgacaa gatccgataa
     3781 aaatagaggg aagagtgata acgtcccctc agaagaagtt gtcaagaaaa tgaaaaatta
     3841 ttggcggcag ctgctgaacg ccaaactgat cacacaacgg aagttcgata atctgactaa
     3901 ggctgaacga ggtggcctgt ctgagttgga taaagccggc ttcatcaaaa ggcagcttgt
     3961 tgagacacgc cagatcacca agcacgtggc ccaaattctc gattcacgca tgaacaccaa
     4021 gtacgatgaa aatgacaaac tgattcgaga ggtgaaagtt attactctga agtctaagct
     4081 ggtttcagat ttcagaaagg actttcagtt ttataaggtg agagagatca acaattacca
     4141 ccatgcgcat gatgcctacc tgaatgcagt ggtaggcact gcacttatca aaaaatatcc
     4201 caagcttgaa tctgaatttg tttacggaga ctataaagtg tacgatgtta ggaaaatgat
     4261 cgcaaagtct gagcaggaaa taggcaaggc caccgctaag tacttctttt acagcaatat
     4321 tatgaatttt ttcaagaccg agattacact ggccaatgga gagattcgga agcgaccact
     4381 tatcgaaaca aacggagaaa caggagaaat cgtgtgggac aagggtaggg atttcgcgac
     4441 agtccggaag gtcctgtcca tgccgcaggt gaacatcgtt aaaaagaccg aagtacagac
     4501 cggaggcttc tccaaggaaa gtatcctccc gaaaaggaac agcgacaagc tgatcgcacg
     4561 caaaaaagat tgggacccca agaaatacgg cggattcgat tctcctacag tcgcttacag
     4621 tgtactggtt gtggccaaag tggagaaagg gaagtctaaa aaactcaaaa gcgtcaagga
     4681 actgctgggc atcacaatca tggagcgatc aagcttcgaa aaaaacccca tcgactttct
     4741 cgaggcgaaa ggatataaag aggtcaaaaa agacctcatc attaagcttc ccaagtactc
     4801 tctctttgag cttgaaaacg gccggaaacg aatgctcgct agtgcgggcg agctgcagaa
     4861 aggtaacgag ctggcactgc cctctaaata cgttaatttc ttgtatctgg ccagccacta
     4921 tgaaaagctc aaaggatctc ccgaagataa tgagcagaag cagctgttcg tggaacaaca
     4981 caaacactac cttgatgaga tcatcgagca aataagcgaa ttctccaaaa gagtgatcct
     5041 cgccgacgct aacctcgata aggtgctttc tgcttacaat aagcacaggg ataagcccat
     5101 cagggagcag gcagaaaaca ttatccactt gtttactctg accaacttgg gcgcgcctgc
     5161 agccttcaag tacttcgaca ccaccataga cagaaagcgg tacacctcta caaaggaggt
     5221 cctggacgcc acactgattc atcagtcaat tacggggctc tatgaaacaa gaatcgacct
     5281 ctctcagctc ggtggagaca gcagggctga ccccaagaag aagaggaagg tgtgagcttg
     5341 gaatggatct tcgatcccga tcgttcaaac atttggcaat aaagtttctt aagattgaat
     5401 cctgttgccg gtcttgcgac gattatcata taatttctgt tgaattacgt taagcatgta
     5461 ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg
     5521 caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta ggataaatta
     5581 tcgcgcgcgg tgtcatctat gttactagat cgggaattgc caagctaatt cttgaagacg
     5641 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     5701 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5761 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataatg
     5821 ggaccgactc gcgcttgaga ccgatatctg acgtaaacct aagagaaaag agcgtttatt
     5881 agaataacgg 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 aacgctcgac
     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 tccgatgagt aaataaacaa
     7141 atacgcaagg ggaacgcatg aaggttatcg ctgtacttaa ccagaaaggc gggtcaggca
     7201 agacgaccat cggaacccat 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 agatcgcgca gctaccagag taaatgagca aatgaataaa
     8041 tgagtagatg aattttagcg gctaaaggag gcggcatgga aaatcaagaa caaccaggca
     8101 ccgacgccgt ggaatgcccc atgtgtggag gaacgggcgg ttggccaggc gtaagcggct
     8161 gggttgtctg ccggccctgc aatggcactg gaacccccaa gcccgaggaa tcggcgtgac
     8221 ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag
     8281 aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc acgccccggt
     8341 gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc
     8401 ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt tttcgttccg
     8461 atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc cgttttccgt
     8521 ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc agacgggcac
     8581 gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt gggattacga cctggtactg
     8641 atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa gggagacaag
     8701 cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg gcgagccgat
     8761 ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac cacgcacgtt
     8821 gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa
     8881 gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag
     8941 atcgagctag ctgattggat gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg
     9001 acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct ctaccgcctg
     9061 gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat ctacgaacgc
     9121 agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct gatcgggtca
     9181 aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc gatcctagtc
     9241 atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg tacggagcag
     9301 atgctagggc aaattgccct agcaggggaa aaaggtcgaa aaggactctt tcctgtggat
     9361 agcacgtaca ttgggaaccc aaagccgtac attgggaacc ggaacccgta cattgggaac
     9421 ccaaagccgt acattgggaa ccggtcacac atgtaagtga ctgatataaa agagaaaaaa
     9481 ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc
     9541 tgtgcatatc tgtctggcca gcgcacagcc gaagagctgc aaaaagcgcc tacccttcgg
     9601 tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc tggccgctca
     9661 aaaatggctg gcctacggcc aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca
     9721 ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga
     9781 aaacctctga cacatgcagc tcccggtgac ggtcacagct tgtctgtaag cggatgccgg
     9841 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     9901 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
     9961 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
    10021 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
    10081 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
    10141 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
    10201 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
    10261 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
    10321 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
    10381 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
    10441 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
    10501 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
    10561 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
    10621 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
    10681 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
    10741 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
    10801 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
    10861 cgttaaggga ttttggtcat gcattctagg tgattatttg ccgactacct tggtgatctc
    10921 gcctttcacg tagtggacaa attcttccaa ctgatctgcg cgcgaggcca agcgatcttc
    10981 ttcttgtcca agataagcct gtctagcttc aagtatgacg ggctgatact gggccggcag
    11041 gcgctccatt gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct
    11101 gtaccaaatg cgggacaacg taagcactac atttcgctca tctccagccc agtcgggcgg
    11161 cgagttccat agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg
    11221 atcaaagagt tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt
    11281 cagcaagata gccagatcaa tgtcgatcgt ggctggctcg aagatacctg caagaatgtc
    11341 attgcgctgc cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat
    11401 gtcgtcgtgc acaacaatgg tgacttctac agcgcggaga atctcgctct ctccagggga
    11461 agccgaagtt tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat caagccttac
    11521 ggtcaccgta accagcaaat caatatcact gtgtggcttc aggccgccat ccactgcgga
    11581 gccgtacaaa tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac
    11641 ctctgatagt tgagtcgata cttcggcgat caccgcttcc ctcatgatgt ttaactttgt
    11701 tttagggcga ctgccctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc
    11761 cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata gactgtaccc caaaaaaaca
    11821 gtcataacaa gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt
    11881 tctggaccag ttgcgtgagc gcatacgcta cttgcattac agcttacgaa ccgaacaggc
    11941 ttatgtccac tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc cggcaacctt
    12001 gggcagcagc gaagtcgagg catttctgtc ctggctggaa cagaacttat tatttccttc
    12061 ctcttttcta cagtatttaa agatacccca agaagctaat tataacaaga cgaactccaa
    12121 ttcactgttc cttgcattct aaaaccttaa ataccagaaa acagcttttt caaagttgtt
    12181 ttcaaagttg gcgtataaca tagtatcgac ggagccgatt ttgaaaccgc ggtgatcaca
    12241 ggcagcaacg ctctgtcatc gttacaatca acatgctacc ctccgcgaga tcatccgtgt
    12301 ttcaaacccg gcagcttagt tgccgttctt ccgaatagca tcggtaacat gagcaaagtc
    12361 tgccgcctta caacggctct cccgctgacg ccgtcccgga ctgatgggct gcctgtatcg
    12421 agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg tggcaggata
    12481 tattgtggtg taacgatatc ggtctcagtc a
//
