LOCUS       sequence-154114-       12054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCas9-SH3 and PmCDA1-SHL in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12054)
  AUTHORS   Nishida K, Arazoe T, Yachie N, Banno S, Kakimoto M, Tabata M, 
            Mochizuki M, Miyabe A, Araki M, Hara KY, Shimatani Z, Kondo A
  TITLE     Targeted nucleotide editing using hybrid prokaryotic and vertebrate 
            adaptive immune systems.
  JOURNAL   Science. 2016 Aug 4. pii: aaf8729.
  PUBMED    27492474
REFERENCE   2  (bases 1 to 12054)
  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: "Science. 
            2016 Aug 4. pii: aaf8729."
COMMENT     Sequence Label: pRS315e_pGal-dCas9-SH3-pGal-PmCDA1-SHL
FEATURES             Location/Qualifiers
     source          1..12054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    70..573
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        610..714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(715..783,784..1575)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(933..952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1746..2334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2488..2505
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2658..2663,2664..2681,2682..2688)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2696..2712
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2701..2723
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2720..2736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2720..2736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2757..2775
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(2853..3476)
                     /codon_start=1
                     /product="cytidine deaminase from the sea lamprey 
                     Petromyzon marinus (Rogozin et al., 2007)"
                     /label=PmCDA1
                     /translation="MTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGER
                     RACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKI
                     LEWYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIF
                     IQSSHNQLNENRWLEKTLKRAEKRRSELSIMIQVKILHTTKSPAV"
     CDS             complement(3489..3512)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        3527..4191
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    3743..3860
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     4155..4178
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             join(4194..4220,4221..4223,4224..6707,6708..6713,6714..6779,
                     6780..6782,6783..8297)
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(4194..4220,4221..4223,4224..6710,6711..6713,
                     6714..8297)
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4194..8297
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8310..8330
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8538..8603
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      8634..8821
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(8689..8713)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     terminator      complement(8858..8903)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(8939..8958)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8940..8958)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8965..8982)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8965..8981)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8974..8996)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9126..9581
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9213..9232)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9423..9444
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9881..10285
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             10298..11392
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(11885..11904)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12004..12026
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1141 ccgaaggagc taaccgcttt ttttcacaac atgggggatc atgtaactcg ccttgatcgt
     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 ggcagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2101 acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc caggggggaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc cgagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctcgaaatt
     2761 aaccctcact aaagggaaca aaagctggta ccgggcccga attctctcct tctcttaggt
     2821 ggcagagcag gtggagggtc gaccatacta gaaacagcag gactcttagt ggtgtggagt
     2881 atttttacct gaatcataat ggacaactcg ctccgtcgtt tttcagctcg cttcaaagtc
     2941 ttctcaagcc atctattctc attcaattga ttgtgcgacg attggatgaa tattttcctg
     3001 caacattggt agtgttcact taccattaca ttcaacccaa ccccgttatc tctgaggttc
     3061 cacagcccaa tttgattcct cgcatttttc tcgtaataga gtttgcaagc ccagattttc
     3121 aaagtgtggc cgttcccccg cagctcctgg ttataccatt ctaagatctt ttcagcgcaa
     3181 tctgcacaag gactccagga tgagtaccaa tttatcgtga attgtccggg gttgtcgcgc
     3241 aggtattctt cgacttttct aatgctaaag atttcggcgt gaatgccacg ttctgtcccg
     3301 ctctgtggtt tattcacagc atagccccaa aaacacgctc tacgttcacc ccgtcgtttt
     3361 aattcaaaga gaacgtagca tctatgcgac acggattttt tgttgttgaa aaactgtttc
     3421 ttaaacgtgt agatgtccaa cttctcatgg attctcacgt actcagcgtc ggtcatcgat
     3481 ccgccgcctt tttcgaactg cgggtggctc caggccacca tactagtttc aaaaattctt
     3541 actttttttt tggatggacg caaagaagtt taataatcat attacatggc attaccacca
     3601 tatacatatc catatacata tccatatcta atcttactta tatgttgtgg aaatgtaaag
     3661 agccccatta tcttagccta aaaaaacctt ctctttggaa ctttcagtaa tacgcttaac
     3721 tgctcattgc tatattgaag tacggattag aagccgccga gcgggtgaca gccctccgaa
     3781 ggaagactct cctccgtgcg tcctcgtctt caccggtcgc gttcctgaaa cgcagatgtg
     3841 cctcgcgccg cactgctccg aacaataaag attctacaat actagctttt atggttatga
     3901 agaggaaaaa ttggcagtaa cctggcccca caaaccttca aatgaacgaa tcaaattaac
     3961 aaccatagga tgataatgcg attagttttt tagccttatt tctggggtaa ttaatcagcg
     4021 aagcgatgat ttttgatcta ttaacagata tataaatgca aaaactgcat aaccacttta
     4081 actaatactt tcaacatttt cggtttgtat tacttcttat tcaaatgtaa taaaagtatc
     4141 aacaaaaaat tgttaatata cctctatact ttaacgtcaa ggagaaaaaa cccatggaca
     4201 agaagtactc cattgggctc gctatcggca caaacagcgt cggttgggcc gtcattacgg
     4261 acgagtacaa ggtgccgagc aaaaaattca aagttctggg caataccgat cgccacagca
     4321 taaagaagaa cctcattggc gccctcctgt tcgactccgg ggagacggcc gaagccacgc
     4381 ggctcaaaag aacagcacgg cgcagatata cccgcagaaa gaatcggatc tgctacctgc
     4441 aggagatctt tagtaatgag atggctaagg tggatgactc tttcttccat aggctggagg
     4501 agtccttttt ggtggaggag gataaaaagc acgagcgcca cccaatcttt ggcaatatcg
     4561 tggacgaggt ggcgtaccat gaaaagtacc caaccatata tcatctgagg aagaagcttg
     4621 tagacagtac tgataaggct gacttgcggt tgatctatct cgcgctggcg catatgatca
     4681 aatttcgggg acacttcctc atcgaggggg acctgaaccc agacaacagc gatgtcgaca
     4741 aactctttat ccaactggtt cagacttaca atcagctttt cgaagagaac ccgatcaacg
     4801 catccggagt tgacgccaaa gcaatcctga gcgctaggct gtccaaatcc cggcggctcg
     4861 aaaacctcat cgcacagctc cctggggaga agaagaacgg cctgtttggt aatcttatcg
     4921 ccctgtcact cgggctgacc cccaacttta aatctaactt cgacctggcc gaagatgcca
     4981 agcttcaact gagcaaagac acctacgatg atgatctcga caatctgctg gcccagatcg
     5041 gcgaccagta cgcagacctt tttttggcgg caaagaacct gtcagacgcc attctgctga
     5101 gtgatattct gcgagtgaac acggagatca ccaaagctcc gctgagcgct agtatgatca
     5161 agcgctatga tgagcaccac caagacttga ctttgctgaa ggcccttgtc agacagcaac
     5221 tgcctgagaa gtacaaggaa attttcttcg atcagtctaa aaatggctac gccggataca
     5281 ttgacggcgg agcaagccag gaggaatttt acaaatttat taagcccatc ttggaaaaaa
     5341 tggacggcac cgaggagctg ctggtaaagc ttaacagaga agatctgttg cgcaaacagc
     5401 gcactttcga caatggaagc atcccccacc agattcacct gggcgaactg cacgctatcc
     5461 tcaggcggca agaggatttc tacccctttt tgaaagataa cagggaaaag attgagaaaa
     5521 tcctcacatt tcggataccc tactatgtag gccccctcgc ccggggaaat tccagattcg
     5581 cgtggatgac tcgcaaatca gaagagacca tcactccctg gaacttcgag gaagtcgtgg
     5641 ataagggggc ctctgcccag tccttcatcg aaaggatgac taactttgat aaaaatctgc
     5701 ctaacgaaaa ggtgcttcct aaacactctc tgctgtacga gtacttcaca gtttataacg
     5761 agctcaccaa ggtcaaatac gtcacagaag ggatgagaaa gccagcattc ctgtctggag
     5821 agcagaagaa agctatcgtg gacctcctct tcaagacgaa ccggaaagtt accgtgaaac
     5881 agctcaaaga agactatttc aaaaagattg aatgtttcga ctctgttgaa atcagcggag
     5941 tggaggatcg cttcaacgca tccctgggaa cgtatcacga tctcctgaaa atcattaaag
     6001 acaaggactt cctggacaat gaggagaacg aggacattct tgaggacatt gtcctcaccc
     6061 ttacgttgtt tgaagatagg gagatgattg aagaacgctt gaaaacttac gctcatctct
     6121 tcgacgacaa agtcatgaaa cagctcaaga ggcgccgata tacaggatgg gggcggctgt
     6181 caagaaaact gatcaatggg atccgagaca agcagagtgg aaagacaatc ctggattttc
     6241 ttaagtccga tggatttgcc aaccggaact tcatgcagtt gatccatgat gactctctca
     6301 cctttaagga ggacatccag aaagcacaag tttctggcca gggggacagt cttcacgagc
     6361 acatcgctaa tcttgcaggt agcccagcta tcaaaaaggg aatactgcag accgttaagg
     6421 tcgtggatga actcgtcaaa gtaatgggaa ggcataagcc cgagaatatc gttatcgaga
     6481 tggcccgaga gaaccaaact acccagaagg gacagaagaa cagtagggaa aggatgaaga
     6541 ggattgaaga gggtataaaa gaactggggt cccaaatcct taaggaacac ccagttgaaa
     6601 acacccagct tcagaatgag aagctctacc tgtactacct gcagaacggc agggacatgt
     6661 acgtggatca ggaactggac atcaatcggc tctccgacta cgacgtggat gctatcgtgc
     6721 cccagtcttt tctcaaagat gattctattg ataataaagt gttgacaaga tccgataaaa
     6781 atagagggaa gagtgataac gtcccctcag aagaagttgt caagaaaatg aaaaattatt
     6841 ggcggcagct gctgaacgcc aaactgatca cacaacggaa gttcgataat ctgactaagg
     6901 ctgaacgagg tggcctgtct gagttggata aagccggctt catcaaaagg cagcttgttg
     6961 agacacgcca gatcaccaag cacgtggccc aaattctcga ttcacgcatg aacaccaagt
     7021 acgatgaaaa tgacaaactg attcgagagg tgaaagttat tactctgaag tctaagctgg
     7081 tctcagattt cagaaaggac tttcagtttt ataaggtgag agagatcaac aattaccacc
     7141 atgcgcatga tgcctacctg aatgcagtgg taggcactgc acttatcaaa aaatatccca
     7201 agcttgaatc tgaatttgtt tacggagact ataaagtgta cgatgttagg aaaatgatcg
     7261 caaagtctga gcaggaaata ggcaaggcca ccgctaagta cttcttttac agcaatatta
     7321 tgaatttttt caagaccgag attacactgg ccaatggaga gattcggaag cgaccactta
     7381 tcgaaacaaa cggagaaaca ggagaaatcg tgtgggacaa gggtagggat ttcgcgacag
     7441 tccggaaggt cctgtccatg ccgcaggtga acatcgttaa aaagaccgaa gtacagaccg
     7501 gaggcttctc caaggaaagt atcctcccga aaaggaacag cgacaagctg atcgcacgca
     7561 aaaaagattg ggaccccaag aaatacggcg gattcgattc tcctacagtc gcttacagtg
     7621 tactggttgt ggccaaagtg gagaaaggga agtctaaaaa actcaaaagc gtcaaggaac
     7681 tgctgggcat cacaatcatg gagcgatcaa gcttcgaaaa aaaccccatc gactttctcg
     7741 aggcgaaagg atataaagag gtcaaaaaag acctcatcat taagcttccc aagtactctc
     7801 tctttgagct tgaaaacggc cggaaacgaa tgctcgctag tgcgggcgag ctgcagaaag
     7861 gtaacgagct ggcactgccc tctaaatacg ttaatttctt gtatctggcc agccactatg
     7921 aaaagctcaa agggtctccc gaagataatg agcagaagca gctgttcgtg gaacaacaca
     7981 aacactacct tgatgagatc atcgagcaaa taagcgaatt ctccaaaaga gtgatcctcg
     8041 ccgacgctaa cctcgataag gtgctttctg cttacaataa gcacagggat aagcccatca
     8101 gggagcaggc agaaaacatt atccacttgt ttactctgac caacttgggc gcgcctgcag
     8161 ccttcaagta cttcgacacc accatagaca gaaagcggta cacctctaca aaggaggtcc
     8221 tggacgccac actgattcat cagtcaatta cggggctcta tgaaacaaga atcgacctct
     8281 ctcagctcgg tggagacagc agggctgacc ccaagaagaa gaggaaggtg ggtggaggag
     8341 gttctggagg tggaggttct gcagagtatg tgcgggccct ctttgacttt aatgggaatg
     8401 atgaagaaga tcttcccttt aagaaaggag acatcctgag aatccgggat aagcctgaag
     8461 agcagtggtg gaatgcagag gacagcgaag gaaagagggg gatgattcct gtcccttacg
     8521 tggagaagta ttccggagac tataaggacc acgacggaga ctacaaggat catgatattg
     8581 attacaaaga cgatgacgat aagtctagag gctccggata aggatcctaa taagcgaatt
     8641 tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat
     8701 tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt
     8761 aggtcaggtt gctttctcag gtatagcatg aggtcgctct tattgaccac acctctaccg
     8821 gcatgccgag caaatgcctg caaatcgctc ccgggcaaaa aaccccctca agacccgttt
     8881 agaggcccca aggggttatg ctatgcatac caagcttccg cggtggagct ccaattcgcc
     8941 ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     9001 ccctggcgtt acccaactta atcgccttgc agcacatccc cccttcgcca gctggcgtaa
     9061 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     9121 gcgcgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     9181 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     9241 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     9301 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     9361 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     9421 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     9481 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     9541 atttaacgcg aattttaaca aaatattaac gtttacaatt tcctgatgcg gtattttctc
     9601 cttacgcatc tgtgcggtat ttcacaccgc atatcgaccc tcgaggagaa cttctagtat
     9661 atccacatac ctaatattat tgccttatta aaaatggaat cggaacaatt acatcaaaat
     9721 ccacattctc ttcaaaatca attgtcctgt acttccttgt tcatgtgtgt tcaaaaacgt
     9781 tatatttata ggataattat actctatttc tcaacaagta attggttgtt tggccgagcg
     9841 gtctaaggcg cctgattcaa gaaatatctt gaccgcagtt aactgtggga atactcaggt
     9901 atcgtaagat gcaagagttc gaatctctta gcaaccatta tttttttcct caacataacg
     9961 agaacacaca ggggcgctat cgcacagaat caaattcgat gactggaaat tttttgttaa
    10021 tttcagaggt cgcctgacgc atataccttt ttcaactgaa aaattgggag aaaaaggaaa
    10081 ggtgagaggc cggaaccggc ttttcatata gaatagagaa gcgttcatga ctaaatgctt
    10141 gcatcacaat acttgaagtt gacaatatta tttaaggacc tattgttttt tccaataggt
    10201 ggttagcaat cgtcttactt tctaactttt cttacctttt acatttcagc aatatatata
    10261 tatatttcaa ggatatacca ttctaatgtc tgcccctatg tctgccccta agaagatcgt
    10321 cgttttgcca ggtgaccacg ttggtcaaga aatcacagcc gaagccatta aggttcttaa
    10381 agctatttct gatgttcgtt ccaatgtcaa gttcgatttc gaaaatcatt taattggtgg
    10441 tgctgctatc gatgctacag gtgtcccact tccagatgag gcgctggaag cctccaagaa
    10501 ggttgatgcc gttttgttag gtgctgtggg tggtcctaaa tggggtaccg gtagtgttag
    10561 acctgaacaa ggtttactaa aaatccgtaa agaacttcaa ttgtacgcca acttaagacc
    10621 atgtaacttt gcatccgact ctcttttaga cttatctcca atcaagccac aatttgctaa
    10681 aggtactgac ttcgttgttg tcagagaatt agtgggaggt atttactttg gtaagagaaa
    10741 ggaagacgat ggtgatggtg tcgcttggga tagtgaacaa tacaccgttc cagaagtgca
    10801 aagaatcaca agaatggccg ctttcatggc cctacaacat gagccaccat tgcctatttg
    10861 gtccttggat aaagctaatg ttttggcctc ttcaagatta tggagaaaaa ctgtggagga
    10921 aaccatcaag aacgagttcc ctacattgaa ggttcaacat caattgattg attctgccgc
    10981 catgatccta gttaagaacc caacccacct aaatggtatt ataatcacca gcaacatgtt
    11041 tggtgatatc atctccgatg aagcctccgt tatcccaggt tccttgggtt tgttgccatc
    11101 tgcgtccttg gcctctttgc cagacaagaa caccgcattt ggtttgtacg aaccatgcca
    11161 cggttctgct ccagatttgc caaagaataa ggttgaccct atcgccacta tcttgtctgc
    11221 tgcaatgatg ttgaaattgt cattgaactt gcctgaagaa ggtaaggcca ttgaagatgc
    11281 agttaaaaag gttttggatg caggtatcag aactggtgat ttaggtggtt ccaacagtac
    11341 caccgaagtc ggtgatgctg tcgccgaaga agttaagaaa atccttgctt aaaaagattc
    11401 tcttttttta tgatatttgt acataaactt tataaatgaa attcataata gaaacgacac
    11461 gaaattacaa aatggaatat gttcataggg tagacgaaac tatatacgca atctacatac
    11521 atttatcaag aaggagaaaa aggaggatag taaaggaata caggtaagca aattgatact
    11581 aatggctcaa cgtgataagg aaaaagaatt gcactttaac attaatattg acaaggagga
    11641 gggcaccaca caaaaagtta ggtgtaacag aaaatcatga aactacgatt cctaatttga
    11701 tattggagga ttttctctaa aaaaaaaaaa atacaacaaa taaaaaacac tcaatgacct
    11761 gaccatttga tggagtttaa gtcaatacct tcttgaacca tttcccataa tggtgaaagt
    11821 tccctcaaga attttactct gtcagaaacg gccttacgac gtagtcgata tggtgcactc
    11881 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
    11941 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
    12001 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcga
//
