LOCUS       sequence-148777-       12005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCas9-PmCDA1 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12005)
  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 12005)
  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-PmCDA1
FEATURES             Location/Qualifiers
     source          1..12005
                     /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"
     protein_bind    2622..2643
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     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"
     promoter        2852..3516
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    3068..3185
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3480..3503
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             join(3519..3545,3546..3548,3549..6032,6033..6038,6039..6104,
                     6105..6107,6108..7622)
                     /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(3519..3545,3546..3548,3549..6035,6036..6038,
                     6039..7622)
                     /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             3519..7622
                     /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             7635..7655
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7863..7928
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7935..8558
                     /codon_start=1
                     /product="cytidine deaminase from the sea lamprey 
                     Petromyzon marinus (Rogozin et al., 2007)"
                     /label=PmCDA1
                     /translation="MTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGER
                     RACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKI
                     LEWYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIF
                     IQSSHNQLNENRWLEKTLKRAEKRRSELSIMIQVKILHTTKSPAV"
     terminator      8589..8776
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(8644..8668)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     terminator      complement(8813..8858)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(8894..8913)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8895..8913)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8920..8937)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8920..8936)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8929..8951)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9077..9532
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9164..9183)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9374..9395
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9832..10236
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             10249..11343
                     /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(11836..11855)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11955..11977
                     /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 ttagctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctcgaaatt
     2761 aaccctcact aaagggaaca aaagctggta ccgggcccga attctctcct tctcttaggt
     2821 ggcagagcag gtggagggtc gaccatacta gtttcaaaaa ttcttacttt ttttttggat
     2881 ggacgcaaag aagtttaata atcatattac atggcattac caccatatac atatccatat
     2941 acatatccat atctaatctt acttatatgt tgtggaaatg taaagagccc cattatctta
     3001 gcctaaaaaa accttctctt tggaactttc agtaatacgc ttaactgctc attgctatat
     3061 tgaagtacgg attagaagcc gccgagcggg tgacagccct ccgaaggaag actctcctcc
     3121 gtgcgtcctc gtcttcaccg gtcgcgttcc tgaaacgcag atgtgcctcg cgccgcactg
     3181 ctccgaacaa taaagattct acaatactag cttttatggt tatgaagagg aaaaattggc
     3241 agtaacctgg ccccacaaac cttcaaatga acgaatcaaa ttaacaacca taggatgata
     3301 atgcgattag ttttttagcc ttatttctgg ggtaattaat cagcgaagcg atgatttttg
     3361 atctattaac agatatataa atgcaaaaac tgcataacca ctttaactaa tactttcaac
     3421 attttcggtt tgtattactt cttattcaaa tgtaataaaa gtatcaacaa aaaattgtta
     3481 atatacctct atactttaac gtcaaggaga aaaaacccat ggacaagaag tactccattg
     3541 ggctcgctat cggcacaaac agcgtcggtt gggccgtcat tacggacgag tacaaggtgc
     3601 cgagcaaaaa attcaaagtt ctgggcaata ccgatcgcca cagcataaag aagaacctca
     3661 ttggcgccct cctgttcgac tccggggaga cggccgaagc cacgcggctc aaaagaacag
     3721 cacggcgcag atatacccgc agaaagaatc ggatctgcta cctgcaggag atctttagta
     3781 atgagatggc taaggtggat gactctttct tccataggct ggaggagtcc tttttggtgg
     3841 aggaggataa aaagcacgag cgccacccaa tctttggcaa tatcgtggac gaggtggcgt
     3901 accatgaaaa gtacccaacc atatatcatc tgaggaagaa gcttgtagac agtactgata
     3961 aggctgactt gcggttgatc tatctcgcgc tggcgcatat gatcaaattt cggggacact
     4021 tcctcatcga gggggacctg aacccagaca acagcgatgt cgacaaactc tttatccaac
     4081 tggttcagac ttacaatcag cttttcgaag agaacccgat caacgcatcc ggagttgacg
     4141 ccaaagcaat cctgagcgct aggctgtcca aatcccggcg gctcgaaaac ctcatcgcac
     4201 agctccctgg ggagaagaag aacggcctgt ttggtaatct tatcgccctg tcactcgggc
     4261 tgacccccaa ctttaaatct aacttcgacc tggccgaaga tgccaagctt caactgagca
     4321 aagacaccta cgatgatgat ctcgacaatc tgctggccca gatcggcgac cagtacgcag
     4381 accttttttt ggcggcaaag aacctgtcag acgccattct gctgagtgat attctgcgag
     4441 tgaacacgga gatcaccaaa gctccgctga gcgctagtat gatcaagcgc tatgatgagc
     4501 accaccaaga cttgactttg ctgaaggccc ttgtcagaca gcaactgcct gagaagtaca
     4561 aggaaatttt cttcgatcag tctaaaaatg gctacgccgg atacattgac ggcggagcaa
     4621 gccaggagga attttacaaa tttattaagc ccatcttgga aaaaatggac ggcaccgagg
     4681 agctgctggt aaagcttaac agagaagatc tgttgcgcaa acagcgcact ttcgacaatg
     4741 gaagcatccc ccaccagatt cacctgggcg aactgcacgc tatcctcagg cggcaagagg
     4801 atttctaccc ctttttgaaa gataacaggg aaaagattga gaaaatcctc acatttcgga
     4861 taccctacta tgtaggcccc ctcgcccggg gaaattccag attcgcgtgg atgactcgca
     4921 aatcagaaga gaccatcact ccctggaact tcgaggaagt cgtggataag ggggcctctg
     4981 cccagtcctt catcgaaagg atgactaact ttgataaaaa tctgcctaac gaaaaggtgc
     5041 ttcctaaaca ctctctgctg tacgagtact tcacagttta taacgagctc accaaggtca
     5101 aatacgtcac agaagggatg agaaagccag cattcctgtc tggagagcag aagaaagcta
     5161 tcgtggacct cctcttcaag acgaaccgga aagttaccgt gaaacagctc aaagaagact
     5221 atttcaaaaa gattgaatgt ttcgactctg ttgaaatcag cggagtggag gatcgcttca
     5281 acgcatccct gggaacgtat cacgatctcc tgaaaatcat taaagacaag gacttcctgg
     5341 acaatgagga gaacgaggac attcttgagg acattgtcct cacccttacg ttgtttgaag
     5401 atagggagat gattgaagaa cgcttgaaaa cttacgctca tctcttcgac gacaaagtca
     5461 tgaaacagct caagaggcgc cgatatacag gatgggggcg gctgtcaaga aaactgatca
     5521 atgggatccg agacaagcag agtggaaaga caatcctgga ttttcttaag tccgatggat
     5581 ttgccaaccg gaacttcatg cagttgatcc atgatgactc tctcaccttt aaggaggaca
     5641 tccagaaagc acaagtttct ggccaggggg acagtcttca cgagcacatc gctaatcttg
     5701 caggtagccc agctatcaaa aagggaatac tgcagaccgt taaggtcgtg gatgaactcg
     5761 tcaaagtaat gggaaggcat aagcccgaga atatcgttat cgagatggcc cgagagaacc
     5821 aaactaccca gaagggacag aagaacagta gggaaaggat gaagaggatt gaagagggta
     5881 taaaagaact ggggtcccaa atccttaagg aacacccagt tgaaaacacc cagcttcaga
     5941 atgagaagct ctacctgtac tacctgcaga acggcaggga catgtacgtg gatcaggaac
     6001 tggacatcaa tcggctctcc gactacgacg tggatgctat cgtgccccag tcttttctca
     6061 aagatgattc tattgataat aaagtgttga caagatccga taaaaataga gggaagagtg
     6121 ataacgtccc ctcagaagaa gttgtcaaga aaatgaaaaa ttattggcgg cagctgctga
     6181 acgccaaact gatcacacaa cggaagttcg ataatctgac taaggctgaa cgaggtggcc
     6241 tgtctgagtt ggataaagcc ggcttcatca aaaggcagct tgttgagaca cgccagatca
     6301 ccaagcacgt ggcccaaatt ctcgattcac gcatgaacac caagtacgat gaaaatgaca
     6361 aactgattcg agaggtgaaa gttattactc tgaagtctaa gctggtctca gatttcagaa
     6421 aggactttca gttttataag gtgagagaga tcaacaatta ccaccatgcg catgatgcct
     6481 acctgaatgc agtggtaggc actgcactta tcaaaaaata tcccaagctt gaatctgaat
     6541 ttgtttacgg agactataaa gtgtacgatg ttaggaaaat gatcgcaaag tctgagcagg
     6601 aaataggcaa ggccaccgct aagtacttct tttacagcaa tattatgaat tttttcaaga
     6661 ccgagattac actggccaat ggagagattc ggaagcgacc acttatcgaa acaaacggag
     6721 aaacaggaga aatcgtgtgg gacaagggta gggatttcgc gacagtccgg aaggtcctgt
     6781 ccatgccgca ggtgaacatc gttaaaaaga ccgaagtaca gaccggaggc ttctccaagg
     6841 aaagtatcct cccgaaaagg aacagcgaca agctgatcgc acgcaaaaaa gattgggacc
     6901 ccaagaaata cggcggattc gattctccta cagtcgctta cagtgtactg gttgtggcca
     6961 aagtggagaa agggaagtct aaaaaactca aaagcgtcaa ggaactgctg ggcatcacaa
     7021 tcatggagcg atcaagcttc gaaaaaaacc ccatcgactt tctcgaggcg aaaggatata
     7081 aagaggtcaa aaaagacctc atcattaagc ttcccaagta ctctctcttt gagcttgaaa
     7141 acggccggaa acgaatgctc gctagtgcgg gcgagctgca gaaaggtaac gagctggcac
     7201 tgccctctaa atacgttaat ttcttgtatc tggccagcca ctatgaaaag ctcaaagggt
     7261 ctcccgaaga taatgagcag aagcagctgt tcgtggaaca acacaaacac taccttgatg
     7321 agatcatcga gcaaataagc gaattctcca aaagagtgat cctcgccgac gctaacctcg
     7381 ataaggtgct ttctgcttac aataagcaca gggataagcc catcagggag caggcagaaa
     7441 acattatcca cttgtttact ctgaccaact tgggcgcgcc tgcagccttc aagtacttcg
     7501 acaccaccat agacagaaag cggtacacct ctacaaagga ggtcctggac gccacactga
     7561 ttcatcagtc aattacgggg ctctatgaaa caagaatcga cctctctcag ctcggtggag
     7621 acagcagggc tgaccccaag aagaagagga aggtgggtgg aggaggttct ggaggtggag
     7681 gttctgcaga gtatgtgcgg gccctctttg actttaatgg gaatgatgaa gaagatcttc
     7741 cctttaagaa aggagacatc ctgagaatcc gggataagcc tgaagagcag tggtggaatg
     7801 cagaggacag cgaaggaaag agggggatga ttcctgtccc ttacgtggag aagtattccg
     7861 gagactataa ggaccacgac ggagactaca aggatcatga tattgattac aaagacgatg
     7921 acgataagtc taggatgacc gacgctgagt acgtgagaat ccatgagaag ttggacatct
     7981 acacgtttaa gaaacagttt ttcaacaaca aaaaatccgt gtcgcataga tgctacgttc
     8041 tctttgaatt aaaacgacgg ggtgaacgta gagcgtgttt ttggggctat gctgtgaata
     8101 aaccacagag cgggacagaa cgtggcattc acgccgaaat ctttagcatt agaaaagtcg
     8161 aagaatacct gcgcgacaac cccggacaat tcacgataaa ttggtactca tcctggagtc
     8221 cttgtgcaga ttgcgctgaa aagatcttag aatggtataa ccaggagctg cgggggaacg
     8281 gccacacttt gaaaatctgg gcttgcaaac tctattacga gaaaaatgcg aggaatcaaa
     8341 ttgggctgtg gaacctcaga gataacgggg ttgggttgaa tgtaatggta agtgaacact
     8401 accaatgttg caggaaaata ttcatccaat cgtcgcacaa tcaattgaat gagaatagat
     8461 ggcttgagaa gactttgaag cgagctgaaa aacgacggag cgagttgtcc attatgattc
     8521 aggtaaaaat actccacacc actaagagtc ctgctgtttc tagaggctcc ggataaggat
     8581 cctaataagc gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa
     8641 taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga
     8701 gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gcatgaggtc gctcttattg
     8761 accacacctc taccggcatg ccgagcaaat gcctgcaaat cgctcccggg caaaaaaccc
     8821 cctcaagacc cgtttagagg ccccaagggg ttatgctatg cataccaagc ttccgcggtg
     8881 gagctccaat tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg
     8941 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     9001 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     9061 cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     9121 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     9181 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     9241 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     9301 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     9361 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     9421 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     9481 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat ttcctgatgc
     9541 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatcgacc ctcgaggaga
     9601 acttctagta tatccacata cctaatatta ttgccttatt aaaaatggaa tcggaacaat
     9661 tacatcaaaa tccacattct cttcaaaatc aattgtcctg tacttccttg ttcatgtgtg
     9721 ttcaaaaacg ttatatttat aggataatta tactctattt ctcaacaagt aattggttgt
     9781 ttggccgagc ggtctaaggc gcctgattca agaaatatct tgaccgcagt taactgtggg
     9841 aatactcagg tatcgtaaga tgcaagagtt cgaatctctt agcaaccatt atttttttcc
     9901 tcaacataac gagaacacac aggggcgcta tcgcacagaa tcaaattcga tgactggaaa
     9961 ttttttgtta atttcagagg tcgcctgacg catatacctt tttcaactga aaaattggga
    10021 gaaaaaggaa aggtgagagg ccggaaccgg cttttcatat agaatagaga agcgttcatg
    10081 actaaatgct tgcatcacaa tacttgaagt tgacaatatt atttaaggac ctattgtttt
    10141 ttccaatagg tggttagcaa tcgtcttact ttctaacttt tcttaccttt tacatttcag
    10201 caatatatat atatatttca aggatatacc attctaatgt ctgcccctat gtctgcccct
    10261 aagaagatcg tcgttttgcc aggtgaccac gttggtcaag aaatcacagc cgaagccatt
    10321 aaggttctta aagctatttc tgatgttcgt tccaatgtca agttcgattt cgaaaatcat
    10381 ttaattggtg gtgctgctat cgatgctaca ggtgtcccac ttccagatga ggcgctggaa
    10441 gcctccaaga aggttgatgc cgttttgtta ggtgctgtgg gtggtcctaa atggggtacc
    10501 ggtagtgtta gacctgaaca aggtttacta aaaatccgta aagaacttca attgtacgcc
    10561 aacttaagac catgtaactt tgcatccgac tctcttttag acttatctcc aatcaagcca
    10621 caatttgcta aaggtactga cttcgttgtt gtcagagaat tagtgggagg tatttacttt
    10681 ggtaagagaa aggaagacga tggtgatggt gtcgcttggg atagtgaaca atacaccgtt
    10741 ccagaagtgc aaagaatcac aagaatggcc gctttcatgg ccctacaaca tgagccacca
    10801 ttgcctattt ggtccttgga taaagctaat gttttggcct cttcaagatt atggagaaaa
    10861 actgtggagg aaaccatcaa gaacgagttc cctacattga aggttcaaca tcaattgatt
    10921 gattctgccg ccatgatcct agttaagaac ccaacccacc taaatggtat tataatcacc
    10981 agcaacatgt ttggtgatat catctccgat gaagcctccg ttatcccagg ttccttgggt
    11041 ttgttgccat ctgcgtcctt ggcctctttg ccagacaaga acaccgcatt tggtttgtac
    11101 gaaccatgcc acggttctgc tccagatttg ccaaagaata aggttgaccc tatcgccact
    11161 atcttgtctg ctgcaatgat gttgaaattg tcattgaact tgcctgaaga aggtaaggcc
    11221 attgaagatg cagttaaaaa ggttttggat gcaggtatca gaactggtga tttaggtggt
    11281 tccaacagta ccaccgaagt cggtgatgct gtcgccgaag aagttaagaa aatccttgct
    11341 taaaaagatt ctcttttttt atgatatttg tacataaact ttataaatga aattcataat
    11401 agaaacgaca cgaaattaca aaatggaata tgttcatagg gtagacgaaa ctatatacgc
    11461 aatctacata catttatcaa gaaggagaaa aaggaggata gtaaaggaat acaggtaagc
    11521 aaattgatac taatggctca acgtgataag gaaaaagaat tgcactttaa cattaatatt
    11581 gacaaggagg agggcaccac acaaaaagtt aggtgtaaca gaaaatcatg aaactacgat
    11641 tcctaatttg atattggagg attttctcta aaaaaaaaaa aatacaacaa ataaaaaaca
    11701 ctcaatgacc tgaccatttg atggagttta agtcaatacc ttcttgaacc atttcccata
    11761 atggtgaaag ttccctcaag aattttactc tgtcagaaac ggccttacga cgtagtcgat
    11821 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
    11881 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
    11941 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
    12001 cgcga
//
