LOCUS       sequence-151412-       11379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCas9-SH3 in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11379)
  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 11379)
  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
FEATURES             Location/Qualifiers
     source          1..11379
                     /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"
     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"
     terminator      7959..8146
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(8014..8038)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     terminator      complement(8183..8228)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(8264..8283)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8265..8283)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8290..8307)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8290..8306)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8299..8321)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8451..8906
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8538..8557)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8748..8769
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9206..9610
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             9623..10717
                     /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(11210..11229)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11329..11351
                     /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 gctcggaatt
     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 tagaggctcc ggataaggat cctaataagc gaatttctta tgatttatga
     7981 tttttattat taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt
     8041 aggttttaaa acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt
     8101 ctcaggtata gcatgaggtc gctcttattg accacacctc taccggcatg ccgagcaaat
     8161 gcctgcaaat cgctcccggg caaaaaaccc cctcaagacc cgtttagagg ccccaagggg
     8221 ttatgctatg cataccaagc ttccgcggtg gagctccaat tcgccctata gtgagtcgta
     8281 ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     8341 acttaatcgc cttgcagcac atcccccctt cgccagctgg cgtaatagcg aagaggcccg
     8401 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcg acgcgccctg
     8461 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     8521 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     8581 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     8641 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     8701 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     8761 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     8821 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     8881 taacaaaata ttaacgttta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     8941 ggtatttcac accgcatatc gaccctcgag gagaacttct agtatatcca catacctaat
     9001 attattgcct tattaaaaat ggaatcggaa caattacatc aaaatccaca ttctcttcaa
     9061 aatcaattgt cctgtacttc cttgttcatg tgtgttcaaa aacgttatat ttataggata
     9121 attatactct atttctcaac aagtaattgg ttgtttggcc gagcggtcta aggcgcctga
     9181 ttcaagaaat atcttgaccg cagttaactg tgggaatact caggtatcgt aagatgcaag
     9241 agttcgaatc tcttagcaac cattattttt ttcctcaaca taacgagaac acacaggggc
     9301 gctatcgcac agaatcaaat tcgatgactg gaaatttttt gttaatttca gaggtcgcct
     9361 gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga gaggccggaa
     9421 ccggcttttc atatagaata gagaagcgtt catgactaaa tgcttgcatc acaatacttg
     9481 aagttgacaa tattatttaa ggacctattg ttttttccaa taggtggtta gcaatcgtct
     9541 tactttctaa cttttcttac cttttacatt tcagcaatat atatatatat ttcaaggata
     9601 taccattcta atgtctgccc ctatgtctgc ccctaagaag atcgtcgttt tgccaggtga
     9661 ccacgttggt caagaaatca cagccgaagc cattaaggtt cttaaagcta tttctgatgt
     9721 tcgttccaat gtcaagttcg atttcgaaaa tcatttaatt ggtggtgctg ctatcgatgc
     9781 tacaggtgtc ccacttccag atgaggcgct ggaagcctcc aagaaggttg atgccgtttt
     9841 gttaggtgct gtgggtggtc ctaaatgggg taccggtagt gttagacctg aacaaggttt
     9901 actaaaaatc cgtaaagaac ttcaattgta cgccaactta agaccatgta actttgcatc
     9961 cgactctctt ttagacttat ctccaatcaa gccacaattt gctaaaggta ctgacttcgt
    10021 tgttgtcaga gaattagtgg gaggtattta ctttggtaag agaaaggaag acgatggtga
    10081 tggtgtcgct tgggatagtg aacaatacac cgttccagaa gtgcaaagaa tcacaagaat
    10141 ggccgctttc atggccctac aacatgagcc accattgcct atttggtcct tggataaagc
    10201 taatgttttg gcctcttcaa gattatggag aaaaactgtg gaggaaacca tcaagaacga
    10261 gttccctaca ttgaaggttc aacatcaatt gattgattct gccgccatga tcctagttaa
    10321 gaacccaacc cacctaaatg gtattataat caccagcaac atgtttggtg atatcatctc
    10381 cgatgaagcc tccgttatcc caggttcctt gggtttgttg ccatctgcgt ccttggcctc
    10441 tttgccagac aagaacaccg catttggttt gtacgaacca tgccacggtt ctgctccaga
    10501 tttgccaaag aataaggttg accctatcgc cactatcttg tctgctgcaa tgatgttgaa
    10561 attgtcattg aacttgcctg aagaaggtaa ggccattgaa gatgcagtta aaaaggtttt
    10621 ggatgcaggt atcagaactg gtgatttagg tggttccaac agtaccaccg aagtcggtga
    10681 tgctgtcgcc gaagaagtta agaaaatcct tgcttaaaaa gattctcttt ttttatgata
    10741 tttgtacata aactttataa atgaaattca taatagaaac gacacgaaat tacaaaatgg
    10801 aatatgttca tagggtagac gaaactatat acgcaatcta catacattta tcaagaagga
    10861 gaaaaaggag gatagtaaag gaatacaggt aagcaaattg atactaatgg ctcaacgtga
    10921 taaggaaaaa gaattgcact ttaacattaa tattgacaag gaggagggca ccacacaaaa
    10981 agttaggtgt aacagaaaat catgaaacta cgattcctaa tttgatattg gaggattttc
    11041 tctaaaaaaa aaaaaataca acaaataaaa aacactcaat gacctgacca tttgatggag
    11101 tttaagtcaa taccttcttg aaccatttcc cataatggtg aaagttccct caagaatttt
    11161 actctgtcag aaacggcctt acgacgtagt cgatatggtg cactctcagt acaatctgct
    11221 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
    11281 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
    11341 tgtgtcagag gttttcaccg tcatcaccga aacgcgcga
//
