LOCUS       sequence-337029-       10687 bp    DNA     circular SYN 14-APR-2025
DEFINITION  pRS415-TEF1p-Cas9-CYC1t; Cas9 expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10687)
  AUTHORS   Xie ZX, Mitchell LA, Liu HM, Li BZ, Liu D, Agmon N, Wu Y, Li X, Zhou
            X, Li B, Xiao WH, Ding MZ, Wang Y, Yuan YJ, Boeke JD
  TITLE     Rapid and Efficient CRISPR/Cas9-Based Mating-Type Switching of 
            Saccharomyces cerevisiae.
  JOURNAL   G3 (Bethesda). 2018 Jan 4;8(1):173-183. doi: 10.1534/g3.117.300347.
  PUBMED    29150593
REFERENCE   2  (bases 1 to 10687)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1534/g3.117"; 
            journalName: "G3 (Bethesda)"; date: "2018-01-4- 4"; volume: "8"; 
            issue: "1"; pages: "173-183"
COMMENT     Sequence Label: pNA0306
FEATURES             Location/Qualifiers
     source          1..10687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        79..183
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(184..252,253..1044)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(402..421)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1215..1803
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1704..1723
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1957..1974
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2091..2112
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2127..2132,2133..2150,2151..2157)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2165..2181
                     /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     2170..2192
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2189..2205
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2189..2205
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2224..2244
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2226..2244
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2262..2660
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     2664..2680
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2664..2680
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2692..6795
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2692..6795
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2692..6795
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6808..6828
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6844..7091
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6860..6878)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(7109..7128)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7110..7128)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7138..7155)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7138..7154)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7147..7169)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7295..7750
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7382..7401)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7592..7613
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8051..8458
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             8459..9553
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(10047..10066)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(10166..10184)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    join(10226..10687,1..42)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 tattaaaaaa tttaaattat aattattttt atagcacgtg atgaaaagga cccaggtggc
       61 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
      121 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
      181 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
      241 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
      301 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
      361 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
      421 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
      481 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
      541 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
      601 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
      661 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
      721 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
      781 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
      841 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtgga
      901 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
      961 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     1021 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     1081 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     1141 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     1201 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     1261 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     1321 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     1381 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     1441 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     1501 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     1561 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     1621 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     1681 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     1741 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     1801 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     1861 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     1921 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     1981 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     2041 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     2101 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     2161 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     2221 cgcgcaatta accctcacta aagggaacaa aagctggagc tcatagcttc aaaatgtttc
     2281 tactcctttt ttactcttcc agattttctc ggactccgcg catcgccgta ccacttcaaa
     2341 acacccaagc acagcatact aaatttcccc tctttcttcc tctagggtgt cgttaattac
     2401 ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc ttcgtcgaaa
     2461 aaggcaataa aaatttttat cacgtttctt tttcttgaaa attttttttt tgattttttt
     2521 ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcttcaat ttctcaagtt
     2581 tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat tagaaagaaa
     2641 gcatagcaat ctaatctaag ttttctagaa ctagtggatc ccccgggaaa aatggacaag
     2701 aagtactcca ttgggctcga tatcggcaca aacagcgtcg gttgggccgt cattacggac
     2761 gagtacaagg tgccgagcaa aaaattcaaa gttctgggca ataccgatcg ccacagcata
     2821 aagaagaacc tcattggcgc cctcctgttc gactccgggg agacggccga agccacgcgg
     2881 ctcaaaagaa cagcacggcg cagatatacc cgcagaaaga atcggatctg ctacctgcag
     2941 gagatcttta gtaatgagat ggctaaggtg gatgactctt tcttccatag gctggaggag
     3001 tcctttttgg tggaggagga taaaaagcac gagcgccacc caatctttgg caatatcgtg
     3061 gacgaggtgg cgtaccatga aaagtaccca accatatatc atctgaggaa gaagcttgta
     3121 gacagtactg ataaggctga cttgcggttg atctatctcg cgctggcgca tatgatcaaa
     3181 tttcggggac acttcctcat cgagggggac ctgaacccag acaacagcga tgtcgacaaa
     3241 ctctttatcc aactggttca gacttacaat cagcttttcg aagagaaccc gatcaacgca
     3301 tccggagttg acgccaaagc aatcctgagc gctaggctgt ccaaatcccg gcggctcgaa
     3361 aacctcatcg cacagctccc tggggagaag aagaacggcc tgtttggtaa tcttatcgcc
     3421 ctgtcactcg ggctgacccc caactttaaa tctaacttcg acctggccga agatgccaag
     3481 cttcaactga gcaaagacac ctacgatgat gatctcgaca atctgctggc ccagatcggc
     3541 gaccagtacg cagacctttt tttggcggca aagaacctgt cagacgccat tctgctgagt
     3601 gatattctgc gagtgaacac ggagatcacc aaagctccgc tgagcgctag tatgatcaag
     3661 cgctatgatg agcaccacca agacttgact ttgctgaagg cccttgtcag acagcaactg
     3721 cctgagaagt acaaggaaat tttcttcgat cagtctaaaa atggctacgc cggatacatt
     3781 gacggcggag caagccagga ggaattttac aaatttatta agcccatctt ggaaaaaatg
     3841 gacggcaccg aggagctgct ggtaaagctt aacagagaag atctgttgcg caaacagcgc
     3901 actttcgaca atggaagcat cccccaccag attcacctgg gcgaactgca cgctatcctc
     3961 aggcggcaag aggatttcta cccctttttg aaagataaca gggaaaagat tgagaaaatc
     4021 ctcacatttc ggatacccta ctatgtaggc cccctcgccc ggggaaattc cagattcgcg
     4081 tggatgactc gcaaatcaga agagaccatc actccctgga acttcgagga agtcgtggat
     4141 aagggggcct ctgcccagtc cttcatcgaa aggatgacta actttgataa aaatctgcct
     4201 aacgaaaagg tgcttcctaa acactctctg ctgtacgagt acttcacagt ttataacgag
     4261 ctcaccaagg tcaaatacgt cacagaaggg atgagaaagc cagcattcct gtctggagag
     4321 cagaagaaag ctatcgtgga cctcctcttc aagacgaacc ggaaagttac cgtgaaacag
     4381 ctcaaagaag actatttcaa aaagattgaa tgtttcgact ctgttgaaat cagcggagtg
     4441 gaggatcgct tcaacgcatc cctgggaacg tatcacgatc tcctgaaaat cattaaagac
     4501 aaggacttcc tggacaatga ggagaacgag gacattcttg aggacattgt cctcaccctt
     4561 acgttgtttg aagataggga gatgattgaa gaacgcttga aaacttacgc tcatctcttc
     4621 gacgacaaag tcatgaaaca gctcaagagg cgccgatata caggatgggg gcggctgtca
     4681 agaaaactga tcaatgggat ccgagacaag cagagtggaa agacaatcct ggattttctt
     4741 aagtccgatg gatttgccaa ccggaacttc atgcagttga tccatgatga ctctctcacc
     4801 tttaaggagg acatccagaa agcacaagtt tctggccagg gggacagtct tcacgagcac
     4861 atcgctaatc ttgcaggtag cccagctatc aaaaagggaa tactgcagac cgttaaggtc
     4921 gtggatgaac tcgtcaaagt aatgggaagg cataagcccg agaatatcgt tatcgagatg
     4981 gcccgagaga accaaactac ccagaaggga cagaagaaca gtagggaaag gatgaagagg
     5041 attgaagagg gtataaaaga actggggtcc caaatcctta aggaacaccc agttgaaaac
     5101 acccagcttc agaatgagaa gctctacctg tactacctgc agaacggcag ggacatgtac
     5161 gtggatcagg aactggacat caatcggctc tccgactacg acgtggatca tatcgtgccc
     5221 cagtcttttc tcaaagatga ttctattgat aataaagtgt tgacaagatc cgataaaaat
     5281 agagggaaga gtgataacgt cccctcagaa gaagttgtca agaaaatgaa aaattattgg
     5341 cggcagctgc tgaacgccaa actgatcaca caacggaagt tcgataatct gactaaggct
     5401 gaacgaggtg gcctgtctga gttggataaa gccggcttca tcaaaaggca gcttgttgag
     5461 acacgccaga tcaccaagca cgtggcccaa attctcgatt cacgcatgaa caccaagtac
     5521 gatgaaaatg acaaactgat tcgagaggtg aaagttatta ctctgaagtc taagctggtc
     5581 tcagatttca gaaaggactt tcagttttat aaggtgagag agatcaacaa ttaccaccat
     5641 gcgcatgatg cctacctgaa tgcagtggta ggcactgcac ttatcaaaaa atatcccaag
     5701 cttgaatctg aatttgttta cggagactat aaagtgtacg atgttaggaa aatgatcgca
     5761 aagtctgagc aggaaatagg caaggccacc gctaagtact tcttttacag caatattatg
     5821 aattttttca agaccgagat tacactggcc aatggagaga ttcggaagcg accacttatc
     5881 gaaacaaacg gagaaacagg agaaatcgtg tgggacaagg gtagggattt cgcgacagtc
     5941 cggaaggtcc tgtccatgcc gcaggtgaac atcgttaaaa agaccgaagt acagaccgga
     6001 ggcttctcca aggaaagtat cctcccgaaa aggaacagcg acaagctgat cgcacgcaaa
     6061 aaagattggg accccaagaa atacggcgga ttcgattctc ctacagtcgc ttacagtgta
     6121 ctggttgtgg ccaaagtgga gaaagggaag tctaaaaaac tcaaaagcgt caaggaactg
     6181 ctgggcatca caatcatgga gcgatcaagc ttcgaaaaaa accccatcga ctttctcgag
     6241 gcgaaaggat ataaagaggt caaaaaagac ctcatcatta agcttcccaa gtactctctc
     6301 tttgagcttg aaaacggccg gaaacgaatg ctcgctagtg cgggcgagct gcagaaaggt
     6361 aacgagctgg cactgccctc taaatacgtt aatttcttgt atctggccag ccactatgaa
     6421 aagctcaaag ggtctcccga agataatgag cagaagcagc tgttcgtgga acaacacaaa
     6481 cactaccttg atgagatcat cgagcaaata agcgaattct ccaaaagagt gatcctcgcc
     6541 gacgctaacc tcgataaggt gctttctgct tacaataagc acagggataa gcccatcagg
     6601 gagcaggcag aaaacattat ccacttgttt actctgacca acttgggcgc gcctgcagcc
     6661 ttcaagtact tcgacaccac catagacaga aagcggtaca cctctacaaa ggaggtcctg
     6721 gacgccacac tgattcatca gtcaattacg gggctctatg aaacaagaat cgacctctct
     6781 cagctcggtg gagacagcag ggctgacccc aagaagaaga ggaaggtgtg atctcttctc
     6841 gagtcatgta attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa
     6901 ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta
     6961 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg
     7021 tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg
     7081 ctttaatttg cggccggtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     7141 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     7201 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     7261 ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc
     7321 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     7381 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     7441 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     7501 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     7561 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     7621 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     7681 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     7741 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     7801 tatcgaccgg tcgaggagaa cttctagtat atctacatac ctaatattat tgccttatta
     7861 aaaatggaat cccaacaatt acatcaaaat ccacattctc ttcaaaatca attgtcctgt
     7921 acttccttgt tcatgtgtgt tcaaaaacgt tatatttata ggataattat actctatttc
     7981 tcaacaagta attggttgtt tggccgagcg gtctaaggcg cctgattcaa gaaatatctt
     8041 gaccgcagtt aactgtggga atactcaggt atcgtaagat gcaagagttc gaatctctta
     8101 gcaaccatta tttttttcct caacataacg agaacacaca ggggcgctat cgcacagaat
     8161 caaattcgat gactggaaat tttttgttaa tttcagaggt cgcctgacgc atataccttt
     8221 ttcaactgaa aaattgggag aaaaaggaaa ggtgagagcg ccggaaccgg cttttcatat
     8281 agaatagaga agcgttcatg actaaatgct tgcatcacaa tacttgaagt tgacaatatt
     8341 atttaaggac ctattgtttt ttccaatagg tggttagcaa tcgtcttact ttctaacttt
     8401 tcttaccttt tacatttcag caatatatat atatatattt caaggatata ccattctaat
     8461 gtctgcccct aagaagatcg tcgttttgcc aggtgaccac gttggtcaag aaatcacagc
     8521 cgaagccatt aaggttctta aagctatttc tgatgttcgt tccaatgtca agttcgattt
     8581 cgaaaatcat ttaattggtg gtgctgctat cgatgctaca ggtgttccac ttccagatga
     8641 ggcgctggaa gcctccaaga aggctgatgc cgttttgtta ggtgctgtgg gtggtcctaa
     8701 atggggtacc ggtagtgtta gacctgaaca aggtttacta aaaatccgta aagaacttca
     8761 attgtacgcc aacttaagac catgtaactt tgcatccgac tctcttttag acttatctcc
     8821 aatcaagcca caatttgcta aaggtactga cttcgttgtt gtcagagaat tagtgggagg
     8881 tatttacttt ggtaagagaa aggaagacga tggtgatggt gtcgcttggg atagtgaaca
     8941 atacaccgtt ccagaagtgc aaagaatcac aagaatggcc gctttcatgg ccctacaaca
     9001 tgagccacca ttgcctattt ggtccttgga taaagctaat gttttggcct cttcaagatt
     9061 atggagaaaa actgtggagg aaaccatcaa gaacgaattc cctacattga aggttcaaca
     9121 tcaattgatt gattctgccg ccatgatcct agttaagaac ccaacccacc taaatggtat
     9181 tataatcacc agcaacatgt ttggtgatat catctccgat gaagcctccg ttatcccagg
     9241 ttccttgggt ttgttgccat ctgcgtcctt ggcctctttg ccagacaaga acaccgcatt
     9301 tggtttgtac gaaccatgcc acggttctgc tccagatttg ccaaagaata aggtcaaccc
     9361 tatcgccact atcttgtctg ctgcaatgat gttgaaattg tcattgaact tgcctgaaga
     9421 aggtaaggcc attgaagatg cagttaaaaa ggttttggat gcaggtatca gaactggtga
     9481 tttaggtggt tccaacagta ccaccgaagt cggtgatgct gtcgccgaag aagttaagaa
     9541 aatccttgct taaaaagatt ctcttttttt atgatatttg tacataaact ttataaatga
     9601 aattcataat agaaacgaca cgaaattaca aaatggaata tgttcatagg gtagacgaaa
     9661 ctatatacgc aatctacata catttatcaa gaaggagaaa aaggaggatg taaaggaata
     9721 caggtaagca aattgatact aatggctcaa cgtgataagg aaaaagaatt gcactttaac
     9781 attaatattg acaaggagga gggcaccaca caaaaagtta ggtgtaacag aaaatcatga
     9841 aactatgatt cctaatttat atattggagg attttctcta aaaaaaaaaa aatacaacaa
     9901 ataaaaaaca ctcaatgacc tgaccatttg atggagttta agtcaatacc ttcttgaacc
     9961 atttcccata atggtgaaag ttccctcaag aattttactc tgtcagaaac ggccttaacg
    10021 acgtagtcga tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
    10081 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
    10141 gcttacagac aagctgtgac aaagggcctc gtgatacgcc tatttttata ggttaatgtc
    10201 atgataataa tggtttctta ggacggatcg cttgcctgta acttacacgc gcctcgtatc
    10261 ttttaatgat ggaataattt gggaatttac tctgtgttta tttattttta tgttttgtat
    10321 ttggatttta gaaagtaaat aaagaaggta gaagagttac ggaatgaaga aaaaaaaata
    10381 aacaaaggtt taaaaaattt caacaaaaag cgtactttac atatatattt attagacaag
    10441 aaaagcagat taaatagata tacattcgat taacgataag taaaatgtaa aatcacagga
    10501 ttttcgtgtg tggtcttcta cacagacaag atgaaacaat tcggcattaa tacctgagag
    10561 caggaagagc aagataaaag gtagtatttg ttggcgatcc ccctagagtc ttttacatct
    10621 tcggaaaaca aaaactattt tttctttaat ttcttttttt actttctatt tttaatttat
    10681 atattta
//
