LOCUS       sequence-157872-       12405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRS416 (URA) + RPR1(TetO) promoter with gRNA locus + Tetracycline 
            Repressor + GalL promoter driven Cas9 for yeast expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12405)
  AUTHORS   Fu BX, St Onge RP, Fire AZ, Smith JD
  TITLE     Distinct patterns of Cas9 mismatch tolerance in vitro and in vivo.
  JOURNAL   Nucleic Acids Res. 2016 May 19. pii: gkw417.
  PUBMED    27198218
REFERENCE   2  (bases 1 to 12405)
  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: "Nucleic 
            Acids Res. 2016 May 19. pii: gkw417."
COMMENT     Sequence Label: pRS416gT-GalL-Cas9
FEATURES             Location/Qualifiers
     source          1..12405
                     /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
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     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     2235..2254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2729..2747
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        2820..2895
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      2908..2970
                     /label=RPR1 terminator
                     /note="transcription terminator for the S. cerevisiae RNase
                     P RNA gene"
     CDS             4103..4726
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(4176..4195)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     5223..5240
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5357..5378
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5393..5398,5399..5416,5417..5423)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5431..5447
                     /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     5436..5458
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5455..5471
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5455..5471
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5490..5510
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5492..5510
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     5919..5942
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     5963..5979
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5963..5979
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             5993..10096
                     /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             5993..10096
                     /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             5993..10096
                     /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             10109..10129
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      10145..10392
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(10161..10179)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(10410..10429)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10411..10429)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10439..10456)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10439..10455)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10448..10470)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10600..11055
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10687..10706)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10897..10918
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(11186..11989)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(11990..12210)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(12236..12255)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12355..12377
                     /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 tttgcacaac 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 gggagtcagg 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 cgtgagctat gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctgg ccttttgctc
     2401 gggggatctg ccaattgaac ataacatggt agttacatat actagtaata tggttcggca
     2461 cacattaaaa gtataaaaac tatctgaatt acgaattaca tatattggtc ataaaaatca
     2521 atcaatcatc gtgtgtttta tatgtctctt atctaagtat aagaatatcc atagttaata
     2581 ttcacttacg ctacctttta acctgtaatc attgtcaaca ggatatgtta acgacccaca
     2641 ttgataaacg ctagtatttc tttttcctct tcttattggc cggctgtctc tatactcccc
     2701 tatagtctgt ttcttttcgt ttcgattgtc cctatcagtg atagagatgg cgcacatggt
     2761 acgctgtggt gctcgcggct gggaacgaaa ctctgggagc tgcgattggc agcggccgcg
     2821 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
     2881 gcaccgagtc ggtgcttttt tctcgagcca tatccaactt ccaatttaat ctttcttttt
     2941 taattttcac ttatttgcga tacagaaaga aaaaagcgat agtaactatt gaattttgtt
     3001 tggatttggt tagattagat atggtttctc tttatattta catgctaaaa atgggctaca
     3061 ccagagatac ataattagat atatatacgc cagtacacct tatcggccca agccttgtcc
     3121 caaggcagcg ttttgttctt ggaaacgctg ccctacacgt tcgctatgct tcaagaactt
     3181 ttctgagcac ttcatgatgc atgtttgttc cttattggtt agctttgatg ttgtgaagtc
     3241 attgacacag tctgtgaaac atctttctac cagattagag tacaaacgca tgaaatcctt
     3301 catttgcttt tgttccacta ctttttggaa ctcttgttgt tctttgctag ctagtcgtgc
     3361 aatgtatgac tttaagattt gtgagcagga agaaaaggga gaatcttcta acgataaacc
     3421 cttgaaaaac tgggtagact acgctatgtt gagttgctac gcaggctgca caattacacg
     3481 agaatgctcc cgcctaggat ttaaggctaa gggacgtgca atgcagacga cagatctaaa
     3541 tgaccgtgtc ggtgaagtgt tcgccaaact tttcggttaa cacatgcagt gatgcacgcg
     3601 cgatggtgct aagttacata tatatatata tgtatagcca tagtgatgtc taagtaacct
     3661 ttatggtata tttcttaatg tggaaagata ctagcgcgcg cacccacaca caagcttcgt
     3721 cttttcttga agaaaagagg aagctcgcta aatgggattc cactttccgt tccctgccag
     3781 ctgatggaaa aaggttagtg gaacgatgaa gaataaaaag agagatccac tgaggtgaaa
     3841 tttcagctga cagcgagttt catgatcgtg atgaacaatg gtaacgagtt gtggctgttg
     3901 ccagggaggg tggttctcaa cttttaatgt atggccaaat cgctacttgg gtttgttata
     3961 taacaaagaa gaaataatga actgattctc ttcctccttc ttgtcctttc ttaattctgt
     4021 tgtaattacc ttcctttgta attttttttg taattattct tcttaataat ccaaacaaac
     4081 acacatatta caatatatta aaatgtctag attagataaa agtaaagtga ttaacagcgc
     4141 attagagctg cttaatgagg tcggaatcga aggtttaaca acccgtaaac tcgcccagaa
     4201 gctaggtgta gagcagccta cattgtattg gcatgtaaaa aataagcggg ctttgctcga
     4261 cgccttagcc attgagatgt tagataggca ccatactcac ttttgccctt tagaagggga
     4321 aagctggcaa gattttttac gtaataacgc taaaagtttt agatgtgctt tactaagtca
     4381 tcgcgatgga gcaaaagtac atttaggtac acggcctaca gaaaaacagt atgaaactct
     4441 cgaaaatcaa ttagcctttt tatgccaaca aggtttttca ctagagaatg cattatatgc
     4501 actcagcgct gtggggcatt ttactttagg ttgcgtattg gaagatcaag agcatcaagt
     4561 cgctaaagaa gaaagggaaa cacctactac tgatagtatg ccgccattat tacgacaagc
     4621 tatcgaatta tttgatcacc aaggtgcaga gccagccttc ttattcggcc ttgaattgat
     4681 catatgcgga ttagaaaaac aacttaaatg tgaaagtggg tcttaagtct gaagaatgaa
     4741 tgatttgatg atttcttttt ccctccattt ttcttactga atatatcaat gatatagact
     4801 tgtatagttt attatttcaa attaagtagc tatatatagt caagataacg tttgtttgac
     4861 acgattacat tattcgtcga catctttttt cagcctgtcg tggtagcaat ttgaggagta
     4921 ttattaattg aataggttca ttttgcgctc gcataaacag ttttcgtcag ggacagtatg
     4981 ttggaatgag tggtaattaa tggtgacatg acatgttata gcaataacct tgatgtttac
     5041 atcgtagttt aatgtacacc ccgcgaattc gttcaagtag gagtgcacca attgcaaagg
     5101 gaaaagctga atgggcagtt cgaaccgcgg tctttcctgc gttatcccct gattctgtgg
     5161 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     5221 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     5281 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     5341 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     5401 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     5461 acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc
     5521 tggagctcgt gacagccctc cgaaggaaga ctctcctccg tgcgtcctcg tcttcaccgg
     5581 tcgcgttcct gaaacgcaga tgtgcctcgc gccgcactgc tccgaacaat aaagattcta
     5641 caatactagc ttttatggtt atgaagagga aaaattggca gtaacctggc cccacaaacc
     5701 ttcaaatgaa cgaatcaaat taacaaccat aggatgataa tgcgattagt tttttagcct
     5761 tatttctggg gtaattaatc agcgaagcga tgatttttga tctattaacg gatatataaa
     5821 tgcaaaaact gcataaccac tttaactaat actttcaaca ttttcggttt gtattacttc
     5881 ttattcaaat gtaataaaag tatcaacaaa aaattgttaa tatacctcta tactttaacg
     5941 tcaaggagaa aaaaccccgg attctagaac tagtggatcc cccgggaaaa aaatggacaa
     6001 gaagtactcc attgggctcg atatcggcac aaacagcgtc ggttgggccg tcattacgga
     6061 cgagtacaag gtgccgagca aaaaattcaa agttctgggc aataccgatc gccacagcat
     6121 aaagaagaac ctcattggcg ccctcctgtt cgactccggg gagacggccg aagccacgcg
     6181 gctcaaaaga acagcacggc gcagatatac ccgcagaaag aatcggatct gctacctgca
     6241 ggagatcttt agtaatgaga tggctaaggt ggatgactct ttcttccata ggctggagga
     6301 gtcctttttg gtggaggagg ataaaaagca cgagcgccac ccaatctttg gcaatatcgt
     6361 ggacgaggtg gcgtaccatg aaaagtaccc aaccatatat catctgagga agaagcttgt
     6421 agacagtact gataaggctg acttgcggtt gatctatctc gcgctggcgc atatgatcaa
     6481 atttcgggga cacttcctca tcgaggggga cctgaaccca gacaacagcg atgtcgacaa
     6541 actctttatc caactggttc agacttacaa tcagcttttc gaagagaacc cgatcaacgc
     6601 atccggagtt gacgccaaag caatcctgag cgctaggctg tccaaatccc ggcggctcga
     6661 aaacctcatc gcacagctcc ctggggagaa gaagaacggc ctgtttggta atcttatcgc
     6721 cctgtcactc gggctgaccc ccaactttaa atctaacttc gacctggccg aagatgccaa
     6781 gcttcaactg agcaaagaca cctacgatga tgatctcgac aatctgctgg cccagatcgg
     6841 cgaccagtac gcagaccttt ttttggcggc aaagaacctg tcagacgcca ttctgctgag
     6901 tgatattctg cgagtgaaca cggagatcac caaagctccg ctgagcgcta gtatgatcaa
     6961 gcgctatgat gagcaccacc aagacttgac tttgctgaag gcccttgtca gacagcaact
     7021 gcctgagaag tacaaggaaa ttttcttcga tcagtctaaa aatggctacg ccggatacat
     7081 tgacggcgga gcaagccagg aggaatttta caaatttatt aagcccatct tggaaaaaat
     7141 ggacggcacc gaggagctgc tggtaaagct taacagagaa gatctgttgc gcaaacagcg
     7201 cactttcgac aatggaagca tcccccacca gattcacctg ggcgaactgc acgctatcct
     7261 caggcggcaa gaggatttct accccttttt gaaagataac agggaaaaga ttgagaaaat
     7321 cctcacattt cggataccct actatgtagg ccccctcgcc cggggaaatt ccagattcgc
     7381 gtggatgact cgcaaatcag aagagaccat cactccctgg aacttcgagg aagtcgtgga
     7441 taagggggcc tctgcccagt ccttcatcga aaggatgact aactttgata aaaatctgcc
     7501 taacgaaaag gtgcttccta aacactctct gctgtacgag tacttcacag tttataacga
     7561 gctcaccaag gtcaaatacg tcacagaagg gatgagaaag ccagcattcc tgtctggaga
     7621 gcagaagaaa gctatcgtgg acctcctctt caagacgaac cggaaagtta ccgtgaaaca
     7681 gctcaaagaa gactatttca aaaagattga atgtttcgac tctgttgaaa tcagcggagt
     7741 ggaggatcgc ttcaacgcat ccctgggaac gtatcacgat ctcctgaaaa tcattaaaga
     7801 caaggacttc ctggacaatg aggagaacga ggacattctt gaggacattg tcctcaccct
     7861 tacgttgttt gaagataggg agatgattga agaacgcttg aaaacttacg ctcatctctt
     7921 cgacgacaaa gtcatgaaac agctcaagag gcgccgatat acaggatggg ggcggctgtc
     7981 aagaaaactg atcaatggga tccgagacaa gcagagtgga aagacaatcc tggattttct
     8041 taagtccgat ggatttgcca accggaactt catgcagttg atccatgatg actctctcac
     8101 ctttaaggag gacatccaga aagcacaagt ttctggccag ggggacagtc ttcacgagca
     8161 catcgctaat cttgcaggta gcccagctat caaaaaggga atactgcaga ccgttaaggt
     8221 cgtggatgaa ctcgtcaaag taatgggaag gcataagccc gagaatatcg ttatcgagat
     8281 ggcccgagag aaccaaacta cccagaaggg acagaagaac agtagggaaa ggatgaagag
     8341 gattgaagag ggtataaaag aactggggtc ccaaatcctt aaggaacacc cagttgaaaa
     8401 cacccagctt cagaatgaga agctctacct gtactacctg cagaacggca gggacatgta
     8461 cgtggatcag gaactggaca tcaatcggct ctccgactac gacgtggatc atatcgtgcc
     8521 ccagtctttt ctcaaagatg attctattga taataaagtg ttgacaagat ccgataaaaa
     8581 tagagggaag agtgataacg tcccctcaga agaagttgtc aagaaaatga aaaattattg
     8641 gcggcagctg ctgaacgcca aactgatcac acaacggaag ttcgataatc tgactaaggc
     8701 tgaacgaggt ggcctgtctg agttggataa agccggcttc atcaaaaggc agcttgttga
     8761 gacacgccag atcaccaagc acgtggccca aattctcgat tcacgcatga acaccaagta
     8821 cgatgaaaat gacaaactga ttcgagaggt gaaagttatt actctgaagt ctaagctggt
     8881 ctcagatttc agaaaggact ttcagtttta taaggtgaga gagatcaaca attaccacca
     8941 tgcgcatgat gcctacctga atgcagtggt aggcactgca cttatcaaaa aatatcccaa
     9001 gcttgaatct gaatttgttt acggagacta taaagtgtac gatgttagga aaatgatcgc
     9061 aaagtctgag caggaaatag gcaaggccac cgctaagtac ttcttttaca gcaatattat
     9121 gaattttttc aagaccgaga ttacactggc caatggagag attcggaagc gaccacttat
     9181 cgaaacaaac ggagaaacag gagaaatcgt gtgggacaag ggtagggatt tcgcgacagt
     9241 ccggaaggtc ctgtccatgc cgcaggtgaa catcgttaaa aagaccgaag tacagaccgg
     9301 aggcttctcc aaggaaagta tcctcccgaa aaggaacagc gacaagctga tcgcacgcaa
     9361 aaaagattgg gaccccaaga aatacggcgg attcgattct cctacagtcg cttacagtgt
     9421 actggttgtg gccaaagtgg agaaagggaa gtctaaaaaa ctcaaaagcg tcaaggaact
     9481 gctgggcatc acaatcatgg agcgatcaag cttcgaaaaa aaccccatcg actttctcga
     9541 ggcgaaagga tataaagagg tcaaaaaaga cctcatcatt aagcttccca agtactctct
     9601 ctttgagctt gaaaacggcc ggaaacgaat gctcgctagt gcgggcgagc tgcagaaagg
     9661 taacgagctg gcactgccct ctaaatacgt taatttcttg tatctggcca gccactatga
     9721 aaagctcaaa gggtctcccg aagataatga gcagaagcag ctgttcgtgg aacaacacaa
     9781 acactacctt gatgagatca tcgagcaaat aagcgaattc tccaaaagag tgatcctcgc
     9841 cgacgctaac ctcgataagg tgctttctgc ttacaataag cacagggata agcccatcag
     9901 ggagcaggca gaaaacatta tccacttgtt tactctgacc aacttgggcg cgcctgcagc
     9961 cttcaagtac ttcgacacca ccatagacag aaagcggtac acctctacaa aggaggtcct
    10021 ggacgccaca ctgattcatc agtcaattac ggggctctat gaaacaagaa tcgacctctc
    10081 tcagctcggt ggagacagca gggctgaccc caagaagaag aggaaggtgt gatctcttct
    10141 cgagtcatgt aattagttat gtcacgctta cattcacgcc ctccccccac atccgctcta
    10201 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt
    10261 atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc
    10321 gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag
    10381 gctttaattt gcggccggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac
    10441 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
    10501 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
    10561 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcga cgcgccctgt agcggcgcat
    10621 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
    10681 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
    10741 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
    10801 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
    10861 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
    10921 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
    10981 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
    11041 taacgtttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
    11101 ccgcataggg taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac
    11161 atgcatttac ttataataca gttttttagt tttgctggcc gcatcttctc aaatatgctt
    11221 cccagcctgc ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata
    11281 gtcctcttcc aacaataata atgtcagatc ctgtagagac cacatcatcc acggttctat
    11341 actgttgacc caatgcgtct cccttgtcat ctaaacccac accgggtgtc ataatcaacc
    11401 aatcgtaacc ttcatctctt ccacccatgt ctctttgagc aataaagccg ataacaaaat
    11461 ctttgtcgct cttcgcaatg tcaacagtac ccttagtata ttctccagta gatagggagc
    11521 ccttgcatga caattctgct aacatcaaaa ggcctctagg ttcctttgtt acttcttctg
    11581 ccgcctgctt caaaccgcta acaatacctg ggcccaccac accgtgtgca ttcgtaatgt
    11641 ctgcccattc tgctattctg tatacacccg cagagtactg caatttgact gtattaccaa
    11701 tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta cttggcggat aatgccttta
    11761 gcggcttaac tgtgccctcc atggaaaaat cagtcaagat atccacatgt gtttttagta
    11821 aacaaatttt gggacctaat gcttcaacta actccagtaa ttccttggtg gtacgaacat
    11881 ccaatgaagc acacaagttt gtttgctttt cgtgcatgat attaaatagc ttggcagcaa
    11941 caggactagg atgagtagca gcacgttcct tatatgtagc tttcgacatg atttatcttc
    12001 gtttcctgca ggtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt
    12061 cttcaacact acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt
    12121 ccttctgttc ggagattacc gaatcaaaaa aatttcaaag aaaccgaaat caaaaaaaag
    12181 aataaaaaaa aaatgatgaa ttgaattgaa aagctgtggt atggtgcact ctcagtacaa
    12241 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
    12301 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
    12361 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcga
//
