LOCUS       sequence-224014-       10663 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Integration Plasmid for galactose inducible Ec86-RT and SpCas9
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10663)
  AUTHORS   Sharon E, Chen SA, Khosla NM, Smith JD, Pritchard JK, Fraser HB
  TITLE     Functional Genetic Variants Revealed by Massively Parallel Precise 
            Genome Editing.
  JOURNAL   Cell. 2018 Sep 18. pii: S0092-8674(18)31118-8. doi: 
            10.1016/j.cell.2018.08.057.
  PUBMED    30245013
REFERENCE   2  (bases 1 to 10663)
  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: "Cell. 2018 
            Sep 18. pii: S0092-8674(18)31118-8. doi: 
            10.1016/j.cell.2018.08.057."
COMMENT     Sequence Label: pZS157 CRISPEY RT/Cas9
FEATURES             Location/Qualifiers
     source          1..10663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     30..52
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(90..108)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        176..280
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(281..349,350..1141)
                     /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(499..518)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1312..1900
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1801..1820
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2054..2071
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2188..2209
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2224..2229,2230..2247,2248..2254)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2262..2278
                     /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     2267..2289
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2286..2302
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2286..2302
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2321..2341
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2323..2341
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      complement(2380..2569)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2581..2599
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(2631..2651)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2664..6767)
                     /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             complement(2664..6767)
                     /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             complement(2664..6767)
                     /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"
     promoter        complement(6800..7241)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(6806..6829)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    7124..7241
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     CDS             7463..7483
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(8676..8695)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8677..8695)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8705..8722)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8705..8721)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8714..8736)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8862..9317
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8949..8968)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9159..9180
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(9577..10239)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(10240..10427)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(10574..10593)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
       61 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
      121 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
      181 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
      241 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
      301 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
      361 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
      421 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
      481 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
      541 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
      601 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
      661 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
      721 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
      781 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
      841 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
      901 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
      961 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     1021 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     1081 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     1141 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     1201 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     1261 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     1321 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     1381 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     1441 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     1501 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     1561 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     1621 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     1681 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     1741 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     1801 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     1861 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     1921 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     1981 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     2041 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     2101 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     2161 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     2221 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     2281 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
     2341 ggaacaaaag ctggagctcc accgcggtga aattaaagcc ttcgagcgtc ccaaaacctt
     2401 ctcaagcaag gttttcagta taatgttaca tgcgtacacg cgtctgtaca gaaaaaaaag
     2461 aaaaatttga aatataaata acgttcttaa tactaacata actataaaaa aataaatagg
     2521 gacctagact tcaggttgtc taactccttc cttttcggtt agagcggatg tggggggagg
     2581 gcgtgaatgt aagcgtgaca taactaatta catgactcga gaagagatca caccttcctc
     2641 ttcttcttgg ggtcagccct gctgtctcca ccgagctgag agaggtcgat tcttgtttca
     2701 tagagccccg taattgactg atgaatcagt gtggcgtcca ggacctcctt tgtagaggtg
     2761 taccgctttc tgtctatggt ggtgtcgaag tacttgaagg ctgcaggcgc gcccaagttg
     2821 gtcagagtaa acaagtggat aatgttttct gcctgctccc tgatgggctt atccctgtgc
     2881 ttattgtaag cagaaagcac cttatcgagg ttagcgtcgg cgaggatcac tcttttggag
     2941 aattcgctta tttgctcgat gatctcatca aggtagtgtt tgtgttgttc cacgaacagc
     3001 tgcttctgct cattatcttc gggagaccct ttgagctttt catagtggct ggccagatac
     3061 aagaaattaa cgtatttaga gggcagtgcc agctcgttac ctttctgcag ctcgcccgca
     3121 ctagcgagca ttcgtttccg gccgttttca agctcaaaga gagagtactt gggaagctta
     3181 atgatgaggt cttttttgac ctctttatat cctttcgcct cgagaaagtc gatggggttt
     3241 ttttcgaagc ttgatcgctc catgattgtg atgcccagca gttccttgac gcttttgagt
     3301 tttttagact tccctttctc cactttggcc acaaccagta cactgtaagc gactgtagga
     3361 gaatcgaatc cgccgtattt cttggggtcc caatcttttt tgcgtgcgat cagcttgtcg
     3421 ctgttccttt tcgggaggat actttccttg gagaagcctc cggtctgtac ttcggtcttt
     3481 ttaacgatgt tcacctgcgg catggacagg accttccgga ctgtcgcgaa atccctaccc
     3541 ttgtcccaca cgatttctcc tgtttctccg tttgtttcga taagtggtcg cttccgaatc
     3601 tctccattgg ccagtgtaat ctcggtcttg aaaaaattca taatattgct gtaaaagaag
     3661 tacttagcgg tggccttgcc tatttcctgc tcagactttg cgatcatttt cctaacatcg
     3721 tacactttat agtctccgta aacaaattca gattcaagct tgggatattt tttgataagt
     3781 gcagtgccta ccactgcatt caggtaggca tcatgcgcat ggtggtaatt gttgatctct
     3841 ctcaccttat aaaactgaaa gtcctttctg aaatctgaga ccagcttaga cttcagagta
     3901 ataactttca cctctcgaat cagtttgtca ttttcatcgt acttggtgtt catgcgtgaa
     3961 tcgagaattt gggccacgtg cttggtgatc tggcgtgtct caacaagctg ccttttgatg
     4021 aagccggctt tatccaactc agacaggcca cctcgttcag ccttagtcag attatcgaac
     4081 ttccgttgtg tgatcagttt ggcgttcagc agctgccgcc aataattttt cattttcttg
     4141 acaacttctt ctgaggggac gttatcactc ttccctctat ttttatcgga tcttgtcaac
     4201 actttattat caatagaatc atctttgaga aaagactggg gcacgatatg atccacgtcg
     4261 tagtcggaga gccgattgat gtccagttcc tgatccacgt acatgtccct gccgttctgc
     4321 aggtagtaca ggtagagctt ctcattctga agctgggtgt tttcaactgg gtgttcctta
     4381 aggatttggg accccagttc ttttataccc tcttcaatcc tcttcatcct ttccctactg
     4441 ttcttctgtc ccttctgggt agtttggttc tctcgggcca tctcgataac gatattctcg
     4501 ggcttatgcc ttcccattac tttgacgagt tcatccacga ccttaacggt ctgcagtatt
     4561 ccctttttga tagctgggct acctgcaaga ttagcgatgt gctcgtgaag actgtccccc
     4621 tggccagaaa cttgtgcttt ctggatgtcc tccttaaagg tgagagagtc atcatggatc
     4681 aactgcatga agttccggtt ggcaaatcca tcggacttaa gaaaatccag gattgtcttt
     4741 ccactctgct tgtctcggat cccattgatc agttttcttg acagccgccc ccatcctgta
     4801 tatcggcgcc tcttgagctg tttcatgact ttgtcgtcga agagatgagc gtaagttttc
     4861 aagcgttctt caatcatctc cctatcttca aacaacgtaa gggtgaggac aatgtcctca
     4921 agaatgtcct cgttctcctc attgtccagg aagtccttgt ctttaatgat tttcaggaga
     4981 tcgtgatacg ttcccaggga tgcgttgaag cgatcctcca ctccgctgat ttcaacagag
     5041 tcgaaacatt caatcttttt gaaatagtct tctttgagct gtttcacggt aactttccgg
     5101 ttcgtcttga agaggaggtc cacgatagct ttcttctgct ctccagacag gaatgctggc
     5161 tttctcatcc cttctgtgac gtatttgacc ttggtgagct cgttataaac tgtgaagtac
     5221 tcgtacagca gagagtgttt aggaagcacc ttttcgttag gcagattttt atcaaagtta
     5281 gtcatccttt cgatgaagga ctgggcagag gcccccttat ccacgacttc ctcgaagttc
     5341 cagggagtga tggtctcttc tgatttgcga gtcatccacg cgaatctgga atttccccgg
     5401 gcgagggggc ctacatagta gggtatccga aatgtgagga ttttctcaat cttttccctg
     5461 ttatctttca aaaaggggta gaaatcctct tgccgcctga ggatagcgtg cagttcgccc
     5521 aggtgaatct ggtgggggat gcttccattg tcgaaagtgc gctgtttgcg caacagatct
     5581 tctctgttaa gctttaccag cagctcctcg gtgccgtcca ttttttccaa gatgggctta
     5641 ataaatttgt aaaattcctc ctggcttgct ccgccgtcaa tgtatccggc gtagccattt
     5701 ttagactgat cgaagaaaat ttccttgtac ttctcaggca gttgctgtct gacaagggcc
     5761 ttcagcaaag tcaagtcttg gtggtgctca tcatagcgct tgatcatact agcgctcagc
     5821 ggagctttgg tgatctccgt gttcactcgc agaatatcac tcagcagaat ggcgtctgac
     5881 aggttctttg ccgccaaaaa aaggtctgcg tactggtcgc cgatctgggc cagcagattg
     5941 tcgagatcat catcgtaggt gtctttgctc agttgaagct tggcatcttc ggccaggtcg
     6001 aagttagatt taaagttggg ggtcagcccg agtgacaggg cgataagatt accaaacagg
     6061 ccgttcttct tctccccagg gagctgtgcg atgaggtttt cgagccgccg ggatttggac
     6121 agcctagcgc tcaggattgc tttggcgtca actccggatg cgttgatcgg gttctcttcg
     6181 aaaagctgat tgtaagtctg aaccagttgg ataaagagtt tgtcgacatc gctgttgtct
     6241 gggttcaggt ccccctcgat gaggaagtgt ccccgaaatt tgatcatatg cgccagcgcg
     6301 agatagatca accgcaagtc agccttatca gtactgtcta caagcttctt cctcagatga
     6361 tatatggttg ggtacttttc atggtacgcc acctcgtcca cgatattgcc aaagattggg
     6421 tggcgctcgt gctttttatc ctcctccacc aaaaaggact cctccagcct atggaagaaa
     6481 gagtcatcca ccttagccat ctcattacta aagatctcct gcaggtagca gatccgattc
     6541 tttctgcggg tatatctgcg ccgtgctgtt cttttgagcc gcgtggcttc ggccgtctcc
     6601 ccggagtcga acaggagggc gccaatgagg ttcttcttta tgctgtggcg atcggtattg
     6661 cccagaactt tgaatttttt gctcggcacc ttgtactcgt ccgtaatgac ggcccaaccg
     6721 acgctgtttg tgccgatatc gagcccaatg gagtacttct tgtccatttt tttcccgggg
     6781 gatccactag ttgttttttc tccttgacgt taaagtatag aggtatatta acaatttttt
     6841 gttgatactt ttatgacatt tgaataagaa gtaatacaaa ctgaaaatgt tgaaagtatt
     6901 agttaaagtg gttatgcagc ttttccattt atatatctgt taatagatca aaaatcatcg
     6961 cttcgctgat taattacccc agaaataagg ctaaaaaact aatcgcatta tcatcctatg
     7021 gttgttaatt tgattcgtta atttgaaggt ttgtggggcc aggttactgc caatttttcc
     7081 tcttcataac cataaaagct agtattgtag aatctttatt gttcggagca gtgcggcgcg
     7141 aggcacatct gcgtttcagg aacgcgaccg gtgaagacga ggacgcacgg aggagagtct
     7201 tccgtcggag ggctgtcgcc cgctcggcgg cttctaatcc gtacttcaat atagcaatga
     7261 gcagttaagc gtattactga aagttccaaa gagaaggttt ttttaggcta agataatggg
     7321 gctctttaca tttccacaac atataagtaa gattagatat ggatatgtat atggtggtaa
     7381 tgccatgtaa tatgattatt aaacttcttt gcgtccatcc aaaaaaaaag taagaatttt
     7441 tgaaaattca atataaatgc caccaaagaa gaaaagaaag gtcaagtccg ctgaatactt
     7501 gaacaccttc agattgagaa atttgggttt gccagtcatg aacaacttgc acgatatgtc
     7561 taaggctacc agaatttctg ttgaaacctt gagattattg atctacaccg ccgatttcag
     7621 atacagaatc tacactgttg aaaagaaggg tccagaaaag agaatgagaa ctatctacca
     7681 accatccaga gaattgaagg ctttacaagg ttgggttttg agaaacatct tggacaagtt
     7741 atcctcctcc ccattttcta tcggttttga aaagcaccaa tccatcttga acaatgccac
     7801 tccacatatt ggtgccaact tcattttgaa catcgacttg gaagatttct tcccatcttt
     7861 gactgctaac aaggttttcg gtgttttcca ttctttgggt tacaacagat tgatctcctc
     7921 tgttttgacc aagatttgct gctacaagaa cttgttgcca caaggtgctc catcttctcc
     7981 aaaattggct aacttgattt gctccaagtt ggactacaga attcaaggtt atgctggttc
     8041 tagaggtttg atatacacta gatacgctga tgacttgacc ttgtctgctc aatctatgaa
     8101 gaaagttgtt aaggccagag acttcttgtt ctccattatt ccatcagaag gtttggtcat
     8161 caactccaaa aagacctgta tttctggtcc aagatcccaa agaaaagtta ccggtttggt
     8221 tatctcccaa gaaaaggttg gtattggtag agaaaagtac aaagaaatca gagccaagat
     8281 ccaccatatt ttctgcggta aatcctccga aatcgaacat gttagaggtt ggttgtcttt
     8341 catcttgtcc gttgattcta agtcccacag aagattgatt acctacatct ctaagttgga
     8401 aaagaagtac ggtaagaacc cattgaacaa ggctaagact tgatttgcca gcttactatc
     8461 cttcttgaaa atatgcactc tatatctttt agttcttaat tgcaacacat agatttgctg
     8521 tataacgaat tttatgctat tttttaaatt tggagttcag tgataaaagt gtcacagcga
     8581 atttcctcac atgtagggac cgaattgttt acaagttctc tgtaccacca tggagacatc
     8641 aaaaattgaa aatctatgga aagatatgca attcgcccta tagtgagtcg tattacgcgc
     8701 gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     8761 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     8821 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc
     8881 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     8941 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     9001 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     9061 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     9121 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     9181 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     9241 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     9301 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     9361 caccgcatag atccgtcgag ttcaagagaa aaaaaaagaa aaagcaaaaa gaaaaaagga
     9421 aagcgcgcct cgttcagaat gacacgtata gaatgatgca ttaccttgtc atcttcagta
     9481 tcatactgtt cgtatacata cttactgaca ttcataggta tacatatata cacatgtata
     9541 tatatcgtat gctgcagctt taaataatcg gtgtcactac ataagaacac ctttggtgga
     9601 gggaacatcg ttggtaccat tgggcgaggt ggcttctctt atggcaaccg caagagcctt
     9661 gaacgcactc tcactacggt gatgatcatt cttgcctcgc agacaatcaa cgtggagggt
     9721 aattctgcta gcctctgcaa agctttcaag aaaatgcggg atcatctcgc aagagagatc
     9781 tcctactttc tccctttgca aaccaagttc gacaactgcg tacggcctgt tcgaaagatc
     9841 taccaccgct ctggaaagtg cctcatccaa aggcgcaaat cctgatccaa acctttttac
     9901 tccacgcacg gcccctaggg cctctttaaa agcttgaccg agagcaatcc cgcagtcttc
     9961 agtggtgtga tggtcgtcta tgtgtaagtc accaatgcac tcaacgatta gcgaccagcc
    10021 ggaatgcttg gccagagcat gtatcatatg gtccagaaac cctatacctg tgtggacgtt
    10081 aatcacttgc gattgtgtgg cctgttctgc tactgcttct gcctcttttt ctgggaagat
    10141 cgagtgctct atcgctaggg gaccaccctt taaagagatc gcaatctgaa tcttggtttc
    10201 atttgtaata cgctttacta gggctttctg ctctgtcatc tttgccttcg tttatcttgc
    10261 ctgctcattt tttagtatat tcttcgaaga aatcacatta ctttatataa tgtataattc
    10321 attatgtgat aatgccaatc gctaagaaaa aaaaagagtc atccgctagg tggaaaaaaa
    10381 aaaatgaaaa tcattaccga ggcataaaaa aatatagagt gtactagagg aggccaagag
    10441 taatagaaaa agaaaattgc gggaaaggac tgtgttatga cttccctgac taatgccgtg
    10501 ttcaaacgat acctggcagt gactcctagc gctcaccaag ctcttaaaac gggaatttat
    10561 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
    10621 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggc
//
