LOCUS       sequence-376550-        9366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Cas9 in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9366)
  AUTHORS   Lopez SC, Crawford KD, Lear SK, Bhattarai-Kline S, Shipman SL
  TITLE     Precise genome editing across kingdoms of life using retron-derived 
            DNA.
  JOURNAL   Nat Chem Biol. 2022 Feb;18(2):199-206. doi: 
            10.1038/s41589-021-00927-y. Epub 2021 Dec 23.
  PUBMED    34949838
REFERENCE   2  (bases 1 to 9366)
  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"; doi: 
            "10.1038/s41589-021-00927-y"; journalName: "Nat Chem Biol"; date: 
            "2022-02"; volume: "18"; issue: "2"; pages: "199-206"
COMMENT     Sequence Label: pSCL.005
FEATURES             Location/Qualifiers
     source          1..9366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      71..526
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(158..177)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     368..389
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(786..1448)
                     /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(1449..1636)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(1783..1802)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1902..1924
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1962..1980)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2048..2152
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2153..2221,2222..3013)
                     /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(2371..2390)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3184..3772
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3673..3692
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3926..3943
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4060..4081
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4096..4101,4102..4119,4120..4126)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4134..4150
                     /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     4139..4161
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4158..4174
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4158..4174
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4193..4213
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4195..4213
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      complement(4252..4441)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     4453..4471
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(4503..4523)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(4536..8639)
                     /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(4536..8639)
                     /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(4536..8639)
                     /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(8672..9113)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(8678..8701)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    8996..9113
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     CDS             9335..9355
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
       61 tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
      121 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
      181 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
      241 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
      301 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
      361 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
      421 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
      481 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt
      541 ttctccttac gcatctgtgc ggtatttcac accgcataga tccgtcgagt tcaagagaaa
      601 aaaaaagaaa aagcaaaaag aaaaaaggaa agcgcgcctc gttcagaatg acacgtatag
      661 aatgatgcat taccttgtca tcttcagtat catactgttc gtatacatac ttactgacat
      721 tcataggtat acatatatac acatgtatat atatcgtatg ctgcagcttt aaataatcgg
      781 tgtcactaca taagaacacc tttggtggag ggaacatcgt tggtaccatt gggcgaggtg
      841 gcttctctta tggcaaccgc aagagccttg aacgcactct cactacggtg atgatcattc
      901 ttgcctcgca gacaatcaac gtggagggta attctgctag cctctgcaaa gctttcaaga
      961 aaatgcggga tcatctcgca agagagatct cctactttct ccctttgcaa accaagttcg
     1021 acaactgcgt acggcctgtt cgaaagatct accaccgctc tggaaagtgc ctcatccaaa
     1081 ggcgcaaatc ctgatccaaa cctttttact ccacgcacgg cccctagggc ctctttaaaa
     1141 gcttgaccga gagcaatccc gcagtcttca gtggtgtgat ggtcgtctat gtgtaagtca
     1201 ccaatgcact caacgattag cgaccagccg gaatgcttgg ccagagcatg tatcatatgg
     1261 tccagaaacc ctatacctgt gtggacgtta atcacttgcg attgtgtggc ctgttctgct
     1321 actgcttctg cctctttttc tgggaagatc gagtgctcta tcgctagggg accacccttt
     1381 aaagagatcg caatctgaat cttggtttca tttgtaatac gctttactag ggctttctgc
     1441 tctgtcatct ttgccttcgt ttatcttgcc tgctcatttt ttagtatatt cttcgaagaa
     1501 atcacattac tttatataat gtataattca ttatgtgata atgccaatcg ctaagaaaaa
     1561 aaaagagtca tccgctaggt ggaaaaaaaa aaatgaaaat cattaccgag gcataaaaaa
     1621 atatagagtg tactagagga ggccaagagt aatagaaaaa gaaaattgcg ggaaaggact
     1681 gtgttatgac ttccctgact aatgccgtgt tcaaacgata cctggcagtg actcctagcg
     1741 ctcaccaagc tcttaaaacg ggaatttatg gtgcactctc agtacaatct gctctgatgc
     1801 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     1861 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     1921 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     1981 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     2041 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2101 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     2161 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     2221 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     2281 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     2341 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     2401 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     2461 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     2521 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     2581 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     2641 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     2701 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     2761 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     2821 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     2881 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2941 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3001 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     3061 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3121 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3181 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3241 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3301 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3361 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3421 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3481 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3541 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3601 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3661 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3721 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3781 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3841 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3901 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     3961 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     4021 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     4081 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     4141 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
     4201 ccctcactaa agggaacaaa agctggagct ccaccgcggt gaaattaaag ccttcgagcg
     4261 tcccaaaacc ttctcaagca aggttttcag tataatgtta catgcgtaca cgcgtctgta
     4321 cagaaaaaaa agaaaaattt gaaatataaa taacgttctt aatactaaca taactataaa
     4381 aaaataaata gggacctaga cttcaggttg tctaactcct tccttttcgg ttagagcgga
     4441 tgtgggggga gggcgtgaat gtaagcgtga cataactaat tacatgactc gagaagagat
     4501 cacaccttcc tcttcttctt ggggtcagcc ctgctgtctc caccgagctg agagaggtcg
     4561 attcttgttt catagagccc cgtaattgac tgatgaatca gtgtggcgtc caggacctcc
     4621 tttgtagagg tgtaccgctt tctgtctatg gtggtgtcga agtacttgaa ggctgcaggc
     4681 gcgcccaagt tggtcagagt aaacaagtgg ataatgtttt ctgcctgctc cctgatgggc
     4741 ttatccctgt gcttattgta agcagaaagc accttatcga ggttagcgtc ggcgaggatc
     4801 actcttttgg agaattcgct tatttgctcg atgatctcat caaggtagtg tttgtgttgt
     4861 tccacgaaca gctgcttctg ctcattatct tcgggagacc ctttgagctt ttcatagtgg
     4921 ctggccagat acaagaaatt aacgtattta gagggcagtg ccagctcgtt acctttctgc
     4981 agctcgcccg cactagcgag cattcgtttc cggccgtttt caagctcaaa gagagagtac
     5041 ttgggaagct taatgatgag gtcttttttg acctctttat atcctttcgc ctcgagaaag
     5101 tcgatggggt ttttttcgaa gcttgatcgc tccatgattg tgatgcccag cagttccttg
     5161 acgcttttga gttttttaga cttccctttc tccactttgg ccacaaccag tacactgtaa
     5221 gcgactgtag gagaatcgaa tccgccgtat ttcttggggt cccaatcttt tttgcgtgcg
     5281 atcagcttgt cgctgttcct tttcgggagg atactttcct tggagaagcc tccggtctgt
     5341 acttcggtct ttttaacgat gttcacctgc ggcatggaca ggaccttccg gactgtcgcg
     5401 aaatccctac ccttgtccca cacgatttct cctgtttctc cgtttgtttc gataagtggt
     5461 cgcttccgaa tctctccatt ggccagtgta atctcggtct tgaaaaaatt cataatattg
     5521 ctgtaaaaga agtacttagc ggtggccttg cctatttcct gctcagactt tgcgatcatt
     5581 ttcctaacat cgtacacttt atagtctccg taaacaaatt cagattcaag cttgggatat
     5641 tttttgataa gtgcagtgcc taccactgca ttcaggtagg catcatgcgc atggtggtaa
     5701 ttgttgatct ctctcacctt ataaaactga aagtcctttc tgaaatctga gaccagctta
     5761 gacttcagag taataacttt cacctctcga atcagtttgt cattttcatc gtacttggtg
     5821 ttcatgcgtg aatcgagaat ttgggccacg tgcttggtga tctggcgtgt ctcaacaagc
     5881 tgccttttga tgaagccggc tttatccaac tcagacaggc cacctcgttc agccttagtc
     5941 agattatcga acttccgttg tgtgatcagt ttggcgttca gcagctgccg ccaataattt
     6001 ttcattttct tgacaacttc ttctgagggg acgttatcac tcttccctct atttttatcg
     6061 gatcttgtca acactttatt atcaatagaa tcatctttga gaaaagactg gggcacgata
     6121 tgatccacgt cgtagtcgga gagccgattg atgtccagtt cctgatccac gtacatgtcc
     6181 ctgccgttct gcaggtagta caggtagagc ttctcattct gaagctgggt gttttcaact
     6241 gggtgttcct taaggatttg ggaccccagt tcttttatac cctcttcaat cctcttcatc
     6301 ctttccctac tgttcttctg tcccttctgg gtagtttggt tctctcgggc catctcgata
     6361 acgatattct cgggcttatg ccttcccatt actttgacga gttcatccac gaccttaacg
     6421 gtctgcagta ttcccttttt gatagctggg ctacctgcaa gattagcgat gtgctcgtga
     6481 agactgtccc cctggccaga aacttgtgct ttctggatgt cctccttaaa ggtgagagag
     6541 tcatcatgga tcaactgcat gaagttccgg ttggcaaatc catcggactt aagaaaatcc
     6601 aggattgtct ttccactctg cttgtctcgg atcccattga tcagttttct tgacagccgc
     6661 ccccatcctg tatatcggcg cctcttgagc tgtttcatga ctttgtcgtc gaagagatga
     6721 gcgtaagttt tcaagcgttc ttcaatcatc tccctatctt caaacaacgt aagggtgagg
     6781 acaatgtcct caagaatgtc ctcgttctcc tcattgtcca ggaagtcctt gtctttaatg
     6841 attttcagga gatcgtgata cgttcccagg gatgcgttga agcgatcctc cactccgctg
     6901 atttcaacag agtcgaaaca ttcaatcttt ttgaaatagt cttctttgag ctgtttcacg
     6961 gtaactttcc ggttcgtctt gaagaggagg tccacgatag ctttcttctg ctctccagac
     7021 aggaatgctg gctttctcat cccttctgtg acgtatttga ccttggtgag ctcgttataa
     7081 actgtgaagt actcgtacag cagagagtgt ttaggaagca ccttttcgtt aggcagattt
     7141 ttatcaaagt tagtcatcct ttcgatgaag gactgggcag aggccccctt atccacgact
     7201 tcctcgaagt tccagggagt gatggtctct tctgatttgc gagtcatcca cgcgaatctg
     7261 gaatttcccc gggcgagggg gcctacatag tagggtatcc gaaatgtgag gattttctca
     7321 atcttttccc tgttatcttt caaaaagggg tagaaatcct cttgccgcct gaggatagcg
     7381 tgcagttcgc ccaggtgaat ctggtggggg atgcttccat tgtcgaaagt gcgctgtttg
     7441 cgcaacagat cttctctgtt aagctttacc agcagctcct cggtgccgtc cattttttcc
     7501 aagatgggct taataaattt gtaaaattcc tcctggcttg ctccgccgtc aatgtatccg
     7561 gcgtagccat ttttagactg atcgaagaaa atttccttgt acttctcagg cagttgctgt
     7621 ctgacaaggg ccttcagcaa agtcaagtct tggtggtgct catcatagcg cttgatcata
     7681 ctagcgctca gcggagcttt ggtgatctcc gtgttcactc gcagaatatc actcagcaga
     7741 atggcgtctg acaggttctt tgccgccaaa aaaaggtctg cgtactggtc gccgatctgg
     7801 gccagcagat tgtcgagatc atcatcgtag gtgtctttgc tcagttgaag cttggcatct
     7861 tcggccaggt cgaagttaga tttaaagttg ggggtcagcc cgagtgacag ggcgataaga
     7921 ttaccaaaca ggccgttctt cttctcccca gggagctgtg cgatgaggtt ttcgagccgc
     7981 cgggatttgg acagcctagc gctcaggatt gctttggcgt caactccgga tgcgttgatc
     8041 gggttctctt cgaaaagctg attgtaagtc tgaaccagtt ggataaagag tttgtcgaca
     8101 tcgctgttgt ctgggttcag gtccccctcg atgaggaagt gtccccgaaa tttgatcata
     8161 tgcgccagcg cgagatagat caaccgcaag tcagccttat cagtactgtc tacaagcttc
     8221 ttcctcagat gatatatggt tgggtacttt tcatggtacg ccacctcgtc cacgatattg
     8281 ccaaagattg ggtggcgctc gtgcttttta tcctcctcca ccaaaaagga ctcctccagc
     8341 ctatggaaga aagagtcatc caccttagcc atctcattac taaagatctc ctgcaggtag
     8401 cagatccgat tctttctgcg ggtatatctg cgccgtgctg ttcttttgag ccgcgtggct
     8461 tcggccgtct ccccggagtc gaacaggagg gcgccaatga ggttcttctt tatgctgtgg
     8521 cgatcggtat tgcccagaac tttgaatttt ttgctcggca ccttgtactc gtccgtaatg
     8581 acggcccaac cgacgctgtt tgtgccgata tcgagcccaa tggagtactt cttgtccatt
     8641 tttttcccgg gggatccact agttgttttt tctccttgac gttaaagtat agaggtatat
     8701 taacaatttt ttgttgatac ttttatgaca tttgaataag aagtaataca aactgaaaat
     8761 gttgaaagta ttagttaaag tggttatgca gcttttccat ttatatatct gttaatagat
     8821 caaaaatcat cgcttcgctg attaattacc ccagaaataa ggctaaaaaa ctaatcgcat
     8881 tatcatccta tggttgttaa tttgattcgt taatttgaag gtttgtgggg ccaggttact
     8941 gccaattttt cctcttcata accataaaag ctagtattgt agaatcttta ttgttcggag
     9001 cagtgcggcg cgaggcacat ctgcgtttca ggaacgcgac cggtgaagac gaggacgcac
     9061 ggaggagagt cttccgtcgg agggctgtcg cccgctcggc ggcttctaat ccgtacttca
     9121 atatagcaat gagcagttaa gcgtattact gaaagttcca aagagaaggt ttttttaggc
     9181 taagataatg gggctcttta catttccaca acatataagt aagattagat atggatatgt
     9241 atatggtggt aatgccatgt aatatgatta ttaaacttct ttgcgtccat ccaaaaaaaa
     9301 agtaagaatt tttgaaaatt caatataaat gccaccaaag aagaaaagaa aggtcaagtc
     9361 cgctga
//
