LOCUS       sequence-358227-       11975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Galactose-Induced Expression vector for Cas9 and I-SceI..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11975)
  AUTHORS   Shively CA, Dong F, Mitra RD
  TITLE     A Suite of New Strain Construction Vectors for Gene Expression 
            Knockdown in Budding Yeast.
  JOURNAL   ACS Synth Biol. 2023 Jan 17. doi: 10.1021/acssynbio.2c00547.
  PUBMED    36650116
REFERENCE   2  (bases 1 to 11975)
  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: "ACS Synth 
            Biol. 2023 Jan 17. doi: 10.1021/acssynbio.2c00547."
COMMENT     Sequence Label: Gal1/10p CAS9 Gal1/10p I-SceI
FEATURES             Location/Qualifiers
     source          1..11975
                     /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"
     CDS             complement(2317..2337)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(2363..2804)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2687..2804
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     rep_origin      2939..3527
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3428..3447
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3681..3698
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3815..3836
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3851..3856,3857..3874,3875..3881)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3889..3905
                     /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     3894..3916
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3913..3929
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3913..3929
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3948..3968
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3950..3968
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     4421..4437
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4421..4437
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             4451..8554
                     /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             4451..8554
                     /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             4451..8554
                     /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             8567..8587
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      8603..8850
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(8619..8637)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(8868..8887)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8869..8887)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8897..8914)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8897..8913)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8906..8928)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9054..9509
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9141..9160)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9351..9372
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9810..10217
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             10218..11312
                     /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(11806..11825)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11925..11947
                     /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 tttatttcag
     1621 gaaagtttcg gaggagatag tgttcggcag tttgtacatc atctgcggga tcaggtacgg
     1681 tttgatcagg ttgtagaaga tcaggtaaga catagaatcg atgtagatga tcggtttgtt
     1741 tttgttgatt tttacgtaac agttcagttg gaatttgtta cgcagaccct taaccaggta
     1801 ttctacttct tcgaaagtga aagactgggt gttcagtacg atcgatttgt tggtagagtt
     1861 tttgttgtaa tcccatttac caccatcatc catgaaccag tatgccagag acatcggggt
     1921 caggtagttt tcaaccaggt tgttcgggat ggtttttttg ttgttaacga tgaacaggtt
     1981 agccagtttg ttgaaagctt ggtgtttgaa agtctgggcg ccccaggtga ttaccaggtt
     2041 acccaggtgg ttaacacgtt cttttttgtg cggcggggac agtacccact gatcgtacag
     2101 cagacatacg tggtccatgt atgctttgtt tttccactcg aactgcatac agtaggtttt
     2161 accttcatca cgagaacgga tgtaagcatc acccaggatc agaccgatac ctgcttcgaa
     2221 ctgttcgatg ttcagttcga tcagctggga tttgtattct ttcagcagtt tagagttcgg
     2281 acccaggttc attacctggt tttttttgat gtttttaacc tttctcttct tttttggcat
     2341 atgcatggat ccggggtttt ttctccttga cgttaaagta tagaggtata ttaacaattt
     2401 tttgttgata cttttattac atttgaataa gaagtaatac aaaccgaaaa tgttgaaagt
     2461 attagttaaa gtggttatgc agtttttgca tttatatatc tgttaataga tcaaaaatca
     2521 tcgcttcgct gattaattac cccagaaata aggctaaaaa actaatcgca ttatcatcct
     2581 atggttgtta atttgattcg ttcatttgaa ggtttgtggg gccaggttac tgccaatttt
     2641 tcctcttcat aaccataaaa gctagtattg tagaatcttt attgttcgga gcagtgcggc
     2701 gcgaggcaca tctgcgtttc aggaacgcga ccggtgaaga cgaggacgca cggaggagag
     2761 tcttccttcg gagggctgtc acccgctcgg cggcttctaa tccggattta aaacttcatt
     2821 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     2881 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2941 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3001 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3061 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3121 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3181 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3241 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3301 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3361 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3421 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3481 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3541 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3601 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3661 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     3721 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     3781 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     3841 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     3901 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
     3961 actaaaggga acaaaagctg gagctcgtga cagccctccg aaggaagact ctcctccgtg
     4021 cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg tgcctcgcgc cgcactgctc
     4081 cgaacaataa agattctaca atactagctt ttatggttat gaagaggaaa aattggcagt
     4141 aacctggccc cacaaacctt caaatgaacg aatcaaatta acaaccatag gatgataatg
     4201 cgattagttt tttagcctta tttctggggt aattaatcag cgaagcgatg atttttgatc
     4261 tattaacgga tatataaatg caaaaactgc ataaccactt taactaatac tttcaacatt
     4321 ttcggtttgt attacttctt attcaaatgt aataaaagta tcaacaaaaa attgttaata
     4381 tacctctata ctttaacgtc aaggagaaaa aaccccggat tctagaacta gtggatcccc
     4441 cgggaaaaaa atggacaaga agtactccat tgggctcgat atcggcacaa acagcgtcgg
     4501 ttgggccgtc attacggacg agtacaaggt gccgagcaaa aaattcaaag ttctgggcaa
     4561 taccgatcgc cacagcataa agaagaacct cattggcgcc ctcctgttcg actccgggga
     4621 gacggccgaa gccacgcggc tcaaaagaac agcacggcgc agatataccc gcagaaagaa
     4681 tcggatctgc tacctgcagg agatctttag taatgagatg gctaaggtgg atgactcttt
     4741 cttccatagg ctggaggagt cctttttggt ggaggaggat aaaaagcacg agcgccaccc
     4801 aatctttggc aatatcgtgg acgaggtggc gtaccatgaa aagtacccaa ccatatatca
     4861 tctgaggaag aagcttgtag acagtactga taaggctgac ttgcggttga tctatctcgc
     4921 gctggcgcat atgatcaaat ttcggggaca cttcctcatc gagggggacc tgaacccaga
     4981 caacagcgat gtcgacaaac tctttatcca actggttcag acttacaatc agcttttcga
     5041 agagaacccg atcaacgcat ccggagttga cgccaaagca atcctgagcg ctaggctgtc
     5101 caaatcccgg cggctcgaaa acctcatcgc acagctccct ggggagaaga agaacggcct
     5161 gtttggtaat cttatcgccc tgtcactcgg gctgaccccc aactttaaat ctaacttcga
     5221 cctggccgaa gatgccaagc ttcaactgag caaagacacc tacgatgatg atctcgacaa
     5281 tctgctggcc cagatcggcg accagtacgc agaccttttt ttggcggcaa agaacctgtc
     5341 agacgccatt ctgctgagtg atattctgcg agtgaacacg gagatcacca aagctccgct
     5401 gagcgctagt atgatcaagc gctatgatga gcaccaccaa gacttgactt tgctgaaggc
     5461 ccttgtcaga cagcaactgc ctgagaagta caaggaaatt ttcttcgatc agtctaaaaa
     5521 tggctacgcc ggatacattg acggcggagc aagccaggag gaattttaca aatttattaa
     5581 gcccatcttg gaaaaaatgg acggcaccga ggagctgctg gtaaagctta acagagaaga
     5641 tctgttgcgc aaacagcgca ctttcgacaa tggaagcatc ccccaccaga ttcacctggg
     5701 cgaactgcac gctatcctca ggcggcaaga ggatttctac ccctttttga aagataacag
     5761 ggaaaagatt gagaaaatcc tcacatttcg gataccctac tatgtaggcc ccctcgcccg
     5821 gggaaattcc agattcgcgt ggatgactcg caaatcagaa gagaccatca ctccctggaa
     5881 cttcgaggaa gtcgtggata agggggcctc tgcccagtcc ttcatcgaaa ggatgactaa
     5941 ctttgataaa aatctgccta acgaaaaggt gcttcctaaa cactctctgc tgtacgagta
     6001 cttcacagtt tataacgagc tcaccaaggt caaatacgtc acagaaggga tgagaaagcc
     6061 agcattcctg tctggagagc agaagaaagc tatcgtggac ctcctcttca agacgaaccg
     6121 gaaagttacc gtgaaacagc tcaaagaaga ctatttcaaa aagattgaat gtttcgactc
     6181 tgttgaaatc agcggagtgg aggatcgctt caacgcatcc ctgggaacgt atcacgatct
     6241 cctgaaaatc attaaagaca aggacttcct ggacaatgag gagaacgagg acattcttga
     6301 ggacattgtc ctcaccctta cgttgtttga agatagggag atgattgaag aacgcttgaa
     6361 aacttacgct catctcttcg acgacaaagt catgaaacag ctcaagaggc gccgatatac
     6421 aggatggggg cggctgtcaa gaaaactgat caatgggatc cgagacaagc agagtggaaa
     6481 gacaatcctg gattttctta agtccgatgg atttgccaac cggaacttca tgcagttgat
     6541 ccatgatgac tctctcacct ttaaggagga catccagaaa gcacaagttt ctggccaggg
     6601 ggacagtctt cacgagcaca tcgctaatct tgcaggtagc ccagctatca aaaagggaat
     6661 actgcagacc gttaaggtcg tggatgaact cgtcaaagta atgggaaggc ataagcccga
     6721 gaatatcgtt atcgagatgg cccgagagaa ccaaactacc cagaagggac agaagaacag
     6781 tagggaaagg atgaagagga ttgaagaggg tataaaagaa ctggggtccc aaatccttaa
     6841 ggaacaccca gttgaaaaca cccagcttca gaatgagaag ctctacctgt actacctgca
     6901 gaacggcagg gacatgtacg tggatcagga actggacatc aatcggctct ccgactacga
     6961 cgtggatcat atcgtgcccc agtcttttct caaagatgat tctattgata ataaagtgtt
     7021 gacaagatcc gataaaaata gagggaagag tgataacgtc ccctcagaag aagttgtcaa
     7081 gaaaatgaaa aattattggc ggcagctgct gaacgccaaa ctgatcacac aacggaagtt
     7141 cgataatctg actaaggctg aacgaggtgg cctgtctgag ttggataaag ccggcttcat
     7201 caaaaggcag cttgttgaga cacgccagat caccaagcac gtggcccaaa ttctcgattc
     7261 acgcatgaac accaagtacg atgaaaatga caaactgatt cgagaggtga aagttattac
     7321 tctgaagtct aagctggtct cagatttcag aaaggacttt cagttttata aggtgagaga
     7381 gatcaacaat taccaccatg cgcatgatgc ctacctgaat gcagtggtag gcactgcact
     7441 tatcaaaaaa tatcccaagc ttgaatctga atttgtttac ggagactata aagtgtacga
     7501 tgttaggaaa atgatcgcaa agtctgagca ggaaataggc aaggccaccg ctaagtactt
     7561 cttttacagc aatattatga attttttcaa gaccgagatt acactggcca atggagagat
     7621 tcggaagcga ccacttatcg aaacaaacgg agaaacagga gaaatcgtgt gggacaaggg
     7681 tagggatttc gcgacagtcc ggaaggtcct gtccatgccg caggtgaaca tcgttaaaaa
     7741 gaccgaagta cagaccggag gcttctccaa ggaaagtatc ctcccgaaaa ggaacagcga
     7801 caagctgatc gcacgcaaaa aagattggga ccccaagaaa tacggcggat tcgattctcc
     7861 tacagtcgct tacagtgtac tggttgtggc caaagtggag aaagggaagt ctaaaaaact
     7921 caaaagcgtc aaggaactgc tgggcatcac aatcatggag cgatcaagct tcgaaaaaaa
     7981 ccccatcgac tttctcgagg cgaaaggata taaagaggtc aaaaaagacc tcatcattaa
     8041 gcttcccaag tactctctct ttgagcttga aaacggccgg aaacgaatgc tcgctagtgc
     8101 gggcgagctg cagaaaggta acgagctggc actgccctct aaatacgtta atttcttgta
     8161 tctggccagc cactatgaaa agctcaaagg gtctcccgaa gataatgagc agaagcagct
     8221 gttcgtggaa caacacaaac actaccttga tgagatcatc gagcaaataa gcgaattctc
     8281 caaaagagtg atcctcgccg acgctaacct cgataaggtg ctttctgctt acaataagca
     8341 cagggataag cccatcaggg agcaggcaga aaacattatc cacttgttta ctctgaccaa
     8401 cttgggcgcg cctgcagcct tcaagtactt cgacaccacc atagacagaa agcggtacac
     8461 ctctacaaag gaggtcctgg acgccacact gattcatcag tcaattacgg ggctctatga
     8521 aacaagaatc gacctctctc agctcggtgg agacagcagg gctgacccca agaagaagag
     8581 gaaggtgtga tctcttctcg agtcatgtaa ttagttatgt cacgcttaca ttcacgccct
     8641 ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
     8701 atttattttt ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     8761 ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag
     8821 gttttgggac gctcgaaggc tttaatttgc ggccggtacc caattcgccc tatagtgagt
     8881 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     8941 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     9001 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc
     9061 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     9121 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     9181 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     9241 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     9301 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     9361 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     9421 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     9481 ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
     9541 tgcggtattt cacaccgcat atcgaccggt cgaggagaac ttctagtata tctacatacc
     9601 taatattatt gccttattaa aaatggaatc ccaacaatta catcaaaatc cacattctct
     9661 tcaaaatcaa ttgtcctgta cttccttgtt catgtgtgtt caaaaacgtt atatttatag
     9721 gataattata ctctatttct caacaagtaa ttggttgttt ggccgagcgg tctaaggcgc
     9781 ctgattcaag aaatatcttg accgcagtta actgtgggaa tactcaggta tcgtaagatg
     9841 caagagttcg aatctcttag caaccattat ttttttcctc aacataacga gaacacacag
     9901 gggcgctatc gcacagaatc aaattcgatg actggaaatt ttttgttaat ttcagaggtc
     9961 gcctgacgca tatacctttt tcaactgaaa aattgggaga aaaaggaaag gtgagagcgc
    10021 cggaaccggc ttttcatata gaatagagaa gcgttcatga ctaaatgctt gcatcacaat
    10081 acttgaagtt gacaatatta tttaaggacc tattgttttt tccaataggt ggttagcaat
    10141 cgtcttactt tctaactttt cttacctttt acatttcagc aatatatata tatatatttc
    10201 aaggatatac cattctaatg tctgccccta agaagatcgt cgttttgcca ggtgaccacg
    10261 ttggtcaaga aatcacagcc gaagccatta aggttcttaa agctatttct gatgttcgtt
    10321 ccaatgtcaa gttcgatttc gaaaatcatt taattggtgg tgctgctatc gatgctacag
    10381 gtgttccact tccagatgag gcgctggaag cctccaagaa ggctgatgcc gttttgttag
    10441 gtgctgtggg tggtcctaaa tggggtaccg gtagtgttag acctgaacaa ggtttactaa
    10501 aaatccgtaa agaacttcaa ttgtacgcca acttaagacc atgtaacttt gcatccgact
    10561 ctcttttaga cttatctcca atcaagccac aatttgctaa aggtactgac ttcgttgttg
    10621 tcagagaatt agtgggaggt atttactttg gtaagagaaa ggaagacgat ggtgatggtg
    10681 tcgcttggga tagtgaacaa tacaccgttc cagaagtgca aagaatcaca agaatggccg
    10741 ctttcatggc cctacaacat gagccaccat tgcctatttg gtccttggat aaagctaatg
    10801 ttttggcctc ttcaagatta tggagaaaaa ctgtggagga aaccatcaag aacgaattcc
    10861 ctacattgaa ggttcaacat caattgattg attctgccgc catgatccta gttaagaacc
    10921 caacccacct aaatggtatt ataatcacca gcaacatgtt tggtgatatc atctccgatg
    10981 aagcctccgt tatcccaggt tccttgggtt tgttgccatc tgcgtccttg gcctctttgc
    11041 cagacaagaa caccgcattt ggtttgtacg aaccatgcca cggttctgct ccagatttgc
    11101 caaagaataa ggtcaaccct atcgccacta tcttgtctgc tgcaatgatg ttgaaattgt
    11161 cattgaactt gcctgaagaa ggtaaggcca ttgaagatgc agttaaaaag gttttggatg
    11221 caggtatcag aactggtgat ttaggtggtt ccaacagtac caccgaagtc ggtgatgctg
    11281 tcgccgaaga agttaagaaa atccttgctt aaaaagattc tcttttttta tgatatttgt
    11341 acataaactt tataaatgaa attcataata gaaacgacac gaaattacaa aatggaatat
    11401 gttcataggg tagacgaaac tatatacgca atctacatac atttatcaag aaggagaaaa
    11461 aggaggatgt aaaggaatac aggtaagcaa attgatacta atggctcaac gtgataagga
    11521 aaaagaattg cactttaaca ttaatattga caaggaggag ggcaccacac aaaaagttag
    11581 gtgtaacaga aaatcatgaa actatgattc ctaatttata tattggagga ttttctctaa
    11641 aaaaaaaaaa atacaacaaa taaaaaacac tcaatgacct gaccatttga tggagtttaa
    11701 gtcaatacct tcttgaacca tttcccataa tggtgaaagt tccctcaaga attttactct
    11761 gtcagaaacg gccttaacga cgtagtcgat atggtgcact ctcagtacaa tctgctctga
    11821 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
    11881 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
    11941 tcagaggttt tcaccgtcat caccgaaacg cgcga
//
