LOCUS       sequence-308874-       10360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Galactose-inducible expression of Cas9 for addressing one target; 
            Contains guide RNA expression cassette with stuffer and KpnI-Pme1 
            restriction sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10360)
  AUTHORS   Degreif D, Kremenovic M, Geiger T, Bertl A
  TITLE     Preloading budding yeast with all-in-one CRISPR/Cas9 vectors for 
            easy and high-efficient genome editing.
  JOURNAL   J Biol Methods. 2018 Sep 12;5(3):e98. doi: 10.14440/jbm.2018.254. 
            eCollection 2018.
  PUBMED    31453248
REFERENCE   2  (bases 1 to 10360)
  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: "J Biol 
            Methods."; date: "2018-09-12"; volume: "5(3):e98. doi"; pages: " 
            10.14440/jbm.2018.254. eCollection 2018"
COMMENT     Sequence Label: pCAS9i
FEATURES             Location/Qualifiers
     source          1..10360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(32..100,101..892)
                     /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(250..269)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1063..1651
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1552..1571
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1805..1822
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1939..1960
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1975..1980,1981..1998,1999..2005)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2013..2029
                     /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     2018..2040
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2037..2053
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2037..2053
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2072..2092
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2074..2092
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2113..2554
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2113..2230
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2525..2548
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2582..6685
                     /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             2582..6685
                     /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             2582..6685
                     /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             6698..6718
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6734..6981
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6750..6768)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        7024..7239
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             7240..8043
                     /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"
     rep_origin      8382..9724
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(9345..9392)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        9785..10053
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        10092..10166
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      10171..10190
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     promoter        join(10287..10360,1..31)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
       61 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
      121 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
      181 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
      241 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
      301 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
      361 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
      421 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
      481 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
      541 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
      601 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
      661 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
      721 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
      781 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
      841 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
      901 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
      961 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     1021 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1081 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1141 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     1201 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     1261 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     1321 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     1381 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     1441 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     1501 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     1561 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     1621 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     1681 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     1741 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     1801 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     1861 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     1921 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     1981 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     2041 aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa
     2101 gctggagctg tacggattag aagccgccga gcgggtgaca gccctccgaa ggaagactct
     2161 cctccgtgcg tcctcgtctt caccggtcgc gttcctgaaa cgcagatgtg cctcgcgccg
     2221 cactgctccg aacaataaag attctacaat actagctttt atggttatga agaggaaaaa
     2281 ttggcagtaa cctggcccca caaaccttca aatgaacgaa tcaaattaac aaccatagga
     2341 tgataatgcg attagttttt tagccttatt tctggggtaa ttaatcagcg aagcgatgat
     2401 ttttgatcta ttaacagata tataaatgca aaaactgcat aaccacttta actaatactt
     2461 tcaacatttt cggtttgtat tacttcttat tcaaatgtaa taaaagtatc aacaaaaaat
     2521 tgttaatata cctctatact ttaacgtcaa ggagaaaaaa ccccgggatc ccccgggaaa
     2581 aatggacaag aagtactcca ttgggctcga tatcggcaca aacagcgtcg gttgggccgt
     2641 cattacggac gagtacaagg tgccgagcaa aaaattcaaa gttctgggca ataccgatcg
     2701 ccacagcata aagaagaacc tcattggcgc cctcctgttc gactccgggg agacggccga
     2761 agccacgcgg ctcaaaagaa cagcacggcg cagatatacc cgcagaaaga atcggatctg
     2821 ctacctgcag gagatcttta gtaatgagat ggctaaggtg gatgactctt tcttccatag
     2881 gctggaggag tcctttttgg tggaggagga taaaaagcac gagcgccacc caatctttgg
     2941 caatatcgtg gacgaggtgg cgtaccatga aaagtaccca accatatatc atctgaggaa
     3001 gaagcttgta gacagtactg ataaggctga cttgcggttg atctatctcg cgctggcgca
     3061 tatgatcaaa tttcggggac acttcctcat cgagggggac ctgaacccag acaacagcga
     3121 tgtcgacaaa ctctttatcc aactggttca gacttacaat cagcttttcg aagagaaccc
     3181 gatcaacgca tccggagttg acgccaaagc aatcctgagc gctaggctgt ccaaatcccg
     3241 gcggctcgaa aacctcatcg cacagctccc tggggagaag aagaacggcc tgtttggtaa
     3301 tcttatcgcc ctgtcactcg ggctgacccc caactttaaa tctaacttcg acctggccga
     3361 agatgccaag cttcaactga gcaaagacac ctacgatgat gatctcgaca atctgctggc
     3421 ccagatcggc gaccagtacg cagacctttt tttggcggca aagaacctgt cagacgccat
     3481 tctgctgagt gatattctgc gagtgaacac ggagatcacc aaagctccgc tgagcgctag
     3541 tatgatcaag cgctatgatg agcaccacca agacttgact ttgctgaagg cccttgtcag
     3601 acagcaactg cctgagaagt acaaggaaat tttcttcgat cagtctaaaa atggctacgc
     3661 cggatacatt gacggcggag caagccagga ggaattttac aaatttatta agcccatctt
     3721 ggaaaaaatg gacggcaccg aggagctgct ggtaaagctt aacagagaag atctgttgcg
     3781 caaacagcgc actttcgaca atggaagcat cccccaccag attcacctgg gcgaactgca
     3841 cgctatcctc aggcggcaag aggatttcta cccctttttg aaagataaca gggaaaagat
     3901 tgagaaaatc ctcacatttc ggatacccta ctatgtaggc cccctcgccc ggggaaattc
     3961 cagattcgcg tggatgactc gcaaatcaga agagaccatc actccctgga acttcgagga
     4021 agtcgtggat aagggggcct ctgcccagtc cttcatcgaa aggatgacta actttgataa
     4081 aaatctgcct aacgaaaagg tgcttcctaa acactctctg ctgtacgagt acttcacagt
     4141 ttataacgag ctcaccaagg tcaaatacgt cacagaaggg atgagaaagc cagcattcct
     4201 gtctggagag cagaagaaag ctatcgtgga cctcctcttc aagacgaacc ggaaagttac
     4261 cgtgaaacag ctcaaagaag actatttcaa aaagattgaa tgtttcgact ctgttgaaat
     4321 cagcggagtg gaggatcgct tcaacgcatc cctgggaacg tatcacgatc tcctgaaaat
     4381 cattaaagac aaggacttcc tggacaatga ggagaacgag gacattcttg aggacattgt
     4441 cctcaccctt acgttgtttg aagataggga gatgattgaa gaacgcttga aaacttacgc
     4501 tcatctcttc gacgacaaag tcatgaaaca gctcaagagg cgccgatata caggatgggg
     4561 gcggctgtca agaaaactga tcaatgggat ccgagacaag cagagtggaa agacaatcct
     4621 ggattttctt aagtccgatg gatttgccaa ccggaacttc atgcagttga tccatgatga
     4681 ctctctcacc tttaaggagg acatccagaa agcacaagtt tctggccagg gggacagtct
     4741 tcacgagcac atcgctaatc ttgcaggtag cccagctatc aaaaagggaa tactgcagac
     4801 cgttaaggtc gtggatgaac tcgtcaaagt aatgggaagg cataagcccg agaatatcgt
     4861 tatcgagatg gcccgagaga accaaactac ccagaaggga cagaagaaca gtagggaaag
     4921 gatgaagagg attgaagagg gtataaaaga actggggtcc caaatcctta aggaacaccc
     4981 agttgaaaac acccagcttc agaatgagaa gctctacctg tactacctgc agaacggcag
     5041 ggacatgtac gtggatcagg aactggacat caatcggctc tccgactacg acgtggatca
     5101 tatcgtgccc cagtcttttc tcaaagatga ttctattgat aataaagtgt tgacaagatc
     5161 cgataaaaat agagggaaga gtgataacgt cccctcagaa gaagttgtca agaaaatgaa
     5221 aaattattgg cggcagctgc tgaacgccaa actgatcaca caacggaagt tcgataatct
     5281 gactaaggct gaacgaggtg gcctgtctga gttggataaa gccggcttca tcaaaaggca
     5341 gcttgttgag acacgccaga tcaccaagca cgtggcccaa attctcgatt cacgcatgaa
     5401 caccaagtac gatgaaaatg acaaactgat tcgagaggtg aaagttatta ctctgaagtc
     5461 taagctggtc tcagatttca gaaaggactt tcagttttat aaggtgagag agatcaacaa
     5521 ttaccaccat gcgcatgatg cctacctgaa tgcagtggta ggcactgcac ttatcaaaaa
     5581 atatcccaag cttgaatctg aatttgttta cggagactat aaagtgtacg atgttaggaa
     5641 aatgatcgca aagtctgagc aggaaatagg caaggccacc gctaagtact tcttttacag
     5701 caatattatg aattttttca agaccgagat tacactggcc aatggagaga ttcggaagcg
     5761 accacttatc gaaacaaacg gagaaacagg agaaatcgtg tgggacaagg gtagggattt
     5821 cgcgacagtc cggaaggtcc tgtccatgcc gcaggtgaac atcgttaaaa agaccgaagt
     5881 acagaccgga ggcttctcca aggaaagtat cctcccgaaa aggaacagcg acaagctgat
     5941 cgcacgcaaa aaagattggg accccaagaa atacggcgga ttcgattctc ctacagtcgc
     6001 ttacagtgta ctggttgtgg ccaaagtgga gaaagggaag tctaaaaaac tcaaaagcgt
     6061 caaggaactg ctgggcatca caatcatgga gcgatcaagc ttcgaaaaaa accccatcga
     6121 ctttctcgag gcgaaaggat ataaagaggt caaaaaagac ctcatcatta agcttcccaa
     6181 gtactctctc tttgagcttg aaaacggccg gaaacgaatg ctcgctagtg cgggcgagct
     6241 gcagaaaggt aacgagctgg cactgccctc taaatacgtt aatttcttgt atctggccag
     6301 ccactatgaa aagctcaaag ggtctcccga agataatgag cagaagcagc tgttcgtgga
     6361 acaacacaaa cactaccttg atgagatcat cgagcaaata agcgaattct ccaaaagagt
     6421 gatcctcgcc gacgctaacc tcgataaggt gctttctgct tacaataagc acagggataa
     6481 gcccatcagg gagcaggcag aaaacattat ccacttgttt actctgacca acttgggcgc
     6541 gcctgcagcc ttcaagtact tcgacaccac catagacaga aagcggtaca cctctacaaa
     6601 ggaggtcctg gacgccacac tgattcatca gtcaattacg gggctctatg aaacaagaat
     6661 cgacctctct cagctcggtg gagacagcag ggctgacccc aagaagaaga ggaaggtgtg
     6721 atctcttctc gagtcatgta attagttatg tcacgcttac attcacgccc tccccccaca
     6781 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
     6841 tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     6901 tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga
     6961 cgctcgaagg ctttaatttg cggaggcgta tcacgaggcc ctttcgtctt caagaattag
     7021 cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc ttgaaatttt
     7081 tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg
     7141 gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa
     7201 ctgcacagaa caaaaacatg caggaaacga agataaatca tgtcgaaagc tacatataag
     7261 gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa
     7321 aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta
     7381 gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat
     7441 ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta
     7501 ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg
     7561 ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca
     7621 ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt
     7681 ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt
     7741 actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac
     7801 atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat
     7861 gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt ctctacagga
     7921 tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag
     7981 ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac
     8041 taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt
     8101 taattatatc agttattacc caattctcat gtttgacagc ttatcatcga tcgtccaact
     8161 gcatggagat gagtcgtggc aagaatacca agagttcctc ggtttgccag ttattaaaag
     8221 actcgtattt ccaaaagact gcaacatact actcagtgca gcttcacaga aacctcattc
     8281 gtttattccc ttgtttgatt cagaagcagg tgggacaggt gaacttttgg attggaactc
     8341 gatttctgac tgggttggaa ggcaagagag ccccgagcct gaacgaagca tctgtgcttc
     8401 attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     8461 gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg
     8521 cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     8581 gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat
     8641 ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc
     8701 atcttaggtt actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt
     8761 tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat
     8821 aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt
     8881 ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg
     8941 tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct
     9001 tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat
     9061 tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa
     9121 cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta
     9181 ggttatatag ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg
     9241 tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt
     9301 tgaaagtgcg tcttcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt
     9361 ctagagaata ggaacttcgg aataggaact tcaaagcgtt tccgaaaacg agcgcttccg
     9421 aaaatgcaac acgagctgcg cacatacagc tcactgttca cgtcgcacct atatctgcgt
     9481 gttgcctgta tatatatata catgagaaga acggcatagt gcgtgtttat gcttaaatgc
     9541 gtacttatat gcgtctattt atgtaggatg aaaggtagtc tagtacctcc tgtgatatta
     9601 tcccattcca cgcggggtat cgtatgcttc cttcagcact accctttagc tgttctatat
     9661 gctgccactc ctcaattgga ttagtctcat ccttcaatgc tatcatttcc tttgatattg
     9721 gatcatacag catgtgacct tttgcaagca attaaatttt gctgatatcg aattcgctgg
     9781 agcttctttg aaaagataat gtatgattat gctttcactc atatttatac agaaacttga
     9841 tgttttcttt cgagtatata caaggtgatt acatgtacgt ttgaagtaca actctagatt
     9901 ttgtagtgcc ctcttgggct agcggtaaag gtgcgcattt tttcacaccc tacaatgttc
     9961 tgttcaaaag attttggtca aacgctgtag aagtgaaagt tggtgcgcat gtttcggcgt
    10021 tcgaaacttc tccgcagtga aagataaatg atcgtttaaa cgacgacatc agcaacatct
    10081 gcgagggtac cgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
    10141 cttgaaaaag tggcaccgag tcggtggtgc tttttttgtt ttttatgtct tcgagtcatg
    10201 taattagtta tgtcacgctc tagagcggcc gccaccgcgg tggagctcca gcttttgttc
    10261 ccggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
    10321 attcaaatat gtatccgctc atgagacaat aaccctgata
//
