LOCUS       sequence-308887-       10765 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Constitutive expression of Cas9 for addressing two targets; Contains
            two 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 10765)
  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 10765)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pCAS9cd
FEATURES             Location/Qualifiers
     source          1..10765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..1343
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(964..1011)
                     /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        1404..1672
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        1711..1785
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      1790..1809
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     promoter        1880..2148
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        2187..2261
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      2266..2285
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     promoter        2353..2457
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2458..2526,2527..3318)
                     /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(2676..2695)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3489..4077
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3978..3997
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4231..4248
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4365..4386
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4401..4406,4407..4424,4425..4431)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4439..4455
                     /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     4444..4466
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4463..4479
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4463..4479
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4498..4518
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4500..4518
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4536..4934
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     4938..4954
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4938..4954
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             4966..9069
                     /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             4966..9069
                     /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             4966..9069
                     /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             9082..9102
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      9118..9365
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(9134..9152)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        9408..9623
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             9624..10427
                     /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"
ORIGIN
        1 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt
       61 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt
      121 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa
      181 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc
      241 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc
      301 gctatttttc taacaaagca tcttaggtta ctttttttct cctttgtgcg ctctataatg
      361 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg
      421 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta
      481 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat
      541 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag
      601 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca
      661 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa
      721 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg
      781 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga
      841 tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc
      901 cggtgcgttt ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc
      961 tctgaagttc ctatactttc tagagaatag gaacttcgga ataggaactt caaagcgttt
     1021 ccgaaaacga gcgcttccga aaatgcaaca cgagctgcgc acatacagct cactgttcac
     1081 gtcgcaccta tatctgcgtg ttgcctgtat atatatatac atgagaagaa cggcatagtg
     1141 cgtgtttatg cttaaatgcg tacttatatg cgtctattta tgtaggatga aaggtagtct
     1201 agtacctcct gtgatattat cccattccac gcggggtatc gtatgcttcc ttcagcacta
     1261 ccctttagct gttctatatg ctgccactcc tcaattggat tagtctcatc cttcaatgct
     1321 atcatttcct ttgatattgg atcatacagc atgtgacctt ttgcaagcaa ttaaattttg
     1381 ctgatatcga attcgctgga gcttctttga aaagataatg tatgattatg ctttcactca
     1441 tatttataca gaaacttgat gttttctttc gagtatatac aaggtgatta catgtacgtt
     1501 tgaagtacaa ctctagattt tgtagtgccc tcttgggcta gcggtaaagg tgcgcatttt
     1561 ttcacaccct acaatgttct gttcaaaaga ttttggtcaa acgctgtaga agtgaaagtt
     1621 ggtgcgcatg tttcggcgtt cgaaacttct ccgcagtgaa agataaatga tcgtttaaac
     1681 gacgacatca gcaacatctg cgagggtacc gttttagagc tagaaatagc aagttaaaat
     1741 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtggtgct ttttttgttt
     1801 tttatgtctt cgagtcatgt aattagttat gtcacgctct agagcggccg ccaccgctga
     1861 tatcgaattc gctggagctt ctttgaaaag ataatgtatg attatgcttt cactcatatt
     1921 tatacagaaa cttgatgttt tctttcgagt atatacaagg tgattacatg tacgtttgaa
     1981 gtacaactct agattttgta gtgccctctt gggctagcgg taaaggtgcg cattttttca
     2041 caccctacaa tgttctgttc aaaagatttt ggtcaaacgc tgtagaagtg aaagttggtg
     2101 cgcatgtttc ggcgttcgaa acttctccgc agtgaaagat aaatgatcgt ttaaacgacg
     2161 acatcagcaa catctgcgag ggtaccgttt tagagctaga aatagcaagt taaaataagg
     2221 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt ggtgcttttt ttgtttttta
     2281 tgtcttcgag tcatgtaaca ccgcggtgga gctccagctt ttgttcccgg tggcactttt
     2341 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2401 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     2461 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     2521 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     2581 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     2641 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     2701 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     2761 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     2821 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     2881 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     2941 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3001 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3061 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3121 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3181 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3241 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3301 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     3361 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3421 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3481 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3541 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3601 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3661 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3721 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3781 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3841 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3901 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3961 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4021 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4081 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4141 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     4201 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     4261 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     4321 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     4381 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     4441 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
     4501 attaaccctc actaaaggga acaaaagctg gagctcatag cttcaaaatg tttctactcc
     4561 ttttttactc ttccagattt tctcggactc cgcgcatcgc cgtaccactt caaaacaccc
     4621 aagcacagca tactaaattt cccctctttc ttcctctagg gtgtcgttaa ttacccgtac
     4681 taaaggtttg gaaaagaaaa aagagaccgc ctcgtttctt tttcttcgtc gaaaaaggca
     4741 ataaaaattt ttatcacgtt tctttttctt gaaaattttt tttttgattt ttttctcttt
     4801 cgatgacctc ccattgatat ttaagttaat aaacggtctt caatttctca agtttcagtt
     4861 tcatttttct tgttctatta caactttttt tacttcttgc tcattagaaa gaaagcatag
     4921 caatctaatc taagttttct agaactagtg gatcccccgg gaaaaatgga caagaagtac
     4981 tccattgggc tcgatatcgg cacaaacagc gtcggttggg ccgtcattac ggacgagtac
     5041 aaggtgccga gcaaaaaatt caaagttctg ggcaataccg atcgccacag cataaagaag
     5101 aacctcattg gcgccctcct gttcgactcc ggggagacgg ccgaagccac gcggctcaaa
     5161 agaacagcac ggcgcagata tacccgcaga aagaatcgga tctgctacct gcaggagatc
     5221 tttagtaatg agatggctaa ggtggatgac tctttcttcc ataggctgga ggagtccttt
     5281 ttggtggagg aggataaaaa gcacgagcgc cacccaatct ttggcaatat cgtggacgag
     5341 gtggcgtacc atgaaaagta cccaaccata tatcatctga ggaagaagct tgtagacagt
     5401 actgataagg ctgacttgcg gttgatctat ctcgcgctgg cgcatatgat caaatttcgg
     5461 ggacacttcc tcatcgaggg ggacctgaac ccagacaaca gcgatgtcga caaactcttt
     5521 atccaactgg ttcagactta caatcagctt ttcgaagaga acccgatcaa cgcatccgga
     5581 gttgacgcca aagcaatcct gagcgctagg ctgtccaaat cccggcggct cgaaaacctc
     5641 atcgcacagc tccctgggga gaagaagaac ggcctgtttg gtaatcttat cgccctgtca
     5701 ctcgggctga cccccaactt taaatctaac ttcgacctgg ccgaagatgc caagcttcaa
     5761 ctgagcaaag acacctacga tgatgatctc gacaatctgc tggcccagat cggcgaccag
     5821 tacgcagacc tttttttggc ggcaaagaac ctgtcagacg ccattctgct gagtgatatt
     5881 ctgcgagtga acacggagat caccaaagct ccgctgagcg ctagtatgat caagcgctat
     5941 gatgagcacc accaagactt gactttgctg aaggcccttg tcagacagca actgcctgag
     6001 aagtacaagg aaattttctt cgatcagtct aaaaatggct acgccggata cattgacggc
     6061 ggagcaagcc aggaggaatt ttacaaattt attaagccca tcttggaaaa aatggacggc
     6121 accgaggagc tgctggtaaa gcttaacaga gaagatctgt tgcgcaaaca gcgcactttc
     6181 gacaatggaa gcatccccca ccagattcac ctgggcgaac tgcacgctat cctcaggcgg
     6241 caagaggatt tctacccctt tttgaaagat aacagggaaa agattgagaa aatcctcaca
     6301 tttcggatac cctactatgt aggccccctc gcccggggaa attccagatt cgcgtggatg
     6361 actcgcaaat cagaagagac catcactccc tggaacttcg aggaagtcgt ggataagggg
     6421 gcctctgccc agtccttcat cgaaaggatg actaactttg ataaaaatct gcctaacgaa
     6481 aaggtgcttc ctaaacactc tctgctgtac gagtacttca cagtttataa cgagctcacc
     6541 aaggtcaaat acgtcacaga agggatgaga aagccagcat tcctgtctgg agagcagaag
     6601 aaagctatcg tggacctcct cttcaagacg aaccggaaag ttaccgtgaa acagctcaaa
     6661 gaagactatt tcaaaaagat tgaatgtttc gactctgttg aaatcagcgg agtggaggat
     6721 cgcttcaacg catccctggg aacgtatcac gatctcctga aaatcattaa agacaaggac
     6781 ttcctggaca atgaggagaa cgaggacatt cttgaggaca ttgtcctcac ccttacgttg
     6841 tttgaagata gggagatgat tgaagaacgc ttgaaaactt acgctcatct cttcgacgac
     6901 aaagtcatga aacagctcaa gaggcgccga tatacaggat gggggcggct gtcaagaaaa
     6961 ctgatcaatg ggatccgaga caagcagagt ggaaagacaa tcctggattt tcttaagtcc
     7021 gatggatttg ccaaccggaa cttcatgcag ttgatccatg atgactctct cacctttaag
     7081 gaggacatcc agaaagcaca agtttctggc cagggggaca gtcttcacga gcacatcgct
     7141 aatcttgcag gtagcccagc tatcaaaaag ggaatactgc agaccgttaa ggtcgtggat
     7201 gaactcgtca aagtaatggg aaggcataag cccgagaata tcgttatcga gatggcccga
     7261 gagaaccaaa ctacccagaa gggacagaag aacagtaggg aaaggatgaa gaggattgaa
     7321 gagggtataa aagaactggg gtcccaaatc cttaaggaac acccagttga aaacacccag
     7381 cttcagaatg agaagctcta cctgtactac ctgcagaacg gcagggacat gtacgtggat
     7441 caggaactgg acatcaatcg gctctccgac tacgacgtgg atcatatcgt gccccagtct
     7501 tttctcaaag atgattctat tgataataaa gtgttgacaa gatccgataa aaatagaggg
     7561 aagagtgata acgtcccctc agaagaagtt gtcaagaaaa tgaaaaatta ttggcggcag
     7621 ctgctgaacg ccaaactgat cacacaacgg aagttcgata atctgactaa ggctgaacga
     7681 ggtggcctgt ctgagttgga taaagccggc ttcatcaaaa ggcagcttgt tgagacacgc
     7741 cagatcacca agcacgtggc ccaaattctc gattcacgca tgaacaccaa gtacgatgaa
     7801 aatgacaaac tgattcgaga ggtgaaagtt attactctga agtctaagct ggtctcagat
     7861 ttcagaaagg actttcagtt ttataaggtg agagagatca acaattacca ccatgcgcat
     7921 gatgcctacc tgaatgcagt ggtaggcact gcacttatca aaaaatatcc caagcttgaa
     7981 tctgaatttg tttacggaga ctataaagtg tacgatgtta ggaaaatgat cgcaaagtct
     8041 gagcaggaaa taggcaaggc caccgctaag tacttctttt acagcaatat tatgaatttt
     8101 ttcaagaccg agattacact ggccaatgga gagattcgga agcgaccact tatcgaaaca
     8161 aacggagaaa caggagaaat cgtgtgggac aagggtaggg atttcgcgac agtccggaag
     8221 gtcctgtcca tgccgcaggt gaacatcgtt aaaaagaccg aagtacagac cggaggcttc
     8281 tccaaggaaa gtatcctccc gaaaaggaac agcgacaagc tgatcgcacg caaaaaagat
     8341 tgggacccca agaaatacgg cggattcgat tctcctacag tcgcttacag tgtactggtt
     8401 gtggccaaag tggagaaagg gaagtctaaa aaactcaaaa gcgtcaagga actgctgggc
     8461 atcacaatca tggagcgatc aagcttcgaa aaaaacccca tcgactttct cgaggcgaaa
     8521 ggatataaag aggtcaaaaa agacctcatc attaagcttc ccaagtactc tctctttgag
     8581 cttgaaaacg gccggaaacg aatgctcgct agtgcgggcg agctgcagaa aggtaacgag
     8641 ctggcactgc cctctaaata cgttaatttc ttgtatctgg ccagccacta tgaaaagctc
     8701 aaagggtctc ccgaagataa tgagcagaag cagctgttcg tggaacaaca caaacactac
     8761 cttgatgaga tcatcgagca aataagcgaa ttctccaaaa gagtgatcct cgccgacgct
     8821 aacctcgata aggtgctttc tgcttacaat aagcacaggg ataagcccat cagggagcag
     8881 gcagaaaaca ttatccactt gtttactctg accaacttgg gcgcgcctgc agccttcaag
     8941 tacttcgaca ccaccataga cagaaagcgg tacacctcta caaaggaggt cctggacgcc
     9001 acactgattc atcagtcaat tacggggctc tatgaaacaa gaatcgacct ctctcagctc
     9061 ggtggagaca gcagggctga ccccaagaag aagaggaagg tgtgatctct tctcgagtca
     9121 tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa
     9181 aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata gttatgttag
     9241 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg
     9301 catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa
     9361 tttgcggagg cgtatcacga ggccctttcg tcttcaagaa ttagcttttc aattcatcat
     9421 ttttttttta ttcttttttt tgatttcggt ttccttgaaa tttttttgat tcggtaatct
     9481 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat
     9541 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa
     9601 catgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc
     9661 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt
     9721 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc
     9781 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca
     9841 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaagacagaa
     9901 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag
     9961 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt
    10021 tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat
    10081 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga
    10141 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg
    10201 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagacgcat
    10261 tgggtcaaca gtatagaacc gtggatgatg tggtctctac aggatctgac attattattg
    10321 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
    10381 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
    10441 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
    10501 tacccaattc tcatgtttga cagcttatca tcgatcgtcc aactgcatgg agatgagtcg
    10561 tggcaagaat accaagagtt cctcggtttg ccagttatta aaagactcgt atttccaaaa
    10621 gactgcaaca tactactcag tgcagcttca cagaaacctc attcgtttat tcccttgttt
    10681 gattcagaag caggtgggac aggtgaactt ttggattgga actcgatttc tgactgggtt
    10741 ggaaggcaag agagccccga gcctg
//
