LOCUS       sequence-308875-       10318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive 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 10318)
  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 10318)
  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: pCAS9c
FEATURES             Location/Qualifiers
     source          1..10318
                     /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        2110..2508
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     2512..2528
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2512..2528
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2540..6643
                     /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             2540..6643
                     /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             2540..6643
                     /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             6656..6676
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6692..6939
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6708..6726)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        6982..7197
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             7198..8001
                     /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      8340..9682
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(9303..9350)
                     /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        9743..10011
                     /label=SNR52 promoter
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_RNA        10050..10124
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      10129..10148
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     promoter        join(10245..10318,1..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 gctggagctc atagcttcaa aatgtttcta ctcctttttt actcttccag attttctcgg
     2161 actccgcgca tcgccgtacc acttcaaaac acccaagcac agcatactaa atttcccctc
     2221 tttcttcctc tagggtgtcg ttaattaccc gtactaaagg tttggaaaag aaaaaagaga
     2281 ccgcctcgtt tctttttctt cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt
     2341 tcttgaaaat tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt
     2401 taataaacgg tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt
     2461 tttttacttc ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttctagaact
     2521 agtggatccc ccgggaaaaa tggacaagaa gtactccatt gggctcgata tcggcacaaa
     2581 cagcgtcggt tgggccgtca ttacggacga gtacaaggtg ccgagcaaaa aattcaaagt
     2641 tctgggcaat accgatcgcc acagcataaa gaagaacctc attggcgccc tcctgttcga
     2701 ctccggggag acggccgaag ccacgcggct caaaagaaca gcacggcgca gatatacccg
     2761 cagaaagaat cggatctgct acctgcagga gatctttagt aatgagatgg ctaaggtgga
     2821 tgactctttc ttccataggc tggaggagtc ctttttggtg gaggaggata aaaagcacga
     2881 gcgccaccca atctttggca atatcgtgga cgaggtggcg taccatgaaa agtacccaac
     2941 catatatcat ctgaggaaga agcttgtaga cagtactgat aaggctgact tgcggttgat
     3001 ctatctcgcg ctggcgcata tgatcaaatt tcggggacac ttcctcatcg agggggacct
     3061 gaacccagac aacagcgatg tcgacaaact ctttatccaa ctggttcaga cttacaatca
     3121 gcttttcgaa gagaacccga tcaacgcatc cggagttgac gccaaagcaa tcctgagcgc
     3181 taggctgtcc aaatcccggc ggctcgaaaa cctcatcgca cagctccctg gggagaagaa
     3241 gaacggcctg tttggtaatc ttatcgccct gtcactcggg ctgaccccca actttaaatc
     3301 taacttcgac ctggccgaag atgccaagct tcaactgagc aaagacacct acgatgatga
     3361 tctcgacaat ctgctggccc agatcggcga ccagtacgca gacctttttt tggcggcaaa
     3421 gaacctgtca gacgccattc tgctgagtga tattctgcga gtgaacacgg agatcaccaa
     3481 agctccgctg agcgctagta tgatcaagcg ctatgatgag caccaccaag acttgacttt
     3541 gctgaaggcc cttgtcagac agcaactgcc tgagaagtac aaggaaattt tcttcgatca
     3601 gtctaaaaat ggctacgccg gatacattga cggcggagca agccaggagg aattttacaa
     3661 atttattaag cccatcttgg aaaaaatgga cggcaccgag gagctgctgg taaagcttaa
     3721 cagagaagat ctgttgcgca aacagcgcac tttcgacaat ggaagcatcc cccaccagat
     3781 tcacctgggc gaactgcacg ctatcctcag gcggcaagag gatttctacc cctttttgaa
     3841 agataacagg gaaaagattg agaaaatcct cacatttcgg ataccctact atgtaggccc
     3901 cctcgcccgg ggaaattcca gattcgcgtg gatgactcgc aaatcagaag agaccatcac
     3961 tccctggaac ttcgaggaag tcgtggataa gggggcctct gcccagtcct tcatcgaaag
     4021 gatgactaac tttgataaaa atctgcctaa cgaaaaggtg cttcctaaac actctctgct
     4081 gtacgagtac ttcacagttt ataacgagct caccaaggtc aaatacgtca cagaagggat
     4141 gagaaagcca gcattcctgt ctggagagca gaagaaagct atcgtggacc tcctcttcaa
     4201 gacgaaccgg aaagttaccg tgaaacagct caaagaagac tatttcaaaa agattgaatg
     4261 tttcgactct gttgaaatca gcggagtgga ggatcgcttc aacgcatccc tgggaacgta
     4321 tcacgatctc ctgaaaatca ttaaagacaa ggacttcctg gacaatgagg agaacgagga
     4381 cattcttgag gacattgtcc tcacccttac gttgtttgaa gatagggaga tgattgaaga
     4441 acgcttgaaa acttacgctc atctcttcga cgacaaagtc atgaaacagc tcaagaggcg
     4501 ccgatataca ggatgggggc ggctgtcaag aaaactgatc aatgggatcc gagacaagca
     4561 gagtggaaag acaatcctgg attttcttaa gtccgatgga tttgccaacc ggaacttcat
     4621 gcagttgatc catgatgact ctctcacctt taaggaggac atccagaaag cacaagtttc
     4681 tggccagggg gacagtcttc acgagcacat cgctaatctt gcaggtagcc cagctatcaa
     4741 aaagggaata ctgcagaccg ttaaggtcgt ggatgaactc gtcaaagtaa tgggaaggca
     4801 taagcccgag aatatcgtta tcgagatggc ccgagagaac caaactaccc agaagggaca
     4861 gaagaacagt agggaaagga tgaagaggat tgaagagggt ataaaagaac tggggtccca
     4921 aatccttaag gaacacccag ttgaaaacac ccagcttcag aatgagaagc tctacctgta
     4981 ctacctgcag aacggcaggg acatgtacgt ggatcaggaa ctggacatca atcggctctc
     5041 cgactacgac gtggatcata tcgtgcccca gtcttttctc aaagatgatt ctattgataa
     5101 taaagtgttg acaagatccg ataaaaatag agggaagagt gataacgtcc cctcagaaga
     5161 agttgtcaag aaaatgaaaa attattggcg gcagctgctg aacgccaaac tgatcacaca
     5221 acggaagttc gataatctga ctaaggctga acgaggtggc ctgtctgagt tggataaagc
     5281 cggcttcatc aaaaggcagc ttgttgagac acgccagatc accaagcacg tggcccaaat
     5341 tctcgattca cgcatgaaca ccaagtacga tgaaaatgac aaactgattc gagaggtgaa
     5401 agttattact ctgaagtcta agctggtctc agatttcaga aaggactttc agttttataa
     5461 ggtgagagag atcaacaatt accaccatgc gcatgatgcc tacctgaatg cagtggtagg
     5521 cactgcactt atcaaaaaat atcccaagct tgaatctgaa tttgtttacg gagactataa
     5581 agtgtacgat gttaggaaaa tgatcgcaaa gtctgagcag gaaataggca aggccaccgc
     5641 taagtacttc ttttacagca atattatgaa ttttttcaag accgagatta cactggccaa
     5701 tggagagatt cggaagcgac cacttatcga aacaaacgga gaaacaggag aaatcgtgtg
     5761 ggacaagggt agggatttcg cgacagtccg gaaggtcctg tccatgccgc aggtgaacat
     5821 cgttaaaaag accgaagtac agaccggagg cttctccaag gaaagtatcc tcccgaaaag
     5881 gaacagcgac aagctgatcg cacgcaaaaa agattgggac cccaagaaat acggcggatt
     5941 cgattctcct acagtcgctt acagtgtact ggttgtggcc aaagtggaga aagggaagtc
     6001 taaaaaactc aaaagcgtca aggaactgct gggcatcaca atcatggagc gatcaagctt
     6061 cgaaaaaaac cccatcgact ttctcgaggc gaaaggatat aaagaggtca aaaaagacct
     6121 catcattaag cttcccaagt actctctctt tgagcttgaa aacggccgga aacgaatgct
     6181 cgctagtgcg ggcgagctgc agaaaggtaa cgagctggca ctgccctcta aatacgttaa
     6241 tttcttgtat ctggccagcc actatgaaaa gctcaaaggg tctcccgaag ataatgagca
     6301 gaagcagctg ttcgtggaac aacacaaaca ctaccttgat gagatcatcg agcaaataag
     6361 cgaattctcc aaaagagtga tcctcgccga cgctaacctc gataaggtgc tttctgctta
     6421 caataagcac agggataagc ccatcaggga gcaggcagaa aacattatcc acttgtttac
     6481 tctgaccaac ttgggcgcgc ctgcagcctt caagtacttc gacaccacca tagacagaaa
     6541 gcggtacacc tctacaaagg aggtcctgga cgccacactg attcatcagt caattacggg
     6601 gctctatgaa acaagaatcg acctctctca gctcggtgga gacagcaggg ctgaccccaa
     6661 gaagaagagg aaggtgtgat ctcttctcga gtcatgtaat tagttatgtc acgcttacat
     6721 tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc
     6781 taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa
     6841 atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg
     6901 cttgagaagg ttttgggacg ctcgaaggct ttaatttgcg gaggcgtatc acgaggccct
     6961 ttcgtcttca agaattagct tttcaattca tcattttttt tttattcttt tttttgattt
     7021 cggtttcctt gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag
     7081 gagcacagac ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc
     7141 ccagtattct taacccaact gcacagaaca aaaacatgca ggaaacgaag ataaatcatg
     7201 tcgaaagcta catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta
     7261 tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc
     7321 aaggaattac tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat
     7381 gtggatatct tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc
     7441 gccaagtaca attttttact cttcgaagac agaaaatttg ctgacattgg taatacagtc
     7501 aaattgcagt actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca
     7561 cacggtgtgg tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca
     7621 aaggaaccta gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact
     7681 ggagaatata ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc
     7741 tttattgctc aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca
     7801 cccggtgtgg gtttagatga caagggagac gcattgggtc aacagtatag aaccgtggat
     7861 gatgtggtct ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga
     7921 agggatgcta aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga
     7981 agatgcggcc agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca
     8041 caaattagag cttcaattta attatatcag ttattaccca attctcatgt ttgacagctt
     8101 atcatcgatc gtccaactgc atggagatga gtcgtggcaa gaataccaag agttcctcgg
     8161 tttgccagtt attaaaagac tcgtatttcc aaaagactgc aacatactac tcagtgcagc
     8221 ttcacagaaa cctcattcgt ttattccctt gtttgattca gaagcaggtg ggacaggtga
     8281 acttttggat tggaactcga tttctgactg ggttggaagg caagagagcc ccgagcctga
     8341 acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc
     8401 aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa gcgctatttt
     8461 accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat
     8521 ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg cgagagcgct
     8581 attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat cccgagagcg
     8641 ctatttttct aacaaagcat cttaggttac tttttttctc ctttgtgcgc tctataatgc
     8701 agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg ctactttggt
     8761 gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac tgattactag
     8821 cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc tataccgatg
     8881 tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc attggtcaga
     8941 aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa atgtttacat
     9001 tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt ttgtctaaag
     9061 agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca agttcaagga
     9121 gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata gcaaagagat
     9181 acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc gttacagtcc
     9241 ggtgcgtttt tggttttttg aaagtgcgtc ttcagagcgc ttttggtttt caaaagcgct
     9301 ctgaagttcc tatactttct agagaatagg aacttcggaa taggaacttc aaagcgtttc
     9361 cgaaaacgag cgcttccgaa aatgcaacac gagctgcgca catacagctc actgttcacg
     9421 tcgcacctat atctgcgtgt tgcctgtata tatatataca tgagaagaac ggcatagtgc
     9481 gtgtttatgc ttaaatgcgt acttatatgc gtctatttat gtaggatgaa aggtagtcta
     9541 gtacctcctg tgatattatc ccattccacg cggggtatcg tatgcttcct tcagcactac
     9601 cctttagctg ttctatatgc tgccactcct caattggatt agtctcatcc ttcaatgcta
     9661 tcatttcctt tgatattgga tcatacagca tgtgaccttt tgcaagcaat taaattttgc
     9721 tgatatcgaa ttcgctggag cttctttgaa aagataatgt atgattatgc tttcactcat
     9781 atttatacag aaacttgatg ttttctttcg agtatataca aggtgattac atgtacgttt
     9841 gaagtacaac tctagatttt gtagtgccct cttgggctag cggtaaaggt gcgcattttt
     9901 tcacacccta caatgttctg ttcaaaagat tttggtcaaa cgctgtagaa gtgaaagttg
     9961 gtgcgcatgt ttcggcgttc gaaacttctc cgcagtgaaa gataaatgat cgtttaaacg
    10021 acgacatcag caacatctgc gagggtaccg ttttagagct agaaatagca agttaaaata
    10081 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtggtgctt tttttgtttt
    10141 ttatgtcttc gagtcatgta attagttatg tcacgctcta gagcggccgc caccgcggtg
    10201 gagctccagc ttttgttccc ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
    10261 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgata
//
