LOCUS       sequence-199888-       11746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single copy yeast plasmid expressing Cas9 from Streptococcus 
            pyogenes (SpCas9), codon optimized for expression in Saccharomyces 
            cerevisiae..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11746)
  AUTHORS   Verwaal R, Buiting-Wiessenhaan N, Dalhuijsen S, Roubos JA
  TITLE     CRISPR/Cpf1 enables fast and simple genome editing of Saccharomyces 
            cerevisiae.
  JOURNAL   Yeast. 2017 Sep 8. doi: 10.1002/yea.3278.
  PUBMED    28886218
REFERENCE   2  (bases 1 to 11746)
  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: "Yeast. 2017
            Sep 8. doi: 10.1002/yea.3278."
COMMENT     Sequence Label: pCSN061
FEATURES             Location/Qualifiers
     source          1..11746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..114
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     106..123
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     107..123
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     133..152
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        133..151
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1166..5269
                     /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             1166..5269
                     /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             5282..5302
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    5651..5684
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            5730..7086
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        5730..6073
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             6074..6883
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(6141..6160)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      6889..7086
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    7134..7167
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(7235..7255)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7235..7253)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7274..7290)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7274..7290)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7287..7309)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7298..7314
                     /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)."
     promoter        complement(join(7322..7328,7329..7346,7347..7352))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7367..7388
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7505..7522)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7676..8264)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7756..7775)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8435..9226,9227..9295))
                     /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     9058..9077
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9296..9400)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    9437..9940
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     9981..9999
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(10037..10059)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     10159..10178
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        10199..10479
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             10480..11154
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(11257..11712)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11394..11415)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     11606..11625
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
       61 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
      121 agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtaccttttc tttttttgcg
      181 gtcaccccca tgtggcgggg aggcagagga gtaggtagag caacgaatcc tactatttat
      241 ccaaattagt ctaggaactc tttttctaga ttttttagat ttgagggcaa gcgctgttaa
      301 cgactcagaa atgtaagcac tacggagtag aacgagaaat ccgccatagg tggaaatcct
      361 agcaaaatct tgcttaccct agctagcctc aggtaagcta gccttagcct gtcaaatttt
      421 tttcaaaatt tggtaagttt ctactagcaa agcaaacacg gttcaacaaa ccgaaaactc
      481 cactcattat acgtggaaac cgaaacaaaa aaacaaaaac caaaatactc gccaatgaga
      541 aagttgctgc gtttctactt tcgaggaaga ggaactgaga ggattgacta cgaaaggggc
      601 aaaaacgagt cgtattctcc cattattgtc tgctaccacg cggtctagta gaataagcaa
      661 ccagtcaacg ctaagacagg taatcaaaat accagtctgc tggctacggg ctagttttta
      721 cctcttttag aacccactgt aaaagtccgt tgtaaagccc gttctcactg ttggcgtttt
      781 tttttttttg gtttagtttc ttatttttca tttttttctt tcatgaccaa aaacaaacaa
      841 atctcgcgat ttgtactgcg gccactgggg cgtggccaaa aaaatgacaa atttagaaac
      901 cttagtttct gatttttcct gttatgagga gatatgataa aaaatattac tgctttattg
      961 tttttttttt atctactgaa atagagaaac ttacccaagg aggaggcaaa aaaaagagta
     1021 tatatacagc agctaccatt cagattttaa tatattcttt tctcttcttc tacactatta
     1081 ttataataat tttactatat tcatttttag cttaaaacct catagaatat tattcttcag
     1141 tcactcgctt aaatacttat caaaaatgga caagaaatac tctattggtt tggatatcgg
     1201 gaccaactcc gtcggttggg ctgtcatcac cgacgaatac aaggttccat ccaagaaatt
     1261 caaggtcttg ggtaacactg acagacactc tatcaagaag aatttgatcg gtgctttgtt
     1321 gttcgactcc ggtgaaaccg ctgaagctac cagattgaag cgtaccgctc gtcgtagata
     1381 cactagacgt aaaaaccgta tttgttactt gcaagaaatc ttttctaacg aaatggccaa
     1441 ggttgacgac tctttcttcc acagattgga agaatctttc ttggttgaag aagacaagaa
     1501 gcacgaaaga catccaatct tcggtaacat cgttgacgaa gttgcttacc acgaaaaata
     1561 ccctaccatc taccatttga gaaagaagtt ggtcgattcc accgacaagg ctgatttgag
     1621 attgatctat ttggccttgg ctcacatgat caagttcaga ggtcacttct tgattgaagg
     1681 tgacttgaac ccagacaact ctgacgtcga caaattgttc atccaattgg tccaaaccta
     1741 caaccaatta ttcgaggaaa acccaattaa cgcttctggt gttgatgcta aggccatctt
     1801 atctgcccgt ttgtccaagt ctagacgttt ggaaaacttg attgctcaat tgcctggtga
     1861 aaagaaaaac ggtttgttcg gtaacttgat cgctttgtcc ttgggtttga ccccaaactt
     1921 caagtccaac ttcgacttgg ctgaagatgc caagttgcaa ttgtccaagg acacctacga
     1981 cgacgactta gacaacttgt tggctcaaat cggtgaccaa tacgccgact tgttcttggc
     2041 tgccaaaaac ttatctgacg ctatcttgtt gtctgacatc ttgagagtta acactgaaat
     2101 taccaaggct ccattgtctg cttctatgat caaaagatac gacgaacacc accaagatct
     2161 gactttgttg aaggctttgg ttagacaaca attgccagaa aagtacaagg aaatcttctt
     2221 cgaccaatcc aaaaatggtt acgccggtta cattgacggt ggtgcttctc aggaagaatt
     2281 ctacaagttc atcaagccaa ttttggaaaa gatggatggt actgaagaat tattggttaa
     2341 gttgaacaga gaagacttat tgagaaagca acgtaccttc gataacggtt ctatcccaca
     2401 ccaaatccac ttgggtgaat tgcacgccat tttgagaaga caggaagatt tctatccatt
     2461 cctaaaggac aacagagaaa agatcgaaaa gatcttaact ttcagaatcc catactacgt
     2521 cggtccattg gccagaggta attctagatt cgcttggatg accagaaagt ctgaagaaac
     2581 catcacccca tggaacttcg aagaagtcgt cgacaagggt gcttctgccc aatctttcat
     2641 cgaaagaatg accaactttg ataagaactt gccaaacgag aaggtcttgc caaagcactc
     2701 tttgttgtac gaatacttca ccgtctacaa cgaattaacc aaggttaaat acgttactga
     2761 aggtatgaga aagccagctt tcctatccgg tgaacaaaag aaggctattg ttgacttgtt
     2821 gtttaagacc aacagaaagg tcactgttaa gcaattgaag gaagactact tcaagaagat
     2881 tgaatgtttc gattccgtcg aaatctccgg tgttgaagac cgtttcaatg cttctttggg
     2941 cacctaccac gatttgttaa agatcatcaa ggacaaggac tttttagata acgaagaaaa
     3001 cgaagacatc ttggaagata tcgttttgac cttgactctt ttcgaggaca gagaaatgat
     3061 tgaagagaga ttgaagacct acgctcactt gttcgacgat aaagttatga agcaactaaa
     3121 gagaagaaga tacactggtt ggggtagatt gtccagaaag ttgattaacg gtatcagaga
     3181 caagcaatcc ggtaagacta ttttagactt tttgaaatcc gatggtttcg ctaacagaaa
     3241 ctttatgcaa ttgattcacg acgattcttt gactttcaag gaagacattc aaaaagccca
     3301 agtctctggt caaggtgatt ctttgcacga acacatcgct aacttggctg gttctccagc
     3361 tattaagaag ggtatcttac aaaccgtcaa ggtcgttgat gaattggtca aagtcatggg
     3421 tagacacaag ccagaaaata ttgtcatcga aatggctaga gaaaaccaaa ctactcaaaa
     3481 gggtcaaaag aactctagag aacgtatgaa gagaattgaa gaaggtatca aggagttggg
     3541 ttctcaaatt ttgaaagaac acccagtcga aaacactcaa ttacaaaacg aaaagctata
     3601 cttgtactac ttgcaaaacg gtcgtgacat gtacgtcgac caagaattgg atatcaacag
     3661 attgtctgac tacgatgtcg atcatatcgt cccacaatcg ttcttgaagg acgattccat
     3721 tgacaacaaa gttttgacta gatctgacaa gaacagaggt aagtctgata acgttccatc
     3781 tgaagaagtt gttaagaaga tgaagaacta ctggagacaa ttgttgaatg ctaagttgat
     3841 cactcaaaga aagttcgaca acttgaccaa ggctgaaaga ggtggtttgt ccgaattgga
     3901 caaagccggt ttcatcaaga gacaattagt cgaaactaga caaatcacca agcatgttgc
     3961 tcaaatcttg gattccagaa tgaacactaa gtacgatgaa aacgacaaac taattagaga
     4021 agttaaggtc atcactttga agtctaagtt ggtttctgac ttcagaaagg acttccaatt
     4081 ttacaaggtc agagaaatca acaactacca tcacgctcac gatgcctact tgaacgctgt
     4141 tgtcggtact gccttaatca aaaagtaccc aaagttggaa tctgaattcg tttacggtga
     4201 ctacaaggtt tacgatgtta gaaagatgat cgccaagtct gaacaagaaa ttggtaaggc
     4261 cactgctaag tacttcttct actctaacat catgaacttt ttcaagactg aaatcacttt
     4321 agctaacggt gaaattagaa agcgtccatt gattgaaacc aatggtgaaa ctggtgaaat
     4381 tgtctgggac aagggtagag atttcgctac cgtcagaaag gttttgtcta tgccacaagt
     4441 taacatcgtc aagaagactg aagttcaaac tggtggtttc tctaaggaat ccattttgcc
     4501 aaagagaaac tctgacaagt tgattgctag aaagaaggac tgggatccta agaagtacgg
     4561 tggtttcgac tctccaactg ttgcttactc cgttttggtc gttgctaagg ttgaaaaggg
     4621 taagtctaag aagttgaagt ctgttaagga attgttgggt atcaccatca tggaaagatc
     4681 ctccttcgaa aagaacccaa tcgacttttt ggaagctaag ggttacaagg aagtcaagaa
     4741 ggatttgatc attaagttac caaaatactc cttgttcgaa ttggaaaacg gtagaaagag
     4801 aatgttggcc tccgctggtg aactacaaaa aggtaacgaa ttggctttac catctaagta
     4861 cgttaacttc ttgtacttgg cttcccacta cgaaaagttg aaaggttccc cagaagacaa
     4921 cgaacaaaag caattgtttg ttgaacaaca caagcactac ttggatgaaa ttattgaaca
     4981 aatctccgaa ttctccaaga gagtcatttt ggctgatgct aacttagata aggttttatc
     5041 cgcttacaac aagcacagag acaaaccaat cagagaacaa gctgaaaaca tcattcattt
     5101 gttcacttta accaacttgg gtgctccagc tgctttcaaa tacttcgaca ctaccattga
     5161 cagaaagaga tacacttcca ccaaagaagt tttagatgct actttgattc accaatctat
     5221 taccggtttg tacgaaacca gaattgactt gtctcaattg ggtggtgatt ccagagctga
     5281 tccaaagaag aagagaaagg tgtaaaggag ttaaaggcaa agttttcttt tctagagccg
     5341 ttcccacaaa taattatacg tatatgcttc ttttcgttta ctatatatct atatttacaa
     5401 gcctttattc actgatgcaa tttgtttcca aatacttttt tggagatctc ataactagat
     5461 atcatgatgg cgcaacttgg cgctatctta attactctgg ctgccaggcc cgtgtagagg
     5521 gccgcaagac cttctgtacg ccatatagtc tctaagaact tgaacaagtt tctagaccta
     5581 ttgccgcctt tcggatcgct attgttgcgg ccgccagctg aagcttcgta cgctgcaggt
     5641 cgacgaattc taccgttcgt ataatgtatg ctatacgaag ttatagatct gtttagcttg
     5701 cctcgtcccc gccgggtcac ccggccagcg acatggaggc ccagaatacc ctccttgaca
     5761 gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt tagcccatac
     5821 atccccatgt ataatcattt gcatccatac attttgatgg ccgcacggcg cgaagcaaaa
     5881 attacggctc ctcgctgcag acctgcgagc agggaaacgc tcccctcaca gacgcgttga
     5941 attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg ccactgaggt
     6001 tcttctttca tatacttcct tttaaaatct tgctaggata cagttctcac atcacatccg
     6061 aacataaaca accatgggta aggaaaagac tcacgtttcg aggccgcgat taaattccaa
     6121 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     6181 gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     6241 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     6301 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     6361 cactgcgatc cccggcaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     6421 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     6481 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     6541 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     6601 ctggaaagaa atgcataagc ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     6661 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     6721 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     6781 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     6841 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagtac tgacaataaa
     6901 aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt agttgttcta
     6961 ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat ctgcccagat
     7021 gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct
     7081 atactgctgt cgattcgata ctaacgccgc catccagtgt cgaaaacgag ctcataactt
     7141 cgtataatgt atgctatacg aacggtagaa ttcgatatca gatccactag tggcctatgc
     7201 ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct
     7261 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     7321 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     7381 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     7441 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     7501 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     7561 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     7621 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     7681 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     7741 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     7801 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     7861 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     7921 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     7981 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     8041 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     8101 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     8161 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     8221 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     8281 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     8341 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     8401 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     8461 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     8521 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     8581 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     8641 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     8701 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     8761 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     8821 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     8881 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     8941 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     9001 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     9061 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     9121 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     9181 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     9241 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     9301 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     9361 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     9421 cacctgggtc cttttcatca cgtgctataa aaataattat aatttaaatt ttttaatata
     9481 aatatataaa ttaaaaatag aaagtaaaaa aagaaattaa agaaaaaata gtttttgttt
     9541 tccgaagatg taaaagactc tagggggatc gccaacaaat actacctttt atcttgctct
     9601 tcctgctctc aggtattaat gccgaattgt ttcatcttgt ctgtgtagaa gaccacacac
     9661 gaaaatcctg tgattttaca ttttacttat cgttaatcga atgtatatct atttaatctg
     9721 cttttcttgt ctaataaata tatatgtaaa gtacgctttt tgttgaaatt ttttaaacct
     9781 ttgtttattt ttttttcttc attccgtaac tcttctacct tctttattta ctttctaaaa
     9841 tccaaataca aaacataaaa ataaataaac acagagtaaa ttcccaaatt attccatcat
     9901 taaaagatac gaggcgcgtg taagttacag gcaagcgatc cgtctaagaa accattatta
     9961 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
    10021 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
    10081 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
    10141 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catagatcac
    10201 gacattacta tatatataat ataggaagca tttaatagaa cagcatcgta atatatgtgt
    10261 actttgcagt tatgacgcca gatggcagta gtggaagata ttctttattg aaaaatagct
    10321 tgtcacctta cgtacaatct tgatccggag cttttctttt tttgccgatt aagaattaat
    10381 tcggtcgaaa aaagaaaagg agagggccaa gagggagggc attggtgact attgagcacg
    10441 tgagtatacg tgattaagca cacaaaggca gcttggagta tgtctgttat taatttcaca
    10501 ggtagttctg gtccattggt gaaagtttgc ggcttgcaga gcacagaggc cgcagaatgt
    10561 gctctagatt ccgatgctga cttgctgggt attatatgtg tgcccaatag aaagagaaca
    10621 attgacccgg ttattgcaag gaaaatttca agtcttgtaa aagcatataa aaatagttca
    10681 ggcactccga aatacttggt tggcgtgttt cgtaatcaac ctaaggagga tgttttggct
    10741 ctggtcaatg attacggcat tgatatcgtc caactgcatg gagatgagtc gtggcaagaa
    10801 taccaagagt tcctcggttt gccagttatt aaaagactcg tatttccaaa agactgcaac
    10861 atactactca gtgcagcttc acagaaacct cattcgttta ttcccttgtt tgattcagaa
    10921 gcaggtggga caggtgaact tttggattgg aactcgattt ctgactgggt tggaaggcaa
    10981 gagagccccg aaagcttaca ttttatgtta gctggtggac tgacgccaga aaatgttggt
    11041 gatgcgctta gattaaatgg cgttattggt gttgatgtaa gcggaggtgt ggagacaaat
    11101 ggtgtaaaag actctaacaa aatagcaaat ttcgtcaaaa atgctaagaa ataggttatt
    11161 actgagtagt atttatttaa gtattgtttg tgcacttgcc gatctatgcg gtgtgaaata
    11221 ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt aatattttgt
    11281 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
    11341 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
    11401 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
    11461 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
    11521 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
    11581 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
    11641 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
    11701 tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaa
//
