LOCUS       sequence-192023-       11742 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 11742)
  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 11742)
  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..11742
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        187..467
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             468..1142
                     /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(1241..1696)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1378..1399)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1590..1609
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1826..1848
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1840..1857
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1841..1857
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1867..1886
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1867..1885
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2900..7003
                     /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             2900..7003
                     /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             7016..7036
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    7385..7418
                     /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            7464..8820
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        7464..7807
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             7808..8617
                     /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(7875..7894)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      8623..8820
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    8868..8901
                     /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(8968..8988)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8968..8986)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(9007..9023)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9007..9023)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9020..9042)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9031..9047
                     /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(9055..9061,9062..9079,9080..9085))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(9238..9255)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9409..9997)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9489..9508)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10168..10959,10960..11028))
                     /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     10791..10810
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11029..11133)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    11170..11673
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     11715..11733
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaacg acattactat atatataata taggaagcat ttaatagaca gcatcgtaat
      241 atatgtgtac tttgcagtta tgacgccaga tggcagtagt ggaagatatt ctttattgaa
      301 aaatagcttg tcaccttacg tacaatcttg atccggagct tttctttttt tgccgattaa
      361 gaattaattc ggtcgaaaaa agaaaaggag agggccaaga gggagggcat tggtgactat
      421 tgagcacgtg agtatacgtg attaagcaca caaaggcagc ttggagtatg tctgttatta
      481 atttcacagg tagttctggt ccattggtga aagtttgcgg cttgcagagc acagaggccg
      541 cagaatgtgc tctagattcc gatgctgact tgctgggtat tatatgtgtg cccaatagaa
      601 agagaacaat tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa
      661 atagttcagg cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg
      721 ttttggctct ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt
      781 ggcaagaata ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag
      841 actgcaacat actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg
      901 attcagaagc aggtgggaca ggtgaacttt tggattggaa ctcgatttct gactgggttg
      961 gaaggcaaga gagccccgaa agcttacatt ttatgttagc tggtggactg acgccagaaa
     1021 atgttggtga tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg
     1081 agacaaatgg tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat
     1141 aggttattac tgagtagtat ttatttaagt attgtttgtg cacttgccta tgcggtgtga
     1201 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt
     1261 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     1321 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     1381 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     1441 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     1501 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     1561 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     1621 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     1681 ccgctacagg gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     1741 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     1801 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
     1861 cgcgcgtaat acgactcact atagggcgaa ttgggtacct tttctttttt tgcggtcacc
     1921 cccatgtggc ggggaggcag aggagtaggt agagcaacga atcctactat ttatccaaat
     1981 tagtctagga actctttttc tagatttttt agatttgagg gcaagcgctg ttaacgactc
     2041 agaaatgtaa gcactacgga gtagaacgag aaatccgcca taggtggaaa tcctagcaaa
     2101 atcttgctta ccctagctag cctcaggtaa gctagcctta gcctgtcaaa tttttttcaa
     2161 aatttggtaa gtttctacta gcaaagcaaa cacggttcaa caaaccgaaa actccactca
     2221 ttatacgtgg aaaccgaaac aaaaaaacaa aaaccaaaat actcgccaat gagaaagttg
     2281 ctgcgtttct actttcgagg aagaggaact gagaggattg actacgaaag gggcaaaaac
     2341 gagtcgtatt ctcccattat tgtctgctac cacgcggtct agtagaataa gcaaccagtc
     2401 aacgctaaga caggtaatca aaataccagt ctgctggcta cgggctagtt tttacctctt
     2461 ttagaaccca ctgtaaaagt ccgttgtaaa gcccgttctc actgttggcg tttttttttt
     2521 tttggtttag tttcttattt ttcatttttt tctttcatga ccaaaaacaa acaaatctcg
     2581 cgatttgtac tgcggccact ggggcgtggc caaaaaaatg acaaatttag aaaccttagt
     2641 ttctgatttt tcctgttatg aggagatatg ataaaaaata ttactgcttt attgtttttt
     2701 ttttatctac tgaaatagag aaacttaccc aaggaggagg caaaaaaaag agtatatata
     2761 cagcagctac cattcagatt ttaatatatt cttttctctt cttctacact attattataa
     2821 taattttact atattcattt ttagcttaaa acctcataga atattattct tcagtcactc
     2881 gcttaaatac ttatcaaaaa tggacaagaa atactctatt ggtttggata tcgggaccaa
     2941 ctccgtcggt tgggctgtca tcaccgacga atacaaggtt ccatccaaga aattcaaggt
     3001 cttgggtaac actgacagac actctatcaa gaagaatttg atcggtgctt tgttgttcga
     3061 ctccggtgaa accgctgaag ctaccagatt gaagcgtacc gctcgtcgta gatacactag
     3121 acgtaaaaac cgtatttgtt acttgcaaga aatcttttct aacgaaatgg ccaaggttga
     3181 cgactctttc ttccacagat tggaagaatc tttcttggtt gaagaagaca agaagcacga
     3241 aagacatcca atcttcggta acatcgttga cgaagttgct taccacgaaa aataccctac
     3301 catctaccat ttgagaaaga agttggtcga ttccaccgac aaggctgatt tgagattgat
     3361 ctatttggcc ttggctcaca tgatcaagtt cagaggtcac ttcttgattg aaggtgactt
     3421 gaacccagac aactctgacg tcgacaaatt gttcatccaa ttggtccaaa cctacaacca
     3481 attattcgag gaaaacccaa ttaacgcttc tggtgttgat gctaaggcca tcttatctgc
     3541 ccgtttgtcc aagtctagac gtttggaaaa cttgattgct caattgcctg gtgaaaagaa
     3601 aaacggtttg ttcggtaact tgatcgcttt gtccttgggt ttgaccccaa acttcaagtc
     3661 caacttcgac ttggctgaag atgccaagtt gcaattgtcc aaggacacct acgacgacga
     3721 cttagacaac ttgttggctc aaatcggtga ccaatacgcc gacttgttct tggctgccaa
     3781 aaacttatct gacgctatct tgttgtctga catcttgaga gttaacactg aaattaccaa
     3841 ggctccattg tctgcttcta tgatcaaaag atacgacgaa caccaccaag atctgacttt
     3901 gttgaaggct ttggttagac aacaattgcc agaaaagtac aaggaaatct tcttcgacca
     3961 atccaaaaat ggttacgccg gttacattga cggtggtgct tctcaggaag aattctacaa
     4021 gttcatcaag ccaattttgg aaaagatgga tggtactgaa gaattattgg ttaagttgaa
     4081 cagagaagac ttattgagaa agcaacgtac cttcgataac ggttctatcc cacaccaaat
     4141 ccacttgggt gaattgcacg ccattttgag aagacaggaa gatttctatc cattcctaaa
     4201 ggacaacaga gaaaagatcg aaaagatctt aactttcaga atcccatact acgtcggtcc
     4261 attggccaga ggtaattcta gattcgcttg gatgaccaga aagtctgaag aaaccatcac
     4321 cccatggaac ttcgaagaag tcgtcgacaa gggtgcttct gcccaatctt tcatcgaaag
     4381 aatgaccaac tttgataaga acttgccaaa cgagaaggtc ttgccaaagc actctttgtt
     4441 gtacgaatac ttcaccgtct acaacgaatt aaccaaggtt aaatacgtta ctgaaggtat
     4501 gagaaagcca gctttcctat ccggtgaaca aaagaaggct attgttgact tgttgtttaa
     4561 gaccaacaga aaggtcactg ttaagcaatt gaaggaagac tacttcaaga agattgaatg
     4621 tttcgattcc gtcgaaatct ccggtgttga agaccgtttc aatgcttctt tgggcaccta
     4681 ccacgatttg ttaaagatca tcaaggacaa ggacttttta gataacgaag aaaacgaaga
     4741 catcttggaa gatatcgttt tgaccttgac tcttttcgag gacagagaaa tgattgaaga
     4801 gagattgaag acctacgctc acttgttcga cgataaagtt atgaagcaac taaagagaag
     4861 aagatacact ggttggggta gattgtccag aaagttgatt aacggtatca gagacaagca
     4921 atccggtaag actattttag actttttgaa atccgatggt ttcgctaaca gaaactttat
     4981 gcaattgatt cacgacgatt ctttgacttt caaggaagac attcaaaaag cccaagtctc
     5041 tggtcaaggt gattctttgc acgaacacat cgctaacttg gctggttctc cagctattaa
     5101 gaagggtatc ttacaaaccg tcaaggtcgt tgatgaattg gtcaaagtca tgggtagaca
     5161 caagccagaa aatattgtca tcgaaatggc tagagaaaac caaactactc aaaagggtca
     5221 aaagaactct agagaacgta tgaagagaat tgaagaaggt atcaaggagt tgggttctca
     5281 aattttgaaa gaacacccag tcgaaaacac tcaattacaa aacgaaaagc tatacttgta
     5341 ctacttgcaa aacggtcgtg acatgtacgt cgaccaagaa ttggatatca acagattgtc
     5401 tgactacgat gtcgatcata tcgtcccaca atcgttcttg aaggacgatt ccattgacaa
     5461 caaagttttg actagatctg acaagaacag aggtaagtct gataacgttc catctgaaga
     5521 agttgttaag aagatgaaga actactggag acaattgttg aatgctaagt tgatcactca
     5581 aagaaagttc gacaacttga ccaaggctga aagaggtggt ttgtccgaat tggacaaagc
     5641 cggtttcatc aagagacaat tagtcgaaac tagacaaatc accaagcatg ttgctcaaat
     5701 cttggattcc agaatgaaca ctaagtacga tgaaaacgac aaactaatta gagaagttaa
     5761 ggtcatcact ttgaagtcta agttggtttc tgacttcaga aaggacttcc aattttacaa
     5821 ggtcagagaa atcaacaact accatcacgc tcacgatgcc tacttgaacg ctgttgtcgg
     5881 tactgcctta atcaaaaagt acccaaagtt ggaatctgaa ttcgtttacg gtgactacaa
     5941 ggtttacgat gttagaaaga tgatcgccaa gtctgaacaa gaaattggta aggccactgc
     6001 taagtacttc ttctactcta acatcatgaa ctttttcaag actgaaatca ctttagctaa
     6061 cggtgaaatt agaaagcgtc cattgattga aaccaatggt gaaactggtg aaattgtctg
     6121 ggacaagggt agagatttcg ctaccgtcag aaaggttttg tctatgccac aagttaacat
     6181 cgtcaagaag actgaagttc aaactggtgg tttctctaag gaatccattt tgccaaagag
     6241 aaactctgac aagttgattg ctagaaagaa ggactgggat cctaagaagt acggtggttt
     6301 cgactctcca actgttgctt actccgtttt ggtcgttgct aaggttgaaa agggtaagtc
     6361 taagaagttg aagtctgtta aggaattgtt gggtatcacc atcatggaaa gatcctcctt
     6421 cgaaaagaac ccaatcgact ttttggaagc taagggttac aaggaagtca agaaggattt
     6481 gatcattaag ttaccaaaat actccttgtt cgaattggaa aacggtagaa agagaatgtt
     6541 ggcctccgct ggtgaactac aaaaaggtaa cgaattggct ttaccatcta agtacgttaa
     6601 cttcttgtac ttggcttccc actacgaaaa gttgaaaggt tccccagaag acaacgaaca
     6661 aaagcaattg tttgttgaac aacacaagca ctacttggat gaaattattg aacaaatctc
     6721 cgaattctcc aagagagtca ttttggctga tgctaactta gataaggttt tatccgctta
     6781 caacaagcac agagacaaac caatcagaga acaagctgaa aacatcattc atttgttcac
     6841 tttaaccaac ttgggtgctc cagctgcttt caaatacttc gacactacca ttgacagaaa
     6901 gagatacact tccaccaaag aagttttaga tgctactttg attcaccaat ctattaccgg
     6961 tttgtacgaa accagaattg acttgtctca attgggtggt gattccagag ctgatccaaa
     7021 gaagaagaga aaggtgtaaa ggagttaaag gcaaagtttt cttttctaga gccgttccca
     7081 caaataatta tacgtatatg cttcttttcg tttactatat atctatattt acaagccttt
     7141 attcactgat gcaatttgtt tccaaatact tttttggaga tctcataact agatatcatg
     7201 atggcgcaac ttggcgctat cttaattact ctggctgcca ggcccgtgta gagggccgca
     7261 agaccttctg tacgccatat agtctctaag aacttgaaca agtttctaga cctattgccg
     7321 cctttcggat cgctattgtt gcggccgcca gctgaagctt cgtacgctgc aggtcgacga
     7381 attctaccgt tcgtataatg tatgctatac gaagttatag atctgtttag cttgcctcgt
     7441 ccccgccggg tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg
     7501 acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc
     7561 atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg
     7621 gctcctcgct gcagacctgc gagcagggaa acgctcccct cacagacgcg ttgaattgtc
     7681 cccacgccgc gcccctgtag agaaatataa aaggttagga tttgccactg aggttcttct
     7741 ttcatatact tccttttaaa atcttgctag gatacagttc tcacatcaca tccgaacata
     7801 aacaaccatg ggtaaggaaa agactcacgt ttcgaggccg cgattaaatt ccaacatgga
     7861 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     7921 ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     7981 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     8041 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     8101 gatccccggc aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     8161 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     8221 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     8281 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     8341 agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     8401 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     8461 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     8521 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     8581 attgcagttt catttgatgc tcgatgagtt tttctaatca gtactgacaa taaaaagatt
     8641 cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt tctattttaa
     8701 tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc agatgcgaag
     8761 ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt cgctatactg
     8821 ctgtcgattc gatactaacg ccgccatcca gtgtcgaaaa cgagctcata acttcgtata
     8881 atgtatgcta tacgaacggt agaattcgaa tcagatccac tagtggccta tgcggccgcc
     8941 accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     9001 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     9061 aggagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgaggt aactcacatt
     9121 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     9181 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     9241 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     9301 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     9361 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     9421 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     9481 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     9541 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     9601 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     9661 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     9721 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     9781 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     9841 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     9901 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     9961 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
    10021 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
    10081 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
    10141 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
    10201 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
    10261 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
    10321 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
    10381 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
    10441 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
    10501 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
    10561 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
    10621 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
    10681 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
    10741 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
    10801 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
    10861 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
    10921 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
    10981 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
    11041 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
    11101 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg
    11161 gtccttttca tcacgtgcta taaaaataat tataatttaa attttttaat ataaatatat
    11221 aaattaaaaa tagaaagtaa aaaaagaaat taaagaaaaa atagtttttg ttttccgaag
    11281 atgtaaaaga ctctaggggg atcgccaaca aatactacct tttatcttgc tcttcctgct
    11341 ctcaggtatt aatgccgaat tgtttcatct tgtctgtgta gaagaccaca cacgaaaatc
    11401 ctgtgatttt acattttact tatcgttaat cgaatgtata tctatttaat ctgcttttct
    11461 tgtctaataa atatatatgt aaagtacgct ttttgttgaa attttttaaa cctttgttta
    11521 tttttttttc ttcattccgt aactcttcta ccttctttat ttactttcta aaatccaaat
    11581 acaaaacata aaaataaata aacacagagt aaattcccaa attattccat cattaaaaga
    11641 tacgaggcgc gtgtaagtta caggcaagcg atccgtccta agaaaccatt attatcatga
    11701 cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
