LOCUS       sequence-288271-       11939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9 gap repair vector - LEU2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11939)
  TITLE     Ellis Lab Yeast CRISPR Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11939)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pWS171
FEATURES             Location/Qualifiers
     source          1..11939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1451..5554
                     /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             1451..5554
                     /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             5567..5587
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5591..5611
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     RBS             6521..6532
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             6539..7252
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      7264..7335
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7351..7378
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        7392..7467
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8179..8586
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             8587..9681
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     rep_origin      9860..10842
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(10900..11715)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(10900..11715)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     11623..11642
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     11707..11726
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtgc ggccgcccct gaattcgcat ctagactgaa
      721 ctggccgata attgcagacg aacggtgagt aaggaaagag tgaggaacta tcgcatacct
      781 gcatttaaag atgccgattt gggcgcgaat cctttatttt ggcttcaccc tcatactatt
      841 atcagggcca gaaaaaggaa gtgtttccct ccttcttgaa ttgatgttac cctcataaag
      901 cacgtggcct cttatcgaga aagaaattac cgtcgctcgt gatttgtttg caaaaagaac
      961 aaaactgaaa aaacccagac acgctcgact tcctgtcatc ctattgattg cagcttccaa
     1021 tttcgtcaca caacaaggtc ctagcgacgg ctcacaggtt ttgtaacaag caatcgaagg
     1081 ttctggaatg gcgggaaagg gtttagtacc acatgctatg atgcccactg tgatctccag
     1141 agcaaagttc gttcgatcgt actgttactc tctctctttc aaacagaatt gtccgaatcg
     1201 tgtgacaaca acagcctgtt ctcacacact cttttcttct aaccaagggg gtggtttagt
     1261 ttagtagaac ctcgtgaaac ttacatttac atatatataa acttgcataa attggtcaat
     1321 gcaagaaata catatttggt cttttctaat tcgtagtttt tcaagttctt agatgctttc
     1381 tttttctctt ttttacagat catcaaggaa gtaattatct actttttaca acaaatataa
     1441 aacaagatct atggacaaga agtattctat cggactggat atcgggacta atagcgtcgg
     1501 gtgggccgtg atcactgacg agtacaaggt gccctctaag aagttcaagg tgctcgggaa
     1561 caccgaccgg cattccatca agaaaaatct gatcggagct ctcctctttg attcagggga
     1621 aaccgctgaa gcaacccgcc tcaagcggac tgctagacgg cggtacacca ggaggaagaa
     1681 ccggatttgt taccttcaag agatattctc caacgaaatg gcaaaggtcg acgacagctt
     1741 cttccatagg ctggaagaat cattcctcgt ggaagaggat aagaagcatg aacggcatcc
     1801 catcttcggt aatatcgtcg acgaggtggc ctatcacgag aaatacccaa ccatctacca
     1861 tcttcgcaaa aagctggtgg actcaaccga caaggcagac ctccggctta tctacctggc
     1921 cctggcccac atgatcaagt tcagaggcca cttcctgatc gagggcgacc tcaatcctga
     1981 caatagcgat gtggataaac tgttcatcca gctggtgcag acttacaacc agctctttga
     2041 agagaacccc atcaatgcaa gcggagtcga tgccaaggcc attctgtcag cccggctgtc
     2101 aaagagccgc agacttgaga atcttatcgc tcagctgccg ggtgaaaaga aaaatggact
     2161 gttcgggaac ctgattgctc tttcacttgg gctgactccc aatttcaagt ctaatttcga
     2221 cctggcagag gatgccaagc tgcaactgtc caaggacacc tatgatgacg atctcgacaa
     2281 cctcctggcc cagatcggtg accaatacgc cgaccttttc cttgctgcta agaatctttc
     2341 tgacgccatc ctgctgtctg acattctccg cgtgaacact gaaatcacca aggcccctct
     2401 ttcagcttca atgattaagc ggtatgatga gcaccaccag gacctgaccc tgcttaaggc
     2461 actcgtccgg cagcagcttc cggagaagta caaggaaatc ttctttgacc agtcaaagaa
     2521 tggatacgcc ggctacatcg acggaggtgc ctcccaagag gaattttata agtttatcaa
     2581 acctatcctt gagaagatgg acggcaccga agagctcctc gtgaaactga atcgggagga
     2641 tctgctgcgg aagcagcgca ctttcgacaa tgggagcatt ccccaccaga tccatcttgg
     2701 ggagcttcac gccatccttc ggcgccaaga ggacttctac ccctttctta aggacaacag
     2761 ggagaagatt gagaaaattc tcactttccg catcccctac tacgtgggac ccctcgccag
     2821 aggaaatagc cggtttgctt ggatgaccag aaagtcagaa gaaactatca ctccctggaa
     2881 cttcgaagag gtggtggaca agggagccag cgctcagtca ttcatcgaac ggatgactaa
     2941 cttcgataag aacctcccca atgagaaggt cctgccgaaa cattccctgc tctacgagta
     3001 ctttaccgtg tacaacgagc tgaccaaggt gaaatatgtc accgaaggga tgaggaagcc
     3061 cgcattcctg tcaggcgaac aaaagaaggc aattgtggac cttctgttca agaccaatag
     3121 aaaggtgacc gtgaagcagc tgaaggagga ctatttcaag aaaattgaat gcttcgactc
     3181 tgtggagatt agcggggtcg aagatcggtt caacgcaagc ctgggtactt accatgatct
     3241 gcttaagatc atcaaggaca aggattttct ggacaatgag gagaacgagg acatccttga
     3301 ggacattgtc ctgactctca ctctgttcga ggaccgggaa atgatcgagg agaggcttaa
     3361 gacctacgcc catctgttcg acgataaagt gatgaagcaa cttaaacgga gaagatatac
     3421 cggatgggga cgccttagcc gcaaactcat caacggaatc cgggacaaac agagcggaaa
     3481 gaccattctt gatttcctta agagcgacgg attcgctaat cgcaacttca tgcaacttat
     3541 ccatgatgat tccctgacct ttaaggagga catccagaag gcccaagtgt ctggacaagg
     3601 tgactcactg cacgagcata tcgcaaatct ggctggttca cccgctatta agaagggtat
     3661 tctccagacc gtgaaagtcg tggacgagct ggtcaaggtg atgggtcgcc ataaaccaga
     3721 gaacattgtc atcgagatgg ccagggaaaa ccagactacc cagaagggac agaagaacag
     3781 cagggagcgg atgaaaagaa ttgaggaagg gattaaggag ctcgggtcac agatccttaa
     3841 agagcacccg gtggaaaaca cccagcttca gaatgagaag ctctatctgt actaccttca
     3901 aaatggacgc gatatgtatg tggaccaaga gcttgatatc aacaggctct cagactacga
     3961 cgtggaccat atcgtccctc agagcttcct caaagacgac tcaattgaca ataaggtgct
     4021 gactcgctca gacaagaacc ggggaaagtc agataacgtg ccctcagagg aagtcgtgaa
     4081 aaagatgaag aactattggc gccagcttct gaacgcaaag ctgatcactc agcggaagtt
     4141 cgacaatctc actaaggctg agaggggcgg actgagcgaa ctggacaaag caggattcat
     4201 taaacggcaa cttgtggaga ctcggcagat tactaaacat gtcgcccaaa tccttgactc
     4261 acgcatgaat accaagtacg acgaaaacga caaacttatc cgcgaggtga aggtgattac
     4321 cctgaagtcc aagctggtca gcgatttcag aaaggacttt caattctaca aagtgcggga
     4381 gatcaataac tatcatcatg ctcatgacgc atatctgaat gccgtggtgg gaaccgccct
     4441 gatcaagaag tacccaaagc tggaaagcga gttcgtgtac ggagactaca aggtctacga
     4501 cgtgcgcaag atgattgcca aatctgagca ggagatcgga aaggccaccg caaagtactt
     4561 cttctacagc aacatcatga atttcttcaa gaccgaaatc acccttgcaa acggtgagat
     4621 ccggaagagg ccgctcatcg agactaatgg ggagactggc gaaatcgtgt gggacaaggg
     4681 cagagatttc gctaccgtgc gcaaagtgct ttctatgcct caagtgaaca tcgtgaagaa
     4741 aaccgaggtg caaaccggag gcttttctaa ggaatcaatc ctccccaagc gcaactccga
     4801 caagctcatt gcaaggaaga aggattggga ccctaagaag tacggcggat tcgattcacc
     4861 aactgtggct tattctgtcc tggtcgtggc taaggtggaa aaaggaaagt ctaagaagct
     4921 caagagcgtg aaggaactgc tgggtatcac cattatggag cgcagctcct tcgagaagaa
     4981 cccaattgac tttctcgaag ccaaaggtta caaggaagtc aagaaggacc ttatcatcaa
     5041 gctcccaaag tatagcctgt tcgaactgga gaatgggcgg aagcggatgc tcgcctccgc
     5101 tggcgaactt cagaagggta atgagctggc tctcccctcc aagtacgtga atttcctcta
     5161 ccttgcaagc cattacgaga agctgaaggg gagccccgag gacaacgagc aaaagcaact
     5221 gtttgtggag cagcataagc attatctgga cgagatcatt gagcagattt ccgagttttc
     5281 taaacgcgtc attctcgctg atgccaacct cgataaagtc cttagcgcat acaataagca
     5341 cagagacaaa ccaattcggg agcaggctga gaatatcatc cacctgttca ccctcaccaa
     5401 tcttggtgcc cctgccgcat tcaagtactt cgacaccacc atcgaccgga aacgctatac
     5461 ctccaccaaa gaagtgctgg acgccaccct catccaccag agcatcaccg gactttacga
     5521 aactcggatt gacctctcac agctcggagg ggatgaggga gctgatccaa aaaagaagag
     5581 aaaggtagat ccaaaaaaga agagaaaggt ataaggatcc taactcgaga ttgaattgaa
     5641 ttgaaatcga tagatcaatt tttttctttt ctctttcccc atcctttacg ctaaaataat
     5701 agtttatttt attttttgaa tattttttat ttatatacgt atatatagac tattatttat
     5761 cttttaatga ttattaagat ttttattaaa aaaaaattcg ctcctctttt aatgccttta
     5821 tgcagttttt ttttcccatt cgatatttct atgtgctgcc aagatatcaa caacagagga
     5881 catatgccct acctccatga tctactgacg tccctgaggc tgcaattcat gtaatggggc
     5941 agtatccgcg gcaagtccta gtgcaatggc ggttttttac cctcgttctg aagaagaggc
     6001 gacgcgggtg cggtcatcac taatgtggaa attggtaaga ctctcgggcc tccgccttta
     6061 ggcggtgctt actctttcat aaaggggctg ttagttatgc aacgatgtgc ttcagtatta
     6121 cattttttgc cttcaacgcc ttgattgttc tatttttgct aataataaat ctatttcatc
     6181 ggactaaaag tccattagtt gtaagcggat ttagctcagt tgggagagcg ccagactgaa
     6241 gaaaaacttc ggtcaagtca tctggaggtc ctgtgttcga tccacagaat tcgcagatgg
     6301 ccggcatggt cccagcctcc tcgctggcgc cggctgggca acaccttcgg gtggcgaatg
     6361 ggacttgaga cggaaagtga aacgtgattt catgcgtcat tttgaacatt ttgtaaatct
     6421 tatttaataa tgtgtgcggc aattcacatt taatttatga atgttttctt aacatcgcgg
     6481 caactcaaga aacggcaggt tcggatctta gctactagag aaagaggaga aatactagat
     6541 gcgtaaaggc gaagagctgt tcactggtgt cgtccctatt ctggtggaac tggatggtga
     6601 tgtcaacggt cataagtttt ccgtgcgtgg cgagggtgaa ggtgacgcaa ctaatggtaa
     6661 actgacgctg aagttcatct gtactactgg taaactgccg gttccttggc cgactctggt
     6721 aacgacgctg acttatggtg ttcagtgctt tgctcgttat ccggaccata tgaagcagca
     6781 tgacttcttc aagtccgcca tgccggaagg ctatgtgcag gaacgcacga tttcctttaa
     6841 ggatgacggc acgtacaaaa cgcgtgcgga agtgaaattt gaaggcgata ccctggtaaa
     6901 ccgcattgag ctgaaaggca ttgactttaa agaggacggc aatatcctgg gccataagct
     6961 ggaatacaat tttaacagcc acaatgttta catcaccgcc gataaacaaa aaaatggcat
     7021 taaagcgaat tttaaaattc gccacaacgt ggaggatggc agcgtgcagc tggctgatca
     7081 ctaccagcaa aacactccaa tcggtgatgg tcctgttctg ctgccagaca atcactatct
     7141 gagcacgcaa agcgttctgt ctaaagatcc gaacgagaaa cgcgatcata tggttctgct
     7201 ggagttcgta accgcagcgg gcatcacgca tggtatggat gaactgtaca aatgaccagg
     7261 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     7321 tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt
     7381 tatacgtctc agttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
     7441 cttgaaaaag tggcaccgag tcggtgcttt ttttattttt tgtcactatt gttatgtaaa
     7501 atgccacctc tgacagtatg gaacgcaaac ttctgtctag tggatagctg gacattccag
     7561 gcagtgcctc tgccgccgga cccctctcgt gattgggtag ctggacatgc ccttgtaaga
     7621 tataacaaga gcctgcctgt ctaatgatct cacggcgaaa gtcggggaga cagcagcggc
     7681 tgcagacatt ataccgcaac aacactaagg tgagataact ccgtacattg actacgcgtt
     7741 cctctagacc ttacttgacc ggatacagtg tctttgacac gtttatgggt tacagcaatc
     7801 acatccaaga ctggctatgc acgaagcaac tcttgagtgt taaaatgttg acccctgtat
     7861 ttgggatgcg ggtagtagat gagtgcagga tatcagcagt tacagagatg ttacgaacca
     7921 ctagtgcact gcagtacatc gaggagaact tctagtatat ctacatacct aatattattg
     7981 ccttattaaa aatggaatcc caacaattac atcaaaatcc acattctctt caaaatcaat
     8041 tgtcctgtac ttccttgttc atgtgtgttc aaaaacgtta tatttatagg ataattatac
     8101 tctatttctc aacaagtaat tggttgtttg gccgagcggt ctaaggcgcc tgattcaaga
     8161 aatatcttga ccgcagttaa ctgtgggaat actcaggtat cgtaagatgc aagagttcga
     8221 atctcttagc aaccattatt tttttcctca acataacgag aacacacagg ggcgctatcg
     8281 cacagaatca aattcgatga ctggaaattt tttgttaatt tcagaggtcg cctgacgcat
     8341 ataccttttt caactgaaaa attgggagaa aaaggaaagg tgagagcgcc ggaaccggct
     8401 tttcatatag aatagagaag cgttcatgac taaatgcttg catcacaata cttgaagttg
     8461 acaatattat ttaaggacct attgtttttt ccaataggtg gttagcaatc gtcttacttt
     8521 ctaacttttc ttacctttta catttcagca atatatatat atatatttca aggatatacc
     8581 attctaatgt ctgcccctaa gaagatcgtc gttttgccag gtgaccacgt tggtcaagaa
     8641 atcacagccg aagccattaa ggttcttaaa gctatttctg atgttcgttc caatgtcaag
     8701 ttcgatttcg aaaatcattt aattggtggt gctgctatcg atgctacagg tgttccactt
     8761 ccagatgagg cgctggaagc ctccaagaag gctgatgccg ttttgttagg tgctgtgggt
     8821 ggtcctaaat ggggaaccgg tagtgttaga cctgaacaag gtttactaaa aatccgtaaa
     8881 gaacttcaat tgtacgccaa cttaagacca tgtaactttg catccgactc tcttttagac
     8941 ttatctccaa tcaagccaca atttgctaaa ggtactgact tcgttgttgt cagagaatta
     9001 gtgggaggta tttactttgg taagagaaag gaggacgatg gtgatggtgt cgcttgggat
     9061 agtgaacaat acaccgttcc agaagtgcaa agaatcacaa gaatggccgc tttcatggcc
     9121 ctacaacatg agccaccatt gcctatttgg tccttggata aagctaatgt tttggcctct
     9181 tcaagattat ggagaaaaac tgtggaggaa accatcaaga acgaatttcc tacattgaag
     9241 gttcaacatc aattgattga ttctgccgcc atgatcctag ttaagaaccc aacccaccta
     9301 aatggtatta taatcaccag caacatgttt ggtgatatca tctccgatga agcctccgtt
     9361 atcccaggtt ccttgggttt gttgccatct gcgtccttgg cctctttgcc agacaagaac
     9421 accgcatttg gtttgtacga accatgccac ggttctgctc cagatttgcc aaagaataag
     9481 gtcaacccta tcgccactat cttgtctgct gcaatgatgt tgaaattgtc attgaacttg
     9541 cctgaagaag gtaaggccat tgaagatgca gttaaaaagg ttttggatgc aggtatcaga
     9601 actggtgatt taggtggttc caacagtacc accgaagtcg gtgatgctgt cgccgaagaa
     9661 gttaagaaaa tccttgctta acctggagga cccttctctt tagactattc tactcttatg
     9721 cacgtaaaaa attctaggaa atatgtatta actaggagta aaataaccgg ctagtggcat
     9781 tcatatagcc gtctgtttac atctacatca cacatttcga gtgtatatct cgcaacgttg
     9841 gcgttaaata ggcaggagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa
     9901 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
     9961 caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa
    10021 tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
    10081 aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac
    10141 aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc
    10201 ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt
    10261 tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc
    10321 ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc
    10381 gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg
    10441 atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta
    10501 cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac
    10561 tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag
    10621 tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca
    10681 gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt
    10741 ttagtagctc gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc
    10801 ttttggtttt caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc
    10861 ccgagcggcc gcgtgttaca accaattaac caattctgat tagaaaaact catcgagcat
    10921 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
    10981 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
    11041 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
    11101 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
    11161 aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
    11221 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga ggcgaaatac
    11281 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
    11341 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
    11401 tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
    11461 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
    11521 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
    11581 cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
    11641 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctggagcaag acgtttcccg
    11701 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
    11761 tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg
    11821 gctttgttga ataaatcgaa cttttgctga gttgaaggat cagtcatgac caaaatccct
    11881 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttc
//
