LOCUS       sequence-169723-       11528 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 in fission yeast using fluoride selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11528)
  AUTHORS   Fernandez R, Berro J
  TITLE     Use of a fluoride channel as a new selection marker for fission 
            yeast plasmids and application to fast genome editing with 
            CRISPR/Cas9.
  JOURNAL   Yeast. 2016 Oct;33(10):549-557. doi: 10.1002/yea.3178. Epub 2016 Sep
            7.
  PUBMED    27327046
REFERENCE   2  (bases 1 to 11528)
  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"; doi: "10.1002/yea.3178"; 
            journalName: "Yeast"; date: "2016-10"; volume: "33"; issue: "10"; 
            pages: "549-557"
COMMENT     Sequence Label: pJB166
FEATURES             Location/Qualifiers
     source          1..11528
                     /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"
     rep_origin      539..1320
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     3494..3516
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3508..3525
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3509..3525
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        complement(3575..3626)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     promoter        complement(3745..4101)
                     /label=rrk1 promoter/leader
                     /note="promoter and leader RNA for the Schizosaccharomyces 
                     pombe rrk1 gene encoding the K-RNA component of RNase P 
                     (Jacobs et al., 2014)"
     primer_bind     complement(4138..4154)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4138..4154)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4151..4173)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4162..4178
                     /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(4186..4192,4193..4210,4211..4216))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4231..4252
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4369..4386)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4540..5128)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4620..4639)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5299..6090,6091..6159))
                     /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     5922..5941
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6160..6264)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6293..7034
                     /label=adh1 promoter
                     /note="promoter for the Schizosaccharomyces pombe adh1 gene
                     encoding alcohol dehydrogenase"
     CDS             7055..11158
                     /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             7055..11158
                     /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             7055..11158
                     /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             11171..11191
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      11207..11454
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(11223..11241)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     11501..11519
                     /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 accataatcg atggccaacc ttccaattca ttaaatcgag tcctccatca gatgatttac
      241 aaggttttgc atagaatccc taagtaagag aagttgttag tgatttcttc aagtttacta
      301 aaatttacat acaaatggat gatctcttct ccattgtttc ctttcttctt gcaaacgcgt
      361 tttacaaaga gatgacatgc tattttctga ttattttttt ctatagtttt ctattttgta
      421 cgtccttggg agtccgaaat gtaaaatcgg ggtatggtta gtagcggcag cgttcattat
      481 ggtatataaa agataaaaag taggggcaag cgagacagaa tgggatacaa gggcatcgtc
      541 tataattata gctaaaaatt gtattttaat ttgtattttt tgtaatttta tttctcattg
      601 ttttacttaa aatgaatgcg aattagagaa aacttcaacg aaatgtcaaa ataagctcag
      661 caaaatatac aattttaggg aaagcgatca gcaacttatt ctccgttgta gattcttttt
      721 cttcataaat tccaacaatg taactgccaa ttgatttttc tctagatgct aataaattga
      781 aactaatgaa tataataaca caggttttga taaattgtaa actcttactc aaaaaaagaa
      841 tgttaaaact aatttaacct ttttataaag gctcatggcg ttaaagagga tagtaaaaat
      901 tctacaattt tttaatttaa tgtcacaatt gtcttgcgaa tataattcta ttaattttcg
      961 ctaactgctg ttttagtact attactttca atatgcctta tatgcaactt taattcatga
     1021 agaatgcaat atttttgcaa tctttcttgt actgagcgct ttttatcatt tagtactcat
     1081 tatttaattt ttcagtaaag ggcaataaat aaatttgtag aagatgcaat gtaatctcct
     1141 ctatcctttt gctcatatgt ttatgagtat acctagtcta gaaaggcttg tatttaaaat
     1201 attattcaat aaaaattcac aatttttacg acatgtgcta ccattcactt aacttcctga
     1261 ttataaaatt ggttcgttta tactaattac ttaagtacct ttaactaaac aaaatgccta
     1321 tatatatatt aatttacaat gagtgtcaga taagtcacta tgtccgagtg gttaaggagt
     1381 tagactcgcc atcgatgagc ttagctacaa atcccactgg ctatatgtat gcatttgtgt
     1441 taaaaaagtt tgtatagatt atttaatcta ctcagcattc tttctctaaa taggaatttg
     1501 ttacttaatg gagaaaaaaa tgtttcgatt tacctagtgt atttgtttgt atactcacgt
     1561 ttaatttcaa acatccattc tatcttgtgt aatttttggc atggtgaaaa agataatcag
     1621 ccttataatc tttacaaaag taagaaattc tgtaaataag ccttaatgcc cttgctttaa
     1681 attaaaatgg ttctttttca tgataatgtt tgcactttgt gaatatattt tagatagttc
     1741 tgtgaggtat aattaagatg ttttagagac ttatacaatt ttgtctttat aaattcttaa
     1801 ttgattttac catcccagtt taactatgct tcgtcggcat ctctgcacat gtcgtgtttt
     1861 cttaccgtat tgtcctacca agaacctctt ttttgcttgg atcgaaatta aaggtttaaa
     1921 agcaaagtta tgctattaac acagtcttac ttttgtataa tgtctatgtt ggggacgcta
     1981 gctcgtctag gacttactgc attaaacaca tacccaggtg caccgttttc tggactttta
     2041 tgggttcaat ttgttggatg cgttattatg ggattttgtc agacagagtc tgttttcttt
     2101 cctcgaccca aacataatgc tacttttttg ctagccatta ctacaggttt ttgtggatct
     2161 cttacaacgt tttcttcttg gatgttgcaa atgtttacag ggatggctaa cttggatcca
     2221 tttgaacgta ggggccgagg ctattcattt ttaagtgtgg tttccgattt tatggtaact
     2281 atgtgcattg cgatgtccag cttaatatgg gggaagcaga ttggaaaaac gacgggacag
     2341 tggagaattg gcaaagttgc atttgcctgg cctatcccgg cgcacacgca tatcgtagtg
     2401 cgggtattac ttttgttact ctcgatttgt ttttttgttg gtgcagcatt ttatacggca
     2461 tatacaacca atgtgacaca tcgtggcata ggttttagtc taatattctc tccttttgct
     2521 gccttgacac gtttgtatct tgcgcgattc cttaattcac ctcaatattt tattccctat
     2581 ggaactcttt gtgccaatgt ctttgctaca cttttgcttt ctatcatgta catgatacca
     2641 caaatcacac actgcactcc tgtgtctaga agtgtgatgt atggaatcca aaacggattt
     2701 tgtgctgtac tctctaccct ttcaacattc tccaacgaac tccacacaat gcctataaaa
     2761 cgagcatata tttattgtat aatttcagtt gctatttcct tttcaatatg tgtcattgtc
     2821 gatggagcta ctgcatgggg tcatggttat actgaaaagt attagtaaaa aaagactaat
     2881 gtaaaatttt tttggttggt tattgaaaaa gtcgatgcct tgtttgcgtt tgttttccta
     2941 ggcgttttat gtcagaaggc atttagaatt agtatacaag tactctttgg taaaatttta
     3001 tgtagcgact aaaatattaa ctattataga taaacacctt gggaataaaa agtaatttgc
     3061 tatagtaatt tattaaacat gctcctacaa cattaccaca atcttttctc ttggattgac
     3121 attgaataag aaaagagtga atttttttag acttgtaatg ataactatgt acaaagccaa
     3181 tgaaagatgt atgtagatga atgtaaaata ccatgtagac aaacaagata aaacttggtt
     3241 ataaacattg gtgttggaac agaataaatt agatgtcaaa aagtttcgtc aatatcacaa
     3301 gctcatcgcg atggtatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     3361 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     3421 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     3481 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttgcatgc
     3541 ctgcaggtcg actctagagg atccccacgg tacccctgtt tcgtcctcac ggactcatca
     3601 gaccggaaag cacatccggt gacagggcac caccgactcg gtgccactca cttttttcaa
     3661 gttgataacg gactagcctt attttaactt gctatttcta gctctaaaac agagagataa
     3721 cccacaagaa ttgagttaag cccattcttc ggtacaggtt atgttttttg gcaacagcaa
     3781 ctattcaaat caactgaaag ttttttagaa ttcaacaact tccaatactc aagtgtccaa
     3841 gctagcaaac tgtttaataa tcgtgaccca acattccttt catgcgcacc ctcagcgtca
     3901 cataaatcaa cactttacaa cacctattca aagcttctat atgccttcgt catttaaatg
     3961 tctaacattt caaaaaagtg tttgctaatg gtaaattctt gagttttctt ggaagcacat
     4021 atataacgag tgtgactgtt ctacgctcct tgctgactag tcatcagtct acgcatgcca
     4081 actaccacca acataagcaa aaggtacccg gggtaccgag ctcgaattcg taatcatgtc
     4141 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4201 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4261 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4321 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4381 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4441 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4501 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4561 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4621 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4681 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4741 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4801 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4861 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4921 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4981 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5041 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5101 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5161 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5221 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5281 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5341 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5401 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5461 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5521 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5581 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5641 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5701 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5761 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5821 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5881 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5941 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6001 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6061 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6121 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6181 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6241 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgccctaca
     6301 acaactaaga aaatggctat catgcggaag ctggtgagaa gaacagcatc gggacaaggg
     6361 aaggaagaac aaagacaaag aaaacaaaag aaagcaattg aaaacaaaac aaaacaattt
     6421 tcattccttc tcttatcatt ccttttcttt tcttttctct cattcaacgc actccatcgt
     6481 atccgtattc ctcttatttt ttctctttct ctatatccat ttctttctct ctaggtgtgt
     6541 cctctctctc tcttcaattt ctctactccg cattccaacg catccttccc ccaacctccc
     6601 atttcctcct tacggcccga tagcgatcgt ctttccctcg ctatcactcg ctaccggccc
     6661 ctcctctgca ccgtaacctc ctacgtattt accatatcat aaagtttttt ccgacgctta
     6721 tcgctgaccc cctgtcgccc tcctattggc ttccggatta tcttcttgtc cataaggtga
     6781 tccatgcttc ctgaagattc ccgaaatgtg tccactttgg cggggaatca ttccatccac
     6841 ttctttctct ctcgctttcc tcattcggcg ctccccttcc gcgtctcatt ggtcttccgc
     6901 tccgtttttg ctttgccgat gttacttggg gagaggtgcg ataatccttt cgcaaaaact
     6961 cggtttgacg cctcccatgg tataaatagt gggtggtgga caggtgcctt cgcttttctt
     7021 taagcaagag aattgctgca ggtcgactct agagatggac aagaagtact ccattgggct
     7081 cgatatcggc acaaacagcg tcggttgggc cgtcattacg gacgagtaca aggtgccgag
     7141 caaaaaattc aaagttctgg gcaataccga tcgccacagc ataaagaaga acctcattgg
     7201 cgccctcctg ttcgactccg gggagacggc cgaagccacg cggctcaaaa gaacagcacg
     7261 gcgcagatat acccgcagaa agaatcggat ctgctacctg caggagatct ttagtaatga
     7321 gatggctaag gtggatgact ctttcttcca taggctggag gagtcctttt tggtggagga
     7381 ggataaaaag cacgagcgcc acccaatctt tggcaatatc gtggacgagg tggcgtacca
     7441 tgaaaagtac ccaaccatat atcatctgag gaagaagctt gtagacagta ctgataaggc
     7501 tgacttgcgg ttgatctatc tcgcgctggc gcatatgatc aaatttcggg gacacttcct
     7561 catcgagggg gacctgaacc cagacaacag cgatgtcgac aaactcttta tccaactggt
     7621 tcagacttac aatcagcttt tcgaagagaa cccgatcaac gcatccggag ttgacgccaa
     7681 agcaatcctg agcgctaggc tgtccaaatc ccggcggctc gaaaacctca tcgcacagct
     7741 ccctggggag aagaagaacg gcctgtttgg taatcttatc gccctgtcac tcgggctgac
     7801 ccccaacttt aaatctaact tcgacctggc cgaagatgcc aagcttcaac tgagcaaaga
     7861 cacctacgat gatgatctcg acaatctgct ggcccagatc ggcgaccagt acgcagacct
     7921 ttttttggcg gcaaagaacc tgtcagacgc cattctgctg agtgatattc tgcgagtgaa
     7981 cacggagatc accaaagctc cgctgagcgc tagtatgatc aagcgctatg atgagcacca
     8041 ccaagacttg actttgctga aggcccttgt cagacagcaa ctgcctgaga agtacaagga
     8101 aattttcttc gatcagtcta aaaatggcta cgccggatac attgacggcg gagcaagcca
     8161 ggaggaattt tacaaattta ttaagcccat cttggaaaaa atggacggca ccgaggagct
     8221 gctggtaaag cttaacagag aagatctgtt gcgcaaacag cgcactttcg acaatggaag
     8281 catcccccac cagattcacc tgggcgaact gcacgctatc ctcaggcggc aagaggattt
     8341 ctaccccttt ttgaaagata acagggaaaa gattgagaaa atcctcacat ttcggatacc
     8401 ctactatgta ggccccctcg cccggggaaa ttccagattc gcgtggatga ctcgcaaatc
     8461 agaagagacc atcactccct ggaacttcga ggaagtcgtg gataaggggg cctctgccca
     8521 gtccttcatc gaaaggatga ctaactttga taaaaatctg cctaacgaaa aggtgcttcc
     8581 taaacactct ctgctgtacg agtacttcac agtttataac gagctcacca aggtcaaata
     8641 cgtcacagaa gggatgagaa agccagcatt cctgtctgga gagcagaaga aagctatcgt
     8701 ggacctcctc ttcaagacga accggaaagt taccgtgaaa cagctcaaag aagactattt
     8761 caaaaagatt gaatgtttcg actctgttga aatcagcgga gtggaggatc gcttcaacgc
     8821 atccctggga acgtatcacg atctcctgaa aatcattaaa gacaaggact tcctggacaa
     8881 tgaggagaac gaggacattc ttgaggacat tgtcctcacc cttacgttgt ttgaagatag
     8941 ggagatgatt gaagaacgct tgaaaactta cgctcatctc ttcgacgaca aagtcatgaa
     9001 acagctcaag aggcgccgat atacaggatg ggggcggctg tcaagaaaac tgatcaatgg
     9061 gatccgagat aagcagtctg gaaagacaat cctggatttt cttaagtccg atggatttgc
     9121 caaccggaac ttcatgcagt tgatccatga tgactctctc acctttaagg aggacatcca
     9181 gaaagcacaa gtttctggcc agggggacag tcttcacgag cacatcgcta atcttgcagg
     9241 tagcccagct atcaaaaagg gaatactgca gaccgttaag gtcgtggatg aactcgtcaa
     9301 agtaatggga aggcataagc ccgagaatat cgttatcgag atggcccgag agaaccaaac
     9361 tacccagaag ggacagaaga acagtaggga aaggatgaag aggattgaag agggtataaa
     9421 agaactgggg tcccaaatcc ttaaggaaca cccagttgaa aacacccagc ttcagaatga
     9481 gaagctctac ctgtactacc tgcagaacgg cagggacatg tacgtggatc aggaactgga
     9541 catcaatcgg ctctccgact acgacgtgga tcatatcgtg ccccagtctt ttctcaaaga
     9601 tgattctatt gataataaag tgttgacaag atccgataaa aatagaggga agagtgataa
     9661 cgtcccctca gaagaagttg tcaagaaaat gaaaaattat tggcggcagc tgctgaacgc
     9721 caaactgatc acacaacgga agttcgataa tctgactaag gctgaacgag gtggcctgtc
     9781 tgagttggat aaagccggct tcatcaaaag gcagcttgtt gagacacgcc agatcaccaa
     9841 gcacgtggcc caaattctcg attcacgcat gaacaccaag tacgatgaaa atgacaaact
     9901 gattcgagag gtgaaagtta ttactctgaa gtctaagctg gtctcagatt tcagaaagga
     9961 ctttcagttt tataaggtga gagagatcaa caattaccac catgcgcatg atgcctacct
    10021 gaatgcagtg gtaggcactg cacttatcaa aaaatatccc aagcttgaat ctgaatttgt
    10081 ttacggagac tataaagtgt acgatgttag gaaaatgatc gcaaagtctg agcaggaaat
    10141 aggcaaggcc accgctaagt acttctttta cagcaatatt atgaattttt tcaagaccga
    10201 gattacactg gccaatggag agattcggaa gcgaccactt atcgaaacaa acggagaaac
    10261 aggagaaatc gtgtgggaca agggtaggga tttcgcgaca gtccggaagg tcctgtccat
    10321 gccgcaggtg aacatcgtta aaaagaccga agtacagacc ggaggcttct ccaaggaaag
    10381 tatcctcccg aaaaggaaca gcgacaagct gatcgcacgc aaaaaagatt gggaccccaa
    10441 gaaatacggc ggattcgatt ctcctacagt cgcttacagt gtactggttg tggccaaagt
    10501 ggagaaaggg aagtctaaaa aactcaaaag cgtcaaggaa ctgctgggca tcacaatcat
    10561 ggagcgatca agcttcgaaa aaaaccccat cgactttctc gaggcgaaag gatataaaga
    10621 ggtcaaaaaa gacctcatca ttaagcttcc caagtactct ctctttgagc ttgaaaacgg
    10681 ccggaaacga atgctcgcta gtgcgggcga gctgcagaaa ggtaacgagc tggcactgcc
    10741 ctctaaatac gttaatttct tgtatctggc cagccactat gaaaagctca aagggtctcc
    10801 cgaagataat gagcagaagc agctgttcgt ggaacaacac aaacactacc ttgatgagat
    10861 catcgagcaa ataagcgaat tctccaaaag agtgatcctc gccgacgcta acctcgataa
    10921 ggtgctttct gcttacaata agcacaggga taagcccatc agggagcagg cagaaaacat
    10981 tatccacttg tttactctga ccaacttggg cgcgcctgca gccttcaagt acttcgacac
    11041 caccatagac agaaagcggt acacctctac aaaggaggtc ctggacgcca cactgattca
    11101 tcagtcaatt acggggctct atgaaacaag aatcgacctc tctcagctcg gtggagacag
    11161 cagggctgac cccaagaaga agaggaaggt gtgatctctt ctcgagtcat gtaattagtt
    11221 atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt
    11281 agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg
    11341 ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt
    11401 atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcgatccc
    11461 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
    11521 ctttcgtc
//
