LOCUS       sequence-173002-       11516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 in fission yeast using fluoride selection and targetting
            pil1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11516)
  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 11516)
  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: pJB172
FEATURES             Location/Qualifiers
     source          1..11516
                     /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      541..1322
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     3496..3518
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3510..3527
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3511..3527
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        complement(3577..3628)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     promoter        complement(3733..4089)
                     /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(4126..4142)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4126..4142)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4139..4161)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4150..4166
                     /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(4174..4180,4181..4198,4199..4204))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4219..4240
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4357..4374)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4528..5116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4608..4627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5287..6078,6079..6147))
                     /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     5910..5929
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6148..6252)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6281..7022
                     /label=adh1 promoter
                     /note="promoter for the Schizosaccharomyces pombe adh1 gene
                     encoding alcohol dehydrogenase"
     CDS             7043..11146
                     /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             7043..11146
                     /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             7043..11146
                     /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             11159..11179
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      11195..11442
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(11211..11229)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     11489..11507
                     /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 accataccat cgatggccaa ccttccaatt cattaaatcg agtcctccat cagatgattt
      241 acaaggtttt gcatagaatc cctaagtaag agaagttgtt agtgatttct tcaagtttac
      301 taaaatttac atacaaatgg atgatctctt ctccattgtt tcctttcttc ttgcaaacgc
      361 gttttacaaa gagatgacat gctattttct gattattttt ttctatagtt ttctattttg
      421 tacgtccttg ggagtccgaa atgtaaaatc ggggtatggt tagtagcggc agcgttcatt
      481 atggtatata aaagataaaa agtaggggca agcgagacag aatgggatac aagggcatcg
      541 tctataatta tagctaaaaa ttgtatttta atttgtattt tttgtaattt tatttctcat
      601 tgttttactt aaaatgaatg cgaattagag aaaacttcaa cgaaatgtca aaataagctc
      661 agcaaaatat acaattttag ggaaagcgat cagcaactta ttctccgttg tagattcttt
      721 ttcttcataa attccaacaa tgtaactgcc aattgatttt tctctagatg ctaataaatt
      781 gaaactaatg aatataataa cacaggtttt gataaattgt aaactcttac tcaaaaaaag
      841 aatgttaaaa ctaatttaac ctttttataa aggctcatgg cgttaaagag gatagtaaaa
      901 attctacaat tttttaattt aatgtcacaa ttgtcttgcg aatataattc tattaatttt
      961 cgctaactgc tgttttagta ctattacttt caatatgcct tatatgcaac tttaattcat
     1021 gaagaatgca atatttttgc aatctttctt gtactgagcg ctttttatca tttagtactc
     1081 attatttaat ttttcagtaa agggcaataa ataaatttgt agaagatgca atgtaatctc
     1141 ctctatcctt ttgctcatat gtttatgagt atacctagtc tagaaaggct tgtatttaaa
     1201 atattattca ataaaaattc acaattttta cgacatgtgc taccattcac ttaacttcct
     1261 gattataaaa ttggttcgtt tatactaatt acttaagtac ctttaactaa acaaaatgcc
     1321 tatatatata ttaatttaca atgagtgtca gataagtcac tatgtccgag tggttaagga
     1381 gttagactcg ccatcgatga gcttagctac aaatcccact ggctatatgt atgcatttgt
     1441 gttaaaaaag tttgtataga ttatttaatc tactcagcat tctttctcta aataggaatt
     1501 tgttacttaa tggagaaaaa aatgtttcga tttacctagt gtatttgttt gtatactcac
     1561 gtttaatttc aaacatccat tctatcttgt gtaatttttg gcatggtgaa aaagataatc
     1621 agccttataa tctttacaaa agtaagaaat tctgtaaata agccttaatg cccttgcttt
     1681 aaattaaaat ggttcttttt catgataatg tttgcacttt gtgaatatat tttagatagt
     1741 tctgtgaggt ataattaaga tgttttagag acttatacaa ttttgtcttt ataaattctt
     1801 aattgatttt accatcccag tttaactatg cttcgtcggc atctctgcac atgtcgtgtt
     1861 ttcttaccgt attgtcctac caagaacctc ttttttgctt ggatcgaaat taaaggttta
     1921 aaagcaaagt tatgctatta acacagtctt acttttgtat aatgtctatg ttggggacgc
     1981 tagctcgtct aggacttact gcattaaaca catacccagg tgcaccgttt tctggacttt
     2041 tatgggttca atttgttgga tgcgttatta tgggattttg tcagacagag tctgttttct
     2101 ttcctcgacc caaacataat gctacttttt tgctagccat tactacaggt ttttgtggat
     2161 ctcttacaac gttttcttct tggatgttgc aaatgtttac agggatggct aacttggatc
     2221 catttgaacg taggggccga ggctattcat ttttaagtgt ggtttccgat tttatggtaa
     2281 ctatgtgcat tgcgatgtcc agcttaatat gggggaagca gattggaaaa acgacgggac
     2341 agtggagaat tggcaaagtt gcatttgcct ggcctatccc ggcgcacacg catatcgtag
     2401 tgcgggtatt acttttgtta ctctcgattt gtttttttgt tggtgcagca ttttatacgg
     2461 catatacaac caatgtgaca catcgtggca taggttttag tctaatattc tctccttttg
     2521 ctgccttgac acgtttgtat cttgcgcgat tccttaattc acctcaatat tttattccct
     2581 atggaactct ttgtgccaat gtctttgcta cacttttgct ttctatcatg tacatgatac
     2641 cacaaatcac acactgcact cctgtgtcta gaagtgtgat gtatggaatc caaaacggat
     2701 tttgtgctgt actctctacc ctttcaacat tctccaacga actccacaca atgcctataa
     2761 aacgagcata tatttattgt ataatttcag ttgctatttc cttttcaata tgtgtcattg
     2821 tcgatggagc tactgcatgg ggtcatggtt atactgaaaa gtattagtaa aaaaagacta
     2881 atgtaaaatt tttttggttg gttattgaaa aagtcgatgc cttgtttgcg tttgttttcc
     2941 taggcgtttt atgtcagaag gcatttagaa ttagtataca agtactcttt ggtaaaattt
     3001 tatgtagcga ctaaaatatt aactattata gataaacacc ttgggaataa aaagtaattt
     3061 gctatagtaa tttattaaac atgctcctac aacattacca caatcttttc tcttggattg
     3121 acattgaata agaaaagagt gaattttttt agacttgtaa tgataactat gtacaaagcc
     3181 aatgaaagat gtatgtagat gaatgtaaaa taccatgtag acaaacaaga taaaacttgg
     3241 ttataaacat tggtgttgga acagaataaa ttagatgtca aaaagtttcg tcaatatcac
     3301 aagctcatcg cgatggtatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     3361 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     3421 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     3481 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagcttgcat
     3541 gcctgcaggt cgactctaga ggatccccac ggtacccctg tttcgtcctc acggactcat
     3601 cagaccggaa agcacatccg gtgacagggc accaccgact cggtgccact cacttttttc
     3661 aagttgataa cggactagcc ttattttaac ttgctatttc tagctctaaa acggacggcg
     3721 gtttgctgag gattcttcgg tacaggttat gttttttggc aacagcaact attcaaatca
     3781 actgaaagtt ttttagaatt caacaacttc caatactcaa gtgtccaagc tagcaaactg
     3841 tttaataatc gtgacccaac attcctttca tgcgcaccct cagcgtcaca taaatcaaca
     3901 ctttacaaca cctattcaaa gcttctatat gccttcgtca tttaaatgtc taacatttca
     3961 aaaaagtgtt tgctaatggt aaattcttga gttttcttgg aagcacatat ataacgagtg
     4021 tgactgttct acgctccttg ctgactagtc atcagtctac gcatgccaac taccaccaac
     4081 ataagcaaaa ggtacccggg gtaccgagct cgaattcgta atcatgtcat agctgtttcc
     4141 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4201 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4261 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4321 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4381 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4441 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4501 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4561 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4621 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4681 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4741 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4801 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4861 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4921 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4981 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5041 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5101 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5161 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5221 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5281 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5341 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5401 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5461 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5521 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5581 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5641 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5701 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5761 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5821 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5881 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5941 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6001 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6061 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6121 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6181 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6241 aggggttccg cgcacatttc cccgaaaagt gccacctgac gccctacaac aactaagaaa
     6301 atggctatca tgcggaagct ggtgagaaga acagcatcgg gacaagggaa ggaagaacaa
     6361 agacaaagaa aacaaaagaa agcaattgaa aacaaaacaa aacaattttc attccttctc
     6421 ttatcattcc ttttcttttc ttttctctca ttcaacgcac tccatcgtat ccgtattcct
     6481 cttatttttt ctctttctct atatccattt ctttctctct aggtgtgtcc tctctctctc
     6541 ttcaatttct ctactccgca ttccaacgca tccttccccc aacctcccat ttcctcctta
     6601 cggcccgata gcgatcgtct ttccctcgct atcactcgct accggcccct cctctgcacc
     6661 gtaacctcct acgtatttac catatcataa agttttttcc gacgcttatc gctgaccccc
     6721 tgtcgccctc ctattggctt ccggattatc ttcttgtcca taaggtgatc catgcttcct
     6781 gaagattccc gaaatgtgtc cactttggcg gggaatcatt ccatccactt ctttctctct
     6841 cgctttcctc attcggcgct ccccttccgc gtctcattgg tcttccgctc cgtttttgct
     6901 ttgccgatgt tacttgggga gaggtgcgat aatcctttcg caaaaactcg gtttgacgcc
     6961 tcccatggta taaatagtgg gtggtggaca ggtgccttcg cttttcttta agcaagagaa
     7021 ttgctgcagg tcgactctag agatggacaa gaagtactcc attgggctcg atatcggcac
     7081 aaacagcgtc ggttgggccg tcattacgga cgagtacaag gtgccgagca aaaaattcaa
     7141 agttctgggc aataccgatc gccacagcat aaagaagaac ctcattggcg ccctcctgtt
     7201 cgactccggg gagacggccg aagccacgcg gctcaaaaga acagcacggc gcagatatac
     7261 ccgcagaaag aatcggatct gctacctgca ggagatcttt agtaatgaga tggctaaggt
     7321 ggatgactct ttcttccata ggctggagga gtcctttttg gtggaggagg ataaaaagca
     7381 cgagcgccac ccaatctttg gcaatatcgt ggacgaggtg gcgtaccatg aaaagtaccc
     7441 aaccatatat catctgagga agaagcttgt agacagtact gataaggctg acttgcggtt
     7501 gatctatctc gcgctggcgc atatgatcaa atttcgggga cacttcctca tcgaggggga
     7561 cctgaaccca gacaacagcg atgtcgacaa actctttatc caactggttc agacttacaa
     7621 tcagcttttc gaagagaacc cgatcaacgc atccggagtt gacgccaaag caatcctgag
     7681 cgctaggctg tccaaatccc ggcggctcga aaacctcatc gcacagctcc ctggggagaa
     7741 gaagaacggc ctgtttggta atcttatcgc cctgtcactc gggctgaccc ccaactttaa
     7801 atctaacttc gacctggccg aagatgccaa gcttcaactg agcaaagaca cctacgatga
     7861 tgatctcgac aatctgctgg cccagatcgg cgaccagtac gcagaccttt ttttggcggc
     7921 aaagaacctg tcagacgcca ttctgctgag tgatattctg cgagtgaaca cggagatcac
     7981 caaagctccg ctgagcgcta gtatgatcaa gcgctatgat gagcaccacc aagacttgac
     8041 tttgctgaag gcccttgtca gacagcaact gcctgagaag tacaaggaaa ttttcttcga
     8101 tcagtctaaa aatggctacg ccggatacat tgacggcgga gcaagccagg aggaatttta
     8161 caaatttatt aagcccatct tggaaaaaat ggacggcacc gaggagctgc tggtaaagct
     8221 taacagagaa gatctgttgc gcaaacagcg cactttcgac aatggaagca tcccccacca
     8281 gattcacctg ggcgaactgc acgctatcct caggcggcaa gaggatttct accccttttt
     8341 gaaagataac agggaaaaga ttgagaaaat cctcacattt cggataccct actatgtagg
     8401 ccccctcgcc cggggaaatt ccagattcgc gtggatgact cgcaaatcag aagagaccat
     8461 cactccctgg aacttcgagg aagtcgtgga taagggggcc tctgcccagt ccttcatcga
     8521 aaggatgact aactttgata aaaatctgcc taacgaaaag gtgcttccta aacactctct
     8581 gctgtacgag tacttcacag tttataacga gctcaccaag gtcaaatacg tcacagaagg
     8641 gatgagaaag ccagcattcc tgtctggaga gcagaagaaa gctatcgtgg acctcctctt
     8701 caagacgaac cggaaagtta ccgtgaaaca gctcaaagaa gactatttca aaaagattga
     8761 atgtttcgac tctgttgaaa tcagcggagt ggaggatcgc ttcaacgcat ccctgggaac
     8821 gtatcacgat ctcctgaaaa tcattaaaga caaggacttc ctggacaatg aggagaacga
     8881 ggacattctt gaggacattg tcctcaccct tacgttgttt gaagataggg agatgattga
     8941 agaacgcttg aaaacttacg ctcatctctt cgacgacaaa gtcatgaaac agctcaagag
     9001 gcgccgatat acaggatggg ggcggctgtc aagaaaactg atcaatggga tccgagataa
     9061 gcagtctgga aagacaatcc tggattttct taagtccgat ggatttgcca accggaactt
     9121 catgcagttg atccatgatg actctctcac ctttaaggag gacatccaga aagcacaagt
     9181 ttctggccag ggggacagtc ttcacgagca catcgctaat cttgcaggta gcccagctat
     9241 caaaaaggga atactgcaga ccgttaaggt cgtggatgaa ctcgtcaaag taatgggaag
     9301 gcataagccc gagaatatcg ttatcgagat ggcccgagag aaccaaacta cccagaaggg
     9361 acagaagaac agtagggaaa ggatgaagag gattgaagag ggtataaaag aactggggtc
     9421 ccaaatcctt aaggaacacc cagttgaaaa cacccagctt cagaatgaga agctctacct
     9481 gtactacctg cagaacggca gggacatgta cgtggatcag gaactggaca tcaatcggct
     9541 ctccgactac gacgtggatc atatcgtgcc ccagtctttt ctcaaagatg attctattga
     9601 taataaagtg ttgacaagat ccgataaaaa tagagggaag agtgataacg tcccctcaga
     9661 agaagttgtc aagaaaatga aaaattattg gcggcagctg ctgaacgcca aactgatcac
     9721 acaacggaag ttcgataatc tgactaaggc tgaacgaggt ggcctgtctg agttggataa
     9781 agccggcttc atcaaaaggc agcttgttga gacacgccag atcaccaagc acgtggccca
     9841 aattctcgat tcacgcatga acaccaagta cgatgaaaat gacaaactga ttcgagaggt
     9901 gaaagttatt actctgaagt ctaagctggt ctcagatttc agaaaggact ttcagtttta
     9961 taaggtgaga gagatcaaca attaccacca tgcgcatgat gcctacctga atgcagtggt
    10021 aggcactgca cttatcaaaa aatatcccaa gcttgaatct gaatttgttt acggagacta
    10081 taaagtgtac gatgttagga aaatgatcgc aaagtctgag caggaaatag gcaaggccac
    10141 cgctaagtac ttcttttaca gcaatattat gaattttttc aagaccgaga ttacactggc
    10201 caatggagag attcggaagc gaccacttat cgaaacaaac ggagaaacag gagaaatcgt
    10261 gtgggacaag ggtagggatt tcgcgacagt ccggaaggtc ctgtccatgc cgcaggtgaa
    10321 catcgttaaa aagaccgaag tacagaccgg aggcttctcc aaggaaagta tcctcccgaa
    10381 aaggaacagc gacaagctga tcgcacgcaa aaaagattgg gaccccaaga aatacggcgg
    10441 attcgattct cctacagtcg cttacagtgt actggttgtg gccaaagtgg agaaagggaa
    10501 gtctaaaaaa ctcaaaagcg tcaaggaact gctgggcatc acaatcatgg agcgatcaag
    10561 cttcgaaaaa aaccccatcg actttctcga ggcgaaagga tataaagagg tcaaaaaaga
    10621 cctcatcatt aagcttccca agtactctct ctttgagctt gaaaacggcc ggaaacgaat
    10681 gctcgctagt gcgggcgagc tgcagaaagg taacgagctg gcactgccct ctaaatacgt
    10741 taatttcttg tatctggcca gccactatga aaagctcaaa gggtctcccg aagataatga
    10801 gcagaagcag ctgttcgtgg aacaacacaa acactacctt gatgagatca tcgagcaaat
    10861 aagcgaattc tccaaaagag tgatcctcgc cgacgctaac ctcgataagg tgctttctgc
    10921 ttacaataag cacagggata agcccatcag ggagcaggca gaaaacatta tccacttgtt
    10981 tactctgacc aacttgggcg cgcctgcagc cttcaagtac ttcgacacca ccatagacag
    11041 aaagcggtac acctctacaa aggaggtcct ggacgccaca ctgattcatc agtcaattac
    11101 ggggctctat gaaacaagaa tcgacctctc tcagctcggt ggagacagca gggctgaccc
    11161 caagaagaag aggaaggtgt gatctcttct cgagtcatgt aattagttat gtcacgctta
    11221 cattcacgcc ctccccccac atccgctcta accgaaaagg aaggagttag acaacctgaa
    11281 gtctaggtcc ctatttattt ttttatagtt atgttagtat taagaacgtt atttatattt
    11341 caaatttttc ttttttttct gtacagacgc gtgtacgcat gtaacattat actgaaaacc
    11401 ttgcttgaga aggttttggg acgctcgaag gctttaattt gcgatccccg tctaagaaac
    11461 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
