LOCUS       sequence-173004-       11530 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 11530)
  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 11530)
  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..11530
                     /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(3747..4103)
                     /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(4140..4156)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4140..4156)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4153..4175)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4164..4180
                     /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(4188..4194,4195..4212,4213..4218))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4233..4254
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4371..4388)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4542..5130)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4622..4641)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5301..6092,6093..6161))
                     /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     5924..5943
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6162..6266)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6295..7036
                     /label=adh1 promoter
                     /note="promoter for the Schizosaccharomyces pombe adh1 gene
                     encoding alcohol dehydrogenase"
     CDS             7057..11160
                     /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             7057..11160
                     /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             7057..11160
                     /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             11173..11193
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      11209..11456
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(11225..11243)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     11503..11521
                     /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 acagagagat
     3721 aacccacaag aattgagtta agcccattct tcggtacagg ttatgttttt tggcaacagc
     3781 aactattcaa atcaactgaa agttttttag aattcaacaa cttccaatac tcaagtgtcc
     3841 aagctagcaa actgtttaat aatcgtgacc caacattcct ttcatgcgca ccctcagcgt
     3901 cacataaatc aacactttac aacacctatt caaagcttct atatgccttc gtcatttaaa
     3961 tgtctaacat ttcaaaaaag tgtttgctaa tggtaaattc ttgagttttc ttggaagcac
     4021 atatataacg agtgtgactg ttctacgctc cttgctgact agtcatcagt ctacgcatgc
     4081 caactaccac caacataagc aaaaggtacc cggggtaccg agctcgaatt cgtaatcatg
     4141 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4201 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4261 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4321 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4381 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4441 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4501 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4561 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4621 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4681 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4741 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4801 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4861 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4921 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4981 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5041 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5101 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5161 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5221 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5281 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5341 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5401 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5461 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5521 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5581 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5641 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5701 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5761 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5821 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5881 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5941 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6001 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6061 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6121 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6181 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6241 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcccta
     6301 caacaactaa gaaaatggct atcatgcgga agctggtgag aagaacagca tcgggacaag
     6361 ggaaggaaga acaaagacaa agaaaacaaa agaaagcaat tgaaaacaaa acaaaacaat
     6421 tttcattcct tctcttatca ttccttttct tttcttttct ctcattcaac gcactccatc
     6481 gtatccgtat tcctcttatt ttttctcttt ctctatatcc atttctttct ctctaggtgt
     6541 gtcctctctc tctcttcaat ttctctactc cgcattccaa cgcatccttc ccccaacctc
     6601 ccatttcctc cttacggccc gatagcgatc gtctttccct cgctatcact cgctaccggc
     6661 ccctcctctg caccgtaacc tcctacgtat ttaccatatc ataaagtttt ttccgacgct
     6721 tatcgctgac cccctgtcgc cctcctattg gcttccggat tatcttcttg tccataaggt
     6781 gatccatgct tcctgaagat tcccgaaatg tgtccacttt ggcggggaat cattccatcc
     6841 acttctttct ctctcgcttt cctcattcgg cgctcccctt ccgcgtctca ttggtcttcc
     6901 gctccgtttt tgctttgccg atgttacttg gggagaggtg cgataatcct ttcgcaaaaa
     6961 ctcggtttga cgcctcccat ggtataaata gtgggtggtg gacaggtgcc ttcgcttttc
     7021 tttaagcaag agaattgctg caggtcgact ctagagatgg acaagaagta ctccattggg
     7081 ctcgatatcg gcacaaacag cgtcggttgg gccgtcatta cggacgagta caaggtgccg
     7141 agcaaaaaat tcaaagttct gggcaatacc gatcgccaca gcataaagaa gaacctcatt
     7201 ggcgccctcc tgttcgactc cggggagacg gccgaagcca cgcggctcaa aagaacagca
     7261 cggcgcagat atacccgcag aaagaatcgg atctgctacc tgcaggagat ctttagtaat
     7321 gagatggcta aggtggatga ctctttcttc cataggctgg aggagtcctt tttggtggag
     7381 gaggataaaa agcacgagcg ccacccaatc tttggcaata tcgtggacga ggtggcgtac
     7441 catgaaaagt acccaaccat atatcatctg aggaagaagc ttgtagacag tactgataag
     7501 gctgacttgc ggttgatcta tctcgcgctg gcgcatatga tcaaatttcg gggacacttc
     7561 ctcatcgagg gggacctgaa cccagacaac agcgatgtcg acaaactctt tatccaactg
     7621 gttcagactt acaatcagct tttcgaagag aacccgatca acgcatccgg agttgacgcc
     7681 aaagcaatcc tgagcgctag gctgtccaaa tcccggcggc tcgaaaacct catcgcacag
     7741 ctccctgggg agaagaagaa cggcctgttt ggtaatctta tcgccctgtc actcgggctg
     7801 acccccaact ttaaatctaa cttcgacctg gccgaagatg ccaagcttca actgagcaaa
     7861 gacacctacg atgatgatct cgacaatctg ctggcccaga tcggcgacca gtacgcagac
     7921 ctttttttgg cggcaaagaa cctgtcagac gccattctgc tgagtgatat tctgcgagtg
     7981 aacacggaga tcaccaaagc tccgctgagc gctagtatga tcaagcgcta tgatgagcac
     8041 caccaagact tgactttgct gaaggccctt gtcagacagc aactgcctga gaagtacaag
     8101 gaaattttct tcgatcagtc taaaaatggc tacgccggat acattgacgg cggagcaagc
     8161 caggaggaat tttacaaatt tattaagccc atcttggaaa aaatggacgg caccgaggag
     8221 ctgctggtaa agcttaacag agaagatctg ttgcgcaaac agcgcacttt cgacaatgga
     8281 agcatccccc accagattca cctgggcgaa ctgcacgcta tcctcaggcg gcaagaggat
     8341 ttctacccct ttttgaaaga taacagggaa aagattgaga aaatcctcac atttcggata
     8401 ccctactatg taggccccct cgcccgggga aattccagat tcgcgtggat gactcgcaaa
     8461 tcagaagaga ccatcactcc ctggaacttc gaggaagtcg tggataaggg ggcctctgcc
     8521 cagtccttca tcgaaaggat gactaacttt gataaaaatc tgcctaacga aaaggtgctt
     8581 cctaaacact ctctgctgta cgagtacttc acagtttata acgagctcac caaggtcaaa
     8641 tacgtcacag aagggatgag aaagccagca ttcctgtctg gagagcagaa gaaagctatc
     8701 gtggacctcc tcttcaagac gaaccggaaa gttaccgtga aacagctcaa agaagactat
     8761 ttcaaaaaga ttgaatgttt cgactctgtt gaaatcagcg gagtggagga tcgcttcaac
     8821 gcatccctgg gaacgtatca cgatctcctg aaaatcatta aagacaagga cttcctggac
     8881 aatgaggaga acgaggacat tcttgaggac attgtcctca cccttacgtt gtttgaagat
     8941 agggagatga ttgaagaacg cttgaaaact tacgctcatc tcttcgacga caaagtcatg
     9001 aaacagctca agaggcgccg atatacagga tgggggcggc tgtcaagaaa actgatcaat
     9061 gggatccgag ataagcagtc tggaaagaca atcctggatt ttcttaagtc cgatggattt
     9121 gccaaccgga acttcatgca gttgatccat gatgactctc tcacctttaa ggaggacatc
     9181 cagaaagcac aagtttctgg ccagggggac agtcttcacg agcacatcgc taatcttgca
     9241 ggtagcccag ctatcaaaaa gggaatactg cagaccgtta aggtcgtgga tgaactcgtc
     9301 aaagtaatgg gaaggcataa gcccgagaat atcgttatcg agatggcccg agagaaccaa
     9361 actacccaga agggacagaa gaacagtagg gaaaggatga agaggattga agagggtata
     9421 aaagaactgg ggtcccaaat ccttaaggaa cacccagttg aaaacaccca gcttcagaat
     9481 gagaagctct acctgtacta cctgcagaac ggcagggaca tgtacgtgga tcaggaactg
     9541 gacatcaatc ggctctccga ctacgacgtg gatcatatcg tgccccagtc ttttctcaaa
     9601 gatgattcta ttgataataa agtgttgaca agatccgata aaaatagagg gaagagtgat
     9661 aacgtcccct cagaagaagt tgtcaagaaa atgaaaaatt attggcggca gctgctgaac
     9721 gccaaactga tcacacaacg gaagttcgat aatctgacta aggctgaacg aggtggcctg
     9781 tctgagttgg ataaagccgg cttcatcaaa aggcagcttg ttgagacacg ccagatcacc
     9841 aagcacgtgg cccaaattct cgattcacgc atgaacacca agtacgatga aaatgacaaa
     9901 ctgattcgag aggtgaaagt tattactctg aagtctaagc tggtctcaga tttcagaaag
     9961 gactttcagt tttataaggt gagagagatc aacaattacc accatgcgca tgatgcctac
    10021 ctgaatgcag tggtaggcac tgcacttatc aaaaaatatc ccaagcttga atctgaattt
    10081 gtttacggag actataaagt gtacgatgtt aggaaaatga tcgcaaagtc tgagcaggaa
    10141 ataggcaagg ccaccgctaa gtacttcttt tacagcaata ttatgaattt tttcaagacc
    10201 gagattacac tggccaatgg agagattcgg aagcgaccac ttatcgaaac aaacggagaa
    10261 acaggagaaa tcgtgtggga caagggtagg gatttcgcga cagtccggaa ggtcctgtcc
    10321 atgccgcagg tgaacatcgt taaaaagacc gaagtacaga ccggaggctt ctccaaggaa
    10381 agtatcctcc cgaaaaggaa cagcgacaag ctgatcgcac gcaaaaaaga ttgggacccc
    10441 aagaaatacg gcggattcga ttctcctaca gtcgcttaca gtgtactggt tgtggccaaa
    10501 gtggagaaag ggaagtctaa aaaactcaaa agcgtcaagg aactgctggg catcacaatc
    10561 atggagcgat caagcttcga aaaaaacccc atcgactttc tcgaggcgaa aggatataaa
    10621 gaggtcaaaa aagacctcat cattaagctt cccaagtact ctctctttga gcttgaaaac
    10681 ggccggaaac gaatgctcgc tagtgcgggc gagctgcaga aaggtaacga gctggcactg
    10741 ccctctaaat acgttaattt cttgtatctg gccagccact atgaaaagct caaagggtct
    10801 cccgaagata atgagcagaa gcagctgttc gtggaacaac acaaacacta ccttgatgag
    10861 atcatcgagc aaataagcga attctccaaa agagtgatcc tcgccgacgc taacctcgat
    10921 aaggtgcttt ctgcttacaa taagcacagg gataagccca tcagggagca ggcagaaaac
    10981 attatccact tgtttactct gaccaacttg ggcgcgcctg cagccttcaa gtacttcgac
    11041 accaccatag acagaaagcg gtacacctct acaaaggagg tcctggacgc cacactgatt
    11101 catcagtcaa ttacggggct ctatgaaaca agaatcgacc tctctcagct cggtggagac
    11161 agcagggctg accccaagaa gaagaggaag gtgtgatctc ttctcgagtc atgtaattag
    11221 ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag
    11281 ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa
    11341 cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca
    11401 ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcgatc
    11461 cccgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
    11521 ccctttcgtc
//
