LOCUS       sequence-234836-       12554 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Cas9 expression controlled by the nmt41 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12554)
  AUTHORS   Hayashi A, Tanaka K
  TITLE     Short-Homology-Mediated CRISPR/Cas9-Based Method for Genome Editing 
            in Fission Yeast.
  JOURNAL   G3 (Bethesda). 2019 Feb 12. pii: g3.118.200976. doi: 
            10.1534/g3.118.200976.
  PUBMED    30755408
REFERENCE   2  (bases 1 to 12554)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "G3 
            (Bethesda). 2019 Feb 12. pii: g3.118.200976. doi: 
            10.1534/g3.118.200976."
COMMENT     Sequence Label: pAH235
FEATURES             Location/Qualifiers
     source          1..12554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..66
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             73..93
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             118..4218
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             118..4218
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4219..4266
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     rep_origin      5352..6133
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     complement(6483..6500)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6483..6499)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6492..6514)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6712..7167
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6799..6818)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7009..7030
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7184..7203)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7303..7325
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7363..7381)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7449..7553
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7554..7622,7623..8414)
                     /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     complement(7772..7791)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8585..9173
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9074..9093
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9327..9344
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9461..9482
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9497..9502,9503..9520,9521..9527)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9535..9551
                     /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)."
     primer_bind     9540..9562
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9559..9575
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9559..9575
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(10024..10818)
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe ura4"
                     /product="orotidine 5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=S. pombe ura4
                     /translation="MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKS
                     EILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVK
                     LQYASGVYKIASWAHITNCHTVPGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSY
                     TEKTLEWFEKHTDFCFGFIAGRRFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIV
                     NCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH"
     promoter        11394..12547
                     /label=nmt1 P41 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring medium strength thiamine-repressible expression"
ORIGIN
        1 gactataagg accacgacgg agactacaag gatcatgata ttgattacaa agacgatgac
       61 gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc agcagccgac
      121 aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc
      181 gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc
      241 atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc
      301 cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg
      361 caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa
      421 gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc
      481 gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg
      541 gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc
      601 aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac
      661 aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac
      721 gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg
      781 gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt
      841 gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc
      901 aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc
      961 ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg
     1021 agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc
     1081 aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag
     1141 ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac
     1201 attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag
     1261 atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag
     1321 cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt
     1381 ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag
     1441 atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc
     1501 gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg
     1561 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg
     1621 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac
     1681 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc
     1741 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag
     1801 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc
     1861 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag
     1921 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc
     1981 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg
     2041 ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg
     2101 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc
     2161 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg
     2221 acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag
     2281 cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag
     2341 gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa
     2401 atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag
     2461 cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa
     2521 aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg
     2581 tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg
     2641 cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag
     2701 aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac
     2761 tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag
     2821 gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg
     2881 gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag
     2941 tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg
     3001 gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac
     3061 cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct
     3121 aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc
     3181 gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc
     3241 atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg
     3301 atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc
     3361 gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca
     3421 ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga
     3481 aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct
     3541 gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag
     3601 ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg
     3661 gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc
     3721 ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag
     3781 ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat
     3841 gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac
     3901 aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg
     3961 gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc
     4021 agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc
     4081 gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg
     4141 ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg
     4201 tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag
     4261 aaaaagtaag atctcccggg taaaaggaat gtctcccttg ccagtactgc tagggttttt
     4321 ctttcaaact atggaagccc attcaagctg catattacga ttttgttttt cgcttttaga
     4381 aagtggttta gatgagataa tagaaaaatt cttgatctcc gacaacgagt acttttattt
     4441 tttttgctaa tcactttact caatattagc tcgaaatcgt agaaacgtag acgggtgcgg
     4501 gataccgagt ggtgtagtta asaattttta taaaccacgt ggcccaaaaa tatgaaccca
     4561 aaacgtttat acatgagtat actttaagaa ggctataccc cttcgtgtta gatgtagttt
     4621 tagctaccca acccgagtct atgagcttga cttcagatgt agaaggcatt aaatcgtttt
     4681 gaatattaat taaaaaacga tgaaaattaa atatttaaaa gcaatcatac gctgaaaatt
     4741 tagtgctgtg gctaatcctt caacatggaa atgccataaa agtgactttg acaaaaaaaa
     4801 aagtatatac aggtagtaaa ctcatctact tcattgactt tgtttacagc atgtggaagg
     4861 aggaatattt attgctaaat cgtagtttaa cattcaataa gtaatactat tgaaattcga
     4921 caagattggc cgcatggatg aaaaagaggc attttgcttt gggagaatta gtccaaatta
     4981 gaactgaaaa aaaaaacttt acgaggcaaa aatgtcggat tgagatcgta aaagttcgct
     5041 cgtcgtcttt tgctttgtga ttgttttcat ggatacatct tgctggatat ttaaatttta
     5101 gtactatgta taagatattc tataaatgtt ttatcaccca aacctgttag cgccttctta
     5161 attctattca atctggcttt tgctctgaga ctacttcttg gactttcact acttgttagt
     5221 tatacggaat ttgtgtaatt agaagtgaaa taatcctttc tattagtaat gcgagctcga
     5281 attcgagtct aactccttaa ccactcggac atagtgactt atctgacact cattgtaaat
     5341 taatatatat ataggcattt tgtttagtta aaggtactta agtaattagt ataaacgaac
     5401 caattttata atcaggaagt taagtgaatg gtagcacatg tcgtaaaaat tgtgaatttt
     5461 tattgaataa tattttaaat acaagccttt ctagactagg tatactcata aacatatatg
     5521 agcaaaagga tagaggagat tacattgcat cttctacaaa tttatttatt gccctttact
     5581 gaaaaattaa ataatgagta ctaaatgata aaaagcgctc agtacaagaa agattgcaaa
     5641 aatattgcat tcttcatgaa ttaaagttgc atataaggca tattgaaagt aatagtacta
     5701 aaacagcagt tagcgaaaat taatagaatt atattcgcaa gacaattgtg acattaaatt
     5761 aaaaaattgt agaattttta ctatcctctt taacgccatg agcctttata aaaaggttaa
     5821 attagtttta acattctttt tttgagtaag agtttacaat ttatcaaaac ctgtgttatt
     5881 atattcatta gtttcaattt attagcatct agagaaaaat caattggcag ttacattgtt
     5941 ggaatttatg aagaaaaaga atctacaacg gagaataagt tgctgatctt tccctaaaat
     6001 tgtatatttt gctgagctta ttttgacatt tcgttgaagt tttctctaat tcgcattcat
     6061 tttaagtaaa acaatgagaa ataaaattac aaaaaataca aattaaaata caatttttag
     6121 ctataattat agacgatgcc cttgtatccc attctgtctc gcttgcccct actttttatc
     6181 ttttatatac cataatgaac gctgccgcta ctaaccatac cccgatttta catttcggac
     6241 tcccaaggac gtacaaaata gaaaactata gaaaaaaata atcagaaaat agcatgtcat
     6301 ctctttgtaa aacgcgtttg caagaagaaa ggaaacaatg gagaagagat catccatttg
     6361 tatgtaaatt ttagtaaact tgaagaaatc actaacaact tctcttactt agggattcta
     6421 tgcaaaacct tgtaaatcat ctgatggagg actcgattta atgaattgga aggttggaat
     6481 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     6541 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     6601 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     6661 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     6721 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     6781 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     6841 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     6901 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     6961 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     7021 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     7081 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     7141 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     7201 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     7261 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     7321 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     7381 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     7441 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     7501 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     7561 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     7621 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     7681 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     7741 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     7801 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     7861 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     7921 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     7981 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     8041 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     8101 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     8161 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     8221 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     8281 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     8341 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     8401 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     8461 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     8521 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     8581 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     8641 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     8701 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     8761 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     8821 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     8881 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     8941 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     9001 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     9061 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     9121 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     9181 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     9241 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     9301 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     9361 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     9421 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     9481 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     9541 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cttgtgatat
     9601 tgacgaaact ttttgacatc taatttattc tgttccaaca ccaatgttta taaccaagtt
     9661 ttatcttgtt tgtctacatg gtattttaca ttcatctaca tacatctttc attggctttg
     9721 tacatagtta tcattacaag tctaaaaaaa ttcactcttt tcttattcaa tgtcaatcca
     9781 agagaaaaga ttgtggtaat gttgtaggag catgtttaat aaattactat agcaaattac
     9841 tttttattcc caaggtgttt atctataata gttaatattt tagtcgctac ataaaatttt
     9901 accaaagagt acttgtatac taattctaaa tgccttctga cataaaacgc ctaggaaaac
     9961 aaacgcaaac aaggcatcga ctttttcaat aaccaaccaa aaaaatttta cattagtctt
    10021 tttttaatgc tgagaaagtc tttgctgata tgccttccaa ccagcttctc tatatctctt
    10081 ggcttcgaca acaggattac gaccagctcc atagactcca cgaccaacaa tgatgatatc
    10141 gctaccgcag tttacaatca cttcttcagg agtacgatat tgctgtccca gcccgtctcc
    10201 tttaacatcc aagccgatac caggggacat agttatgtag tcgctttgaa ggttaggaaa
    10261 tcgacgacca gctataaagc caaagcaaaa atcggtatgc ttctcaaacc attctaaggt
    10321 tttctctgtg taggaaccag tagccaaaga gcctttggaa gacatttcag ccaaaagcaa
    10381 gagaccacgt cccaaaggta aaccaacttc tttgaggcct tgtataatac cctcgcctgg
    10441 cactgtatgg caatttgtga tatgagccca agaagcaatt ttgtacacac cagatgcata
    10501 ttgtagcttg acggtatttc caatgtctgc gaatttgcga tcctcaaaga taagaaaacg
    10561 atgcttttta cctaaggcca ccagtttttc taccatatcc tggtcgaaat cctcgacaac
    10621 gtcaatatgt gtcttgataa cacagacata gggtccaatt ttatctacca attctaagat
    10681 ttcggatttc ttcgtcaaat cgaccgcgac tgacaagttg ctttgctttt cttccatcaa
    10741 agccaacaat tccttggcaa tgggattttt catcccctca gctctagctg aatagctttg
    10801 aaatactcta gcatccataa ctttgctttt aaacctttaa tttcgatcca agcaaaaaag
    10861 aggttcttgg taggacaata cggtaagaaa acacgacatg tgcagagatg ccgacgaagc
    10921 atagttaaac tgggatggta aaatcaatta agaatttata aagacaaaat tgtataagtc
    10981 tctaaaacat cttaattata cctcacagaa ctatctaaaa tatattcaca aagtgcaaac
    11041 attatcatga aaaagaacca ttttaattta aagcaagggc attaaggctt atttacagaa
    11101 tttcttactt ttgtaaagat tataaggctg attatctttt tcaccatgcc aaaaattaca
    11161 caagatagaa tggatgtttg aaattaaacg tgagtataca aacaaataca ctaggtaaat
    11221 cgaaacattt ttttctccat taagtaacaa attcctattt agagaaagaa tgctgagtag
    11281 attaaataat ctatacaaac ttttttaaca caaatgcata catatagcca gtgggatttg
    11341 tagctaagct tgcatgcctg caggtcgatc gactctagag gatcagaaaa ttatcgccat
    11401 aaaagacaga ataagtcatc agcggttgtt tcatttccta tatttttttt ttattttttt
    11461 attttttaat aagggaaaat ttaacgtcta aggatacaga agattgttag cacattaaag
    11521 taataaaggc ttaagtagta agtgccttag catgttattg tatttcaaag gacataatct
    11581 aaaataataa caatatcatt tctcacaagt tattcaattt tctttttttt ttctaataat
    11641 atcaagaatg tattatttgt ttgacataag tcaactaatt tatttaatat gctggattaa
    11701 tcttgcagac atgtaaatta acaagtttta gtcaaataac gttgaagttt caatgaactc
    11761 aaataatttc tctttttttt tatataacca taatctgatt tatattttcc gcagggatca
    11821 actgaagtta tgacatttgg attggatcac ttataacctt ggtcgccaaa taatacaaaa
    11881 atcagcgtta taaaacaaag aaggtttttg ttaagaaatt aatcctcttt cttgataaga
    11941 aagttgaacc gaaattgcag atactgatat atgaaaataa tacccacaat tttgggaata
    12001 gcgcaagcct caatttaaac aataggtgag gacacatgat aatgacctca atgattgtta
    12061 gaagaaaaga gcctcattac aaaatcgaaa aatgaatggt tgggtacaag tttccaaaac
    12121 atggtaaagt ggactttgcg tatgagacgt aaatagaaaa aaacacttgt tatatgtttt
    12181 ctagaattat tgttgtctct ttatggttgg atgatgcaaa atagtaattt cggttagttg
    12241 ctgtaaaaca ccacgagaca aatagatatg gatatttatt aaatcaggaa aaacgtaact
    12301 ctcggctact ggatggttca gtcacccaac gattactggg gagagaaaac agggcaaaag
    12361 caaagcttaa aggaatccga ttgtcattcg gcaatgtgca gcgaaactaa aaaccggata
    12421 atggacctgt taatcgaaac attgaagata aaggaagagg aatcctggca tatcatcaat
    12481 tgaataagtt gaattaatta tttcaatctc attctcactt tctgacttat agtcgctttg
    12541 ttaaatctcg aatg
//
