LOCUS       sequence-234796-       15430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of both nmt41p-cas9 and rrk1p-gRNA (LEU2 marker) for 
            CRISPR genome editing in fission yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15430)
  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 15430)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "G3 
            (Bethesda). 2019 Feb 12. pii: g3.118.200976. doi: 
            10.1534/g3.118.200976."
COMMENT     Sequence Label: pAH237
FEATURES             Location/Qualifiers
     source          1..15430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        107..158
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     promoter        1064..2216
                     /label=nmt1 P41 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring medium strength thiamine-repressible expression"
     CDS             2224..2289
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2296..2316
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2341..6441
                     /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             2341..6441
                     /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             6442..6489
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     rep_origin      7635..8418
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     complement(8768..8785)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8768..8784)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8777..8799)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8997..9452
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9084..9103)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9294..9315
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9469..9488)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9588..9610
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9648..9666)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9734..9838
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9839..9907,9908..10699)
                     /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(10057..10076)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10870..11458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11359..11378
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11612..11629
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11746..11767
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11782..11787,11788..11805,11806..11812)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11820..11836
                     /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     11825..11847
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11844..11860
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11844..11860
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(12361..13455)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(13456..13861)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
ORIGIN
        1 caccgggtct tcgagaagac ctgttttaga gctagaaata gcaagttaaa ataaggctag
       61 tccgttatca acttgaaaaa agtgagtggc accgagtcgg tggtgccctg tcaccggatg
      121 tgctttccgg tctgatgagt ccgtgaggac gaaacaggta cggttatttg ttagcaatgg
      181 taggagaaaa ttagggattg tttaccaaaa caacgtgaac aaatagttat agttttaatt
      241 taactcatga acatattatc ggaaaaaatc tactcattta tgtctgggtt tagctccctt
      301 tatggtttcg ttttgcagta gaatacacta tggtatttat tttgtgcact acaaaccgtg
      361 ctttttatgt gaacgttttg atgaaaacgt tctagagatt gatttattat tcaatgttaa
      421 agtaactcat tttgctatat gtatcttttt ttcagcgaac tttcgttact gagcagttca
      481 agcagtgatt tggcattatg gtatcaataa agacatatca ttaaggcaag gctacggaaa
      541 gaaaaaagaa ggtttttaaa ttaatataca acaaattaac cgtgatcgga cagtatattt
      601 gatgcaaaga tataatgaaa gttgggaaaa taagaatata gcactcaaat gattgcctta
      661 caaaagcatt gctttctcta caggtaaggt tgaagtgcct ttacggtatc ttcaaaaacc
      721 gcatcgggat cttgctcggc atcgatggtt atcagacgat tttgtgattt cagatattcg
      781 acgacaggca tgctggcttt agtatatgta acaaagcgtt tcttgatcga ctcaatgttg
      841 tcatcagaac ggccactagt cttaccacga tgaataagac gcttcaacat ggtttcctga
      901 ccacaacgga aataaagagc aaacttagcg gggcagaccg acttttcaaa accttcgcat
      961 tgatccatct cacgggggaa accgtcaatc aagaacttgt caattccttt atcatggcat
     1021 tccttcattt tagtctctaa aagggatata gttatttcca tggtcgccat aaaagacaga
     1081 ataagtcatc agcggttgtt tcatttccta tatttttttt ttattttttt attttttaat
     1141 aagggaaaat ttaacgtcta aggatacaga agattgttag cacattaaag taataaaggc
     1201 ttaagtagta agtgccttag catgttattg tatttcaaag gacataatct aaaataataa
     1261 caatatcatt tctcacaagt tattcaattt tctttttttt ttctaataat atcaagaatg
     1321 tattatttgt ttgacataag tcaactaatt tatttaatat gctggattaa tcttgcagac
     1381 atgtaaatta acaagtttta gtcaaataac gttgaagttt caatgaactc aaataatttc
     1441 tctttttttt atataaccat aatctgattt atattttccg cagggatcaa ctgaagttat
     1501 gacatttgga ttggatcact tataaccttg gtcgccaaat aatacaaaaa tcagcgttat
     1561 aaaacaaaga aggtttttgt taagaaatta atcctctttc ttgataagaa agttgaaccg
     1621 aaattgcaga tactgatata tgaaaataat acccacaatt ttgggaatag cgcaagcctc
     1681 aatttaaaca ataggtgagg acacatgata atgacctcaa tgattgttag aagaaaagag
     1741 cctcattaca aaatcgaaaa atgaatggtt gggtacaagt ttccaaaaca tggtaaagtg
     1801 gactttgcgt atgagacgta aatagaaaaa aacacttgtt atatgttttc tagaattatt
     1861 gttgtctctt tatggttgga tgatgcaaaa tagtaatttc ggttagttgc tgtaaaacac
     1921 cacgagacaa atagatatgg atatttatta aatcaggaaa aacgtaactc tcggctactg
     1981 gatggttcag tcacccaacg attactgggg agagaaaaca gggcaaaagc aaagcttaaa
     2041 ggaatccgat tgtcattcgg caatgtgcag cgaaactaaa aaccggataa tggacctgtt
     2101 aatcgaaaca ttgaagataa aggaagagga atcctggcat atcatcaatt gaataagttg
     2161 aattaattat ttcaatctca ttctcacttt ctgacttata gtcgctttgt taaatctcga
     2221 atggactata aggaccacga cggagactac aaggatcatg atattgatta caaagacgat
     2281 gacgataaga tggccccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc
     2341 gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc
     2401 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
     2461 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
     2521 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
     2581 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
     2641 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
     2701 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
     2761 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
     2821 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
     2881 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
     2941 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
     3001 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg
     3061 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
     3121 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
     3181 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
     3241 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
     3301 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
     3361 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
     3421 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
     3481 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
     3541 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
     3601 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
     3661 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
     3721 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
     3781 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
     3841 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
     3901 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
     3961 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
     4021 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
     4081 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
     4141 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
     4201 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
     4261 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
     4321 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
     4381 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
     4441 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
     4501 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
     4561 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
     4621 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
     4681 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
     4741 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
     4801 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc
     4861 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac
     4921 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
     4981 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
     5041 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
     5101 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
     5161 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
     5221 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
     5281 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
     5341 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
     5401 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
     5461 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
     5521 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc
     5581 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
     5641 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc
     5701 agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat
     5761 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
     5821 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
     5881 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
     5941 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag
     6001 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
     6061 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
     6121 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
     6181 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc
     6241 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
     6301 gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag
     6361 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
     6421 ctgtctcagc tgggaggcga caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa
     6481 aagaaaaagt aagatctccc gggtaaaagg aatgtctccc ttgccagtac tgctagggtt
     6541 tttctttcaa actatggaag cccattcaag ctgcatatta cgattttgtt tttcgctttt
     6601 agaaagtggt ttagatgaga taatagaaaa attcttgatc tccgacaacg agtactttta
     6661 ttttttttgc taatcacttt actcaatatt agctcgaaat cgtagaaacg tagacgggtg
     6721 cgggataccg agtggtgtag ttaagaattt ttataaacca cgtggcccaa aaatatgaac
     6781 ccaaaacgtt tatacatgag tatactttaa gaaggctata ccccttcgtg ttagatgtag
     6841 ttttagctac ccaacccgag tctatgagct tgacttcaga tgtagaaggc attaaatcgt
     6901 tttgaatatt aattaaaaaa cgatgaaaat taaatattta aaagcaatca tacgctgaaa
     6961 atttagtgct gtggctaatc cttcaacatg gaaatgccat aaaagtgact ttgacaaaaa
     7021 aaaagtatat acaggtagta aactcatcta cttcattgac tttgtttaca gcatgtggaa
     7081 ggaggaatat ttattgctaa atcgtagttt aacattcaat aagtaatact attgaaattc
     7141 gacaagattg gccgcatgga tgaaaaagag gcattttgct ttgggagaat tagtccaaat
     7201 tagaactgaa aaaaaaaact ttacgaggca aaaatgtcgg attgagatcg taaaagttcg
     7261 ctcgtcgtct tttgctttgt gattgttttc atggatacat cttgctggat atttaaattt
     7321 tagtactatg tataagatat tctataaatg ttttatcacc caaacctgtt agcgccttct
     7381 taattctatt caatctggct tttgctctga gactacttct tggactttca ctacttgtta
     7441 gttatacgga atttgtgtaa ttagaagtga aataatcctt tctattagta atgccatggg
     7501 cacaatttta ccatctttga tatattcttt aataaggttt ccgtatttgc tgccaggacg
     7561 cgaattcgag tctaactcct taaccactcg gacatagtga cttatctgac actcattgta
     7621 aattaatata tatataggca ttttgtttag ttaaaggtac ttaagtaatt agtataaacg
     7681 aaccaatttt ataatcagga agttaagtga atggtagcac atgtcgtaaa aattgtgaat
     7741 ttttattgaa taatatttta aatacaagcc tttctagact aggtatactc ataaacatat
     7801 atgagcaaaa ggatagagga gattacattg catcttctac aaatttattt attgcccttt
     7861 actgaaaaat taaataatga gtactaaatg ataaaaagcg ctcagtacaa gaaagattgc
     7921 aaaaatattg cattcttcat gaattaaagt tgcatataag gcatattgaa agtaatagta
     7981 ctaaaacagc agttagcgaa aattaataga attatattcg caagacaatt gtgacattaa
     8041 attaaaaaat tgtagaattt ttactatcct ctttaacgcc atgagccttt ataaaaaggt
     8101 taaattagtt ttaacattct ttttttgagt aagagtttac aatttatcaa aacctgtgtt
     8161 attatattca ttagtttcaa tttattagca tctagagaaa aatcaattgg cagttacatt
     8221 gttggaattt atgaagaaaa agaatctaca acggagaata agttgctgat cgctttccct
     8281 aaaattgtat attttgctga gcttattttg acatttcgtt gaagttttct ctaattcgca
     8341 ttcattttaa gtaaaacaat gagaaataaa attacaaaaa atacaaatta aaatacaatt
     8401 tttagctata attatagacg atgcccttgt atcccattct gtctcgcttg cccctacttt
     8461 ttatctttta tataccataa tgaacgctgc cgctactaac cataccccga ttttacattt
     8521 cggactccca aggacgtaca aaatagaaaa ctatagaaaa aaataatcag aaaatagcat
     8581 gtcatctctt tgtaaaacgc gtttgcaaga agaaaggaaa caatggagaa gagatcatcc
     8641 atttgtatgt aaattttagt aaacttgaag aaatcactaa caacttctct tacttaggga
     8701 ttctatgcaa aaccttgtaa atcatctgat ggaggactcg atttaatgaa ttggaaggtt
     8761 ggaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     8821 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     8881 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt
     8941 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     9001 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     9061 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     9121 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     9181 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     9241 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     9301 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     9361 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     9421 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     9481 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     9541 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     9601 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     9661 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     9721 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     9781 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     9841 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     9901 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     9961 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
    10021 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
    10081 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
    10141 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
    10201 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
    10261 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
    10321 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
    10381 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
    10441 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
    10501 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
    10561 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
    10621 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
    10681 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
    10741 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
    10801 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
    10861 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
    10921 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
    10981 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
    11041 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
    11101 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
    11161 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
    11221 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
    11281 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
    11341 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
    11401 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
    11461 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
    11521 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
    11581 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
    11641 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
    11701 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
    11761 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
    11821 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgt
    11881 cgatcgacta cgtcgttaag gccgtttctg acagagtaaa attcttgagg gaactttcac
    11941 cattatggga aatggttcaa gaaggtattg acttaaactc catcaaatgg tcaggtcatt
    12001 gagtgttttt tatttgttgt attttttttt ttttagagaa aatcctccaa tatataaatt
    12061 aggaatcata gtttcatgat tttctgttac acctaacttt ttgtgtggtg ccctcctcct
    12121 tgtcaatatt aatgttaaag tgcaattctt tttccttatc acgttgagcc attagtatca
    12181 atttgcttac ctgtattcct ttacatcctc ctttttctcc ttcttgataa atgtatgtag
    12241 attgcgtata tagtttcgtc taccctatga acatattcca ttttgtaatt tcgtgtcgtt
    12301 tctattatga atttcattta taaagtttat gtacaaatat cataaaaaaa gagaatcttt
    12361 ttaagcaagg attttcttaa cttcttcggc gacagcatca ccgacttcgg tggtactgtt
    12421 ggaaccacct aaatcaccag ttctgatacc tgcatccaaa acctttttaa ctgcatcttc
    12481 aatggcctta ccttcttcag gcaagttcaa tgacaatttc aacatcattg cagcagacaa
    12541 gatagtggcg atagggttga ccttattctt tggcaaatct ggagcagaac cgtggcatgg
    12601 ttcgtacaaa ccaaatgcgg tgttcttgtc tggcaaagag gccaaggacg cagatggcaa
    12661 caaacccaag gaacctggga taacggaggc ttcatcggag atgatatcac caaacatgtt
    12721 gctggtgatt ataataccat ttaggtgggt tgggttctta actaggatca tggcggcaga
    12781 atcaatcaat tgatgttgaa ccttcaatgt agggaattcg ttcttgatgg tttcctccac
    12841 agtttttctc cataatcttg aagaggccaa aacattagct ttatccaagg accaaatagg
    12901 caatggtggc tcatgttgta gggccatgaa agcggccatt cttgtgattc tttgcacttc
    12961 tggaacggtg tattgttcac tatcccaagc gacaccatca ccatcatctt cctttctctt
    13021 accaaagtaa atacctccca ctaattctct gacaacaacg aagtcagtac ctttagcaaa
    13081 ttgtggcttg attggagata agtctaaaag agagtcggat gcaaagttac atggtcttaa
    13141 gttggcgtac aattgaagtt ctttacggat ttttagtaaa ccttgttcag gtctaacact
    13201 accggtaccc catttaggac cacccacagc acctaacaaa acggcatcag ccttcttgga
    13261 ggcttccagc gcctcatctg gaagtggaac acctgtagca tcgatagcag caccaccaat
    13321 taaatgattt tcgaaatcga acttgacatt ggaacgaaca tcagaaatag ctttaagaac
    13381 cttaatggct tcggctgtga tttcttgacc aacgtggtca cctggcaaaa cgacgatctt
    13441 cttaggggca gacattagaa tggtatatcc ttgaaatata tatatatatt gctgaaatgt
    13501 aaaaggtaag aaaagttaga aagtaagacg attgctaacc acctattgga aaaaacaata
    13561 ggtccttaaa taatattgtc aacttcaagt attgtgatgc aagcatttag tcatgaacgc
    13621 ttctctattc tatatgaaaa gccggttccg gcgctctcac ctttcctttt tctcccaatt
    13681 tttcagttga aaaaggtata tgcgtcaggc gacctctgaa attaacaaaa aatttccagt
    13741 catcgaattt gattctgtgc gatagcgccc ctgtgtgttc tcgttatgtt gaggaaaaaa
    13801 ataatggttg ctaagagatt cgaactcttg catcttacga tacctgagta ttcccacagt
    13861 taactgcggt caagatattt cttgaatcag gcgccttaga ccgctcggcc aaacaaccaa
    13921 ttacttgttg agaaatagag tataattatc ctataaatat aacgtttttg aacacacatg
    13981 aacaaggaag tacaggacaa ttgattttga agagaatgtg gattttgatg taattgttgg
    14041 gattccattt ttaataaggc aataatatta ggtatgtaga tatactagaa gttctcctcg
    14101 acaagcttgc atgcctgcag ggaaatcata acatagtata ataggaatca tatgacattt
    14161 actcttgcat agcaatatcc tcttcttcgc catcagcaat ccgacgagca tatgcttcag
    14221 cctcttgcaa aagctcatca ggaacgggtg tatgaatacc gtttgtttca acaccaaccc
    14281 aagtcatctc aatttcatgc tctttgttgt tttcttcgtc atgagttgcg tataatctaa
    14341 aaaagagtga attattagca aatagaatga aaaaaaattg tactcaacaa agcaaaacgt
    14401 acattctagc agcttcttta accgcatctt tcatttttaa tgaggaaaaa ttcagttttt
    14461 ctaattcgtt tcgagctact tgccgacctt taccagctgc agcaccgttg taaccccaat
    14521 aaacaccatt aggttcaacc atatacaaat gtggaccttc tgagtcataa gtagcaacaa
    14581 aagacataac accgaatgga cggacgctag aatagcaagt gaaaagttga acgtagttcc
    14641 ccaagcgatc agcaataact gttccaggga tgggagatcc gtagttatct ctccatgagg
    14701 tggcttcgtc ccttgcacgt tttacgatgt gttggccatc aggaataaag ccagtagtag
    14761 caataccaat gtgtcgatcg accgacccaa ttctgttatt tactcgcggc ttaagtagtt
    14821 tgctagtaac taccttttct agagctagaa taactccatc ttcacatttt attccaatgc
    14881 aagtactaaa tacaatgtta aaatcaatcg ataaggaaat tcaatccacc ttaccttgca
    14941 ttttcgacgg ctttatacgc gtactccgct tggaataaac gaccatccgg ggagaagaat
    15001 aatcctaaat cgtatccagt accaattgaa ctctgtttgt taatgtgatg tgctactcct
    15061 tgctaactta ccatttgctt atgttggtgg tagttggcat gcgtagactg atgactagtc
    15121 agcaaggagc gtagaacagt cacactcgtt atatatgtgc ttccaagaaa actcaagaat
    15181 ttaccattag caaacacttt tttgaaatgt tagacattta aatgacgaag gcatatagaa
    15241 gctttgaata ggtgttgtaa agtgttgatt tatgtgacgc tgagggtgcg catgaaagga
    15301 atgttgggtc acgattatta aacagtttgc tagcttggac acttgagtat tggaagttgt
    15361 tgaattctaa aaaactttca gttgatttga atagttgctg ttgccaaaaa acataacctg
    15421 taccaaagaa
//
