LOCUS       sequence-449152-       10950 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 and gRNA targeting KU70 gene in K. phaffii..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10950)
  AUTHORS   Strucko T, Gadar-Lopez AE, Frohling FB, Frost ET, Iversen EF, Olsson
            H, Jarczynska ZD, Mortensen UH
  TITLE     Oligonucleotide-based CRISPR-Cas9 toolbox for efficient engineering 
            of Komagataella phaffii.
  JOURNAL   FEMS Yeast Res. 2024 Aug 23:foae026. doi: 10.1093/femsyr/foae026.
  PUBMED    39179418
REFERENCE   2  (bases 1 to 10950)
  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: "FEMS Yeast 
            Res. 2024 Aug 23:foae026. doi: 10.1093/femsyr/foae026."
COMMENT     Sequence Label: pDIV270
FEATURES             Location/Qualifiers
     source          1..10950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2346..2366
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      2382..2629
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2398..2416)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    2678..2711
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2766..3109
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      3688..3885
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    3948..3981
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(4051..4084)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(4554..4571)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4554..4570)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4669..4686)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4840..5428)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4920..4939)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5550..6341,6342..6410))
                     /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     6173..6192
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6411..6502)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6544..6563
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6569..6588
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    6795..6828
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_RNA        8296..8371
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8694..9170
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             join(9180..10950,1..2333)
                     /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             join(9180..10950,1..2333)
                     /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             join(9180..10950,1..2333)
                     /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"
ORIGIN
        1 tgggaacgta tcacgatctc ctgaaaatca ttaaagacaa ggacttcctg gacaatgagg
       61 agaacgagga cattcttgag gacattgtcc tcacccttac gttgtttgaa gatagggaga
      121 tgattgaaga acgcttgaaa acttacgctc atctcttcga cgacaaagtc atgaaacagc
      181 tcaagaggcg ccgatataca ggatgggggc ggctgtcaag aaaactgatc aatgggatcc
      241 gagacaagca gagtggaaag acaatcctgg attttcttaa gtccgatgga tttgccaacc
      301 ggaacttcat gcagttgatc catgatgact ctctcacctt taaggaggac atccagaaag
      361 cacaagtttc tggccagggg gacagtcttc acgagcacat cgctaatctt gcaggtagcc
      421 cagctatcaa aaagggaata ctgcagaccg ttaaggtcgt ggatgaactc gtcaaagtaa
      481 tgggaagaca taagcccgag aatatcgtta tcgagatggc ccgagagaac caaactaccc
      541 agaagggaca gaagaacagt agggaaagga tgaagaggat tgaagagggt ataaaagaac
      601 tggggtccca aatccttaag gaacacccag ttgaaaacac ccagcttcag aatgagaagc
      661 tctacctgta ctacctgcag aacggcaggg acatgtacgt ggatcaggaa ctggacatca
      721 atcggctctc cgactacgac gtggatcata tcgtgcccca gtcttttctc aaagatgatt
      781 ctattgataa taaagtgttg acaagatccg ataaaaatag agggaagagt gataacgtcc
      841 cctcagaaga agttgtcaag aaaatgaaaa attattggcg gcagctgctg aacgccaaac
      901 tgatcacaca acggaagttc gataatctga ctaaggctga acgaggtggc ctgtctgagt
      961 tggataaagc cggcttcatc aaaaggcagc ttgttgagac acgccagatc accaagcacg
     1021 tggcccaaat tctcgattca cgcatgaaca ccaagtacga tgaaaatgac aaactgattc
     1081 gagaggtgaa agttattact ctgaagtcta agctggtctc agatttcaga aaggactttc
     1141 agttttataa ggtgagagag atcaacaatt accaccatgc gcatgatgcc tacctgaatg
     1201 cagtggtagg cactgcactt atcaaaaaat atcccaagct tgaatctgaa tttgtttacg
     1261 gagactataa agtgtacgat gttaggaaaa tgatcgcaaa gtctgagcag gaaataggca
     1321 aggccaccgc taagtacttc ttttacagca atattatgaa ttttttcaag accgagatta
     1381 cactggccaa tggagagatt cggaagcgac cacttatcga aacaaacgga gaaacaggag
     1441 aaatcgtgtg ggacaagggt agggatttcg cgacagtccg gaaggtcctg tccatgccgc
     1501 aggtgaacat cgttaaaaag accgaagtac agaccggagg cttctccaag gaaagtatcc
     1561 tcccgaaaag gaacagcgac aagctgatcg cacgcaaaaa agattgggac cccaagaaat
     1621 acggcggatt cgattctcct acagtcgctt acagtgtact ggttgtggcc aaagtggaga
     1681 aagggaagtc taaaaaactc aaaagcgtca aggaactgct gggcatcaca atcatggagc
     1741 gatcaagctt cgaaaaaaac cccatcgact ttctcgaggc gaaaggatat aaagaggtca
     1801 aaaaagacct catcattaag cttcccaagt actctctctt tgagcttgaa aacggccgga
     1861 aacgaatgct cgctagtgcg ggcgagctgc agaaaggtaa cgagctggca ctgccctcta
     1921 aatacgttaa tttcttgtat ctggccagcc actatgaaaa gctcaaaggg tctcccgaag
     1981 ataatgagca gaagcagctg ttcgtggaac aacacaaaca ctaccttgat gagatcatcg
     2041 agcaaataag cgaattctcc aaaagagtga tcctcgccga cgctaacctc gataaggtgc
     2101 tttctgctta caataagcac agggataagc ccatcaggga gcaggcagaa aacattatcc
     2161 acttgtttac tctgaccaac ttgggcgcgc ctgcagcctt caagtacttc gacaccacca
     2221 tagacagaaa gcggtacacc tctacaaagg aggtcctgga cgccacactg attcatcagt
     2281 caattacggg gctctatgaa acaagaatcg acctctctca gctcggtgga gacagcaggg
     2341 ctgaccccaa gaagaagagg aaggtgtgat ctcttctcga gtcatgtaat tagttatgtc
     2401 acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca
     2461 acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt
     2521 tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact
     2581 gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgcg gccggtacat
     2641 cgcgtgcatt caacgctgca ggtcgacaac ccttaatata acttcgtata atgtatgcta
     2701 tacgaagtta ttaggtctag agatctgttt agcttgcctc gtccccgccg ggtcacccgg
     2761 ccagcgacat ggaggcccag aataccctcc ttgacagtct tgacgtgcgc agctcagggg
     2821 catgatgtga ctgtcgcccg tacatttagc ccatacatcc ccatgtataa tcatttgcat
     2881 ccatacattt tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct
     2941 gcgagcaggg aaacgctccc ctcacagacg cgttgaattg tccccacgcc gcgcccctgt
     3001 agagaaatat aaaaggttag gatttgccac tgaggttctt ctttcatata cttcctttta
     3061 aaatcttgct aggatacagt tctcacatca catccgaaca taaacaacca tgggaaccac
     3121 cttggatgac actgcttatc gataccgaac atctgtacct ggtgatgctg aagctatcga
     3181 agcccttgac gggtcattca caactgacac cgtctttaga gttacagcta ctggggatgg
     3241 gtttacattg cgagaagtac cagtagatcc tcctttgacc aaagtattcc cagatgacga
     3301 aagtgacgat gaatccgatg acggtgagga cggtgaccca gactcaagaa catttgtcgc
     3361 ttatggagac gacggtgatt tggccggttt cgtagttgta tcctattctg gatggaaccg
     3421 aagacttact gtcgaggata tagaggttgc cccagaacat cgtgggcacg gtgttgggcg
     3481 agcccttatg ggattagcta cagaatttgc ccgtgaacgt ggggctggac atttatggtt
     3541 agaagtcacc aatgttaatg caccagctat acatgcatat cgaagaatgg ggttcacact
     3601 ttgtggattg gatacagctt tatacgatgg tacagcctca gacggagaac aagctttata
     3661 catgtctatg ccttgtccat aatcagtact gacaataaaa agattcttgt tttcaagaac
     3721 ttgtcatttg tatagttttt ttatattgta gttgttctat tttaatcaaa tgttagcgtg
     3781 atttatattt tttttcgcct cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa
     3841 gtaatatcat gcgttaatcg tatgtgaatg ctggtcgcta tactgctgtc gattcgatac
     3901 taacgccgcc atccagtgtc gaaaacgagc tctcgagaac ccttaatata acttcgtata
     3961 atgtatgcta tacgaagtta ttaggtgata tcagatccac tagtggccat taagacctca
     4021 gcgcggccgc aaatttaaat aaaatgaagt gaagttccta tactttctag agaataggaa
     4081 cttctatagt gagtcgaata agggcgacac aaaatttatt ctaaatgcat aataaatact
     4141 gataacatct tatagtttgt attatatttt gtattatcgt tgacatgtat aattttgata
     4201 tcaaaaactg attttccctt tattattttc gagatttatt ttcttaattc tctttaacaa
     4261 actagaaata ttgtatatac aaaaaatcat aaataataga tgaatagttt aattataggt
     4321 gttcatcaat cgaaaaagca acgtatctta tttaaagtgc gttgcttttt tctcatttat
     4381 aaggttaaat aattctcata tatcaagcaa agtgacaggc gcccttaaat attctgacaa
     4441 atgctctttc cctaaactcc ccccataaaa aaacccgccg aagcgggttt ttacgttatt
     4501 tgcggattaa cgattactcg ttatcagaac cgcccagggg gcccgagctt aagactggcc
     4561 gtcgttttac aacacagaaa gagtttgtag aaacgcaaaa aggccatccg tcaggggcct
     4621 tctgcttagt ttgatgcctg gcagttccct actctcgcct tccgcttcct cgctcactga
     4681 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4741 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4801 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4861 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4921 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4981 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5041 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5101 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5161 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5221 gtatgtaggc ggtgctacag agttcttgaa gtggtgggct aactacggct acactagaag
     5281 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5341 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5401 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5461 cgctcagtgg aacgacgcgc gcgtaactca cgttaaggga ttttggtcat gagcttgcgc
     5521 cgtcccgtca agtcagcgta atgctctgct taccaatgct taatcagtga ggcacctatc
     5581 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5641 acgatacggg agggcttacc atctggcccc agcgctgcga tgataccgcg agaaccacgc
     5701 tcaccggctc cggatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5761 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5821 agtagttcgc cagttaatag tttgcgcaac gttgttgcca tcgctacagg catcgtggtg
     5881 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5941 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6001 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6061 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6121 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6181 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6241 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6301 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6361 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat attcttcctt
     6421 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6481 tgtatttaga aaaataaaca aataggggtc agtgttacaa ccaattaacc aattctgaac
     6541 attatcgcga gcccatttat acctgaatat ggctcataac accccttgtt tgcctggcgg
     6601 cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc
     6661 cgatggtagt gtggggactc cccatgcgag agtagggaac tgccaggcat caaataaaac
     6721 gaaaggctca gtcgaaagac tgggcctttc gcccgggcta attatggggt gtcgccctta
     6781 ttcgactcta tagtgaagtt cctattctct agaaagtata ggaacttctg aagtggggat
     6841 ttaaatgcgg ccgcgctgag ggtttaatgg gcttagttag ccttgcttgt gactttgcag
     6901 ctttatccat gaacggatca tacgattgta tttctgaatt tgtcatagtg actttaagtt
     6961 gttctttatt tggtatgatt ctatttttta ggactttcta aaaaatcaat ttaaattttg
     7021 gaaaaccatt tgtggttttt cttaaatgat tgcaaaatta ttaaaaaatc gttaaaaaaa
     7081 tattcagtgt aaaaagacta gcattcatga tctcaacatc tttggataat atcagaatga
     7141 gaaagaacag atacgcagta cgttttttgg tgagctcttt gcacttcttt agttctttcc
     7201 atcaatatca gttttttaaa cttttaggac taaaagtgat gtttaacttc aaaatgttta
     7261 aaattttgtt cttcccgacg ttcattaaga atactaatac actttaataa ttattttaat
     7321 attttgttct aaataatgac ttttaattaa aaaagataaa atataaaaac atcataataa
     7381 ctcaccagag gttaagaaca aaaaaacaaa ttagatatct gctaatccaa tatagttaaa
     7441 tcaatctttc cttggtataa tgggtatatt acatatattt caaggaccga cactcctacc
     7501 aaatatctaa aatttaccat attaacataa catgtatata aacgtcaaat cataatcagc
     7561 actagcggaa tgcgtgcgat taactctggc aaccagtaac acgcttaagg tttggaacaa
     7621 cactaaacta ccttgcggta ctaccattga cactacacat ccttaattcc aatcctgtct
     7681 ggcctccttc accttttaac catcttgccc attccaactc gtgtcagatt gcgtatcaag
     7741 tgaaaaaaaa aaaattttaa atctttaacc caatcaggta ataactgtcg cctcttttat
     7801 ctgccgcact gcatgaggtg tccccttagt gggaaagagt actgagccaa ccctggagga
     7861 cagcaaggga aaaataccta caacttgctt cataatggtc gtaaaaacaa tccttgtcgg
     7921 atataagtgt tgtagactgt cccttatcct ctgcgatgtt cttcctctca aagtttgcga
     7981 tttctctcta tcagaattgc catcaagaga ctcaggacta atttcgcagt cccacacgca
     8041 ctcgtacatg attggctgaa atttccctaa agaatttctt tttcacgaaa attttttttt
     8101 tacacaagat tttcagcaga tataaaatgg agagcaggac ctccgctgtg actcttcttt
     8161 tttttctttt attctcacta catacatttt agttattcgc caacgcatca ttggtctagt
     8221 ggtagaattc atcgttgcca tcgatgaggc ccgtgttcga ttcacggatg atgcaaagca
     8281 atacgacatc cacgagtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
     8341 tcaacttgaa aaagtggcac cgagtcggtg cttttgcatc attggtctag tggtagaatt
     8401 catcgttgcc atcgatgagg cccgtgttcg attcacggat gatgcatcaa gaggatgtca
     8461 gaatgccatt tgcctgagag atgcaggctt catttttgat acttttttat ttgtaaccta
     8521 tatagtatag gatttttttt gtcattttgt ttcttctcgt acgagcttgc tcctgatcag
     8581 cctatctcgc agctgatgaa tatcttgtgg taggggtttg ggaaaatcat tcgagtttga
     8641 tgtttttctt ggtatttccc actcctcttc agagtacaga atcgcgtgca ttctttttgt
     8701 agaaatgtct tggtgtcctc gtccaatcag gtagccatct ctgaaatatc tggctccgtt
     8761 gcaactccga acgacctgct ggcaacgtaa aattctccgg ggtaaaactt aaatgtggag
     8821 taatggaacc agaaacgtct cttcccttct ctctccttcc accgcccgtt accgtcccta
     8881 ggaaatttta ctctgctgga gagcttcttc tacggccccc ttgcagcaat gctcttccca
     8941 gcattacgtt gcgggtaaaa cggaggtcgt gtacccgacc tagcagccca gggatggaaa
     9001 agtcccggcc gtcgctggca ataatagcgg gcggacgcat gtcatgagat tattggaaac
     9061 caccagaatc gaatataaaa ggcgaacacc tttcccaatt ttggtttctc ctgacccaaa
     9121 gactttaaat ttaatttatt tgtccctatt tcaatcaatt gaacaactat caaaacacaa
     9181 tggacaagaa gtactccatt gggctcgata tcggcacaaa cagcgtcggt tgggccgtca
     9241 ttacggacga gtacaaggtg ccgagcaaaa aattcaaagt tctgggcaat accgatcgcc
     9301 acagcataaa gaagaacctc attggcgccc tcctgttcga ctccggggag acggccgaag
     9361 ccacgcggct caaaagaaca gcacggcgca gatatacccg cagaaagaat cggatctgct
     9421 acctgcagga gatctttagt aatgagatgg ctaaggtgga tgactctttc ttccataggc
     9481 tggaggagtc ctttttggtg gaggaggata aaaagcacga gcgccaccca atctttggca
     9541 atatcgtgga cgaggtggcg taccatgaaa agtacccaac catatatcat ctgaggaaga
     9601 agcttgtaga cagtactgat aaggctgact tgcggttgat ctatctcgcg ctggcgcata
     9661 tgatcaaatt tcggggacac ttcctcatcg agggggacct gaacccagac aacagcgatg
     9721 tcgacaaact ctttatccaa ctggttcaga cttacaatca gcttttcgaa gagaacccga
     9781 tcaacgcatc cggagttgac gccaaagcaa tcctgagcgc taggctgtcc aaatcccggc
     9841 ggctcgaaaa cctcatcgca cagctccctg gggagaagaa gaacggcctg tttggtaatc
     9901 ttatcgccct gtcactcggg ctgaccccca actttaaatc taacttcgac ctggccgaag
     9961 atgccaagct tcaactgagc aaagacacct acgatgatga tctcgacaat ctgctggccc
    10021 agatcggcga ccagtacgca gacctttttt tggcggcaaa gaacctgtca gacgccattc
    10081 tgctgagtga tattctgcga gtgaacacgg agatcaccaa agctccgctg agcgctagta
    10141 tgatcaagcg ctatgatgag caccaccaag acttgacttt gctgaaggcc cttgtcagac
    10201 agcaactgcc tgagaagtac aaggaaattt tcttcgatca gtctaaaaat ggctacgccg
    10261 gatacattga cggcggagca agccaggagg aattttacaa atttattaag cccatcttgg
    10321 aaaaaatgga cggcaccgag gagctgctgg taaagcttaa cagagaagat ctgttgcgca
    10381 aacagcgcac tttcgacaat ggaagcatcc cccaccagat tcacctgggc gaactgcacg
    10441 ctatcctcag gcggcaagag gatttctacc cctttttgaa agataacagg gaaaagattg
    10501 agaaaatcct cacatttcgg ataccctact atgtaggccc cctcgcccgg ggaaattcca
    10561 gattcgcgtg gatgactcgc aaatcagaag agaccatcac tccctggaac ttcgaggaag
    10621 tcgtggataa gggggcctct gcccagtcct tcatcgaaag gatgactaac tttgataaaa
    10681 atctgcctaa cgaaaaggtg cttcctaaac actctctgct gtacgagtac ttcacagttt
    10741 ataacgagct caccaaggtc aaatacgtca cagaagggat gagaaagcca gcattcctgt
    10801 ctggagagca gaagaaagct atcgtggacc tcctcttcaa gacgaaccgg aaagttaccg
    10861 tgaaacagct caaagaagac tatttcaaaa agattgaatg tttcgactct gttgaaatca
    10921 gcggagtgga ggatcgcttc aacgcatccc
//
