LOCUS       sequence-449151-        9848 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 in a backbone compatible with K. phaffii..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9848)
  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 9848)
  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: pDIV151
FEATURES             Location/Qualifiers
     source          1..9848
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1315..1791)
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     protein_bind    complement(2555..2588)
                     /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(2795..2814)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2820..2839)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        2881..2972
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2973..3041,3042..3833)
                     /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(3191..3210)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3955..4543
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4444..4463
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4697..4714
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4812..4829
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4813..4829
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5299..5332
                     /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)"
     protein_bind    complement(5402..5435)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      complement(5498..5695)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(6274..6617)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     protein_bind    complement(6672..6705)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      complement(6754..7001)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     6967..6985
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(7017..7037)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(7050..9848,1..1305))
                     /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             complement(join(7050..9848,1..1305))
                     /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             complement(join(7050..9848,1..1305))
                     /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 atctttcaaa aaggggtaga aatcctcttg ccgcctgagg atagcgtgca gttcgcccag
       61 gtgaatctgg tgggggatgc ttccattgtc gaaagtgcgc tgtttgcgca acagatcttc
      121 tctgttaagc tttaccagca gctcctcggt gccgtccatt ttttccaaga tgggcttaat
      181 aaatttgtaa aattcctcct ggcttgctcc gccgtcaatg tatccggcgt agccattttt
      241 agactgatcg aagaaaattt ccttgtactt ctcaggcagt tgctgtctga caagggcctt
      301 cagcaaagtc aagtcttggt ggtgctcatc atagcgcttg atcatactag cgctcagcgg
      361 agctttggtg atctccgtgt tcactcgcag aatatcactc agcagaatgg cgtctgacag
      421 gttctttgcc gccaaaaaaa ggtctgcgta ctggtcgccg atctgggcca gcagattgtc
      481 gagatcatca tcgtaggtgt ctttgctcag ttgaagcttg gcatcttcgg ccaggtcgaa
      541 gttagattta aagttggggg tcagcccgag tgacagggcg ataagattac caaacaggcc
      601 gttcttcttc tccccaggga gctgtgcgat gaggttttcg agccgccggg atttggacag
      661 cctagcgctc aggattgctt tggcgtcaac tccggatgcg ttgatcgggt tctcttcgaa
      721 aagctgattg taagtctgaa ccagttggat aaagagtttg tcgacatcgc tgttgtctgg
      781 gttcaggtcc ccctcgatga ggaagtgtcc ccgaaatttg atcatatgcg ccagcgcgag
      841 atagatcaac cgcaagtcag ccttatcagt actgtctaca agcttcttcc tcagatgata
      901 tatggttggg tacttttcat ggtacgccac ctcgtccacg atattgccaa agattgggtg
      961 gcgctcgtgc tttttatcct cctccaccaa aaaggactcc tccagcctat ggaagaaaga
     1021 gtcatccacc ttagccatct cattactaaa gatctcctgc aggtagcaga tccgattctt
     1081 tctgcgggta tatctgcgcc gtgctgttct tttgagccgc gtggcttcgg ccgtctcccc
     1141 ggagtcgaac aggagggcgc caatgaggtt cttctttatg ctgtggcgat cggtattgcc
     1201 cagaactttg aattttttgc tcggcacctt gtactcgtcc gtaatgacgg cccaaccgac
     1261 gctgtttgtg ccgatatcga gcccaatgga gtacttcttg tccattgtgt tttgatagtt
     1321 gttcaattga ttgaaatagg gacaaataaa ttaaatttaa agtctttggg tcaggagaaa
     1381 ccaaaattgg gaaaggtgtt cgccttttat attcgattct ggtggtttcc aataatctca
     1441 tgacatgcgt ccgcccgcta ttattgccag cgacggccgg gacttttcca tccctgggct
     1501 gctaggtcgg gtacacgacc tccgttttac ccgcaacgta atgctgggaa gagcattgct
     1561 gcaagggggc cgtagaagaa gctctccagc agagtaaaat ttcctaggga cggtaacggg
     1621 cggtggaagg agagagaagg gaagagacgt ttctggttcc attactccac atttaagttt
     1681 taccccggag aattttacgt tgccagcagg tcgttcggag ttgcaacgga gccagatatt
     1741 tcagagatgg ctacctgatt ggacgaggac accaagacat ttctacaaaa agaatgcacg
     1801 cgatcgcacg cattccgcta gtgctgatta tgatttgacg tttatataca tgttatgtta
     1861 atatggtaaa ttttagatat ttggtaggag tgtcggtcct tgaaatatat gtaatatacc
     1921 cattatacca aggaaagatt gatttaacta tattggatta gcagatatct aatttgtttt
     1981 tttgttctta acctctggtg agttattatg atgtttttat attttatctt ttttaattaa
     2041 aagtcattat ttagaacaaa atattaaaat aattattaaa gtgtattagt attcttaatg
     2101 aacgtcggga agaacaaaat tttaaacatt ttgaagttaa acatcacttt tagtcctaaa
     2161 agtttaaaaa actgatattg atggaaagaa ctaaagaagt gcaaagagct caccaaaaaa
     2221 cgtactgcgt atctgttctt tctcattctg atattatcca aagatgttga gatcatgaat
     2281 gctagtcttt ttacactgaa tattttttta acgatttttt aataattttg caatcattta
     2341 agaaaaacca caaatggttt tccaaaattt aaattgattt tttagaaagt cctaaaaaat
     2401 agaatcatac caaataaaga acaacttaaa gtcactatga caaattcaga aatacaatcg
     2461 tatgatccgt tcatggataa agctgcaaag tcacaagcaa ggctaactaa gcccattaaa
     2521 ccctcagcgc ggccgcattt aaatccccac ttcagaagtt cctatacttt ctagagaata
     2581 ggaacttcac tatagagtcg aataagggcg acaccccata attagcccgg gcgaaaggcc
     2641 cagtctttcg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcatg
     2701 gggagtcccc acactaccat cggcgctacg gcgtttcact tctgagttcg gcatggggtc
     2761 aggtgggacc accgcgctac tgccgccagg caaacaaggg gtgttatgag ccatattcag
     2821 gtataaatgg gctcgcgata atgttcagaa ttggttaatt ggttgtaaca ctgaccccta
     2881 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2941 aaatgcttca ataatattga aaaaggaaga atatgagtat tcaacatttc cgtgtcgccc
     3001 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3061 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3121 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3181 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3241 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3301 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3361 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3421 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3481 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc gatggcaaca acgttgcgca
     3541 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3601 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3661 ctgataaatc cggagccggt gagcgtggtt ctcgcggtat catcgcagcg ctggggccag
     3721 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3781 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taagcagagc
     3841 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac
     3901 gcgcgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3961 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4021 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4081 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttagcccacc acttcaagaa
     4141 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4201 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4261 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4321 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4381 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4441 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4501 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4561 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4621 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4681 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaaggcgaga gtagggaact
     4741 gccaggcatc aaactaagca gaaggcccct gacggatggc ctttttgcgt ttctacaaac
     4801 tctttctgtg ttgtaaaacg acggccagtc ttaagctcgg gccccctggg cggttctgat
     4861 aacgagtaat cgttaatccg caaataacgt aaaaacccgc ttcggcgggt ttttttatgg
     4921 ggggagttta gggaaagagc atttgtcaga atatttaagg gcgcctgtca ctttgcttga
     4981 tatatgagaa ttatttaacc ttataaatga gaaaaaagca acgcacttta aataagatac
     5041 gttgcttttt cgattgatga acacctataa ttaaactatt catctattat ttatgatttt
     5101 ttgtatatac aatatttcta gtttgttaaa gagaattaag aaaataaatc tcgaaaataa
     5161 taaagggaaa atcagttttt gatatcaaaa ttatacatgt caacgataat acaaaatata
     5221 atacaaacta taagatgtta tcagtattta ttatgcattt agaataaatt ttgtgtcgcc
     5281 cttattcgac tcactataga agttcctatt ctctagaaag tataggaact tcacttcatt
     5341 ttatttaaat ttgcggccgc gctgaggtct taatggccac tagtggatct gatatcacct
     5401 aataacttcg tatagcatac attatacgaa gttatattaa gggttctcga gagctcgttt
     5461 tcgacactgg atggcggcgt tagtatcgaa tcgacagcag tatagcgacc agcattcaca
     5521 tacgattaac gcatgatatt actttctgcg cacttaactt cgcatctggg cagatgatgt
     5581 cgaggcgaaa aaaaatataa atcacgctaa catttgatta aaatagaaca actacaatat
     5641 aaaaaaacta tacaaatgac aagttcttga aaacaagaat ctttttattg tcagtactga
     5701 ttatggacaa ggcatagaca tgtataaagc ttgttctccg tctgaggctg taccatcgta
     5761 taaagctgta tccaatccac aaagtgtgaa ccccattctt cgatatgcat gtatagctgg
     5821 tgcattaaca ttggtgactt ctaaccataa atgtccagcc ccacgttcac gggcaaattc
     5881 tgtagctaat cccataaggg ctcgcccaac accgtgccca cgatgttctg gggcaacctc
     5941 tatatcctcg acagtaagtc ttcggttcca tccagaatag gatacaacta cgaaaccggc
     6001 caaatcaccg tcgtctccat aagcgacaaa tgttcttgag tctgggtcac cgtcctcacc
     6061 gtcatcggat tcatcgtcac tttcgtcatc tgggaatact ttggtcaaag gaggatctac
     6121 tggtacttct cgcaatgtaa acccatcccc agtagctgta actctaaaga cggtgtcagt
     6181 tgtgaatgac ccgtcaaggg cttcgatagc ttcagcatca ccaggtacag atgttcggta
     6241 tcgataagca gtgtcatcca aggtggttcc catggttgtt tatgttcgga tgtgatgtga
     6301 gaactgtatc ctagcaagat tttaaaagga agtatatgaa agaagaacct cagtggcaaa
     6361 tcctaacctt ttatatttct ctacaggggc gcggcgtggg gacaattcaa cgcgtctgtg
     6421 aggggagcgt ttccctgctc gcaggtctgc agcgaggagc cgtaattttt gcttcgcgcc
     6481 gtgcggccat caaaatgtat ggatgcaaat gattatacat ggggatgtat gggctaaatg
     6541 tacgggcgac agtcacatca tgcccctgag ctgcgcacgt caagactgtc aaggagggta
     6601 ttctgggcct ccatgtcgct ggccgggtga cccggcgggg acgaggcaag ctaaacagat
     6661 ctctagacct aataacttcg tatagcatac attatacgaa gttatattaa gggttgtcga
     6721 cctgcagcgt tgaatgcacg cgatgtaccg gccgcaaatt aaagccttcg agcgtcccaa
     6781 aaccttctca agcaaggttt tcagtataat gttacatgcg tacacgcgtc tgtacagaaa
     6841 aaaaagaaaa atttgaaata taaataacgt tcttaatact aacataacta taaaaaaata
     6901 aatagggacc tagacttcag gttgtctaac tccttccttt tcggttagag cggatgtggg
     6961 gggagggcgt gaatgtaagc gtgacataac taattacatg actcgagaag agatcacacc
     7021 ttcctcttct tcttggggtc agccctgctg tctccaccga gctgagagag gtcgattctt
     7081 gtttcataga gccccgtaat tgactgatga atcagtgtgg cgtccaggac ctcctttgta
     7141 gaggtgtacc gctttctgtc tatggtggtg tcgaagtact tgaaggctgc aggcgcgccc
     7201 aagttggtca gagtaaacaa gtggataatg ttttctgcct gctccctgat gggcttatcc
     7261 ctgtgcttat tgtaagcaga aagcacctta tcgaggttag cgtcggcgag gatcactctt
     7321 ttggagaatt cgcttatttg ctcgatgatc tcatcaaggt agtgtttgtg ttgttccacg
     7381 aacagctgct tctgctcatt atcttcggga gaccctttga gcttttcata gtggctggcc
     7441 agatacaaga aattaacgta tttagagggc agtgccagct cgttaccttt ctgcagctcg
     7501 cccgcactag cgagcattcg tttccggccg ttttcaagct caaagagaga gtacttggga
     7561 agcttaatga tgaggtcttt tttgacctct ttatatcctt tcgcctcgag aaagtcgatg
     7621 gggttttttt cgaagcttga tcgctccatg attgtgatgc ccagcagttc cttgacgctt
     7681 ttgagttttt tagacttccc tttctccact ttggccacaa ccagtacact gtaagcgact
     7741 gtaggagaat cgaatccgcc gtatttcttg gggtcccaat cttttttgcg tgcgatcagc
     7801 ttgtcgctgt tccttttcgg gaggatactt tccttggaga agcctccggt ctgtacttcg
     7861 gtctttttaa cgatgttcac ctgcggcatg gacaggacct tccggactgt cgcgaaatcc
     7921 ctacccttgt cccacacgat ttctcctgtt tctccgtttg tttcgataag tggtcgcttc
     7981 cgaatctctc cattggccag tgtaatctcg gtcttgaaaa aattcataat attgctgtaa
     8041 aagaagtact tagcggtggc cttgcctatt tcctgctcag actttgcgat cattttccta
     8101 acatcgtaca ctttatagtc tccgtaaaca aattcagatt caagcttggg atattttttg
     8161 ataagtgcag tgcctaccac tgcattcagg taggcatcat gcgcatggtg gtaattgttg
     8221 atctctctca ccttataaaa ctgaaagtcc tttctgaaat ctgagaccag cttagacttc
     8281 agagtaataa ctttcacctc tcgaatcagt ttgtcatttt catcgtactt ggtgttcatg
     8341 cgtgaatcga gaatttgggc cacgtgcttg gtgatctggc gtgtctcaac aagctgcctt
     8401 ttgatgaagc cggctttatc caactcagac aggccacctc gttcagcctt agtcagatta
     8461 tcgaacttcc gttgtgtgat cagtttggcg ttcagcagct gccgccaata atttttcatt
     8521 ttcttgacaa cttcttctga ggggacgtta tcactcttcc ctctattttt atcggatctt
     8581 gtcaacactt tattatcaat agaatcatct ttgagaaaag actggggcac gatatgatcc
     8641 acgtcgtagt cggagagccg attgatgtcc agttcctgat ccacgtacat gtccctgccg
     8701 ttctgcaggt agtacaggta gagcttctca ttctgaagct gggtgttttc aactgggtgt
     8761 tccttaagga tttgggaccc cagttctttt ataccctctt caatcctctt catcctttcc
     8821 ctactgttct tctgtccctt ctgggtagtt tggttctctc gggccatctc gataacgata
     8881 ttctcgggct tatgtcttcc cattactttg acgagttcat ccacgacctt aacggtctgc
     8941 agtattccct ttttgatagc tgggctacct gcaagattag cgatgtgctc gtgaagactg
     9001 tccccctggc cagaaacttg tgctttctgg atgtcctcct taaaggtgag agagtcatca
     9061 tggatcaact gcatgaagtt ccggttggca aatccatcgg acttaagaaa atccaggatt
     9121 gtctttccac tctgcttgtc tcggatccca ttgatcagtt ttcttgacag ccgcccccat
     9181 cctgtatatc ggcgcctctt gagctgtttc atgactttgt cgtcgaagag atgagcgtaa
     9241 gttttcaagc gttcttcaat catctcccta tcttcaaaca acgtaagggt gaggacaatg
     9301 tcctcaagaa tgtcctcgtt ctcctcattg tccaggaagt ccttgtcttt aatgattttc
     9361 aggagatcgt gatacgttcc cagggatgcg ttgaagcgat cctccactcc gctgatttca
     9421 acagagtcga aacattcaat ctttttgaaa tagtcttctt tgagctgttt cacggtaact
     9481 ttccggttcg tcttgaagag gaggtccacg atagctttct tctgctctcc agacaggaat
     9541 gctggctttc tcatcccttc tgtgacgtat ttgaccttgg tgagctcgtt ataaactgtg
     9601 aagtactcgt acagcagaga gtgtttagga agcacctttt cgttaggcag atttttatca
     9661 aagttagtca tcctttcgat gaaggactgg gcagaggccc ccttatccac gacttcctcg
     9721 aagttccagg gagtgatggt ctcttctgat ttgcgagtca tccacgcgaa tctggaattt
     9781 ccccgggcga gggggcctac atagtagggt atccgaaatg tgaggatttt ctcaatcttt
     9841 tccctgtt
//
