LOCUS       sequence-269032-        8090 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hCas9 under control of pPFK300 for direct cloning of HH-sgRNA-HDV 
            PCR products for episomal expression in P. pastoris and selection on
            NTC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8090)
  AUTHORS   Gassler T, Heistinger L, Mattanovich D, Gasser B, Prielhofer R
  TITLE     CRISPR/Cas9-Mediated Homology-Directed Genome Editing in Pichia 
            pastoris.
  JOURNAL   Methods Mol Biol. 2019;1923:211-225. doi: 
            10.1007/978-1-4939-9024-5_9.
  PUBMED    30737742
REFERENCE   2  (bases 1 to 8090)
  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: "Methods Mol
            Biol."; date: "2019"; volume: "1923:211-225. doi"; pages: " 
            10.1007/978-1-4939-9024-5_9"
COMMENT     Sequence Label: BB3cN_pGAP_23*_pPFK300_Cas9
FEATURES             Location/Qualifiers
     source          1..8090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(62..650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(142..161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        737..1213
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             1980..6083
                     /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             1980..6083
                     /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             1980..6083
                     /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             6096..6116
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6188..6435
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6204..6222)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     gene            6455..7574
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        6455..6798
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             6799..7371
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      7377..7574
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_feature    7586..8089
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 cgctgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
       61 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      241 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      481 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      601 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      661 tgatcttttc tacggggtgg agcgcaaaaa accccgcccc tgacagggcg gggttttttc
      721 gcgatcggag gatccttttt tgtagaaatg tcttggtgtc ctcgtccaat caggtagcca
      781 tctctgaaat atctggctcc gttgcaactc cgaacgacct gctggcaacg taaaattctc
      841 cggggtaaaa cttaaatgtg gagtaatgga accagaaacg tctcttccct tctctctcct
      901 tccaccgccc gttaccgtcc ctaggaaatt ttactctgct ggagagcttc ttctacggcc
      961 cccttgcagc aatgctcttc ccagcattac gttgcgggta aaacggaggt cgtgtacccg
     1021 acctagcagc ccagggatgg aaaagtcccg gccgtcgctg gcaataatag cgggcggacg
     1081 catgtcatga gattattgga aaccaccaga atcgaatata aaaggcgaac acctttccca
     1141 attttggttt ctcctgaccc aaagacttta aatttaattt atttgtccct atttcaatca
     1201 attgaacaac tatcaaaaca ccatggcgtc ttcggatccc tcgaggaaga cgcgctttta
     1261 gtgtacatct gataatatag tactaccacg tatgataatg tagagaatag gcttccttgt
     1321 cgagtgtgtt tgcagttttc ttgagtttca aggtttaaat gctggtatat tagttcatcg
     1381 aaggtttcag ccaatagcac cttaaatcaa tcaaactaat tcgactctta cgaaagagcc
     1441 tactgtgttt agtatcgaag tcgtttacct ttcatgttga atagcttcct ctctgaccct
     1501 aacatttcaa gatcctccta aagttacccg gattgtgaaa ttctaatgat ccacctgccc
     1561 aatgcatttt ttctttattc agtttacctt ttttacctaa tatacgagct tgttaaagta
     1621 agtggcactg caatactagg cttattgttg atattatgat gaatcgtttt cacaaacttg
     1681 atttcctgtg aactcaccat gtactaagga aaaaaacatg catcaccatc tgaatatttg
     1741 accgctccgg ggagcctgcc attaggcaaa caccaactag gaacctacac caagttatga
     1801 aggagcttgg aaaggattat aaaaaatgcg attacctaat acacacacga tgcctgagcg
     1861 gttaatgtga aatttcccgg ccatcgatcg ccccacaatc agactatttt cgctcataca
     1921 ataaaaagtc tcctgtttcc acccgagtat caccagaaca aagtaacaaa tacaaaacca
     1981 tggacaagaa gtactccatt gggctcgata tcggcacaaa cagcgtcggt tgggccgtca
     2041 ttacggacga gtacaaggtg ccgagcaaaa aattcaaagt tctgggcaat accgatcgcc
     2101 acagcataaa gaagaacctc attggcgccc tcctgttcga ctccggggag acggccgaag
     2161 ccacgcggct caaaagaaca gcacggcgca gatatacccg cagaaagaat cggatctgct
     2221 acctgcagga gatctttagt aatgagatgg ctaaggtgga tgactctttc ttccataggc
     2281 tggaggagtc ctttttggtg gaggaggata aaaagcacga gcgccaccca atctttggca
     2341 atatcgtgga cgaggtggcg taccatgaaa agtacccaac catatatcat ctgaggaaga
     2401 agcttgtaga cagtactgat aaggctgact tgcggttgat ctatctcgcg ctggcgcata
     2461 tgatcaaatt tcggggacac ttcctcatcg agggggacct gaacccagac aacagcgatg
     2521 tcgacaaact ctttatccaa ctggttcaga cttacaatca gcttttcgaa gagaacccga
     2581 tcaacgcatc cggagttgac gccaaagcaa tcctgagcgc taggctgtcc aaatcccggc
     2641 ggctcgaaaa cctcatcgca cagctccctg gggagaagaa gaacggcctg tttggtaatc
     2701 ttatcgccct gtcactcggg ctgaccccca actttaaatc taacttcgac ctggccgaag
     2761 atgccaagct tcaactgagc aaagacacct acgatgatga tctcgacaat ctgctggccc
     2821 agatcggcga ccagtacgca gacctttttt tggcggcaaa gaacctgtca gacgccattc
     2881 tgctgagtga tattctgcga gtgaacacgg agatcaccaa agctccgctg agcgctagta
     2941 tgatcaagcg ctatgatgag caccaccaag acttgacttt gctgaaggcc cttgtcagac
     3001 agcaactgcc tgagaagtac aaggaaattt tcttcgatca gtctaaaaat ggctacgccg
     3061 gatacattga cggcggagca agccaggagg aattttacaa atttattaag cccatcttgg
     3121 aaaaaatgga cggcaccgag gagctgctgg taaagcttaa cagagaagat ctgttgcgca
     3181 aacagcgcac tttcgacaat ggaagcatcc cccaccagat tcacctgggc gaactgcacg
     3241 ctatcctcag gcggcaagag gatttctacc cctttttgaa agataacagg gaaaagattg
     3301 agaaaatcct cacatttcgg ataccctact atgtaggccc cctcgcccgg ggaaattcca
     3361 gattcgcgtg gatgactcgc aaatcagaag agactatcac tccctggaac ttcgaggaag
     3421 tcgtggataa gggggcctct gcccagtcct tcatcgaaag gatgactaac tttgataaaa
     3481 atctgcctaa cgaaaaggtg cttcctaaac actctctgct gtacgagtac ttcacagttt
     3541 ataacgagct caccaaggtc aaatacgtca cagaagggat gagaaagcca gcattcctgt
     3601 ctggagagca gaagaaagct atcgtggacc tcctcttcaa gacgaaccgg aaagttaccg
     3661 tgaaacagct caaagaggac tatttcaaaa agattgaatg tttcgactct gttgaaatca
     3721 gcggagtgga ggatcgcttc aacgcatccc tgggaacgta tcacgatctc ctgaaaatca
     3781 ttaaagacaa ggacttcctg gacaatgagg agaacgagga cattcttgag gacattgtcc
     3841 tcacccttac gttgtttgaa gatagggaga tgattgaaga acgcttgaaa acttacgctc
     3901 atctcttcga cgacaaagtc atgaaacagc tcaagaggcg ccgatataca ggatgggggc
     3961 ggctgtcaag aaaactgatc aatgggatcc gagacaagca gagtggaaag acaatcctgg
     4021 attttcttaa gtccgatgga tttgccaacc ggaacttcat gcagttgatc catgatgact
     4081 ctctcacctt taaggaggac atccagaaag cacaagtttc tggccagggg gacagccttc
     4141 acgagcacat cgctaatctt gcaggtagcc cagctatcaa aaagggaata ctgcagaccg
     4201 ttaaggtcgt ggatgaactc gtcaaagtaa tgggaaggca taagcccgag aatatcgtta
     4261 tcgagatggc ccgagagaac caaactaccc agaagggaca gaagaacagt agggaaagga
     4321 tgaagaggat tgaagagggt ataaaagaac tggggtccca aatccttaag gaacacccag
     4381 ttgaaaacac ccagcttcag aatgagaagc tctacctgta ctacctgcag aacggcaggg
     4441 acatgtacgt ggatcaggaa ctggacatca atcggctctc cgactacgac gtggatcata
     4501 tcgtgcccca gtcttttctc aaagatgatt ctattgataa taaagtgttg acaagatccg
     4561 ataaaaatag agggaagagt gataacgtcc cctcagaaga agttgtcaag aaaatgaaaa
     4621 attattggcg gcagctgctg aacgccaaac tgatcacaca acggaagttc gataatctga
     4681 ctaaggctga acgaggtggc ctgtctgagt tggataaagc cggcttcatc aaaaggcagc
     4741 ttgttgagac acgccagatc accaagcacg tggcccaaat tctcgattca cgcatgaaca
     4801 ccaagtacga tgaaaatgac aaactgattc gagaggtgaa agttattact ctgaagtcta
     4861 agctcgtctc agatttcaga aaggactttc agttttataa ggtgagagag atcaacaatt
     4921 accaccatgc gcatgatgcc tacctgaatg cagtggtagg cactgcactt atcaaaaaat
     4981 atcccaagct tgaatctgaa tttgtttacg gagactataa agtgtacgat gttaggaaaa
     5041 tgatcgcaaa gtctgagcag gaaataggca aggccaccgc taagtacttc ttttacagca
     5101 atattatgaa ttttttcaag accgagatta cactggccaa tggagagatt cggaagcgac
     5161 cacttatcga aacaaacgga gaaacaggag aaatcgtgtg ggacaagggt agggatttcg
     5221 cgacagtccg gaaggtcctg tccatgccgc aggtgaacat cgttaaaaag accgaagtac
     5281 agaccggagg cttctccaag gaaagtatcc tcccgaaaag gaacagcgac aagctgatcg
     5341 cacgcaaaaa agattgggac cccaagaaat acggcggatt cgattctcct acagtcgctt
     5401 acagtgtact ggttgtggcc aaagtggaga aagggaagtc taaaaaactc aaaagcgtca
     5461 aggaactgct gggcatcaca atcatggagc gatcaagctt cgaaaaaaac cccatcgact
     5521 ttctcgaggc gaaaggatat aaagaggtca aaaaagacct catcattaag cttcccaagt
     5581 actctctctt tgagcttgaa aacggccgga aacgaatgct cgctagtgcg ggcgagctgc
     5641 agaaaggtaa cgagctggca ctgccctcta aatacgttaa tttcttgtat ctggccagcc
     5701 actatgaaaa gctcaaaggg tcccccgaag ataatgagca gaagcagctg ttcgtggaac
     5761 aacacaaaca ctaccttgat gagatcatcg agcaaataag cgaattctcc aaaagagtga
     5821 tcctcgccga cgctaacctc gataaggtgc tttctgctta caataagcac agggataagc
     5881 ccatcaggga gcaggcagaa aacattatcc acttgtttac tctgaccaac ttgggcgcgc
     5941 ctgcagcctt caagtacttc gacaccacca tagacagaaa gcggtacacc tctacaaagg
     6001 aggtcctgga cgccacactg attcatcagt caattacggg gctctatgaa acaagaatcg
     6061 acctctctca gctcggtgga gacagcaggg ctgaccccaa gaagaagagg aaggtgtgag
     6121 cttcgtccga cggcggcccg acgggtccga ggcctcggag atccgtcccc cttttccttt
     6181 gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct
     6241 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata
     6301 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga
     6361 cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg
     6421 aaggctttaa tttgcaagct ggcgctaatt catggacatg gaggcccaga ataccctcct
     6481 tgacagtctt gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt acatttagcc
     6541 catacatccc catgtataat catttgcatc catacatttt gatggccgca cggcgcgaag
     6601 caaaaattac ggctcctcgc tgcagacctg cgagcaggga aacgctcccc tcacagacgc
     6661 gttgaattgt ccccacgccg cgcccctgta gagaaatata aaaggttagg atttgccact
     6721 gaggttcttc tttcatatac ttccttttaa aatcttgcta ggatacagtt ctcacatcac
     6781 atccgaacat aaacaaccat gggtaccact cttgacgaca cggcttaccg gtaccgcacc
     6841 agtgtcccgg gggacgccga ggccatcgag gcactggatg ggtccttcac caccgacacc
     6901 gtttttcgcg tcaccgccac cggggacggc ttcaccctgc gggaggtgcc ggtggacccg
     6961 cccctgacca aggtgttccc cgacgacgaa tcggacgacg aatcggacga cggggaggac
     7021 ggcgacccgg actcccggac gttcgtcgcg tacggggacg acggcgacct ggcgggcttc
     7081 gtggtcgtct cgtactccgg ctggaaccgc cggctgaccg tcgaggacat cgaggtcgcc
     7141 ccggagcacc gggggcacgg ggtcgggcgc gcgttgatgg ggctcgcgac ggagttcgcc
     7201 cgcgagcggg gcgccgggca cctctggctg gaggtcacca acgtcaacgc accggcgatc
     7261 cacgcgtacc ggcggatggg gttcaccctc tgcggcctgg acaccgccct gtacgacggc
     7321 accgcctcgg acggcgagca ggcgctctac atgagcatgc cctgccccta atcagtactg
     7381 acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt tatattgtag
     7441 ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc gacatcatct
     7501 gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc
     7561 tggtcgctat actggcttcc ttttcatcac gtgctataaa aataattata atttaaattt
     7621 tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag
     7681 tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta
     7741 tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaac
     7801 accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta
     7861 tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt
     7921 tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac
     7981 tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta
     8041 ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcg
//
