LOCUS       sequence-206061-        8327 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
            G418.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8327)
  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 8327)
  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: BB3cK_pGAP_23*_pPFK300_Cas9
FEATURES             Location/Qualifiers
     source          1..8327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(70..658)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(150..169)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        745..1221
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             1988..6091
                     /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             1988..6091
                     /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             1988..6091
                     /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             6104..6124
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6196..6443
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6212..6230)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     gene            6463..7819
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        6463..6806
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             6807..7616
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(6874..6893)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      7622..7819
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_feature    join(7831..8327,1..7)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 agcgatcgcg ctgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
       61 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      121 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
      181 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
      241 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
      301 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
      361 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
      421 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
      481 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
      541 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
      601 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
      661 agatcctttg atcttttcta cggggtggag cgcaaaaaac cccgcccctg acagggcggg
      721 gttttttcgc gatcggagga tccttttttg tagaaatgtc ttggtgtcct cgtccaatca
      781 ggtagccatc tctgaaatat ctggctccgt tgcaactccg aacgacctgc tggcaacgta
      841 aaattctccg gggtaaaact taaatgtgga gtaatggaac cagaaacgtc tcttcccttc
      901 tctctccttc caccgcccgt taccgtccct aggaaatttt actctgctgg agagcttctt
      961 ctacggcccc cttgcagcaa tgctcttccc agcattacgt tgcgggtaaa acggaggtcg
     1021 tgtacccgac ctagcagccc agggatggaa aagtcccggc cgtcgctggc aataatagcg
     1081 ggcggacgca tgtcatgaga ttattggaaa ccaccagaat cgaatataaa aggcgaacac
     1141 ctttcccaat tttggtttct cctgacccaa agactttaaa tttaatttat ttgtccctat
     1201 ttcaatcaat tgaacaacta tcaaaacacc atggcgtctt cggatccctc gaggaagacg
     1261 cgcttttagt gtacatctga taatatagta ctaccacgta tgataatgta gagaataggc
     1321 ttccttgtcg agtgtgtttg cagttttctt gagtttcaag gtttaaatgc tggtatatta
     1381 gttcatcgaa ggtttcagcc aatagcacct taaatcaatc aaactaattc gactcttacg
     1441 aaagagccta ctgtgtttag tatcgaagtc gtttaccttt catgttgaat agcttcctct
     1501 ctgaccctaa catttcaaga tcctcctaaa gttacccgga ttgtgaaatt ctaatgatcc
     1561 acctgcccaa tgcatttttt ctttattcag tttacctttt ttacctaata tacgagcttg
     1621 ttaaagtaag tggcactgca atactaggct tattgttgat attatgatga atcgttttca
     1681 caaacttgat ttcctgtgaa ctcaccatgt actaaggaaa aaaacatgca tcaccatctg
     1741 aatatttgac cgctccgggg agcctgccat taggcaaaca ccaactagga acctacacca
     1801 agttatgaag gagcttggaa aggattataa aaaatgcgat tacctaatac acacacgatg
     1861 cctgagcggt taatgtgaaa tttcccggcc atcgatcgcc ccacaatcag actattttcg
     1921 ctcatacaat aaaaagtctc ctgtttccac ccgagtatca ccagaacaaa gtaacaaata
     1981 caaaaccatg gacaagaagt actccattgg gctcgatatc ggcacaaaca gcgtcggttg
     2041 ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc tgggcaatac
     2101 cgatcgccac agcataaaga agaacctcat tggcgccctc ctgttcgact ccggggagac
     2161 ggccgaagcc acgcggctca aaagaacagc acggcgcaga tatacccgca gaaagaatcg
     2221 gatctgctac ctgcaggaga tctttagtaa tgagatggct aaggtggatg actctttctt
     2281 ccataggctg gaggagtcct ttttggtgga ggaggataaa aagcacgagc gccacccaat
     2341 ctttggcaat atcgtggacg aggtggcgta ccatgaaaag tacccaacca tatatcatct
     2401 gaggaagaag cttgtagaca gtactgataa ggctgacttg cggttgatct atctcgcgct
     2461 ggcgcatatg atcaaatttc ggggacactt cctcatcgag ggggacctga acccagacaa
     2521 cagcgatgtc gacaaactct ttatccaact ggttcagact tacaatcagc ttttcgaaga
     2581 gaacccgatc aacgcatccg gagttgacgc caaagcaatc ctgagcgcta ggctgtccaa
     2641 atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga acggcctgtt
     2701 tggtaatctt atcgccctgt cactcgggct gacccccaac tttaaatcta acttcgacct
     2761 ggccgaagat gccaagcttc aactgagcaa agacacctac gatgatgatc tcgacaatct
     2821 gctggcccag atcggcgacc agtacgcaga cctttttttg gcggcaaaga acctgtcaga
     2881 cgccattctg ctgagtgata ttctgcgagt gaacacggag atcaccaaag ctccgctgag
     2941 cgctagtatg atcaagcgct atgatgagca ccaccaagac ttgactttgc tgaaggccct
     3001 tgtcagacag caactgcctg agaagtacaa ggaaattttc ttcgatcagt ctaaaaatgg
     3061 ctacgccgga tacattgacg gcggagcaag ccaggaggaa ttttacaaat ttattaagcc
     3121 catcttggaa aaaatggacg gcaccgagga gctgctggta aagcttaaca gagaagatct
     3181 gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc caccagattc acctgggcga
     3241 actgcacgct atcctcaggc ggcaagagga tttctacccc tttttgaaag ataacaggga
     3301 aaagattgag aaaatcctca catttcggat accctactat gtaggccccc tcgcccgggg
     3361 aaattccaga ttcgcgtgga tgactcgcaa atcagaagag actatcactc cctggaactt
     3421 cgaggaagtc gtggataagg gggcctctgc ccagtccttc atcgaaagga tgactaactt
     3481 tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt acgagtactt
     3541 cacagtttat aacgagctca ccaaggtcaa atacgtcaca gaagggatga gaaagccagc
     3601 attcctgtct ggagagcaga agaaagctat cgtggacctc ctcttcaaga cgaaccggaa
     3661 agttaccgtg aaacagctca aagaggacta tttcaaaaag attgaatgtt tcgactctgt
     3721 tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc acgatctcct
     3781 gaaaatcatt aaagacaagg acttcctgga caatgaggag aacgaggaca ttcttgagga
     3841 cattgtcctc acccttacgt tgtttgaaga tagggagatg attgaagaac gcttgaaaac
     3901 ttacgctcat ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc gatatacagg
     3961 atgggggcgg ctgtcaagaa aactgatcaa tgggatccga gacaagcaga gtggaaagac
     4021 aatcctggat tttcttaagt ccgatggatt tgccaaccgg aacttcatgc agttgatcca
     4081 tgatgactct ctcaccttta aggaggacat ccagaaagca caagtttctg gccaggggga
     4141 cagccttcac gagcacatcg ctaatcttgc aggtagccca gctatcaaaa agggaatact
     4201 gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata agcccgagaa
     4261 tatcgttatc gagatggccc gagagaacca aactacccag aagggacaga agaacagtag
     4321 ggaaaggatg aagaggattg aagagggtat aaaagaactg gggtcccaaa tccttaagga
     4381 acacccagtt gaaaacaccc agcttcagaa tgagaagctc tacctgtact acctgcagaa
     4441 cggcagggac atgtacgtgg atcaggaact ggacatcaat cggctctccg actacgacgt
     4501 ggatcatatc gtgccccagt cttttctcaa agatgattct attgataata aagtgttgac
     4561 aagatccgat aaaaatagag ggaagagtga taacgtcccc tcagaagaag ttgtcaagaa
     4621 aatgaaaaat tattggcggc agctgctgaa cgccaaactg atcacacaac ggaagttcga
     4681 taatctgact aaggctgaac gaggtggcct gtctgagttg gataaagccg gcttcatcaa
     4741 aaggcagctt gttgagacac gccagatcac caagcacgtg gcccaaattc tcgattcacg
     4801 catgaacacc aagtacgatg aaaatgacaa actgattcga gaggtgaaag ttattactct
     4861 gaagtctaag ctcgtctcag atttcagaaa ggactttcag ttttataagg tgagagagat
     4921 caacaattac caccatgcgc atgatgccta cctgaatgca gtggtaggca ctgcacttat
     4981 caaaaaatat cccaagcttg aatctgaatt tgtttacgga gactataaag tgtacgatgt
     5041 taggaaaatg atcgcaaagt ctgagcagga aataggcaag gccaccgcta agtacttctt
     5101 ttacagcaat attatgaatt ttttcaagac cgagattaca ctggccaatg gagagattcg
     5161 gaagcgacca cttatcgaaa caaacggaga aacaggagaa atcgtgtggg acaagggtag
     5221 ggatttcgcg acagtccgga aggtcctgtc catgccgcag gtgaacatcg ttaaaaagac
     5281 cgaagtacag accggaggct tctccaagga aagtatcctc ccgaaaagga acagcgacaa
     5341 gctgatcgca cgcaaaaaag attgggaccc caagaaatac ggcggattcg attctcctac
     5401 agtcgcttac agtgtactgg ttgtggccaa agtggagaaa gggaagtcta aaaaactcaa
     5461 aagcgtcaag gaactgctgg gcatcacaat catggagcga tcaagcttcg aaaaaaaccc
     5521 catcgacttt ctcgaggcga aaggatataa agaggtcaaa aaagacctca tcattaagct
     5581 tcccaagtac tctctctttg agcttgaaaa cggccggaaa cgaatgctcg ctagtgcggg
     5641 cgagctgcag aaaggtaacg agctggcact gccctctaaa tacgttaatt tcttgtatct
     5701 ggccagccac tatgaaaagc tcaaagggtc ccccgaagat aatgagcaga agcagctgtt
     5761 cgtggaacaa cacaaacact accttgatga gatcatcgag caaataagcg aattctccaa
     5821 aagagtgatc ctcgccgacg ctaacctcga taaggtgctt tctgcttaca ataagcacag
     5881 ggataagccc atcagggagc aggcagaaaa cattatccac ttgtttactc tgaccaactt
     5941 gggcgcgcct gcagccttca agtacttcga caccaccata gacagaaagc ggtacacctc
     6001 tacaaaggag gtcctggacg ccacactgat tcatcagtca attacggggc tctatgaaac
     6061 aagaatcgac ctctctcagc tcggtggaga cagcagggct gaccccaaga agaagaggaa
     6121 ggtgtgagct tcgtccgacg gcggcccgac gggtccgagg cctcggagat ccgtccccct
     6181 tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc cctcccccca
     6241 catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt
     6301 tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc
     6361 tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg
     6421 gacgctcgaa ggctttaatt tgcaagctgg cgctaattca tggacatgga ggcccagaat
     6481 accctccttg acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac
     6541 atttagccca tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg
     6601 gcgcgaagca aaaattacgg ctcctcgctg cagacgtgcg agcagggaaa cgctcccctc
     6661 acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat
     6721 ttgccactga ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct
     6781 cacatcacat ccgaacataa acaaccatgg gtaaggaaaa gactcacgtt tcgaggccgc
     6841 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     6901 ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca gagttgtttc
     6961 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     7021 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     7081 catggttact caccactgcg atccccggca aaacagcatt ccaggtatta gaagaatatc
     7141 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     7201 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     7261 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     7321 ctgttgaaca agtctggaaa gaaatgcata agcttttgcc attctcaccg gattcagtcg
     7381 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     7441 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     7501 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     7561 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     7621 tactgacaat aaaaagattc ttgttttcaa gaacttgtca tttgtatagt ttttttatat
     7681 tgtagttgtt ctattttaat caaatgttag cgtgatttat attttttttc gcctcgacat
     7741 catctgccca gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca atcgtatgtg
     7801 aatgctggtc gctatactgg cttccttttc atcacgtgct ataaaaataa ttataattta
     7861 aattttttaa tataaatata taaattaaaa atagaaagta aaaaaagaaa ttaaagaaaa
     7921 aatagttttt gttttccgaa gatgtaaaag actctagggg gatcgccaac aaatactacc
     7981 ttttatcttg ctcttcctgc tctcaggtat taatgccgaa ttgtttcatc ttgtctgtgt
     8041 agaacaccac acacgaaaat cctgtgattt tacattttac ttatcgttaa tcgaatgtat
     8101 atctatttaa tctgcttttc ttgtctaata aatatatatg taaagtacgc tttttgttga
     8161 aattttttaa acctttgttt attttttttt cttcattccg taactcttct accttcttta
     8221 tttactttct aaaatccaaa tacaaaacat aaaaataaat aaacacagag taaattccca
     8281 aattattcca tcattaaaag atacgaggcg cgtgtaagtt acaggca
//
