LOCUS       sequence-206071-        8516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hCas9 under control of Tef1 for direct cloning of HH-sgRNA-HDV PCR 
            products and episomal expression in P. pastoris and G418 selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8516)
  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 8516)
  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*_pTEF_Cas9
FEATURES             Location/Qualifiers
     source          1..8516
                     /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"
     promoter        1763..2169
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             2177..6280
                     /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             2177..6280
                     /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             2177..6280
                     /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             6293..6313
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      6385..6632
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6401..6419)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     gene            6652..8008
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        6652..6995
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             6996..7805
                     /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(7063..7082)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      7811..8008
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_feature    join(8020..8516,1..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 agcacacacc atagcttcaa aatgtttcta ctcctttttt
     1801 actcttccag attttctcgg actccgcgca tcgccgtacc acttcaaaac acccaagcac
     1861 agcatactaa atttcccctc tttcttcctc tagggtgtcg ttaattaccc gtactaaagg
     1921 tttggaaaag aaaaaagcga ccgcctcgtt tctttttctt cgtcgaaaaa ggcaataaaa
     1981 atttttatca cgtttctttt tcttgaaaat tttttttttt gatttttttc tctttcgatg
     2041 acctcccatt gatatttaag ttaataaacg gtgttcaatt tctcaagttt cagtttcatt
     2101 tttcttgttc tattacaact ttttttactt cttgctcatt agaaagaaag catagcaatc
     2161 taatctaagg gcggtcatgg acaagaagta ctccattggg ctcgatatcg gcacaaacag
     2221 cgtcggttgg gccgtcatta cggacgagta caaggtgccg agcaaaaaat tcaaagttct
     2281 gggcaatacc gatcgccaca gcataaagaa gaacctcatt ggcgccctcc tgttcgactc
     2341 cggggagacg gccgaagcca cgcggctcaa aagaacagca cggcgcagat atacccgcag
     2401 aaagaatcgg atctgctacc tgcaggagat ctttagtaat gagatggcta aggtggatga
     2461 ctctttcttc cataggctgg aggagtcctt tttggtggag gaggataaaa agcacgagcg
     2521 ccacccaatc tttggcaata tcgtggacga ggtggcgtac catgaaaagt acccaaccat
     2581 atatcatctg aggaagaagc ttgtagacag tactgataag gctgacttgc ggttgatcta
     2641 tctcgcgctg gcgcatatga tcaaatttcg gggacacttc ctcatcgagg gggacctgaa
     2701 cccagacaac agcgatgtcg acaaactctt tatccaactg gttcagactt acaatcagct
     2761 tttcgaagag aacccgatca acgcatccgg agttgacgcc aaagcaatcc tgagcgctag
     2821 gctgtccaaa tcccggcggc tcgaaaacct catcgcacag ctccctgggg agaagaagaa
     2881 cggcctgttt ggtaatctta tcgccctgtc actcgggctg acccccaact ttaaatctaa
     2941 cttcgacctg gccgaagatg ccaagcttca actgagcaaa gacacctacg atgatgatct
     3001 cgacaatctg ctggcccaga tcggcgacca gtacgcagac ctttttttgg cggcaaagaa
     3061 cctgtcagac gccattctgc tgagtgatat tctgcgagtg aacacggaga tcaccaaagc
     3121 tccgctgagc gctagtatga tcaagcgcta tgatgagcac caccaagact tgactttgct
     3181 gaaggccctt gtcagacagc aactgcctga gaagtacaag gaaattttct tcgatcagtc
     3241 taaaaatggc tacgccggat acattgacgg cggagcaagc caggaggaat tttacaaatt
     3301 tattaagccc atcttggaaa aaatggacgg caccgaggag ctgctggtaa agcttaacag
     3361 agaagatctg ttgcgcaaac agcgcacttt cgacaatgga agcatccccc accagattca
     3421 cctgggcgaa ctgcacgcta tcctcaggcg gcaagaggat ttctacccct ttttgaaaga
     3481 taacagggaa aagattgaga aaatcctcac atttcggata ccctactatg taggccccct
     3541 cgcccgggga aattccagat tcgcgtggat gactcgcaaa tcagaagaga ctatcactcc
     3601 ctggaacttc gaggaagtcg tggataaggg ggcctctgcc cagtccttca tcgaaaggat
     3661 gactaacttt gataaaaatc tgcctaacga aaaggtgctt cctaaacact ctctgctgta
     3721 cgagtacttc acagtttata acgagctcac caaggtcaaa tacgtcacag aagggatgag
     3781 aaagccagca ttcctgtctg gagagcagaa gaaagctatc gtggacctcc tcttcaagac
     3841 gaaccggaaa gttaccgtga aacagctcaa agaggactat ttcaaaaaga ttgaatgttt
     3901 cgactctgtt gaaatcagcg gagtggagga tcgcttcaac gcatccctgg gaacgtatca
     3961 cgatctcctg aaaatcatta aagacaagga cttcctggac aatgaggaga acgaggacat
     4021 tcttgaggac attgtcctca cccttacgtt gtttgaagat agggagatga ttgaagaacg
     4081 cttgaaaact tacgctcatc tcttcgacga caaagtcatg aaacagctca agaggcgccg
     4141 atatacagga tgggggcggc tgtcaagaaa actgatcaat gggatccgag acaagcagag
     4201 tggaaagaca atcctggatt ttcttaagtc cgatggattt gccaaccgga acttcatgca
     4261 gttgatccat gatgactctc tcacctttaa ggaggacatc cagaaagcac aagtttctgg
     4321 ccagggggac agccttcacg agcacatcgc taatcttgca ggtagcccag ctatcaaaaa
     4381 gggaatactg cagaccgtta aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa
     4441 gcccgagaat atcgttatcg agatggcccg agagaaccaa actacccaga agggacagaa
     4501 gaacagtagg gaaaggatga agaggattga agagggtata aaagaactgg ggtcccaaat
     4561 ccttaaggaa cacccagttg aaaacaccca gcttcagaat gagaagctct acctgtacta
     4621 cctgcagaac ggcagggaca tgtacgtgga tcaggaactg gacatcaatc ggctctccga
     4681 ctacgacgtg gatcatatcg tgccccagtc ttttctcaaa gatgattcta ttgataataa
     4741 agtgttgaca agatccgata aaaatagagg gaagagtgat aacgtcccct cagaagaagt
     4801 tgtcaagaaa atgaaaaatt attggcggca gctgctgaac gccaaactga tcacacaacg
     4861 gaagttcgat aatctgacta aggctgaacg aggtggcctg tctgagttgg ataaagccgg
     4921 cttcatcaaa aggcagcttg ttgagacacg ccagatcacc aagcacgtgg cccaaattct
     4981 cgattcacgc atgaacacca agtacgatga aaatgacaaa ctgattcgag aggtgaaagt
     5041 tattactctg aagtctaagc tcgtctcaga tttcagaaag gactttcagt tttataaggt
     5101 gagagagatc aacaattacc accatgcgca tgatgcctac ctgaatgcag tggtaggcac
     5161 tgcacttatc aaaaaatatc ccaagcttga atctgaattt gtttacggag actataaagt
     5221 gtacgatgtt aggaaaatga tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa
     5281 gtacttcttt tacagcaata ttatgaattt tttcaagacc gagattacac tggccaatgg
     5341 agagattcgg aagcgaccac ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga
     5401 caagggtagg gatttcgcga cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt
     5461 taaaaagacc gaagtacaga ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa
     5521 cagcgacaag ctgatcgcac gcaaaaaaga ttgggacccc aagaaatacg gcggattcga
     5581 ttctcctaca gtcgcttaca gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa
     5641 aaaactcaaa agcgtcaagg aactgctggg catcacaatc atggagcgat caagcttcga
     5701 aaaaaacccc atcgactttc tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat
     5761 cattaagctt cccaagtact ctctctttga gcttgaaaac ggccggaaac gaatgctcgc
     5821 tagtgcgggc gagctgcaga aaggtaacga gctggcactg ccctctaaat acgttaattt
     5881 cttgtatctg gccagccact atgaaaagct caaagggtcc cccgaagata atgagcagaa
     5941 gcagctgttc gtggaacaac acaaacacta ccttgatgag atcatcgagc aaataagcga
     6001 attctccaaa agagtgatcc tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa
     6061 taagcacagg gataagccca tcagggagca ggcagaaaac attatccact tgtttactct
     6121 gaccaacttg ggcgcgcctg cagccttcaa gtacttcgac accaccatag acagaaagcg
     6181 gtacacctct acaaaggagg tcctggacgc cacactgatt catcagtcaa ttacggggct
     6241 ctatgaaaca agaatcgacc tctctcagct cggtggagac agcagggctg accccaagaa
     6301 gaagaggaag gtgtgagctt cgtccgacgg cggcccgacg ggtccgaggc ctcggagatc
     6361 cgtccccctt ttcctttgtc gatatcatgt aattagttat gtcacgctta cattcacgcc
     6421 ctccccccac atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc
     6481 ctatttattt ttttatagtt atgttagtat taagaacgtt atttatattt caaatttttc
     6541 ttttttttct gtacagacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga
     6601 aggttttggg acgctcgaag gctttaattt gcaagctggc gctaattcat ggacatggag
     6661 gcccagaata ccctccttga cagtcttgac gtgcgcagct caggggcatg atgtgactgt
     6721 cgcccgtaca tttagcccat acatccccat gtataatcat ttgcatccat acattttgat
     6781 ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc agacgtgcga gcagggaaac
     6841 gctcccctca cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag aaatataaaa
     6901 ggttaggatt tgccactgag gttcttcttt catatacttc cttttaaaat cttgctagga
     6961 tacagttctc acatcacatc cgaacataaa caaccatggg taaggaaaag actcacgttt
     7021 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
     7081 ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag
     7141 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
     7201 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
     7261 ctgatgatgc atggttactc accactgcga tccccggcaa aacagcattc caggtattag
     7321 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
     7381 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc
     7441 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
     7501 atggctggcc tgttgaacaa gtctggaaag aaatgcataa gcttttgcca ttctcaccgg
     7561 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
     7621 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     7681 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     7741 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     7801 tctaatcagt actgacaata aaaagattct tgttttcaag aacttgtcat ttgtatagtt
     7861 tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata ttttttttcg
     7921 cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa
     7981 tcgtatgtga atgctggtcg ctatactggc ttccttttca tcacgtgcta taaaaataat
     8041 tataatttaa attttttaat ataaatatat aaattaaaaa tagaaagtaa aaaaagaaat
     8101 taaagaaaaa atagtttttg ttttccgaag atgtaaaaga ctctaggggg atcgccaaca
     8161 aatactacct tttatcttgc tcttcctgct ctcaggtatt aatgccgaat tgtttcatct
     8221 tgtctgtgta gaacaccaca cacgaaaatc ctgtgatttt acattttact tatcgttaat
     8281 cgaatgtata tctatttaat ctgcttttct tgtctaataa atatatatgt aaagtacgct
     8341 ttttgttgaa attttttaaa cctttgttta tttttttttc ttcattccgt aactcttcta
     8401 ccttctttat ttactttcta aaatccaaat acaaaacata aaaataaata aacacagagt
     8461 aaattcccaa attattccat cattaaaaga tacgaggcgc gtgtaagtta caggca
//
