LOCUS       sequence-327844-       14760 bp    DNA     circular SYN 15-APR-2025
DEFINITION  (also pMST621-BB3_gpyrG1_cas9) CRISPR/Cas9 plasmid with gRNA for 
            pyrG1, Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14760)
  AUTHORS   Sarkari P, Marx H, Blumhoff ML, Mattanovich D, Sauer M, Steiger MG
  TITLE     An efficient tool for metabolic pathway construction and gene 
            integration for Aspergillus niger.
  JOURNAL   Bioresour Technol. 2017 May 4. pii: S0960-8524(17)30643-0. doi: 
            10.1016/j.biortech.2017.05.004.
  PUBMED    28533066
REFERENCE   2  (bases 1 to 14760)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Bioresour 
            Technol. 2017 May 4. pii: S0960-8524(17)30643-0. doi: 
            10.1016/j.biortech.2017.05.004."
COMMENT     Sequence Label: pCAS_gpyrG1
FEATURES             Location/Qualifiers
     source          1..14760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1347..5450
                     /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             1347..5450
                     /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             1347..5450
                     /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             5463..5483
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5491..6264
                     /gene="trpC"
                     /label=trpC terminator
                     /note="transcription terminator from the Aspergillus 
                     nidulans trpC gene (Punt et al., 1987)"
     regulatory      6420..6429
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7013..8038
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             7013..8038
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     rep_origin      complement(11618..12206)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11698..11717)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     ncRNA           13799..13835
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     misc_RNA        13856..13931
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           13936..14003
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     terminator      join(14019..14760,1..32)
                     /gene="trpC"
                     /label=trpC terminator
                     /note="transcription terminator from the Aspergillus 
                     nidulans trpC gene (Punt et al., 1987)"
ORIGIN
        1 ctgtgtaagc gcccactcca catctccact cgctccgggg agaaccgaag aggatgggaa
       61 ccccgaccat ggcctgcggc cgcagctatc agtcctctca gctgacgccc ctcacgaacc
      121 ctcgcactct cccgaaatgc tcttcggtgt ccatggcgca tcatctcagc agcatcccat
      181 gtcaaaccat ggctttgggc ccacggactc tgtggccctg ccgcaacatc accatcacca
      241 tcgactcccg ccccatgcag cgctgcgcgc cccagggcgt catggagtga atgtggaatc
      301 tcctcctttg ccctcgggcc cgccgagtgt ggtgggccag cctggaatgc ctgatccagc
      361 ccccaggcct cgaggaccaa aactgaagtt tactcccgaa gaggacgctc tactggtgga
      421 gttgaaggaa aacaagaact tgacgtggaa gcaaattgca gacttcttcc cgggccgaac
      481 gagcggtacc ttgcaagtcc gatactgcac caagctgaag gctaaggatg tagcttggag
      541 tgacgaaatg gttcgatttg ctcctgatgt atttctacgc ctgtctcaca catgctaatg
      601 aagggaatag gtacaaaggc tgcagcgggc aatgcacgag tacgagaacg atcggtggcg
      661 catcattgca gggaaggttg gaaatggctt caccccagct gcttgccgcg agaaagccat
      721 gcagctccat gagtaaaagc gttggggaat tttcatattt atatctactg tcgccagatt
      781 cggccctgct tggaccctct gatctcctta ctctccatat tggttcaaat gtcgggtcac
      841 cgatagggct ggtggtgcag gcttgttgta ggcacgggag gatgatcagc ataactctga
      901 gtcactatag ggacgggttg atgtaaggta ttaagtgatg tatgataatt cattttagcc
      961 cgggggaaca tatggcgccg gcatttgttc gttcgcaatg aaccgacact agcgtccgct
     1021 ctcgcagttt agcaccggct gatcccgggc tgaacgcggc cattgctcgg ccggggcatg
     1081 tgttccttat ctacggcaga ccgcagatga ccactggagc agattataga ccctaagccc
     1141 taagccggac acccaatcga gtaggtctgc ggaccaggtc actgcgggca gccggagaag
     1201 ctccgcaacc aatcaatccc cggcgctgac taagggcagg cgaccacggg ccgaagcggc
     1261 ttcaaactca cctcaacctc caaactccct catctccaaa cgtccttgcc ttgtctgccg
     1321 tcattgcaac ccacccacca ggacacatgg acaagaagta ctccattggg ctcgatatcg
     1381 gcacaaacag cgtcggttgg gccgtcatta cggacgagta caaggtgccg agcaaaaaat
     1441 tcaaagttct gggcaatacc gatcgccaca gcataaagaa gaacctcatt ggcgccctcc
     1501 tgttcgactc cggggagacg gccgaagcca cgcggctcaa aagaacagca cggcgcagat
     1561 atacccgcag aaagaatcgg atctgctacc tgcaggagat ctttagtaat gagatggcta
     1621 aggtggatga ctctttcttc cataggctgg aggagtcctt tttggtggag gaggataaaa
     1681 agcacgagcg ccacccaatc tttggcaata tcgtggacga ggtggcgtac catgaaaagt
     1741 acccaaccat atatcatctg aggaagaagc ttgtagacag tactgataag gctgacttgc
     1801 ggttgatcta tctcgcgctg gcgcatatga tcaaatttcg gggacacttc ctcatcgagg
     1861 gggacctgaa cccagacaac agcgatgtcg acaaactctt tatccaactg gttcagactt
     1921 acaatcagct tttcgaagag aacccgatca acgcatccgg agttgacgcc aaagcaatcc
     1981 tgagcgctag gctgtccaaa tcccggcggc tcgaaaacct catcgcacag ctccctgggg
     2041 agaagaagaa cggcctgttt ggtaatctta tcgccctgtc actcgggctg acccccaact
     2101 ttaaatctaa cttcgacctg gccgaagatg ccaagcttca actgagcaaa gacacctacg
     2161 atgatgatct cgacaatctg ctggcccaga tcggcgacca gtacgcagac ctttttttgg
     2221 cggcaaagaa cctgtcagac gccattctgc tgagtgatat tctgcgagtg aacacggaga
     2281 tcaccaaagc tccgctgagc gctagtatga tcaagcgcta tgatgagcac caccaagact
     2341 tgactttgct gaaggccctt gtcagacagc aactgcctga gaagtacaag gaaattttct
     2401 tcgatcagtc taaaaatggc tacgccggat acattgacgg cggagcaagc caggaggaat
     2461 tttacaaatt tattaagccc atcttggaaa aaatggacgg caccgaggag ctgctggtaa
     2521 agcttaacag agaagatctg ttgcgcaaac agcgcacttt cgacaatgga agcatccccc
     2581 accagattca cctgggcgaa ctgcacgcta tcctcaggcg gcaagaggat ttctacccct
     2641 ttttgaaaga taacagggaa aagattgaga aaatcctcac atttcggata ccctactatg
     2701 taggccccct cgcccgggga aattccagat tcgcgtggat gactcgcaaa tcagaagaga
     2761 ctatcactcc ctggaacttc gaggaagtcg tggataaggg ggcctctgcc cagtccttca
     2821 tcgaaaggat gactaacttt gataaaaatc tgcctaacga aaaggtgctt cctaaacact
     2881 ctctgctgta cgagtacttc acagtttata acgagctcac caaggtcaaa tacgtcacag
     2941 aagggatgag aaagccagca ttcctgtctg gagagcagaa gaaagctatc gtggacctcc
     3001 tcttcaagac gaaccggaaa gttaccgtga aacagctcaa agaggactat ttcaaaaaga
     3061 ttgaatgttt cgactctgtt gaaatcagcg gagtggagga tcgcttcaac gcatccctgg
     3121 gaacgtatca cgatctcctg aaaatcatta aagacaagga cttcctggac aatgaggaga
     3181 acgaggacat tcttgaggac attgtcctca cccttacgtt gtttgaagat agggagatga
     3241 ttgaagaacg cttgaaaact tacgctcatc tcttcgacga caaagtcatg aaacagctca
     3301 agaggcgccg atatacagga tgggggcggc tgtcaagaaa actgatcaat gggatccgag
     3361 acaagcagag tggaaagaca atcctggatt ttcttaagtc cgatggattt gccaaccgga
     3421 acttcatgca gttgatccat gatgactctc tcacctttaa ggaggacatc cagaaagcac
     3481 aagtttctgg ccagggggac agccttcacg agcacatcgc taatcttgca ggtagcccag
     3541 ctatcaaaaa gggaatactg cagaccgtta aggtcgtgga tgaactcgtc aaagtaatgg
     3601 gaaggcataa gcccgagaat atcgttatcg agatggcccg agagaaccaa actacccaga
     3661 agggacagaa gaacagtagg gaaaggatga agaggattga agagggtata aaagaactgg
     3721 ggtcccaaat ccttaaggaa cacccagttg aaaacaccca gcttcagaat gagaagctct
     3781 acctgtacta cctgcagaac ggcagggaca tgtacgtgga tcaggaactg gacatcaatc
     3841 ggctctccga ctacgacgtg gatcatatcg tgccccagtc ttttctcaaa gatgattcta
     3901 ttgataataa agtgttgaca agatccgata aaaatagagg gaagagtgat aacgtcccct
     3961 cagaagaagt tgtcaagaaa atgaaaaatt attggcggca gctgctgaac gccaaactga
     4021 tcacacaacg gaagttcgat aatctgacta aggctgaacg aggtggcctg tctgagttgg
     4081 ataaagccgg cttcatcaaa aggcagcttg ttgagacacg ccagatcacc aagcacgtgg
     4141 cccaaattct cgattcacgc atgaacacca agtacgatga aaatgacaaa ctgattcgag
     4201 aggtgaaagt tattactctg aagtctaagc tcgtctcaga tttcagaaag gactttcagt
     4261 tttataaggt gagagagatc aacaattacc accatgcgca tgatgcctac ctgaatgcag
     4321 tggtaggcac tgcacttatc aaaaaatatc ccaagcttga atctgaattt gtttacggag
     4381 actataaagt gtacgatgtt aggaaaatga tcgcaaagtc tgagcaggaa ataggcaagg
     4441 ccaccgctaa gtacttcttt tacagcaata ttatgaattt tttcaagacc gagattacac
     4501 tggccaatgg agagattcgg aagcgaccac ttatcgaaac aaacggagaa acaggagaaa
     4561 tcgtgtggga caagggtagg gatttcgcga cagtccggaa ggtcctgtcc atgccgcagg
     4621 tgaacatcgt taaaaagacc gaagtacaga ccggaggctt ctccaaggaa agtatcctcc
     4681 cgaaaaggaa cagcgacaag ctgatcgcac gcaaaaaaga ttgggacccc aagaaatacg
     4741 gcggattcga ttctcctaca gtcgcttaca gtgtactggt tgtggccaaa gtggagaaag
     4801 ggaagtctaa aaaactcaaa agcgtcaagg aactgctggg catcacaatc atggagcgat
     4861 caagcttcga aaaaaacccc atcgactttc tcgaggcgaa aggatataaa gaggtcaaaa
     4921 aagacctcat cattaagctt cccaagtact ctctctttga gcttgaaaac ggccggaaac
     4981 gaatgctcgc tagtgcgggc gagctgcaga aaggtaacga gctggcactg ccctctaaat
     5041 acgttaattt cttgtatctg gccagccact atgaaaagct caaagggtcc cccgaagata
     5101 atgagcagaa gcagctgttc gtggaacaac acaaacacta ccttgatgag atcatcgagc
     5161 aaataagcga attctccaaa agagtgatcc tcgccgacgc taacctcgat aaggtgcttt
     5221 ctgcttacaa taagcacagg gataagccca tcagggagca ggcagaaaac attatccact
     5281 tgtttactct gaccaacttg ggcgcgcctg cagccttcaa gtacttcgac accaccatag
     5341 acagaaagcg gtacacctct acaaaggagg tcctggacgc cacactgatt catcagtcaa
     5401 ttacggggct ctatgaaaca agaatcgacc tctctcagct cggtggagac agcagggctg
     5461 accccaagaa gaagaggaag gtgtgagctt gatccactta acgttactga aatcatcaaa
     5521 cagcttgacg aatctggata taagatcgtt ggtgtcgatg tcagctccgg agttgagaca
     5581 aatggtgttc aggatctcga taagatacgt tcatttgtcc aagcagcaaa gagtgccttc
     5641 tagtgattta atagctccat gtcaacaaga ataaaacgcg tttcgggttt acctcttcca
     5701 gatacagctc atctgcaatg cattaatgca ttggacctcg caaccctagt acgcccttca
     5761 ggctccggcg aagcagaaga atagcttagc agagtctatt ttcattttcg ggagacgaga
     5821 tcaagcagat caacggtcgt caagagtcct acgagactga ggaatccgct cttggctcca
     5881 cgcgactata tatttgtctc taattgtact ttgacatgct cctcttcttt actctgatag
     5941 cttgactatg aaaattccgt caccagcccc tgggttcgca aagataattg cactgtttct
     6001 tccttgaact ctcaagccta caggacacac attcatcgta ggtataaacc tcgaaaatca
     6061 ttcctactaa gatgggtata caatagtaac catgcatggt tgcctagtga atgctccgta
     6121 acacccaata cgccggccga aactttttta caactctcct atgagtcgtt tacccagaat
     6181 gcacaggtac acttgtttag aggtaatcct tctttctaga agtcctcgtg tactgtgtaa
     6241 gcgcccactc cacatctcca ctcgctaatt catgaataac ggtgagacta gcggccggtc
     6301 cccttatccc agctgttcca cgttggcctg cccctcagtt agcgctcaac tcaatgcccc
     6361 tcactggcga ggcgagggca aggatggagg ggcagcatcg cctgagttgg agcaaagcgg
     6421 ccgccatggg agcagcgaac caacggaggg atgccgtgct ttgtcgtggc tgctgtggcc
     6481 aatccgggcc cttggttggc tcacagagcg ttgctgtgag tccatgagct attattgcta
     6541 ggtacagtat agagagagga gagagagaga gagagagaga ggggaaaaaa ggtgaggttg
     6601 aagtgagaaa aaaaaaaaaa aaaatccaac cactgacggc tgccggctct gccacccccc
     6661 tccctccacc ccagaccacc tgcacactca gcgcgcagca tcacctaatc ttggctcgcc
     6721 ttcccgcagc tcaggttgtt ttttttttct ctctccctcg tcgaagccgc ccttgttccc
     6781 ttatttattt ccctctccat ccttgtctgc ctttggtcca tctgcccctt tgtctgcatc
     6841 tcttttgcac gcatcgcctt atcgtcgtct cttttttcac tcacgggagc ttgacgatga
     6901 cctgactcgt gagcctcacc tgctgatttc tctccccccc tcccgaccgg cttgactttt
     6961 gtttctcctc cagtacctta tcgcgaagcc ggaagaacct cttaacctct agatgaaaaa
     7021 gcctgaactc accgcgacgt ctgtcgagaa gttcctgatc gaaaagttcg acagcgtctc
     7081 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     7141 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     7201 ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt
     7261 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     7321 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc
     7381 tgcggccgat ctcagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     7441 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     7501 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     7561 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     7621 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     7681 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     7741 gcagcagacg cgctacttcg agcggaggca cccggagctt gcaggatcgc cgcggctccg
     7801 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     7861 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     7921 tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga
     7981 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagat
     8041 gcatggcttt cgtgaccggg cttcaaacaa tgatgtgcga tggtgtggtt cccggttggc
     8101 ggagtctttg tctactttgg ttgtctgtcg caggtcggta gaccgcaaat gagcaactga
     8161 tggattgttg ccagcgatac tataattcac atggatggtc tttgtcgatc agtagctagt
     8221 gagagagaga gaacatctat ccacaatgtc gagtgtctat tagacatact ccgagaataa
     8281 agtcaactgt gtctgtgatc taaagatcga ttcggcagtc gagtagcgta taacaactcc
     8341 gagtaccagc gaaagcacgt cgtgacagga gcagggcttt gccaactgcg caaccttgct
     8401 tgaatgagga tacacggggt gcaacatggc tgtactgatc catcgcaacc aaaatttctg
     8461 tttatagatc aagctggtag attccaatta ctccacctct tgcgcttctc catgacatgt
     8521 aagtgcacgt ggaaaccata cccaaattgc ctacagctgc ggagcatgag cctatggcga
     8581 tcagtctggt catgttaacc agcctgtgct ctgacgttaa tgcagaatag cttatgctgc
     8641 agacgcgtta cggcggagat gctgcagacg cgttacgcgg agatgctgca gacgcgttac
     8701 gtatcggatc cagaattcgt gatatctgaa ttcgtcgaca agcttatttt ttgtatactg
     8761 ttttgtgata gcacgaagtt tttccacggt atcttgttaa aaatatatat ttgtggcggg
     8821 cttacctaca tcaaattaat aagagactaa ttataaacta aacacacaag caagctactt
     8881 tagggtaaaa gtttataaat gcttttgacg tataaacgtt gcttgtattt attattacaa
     8941 ttaaaggtgg atagaaaacc tagagactag ttagaaacta atctcaggtt tgcgttaaac
     9001 taaatcagag cccgagaggt taacagaacc tagaagggga ctagatatcc gggtagggaa
     9061 acaaaaaaaa aaaacaagac agccacatat tagggagact agttagaagc tagttccagg
     9121 actaggaaaa taaaagacaa tgataccaca gtctagttga caactagata gattctagat
     9181 tgaggccaaa gtctctgaga tccaggttag ttgcaactaa tactagttag tatctagtct
     9241 cctataactc tgaagctaga ataacttact actattatcc tcaccactgt tcagctgcgc
     9301 aaacggagtg attgcaaggt gttcagagac tagttattga ctagtcagtg actagcaata
     9361 actaacaagg tattaaccta ccatgtctgc catcaccctg cacttcctcg ggctcagcag
     9421 ccttttcctc ctcattttca tgctcatttt ccttgtttaa gactgtgact agtcaaagac
     9481 tagtccagaa ccacaaagga gaaatgtctt accactttct tcattgcttg tctcttttgc
     9541 attatccatg tctgcaacta gttagagtct agttagtgac tagtccgacg aggacttgct
     9601 tgtctccgga ttgttggagg aactctccag ggcctcaaga tccacaacag agccttctag
     9661 atgactggtc aataactagt tggtctttgt ctgagtctga cttacgaggt tgcatactcg
     9721 ctccctttgc ctcgtcaatc gatgagaaaa agcgccaaaa ctcgcaatat ggctttgaac
     9781 cacacggtgc tgagactagt tagaatctag tcccaaacta gcttggatag cttacctttg
     9841 ccctttgcgt tgcgacaggt cttgcagggt atggttcctt tctcaccagc tgatttagct
     9901 gccttgctac cctcacggcg gatctgcata aagagtggct agaggttata aattagcact
     9961 gatcctaggt acggggctga atgtaacttg cctttccttt ctcatcgcgc ggcaagacag
    10021 gcttgctcaa attcctacca gtcacagggg tatgcacggc gtacggacca cttgaactag
    10081 tcacagatta gttagcaact agtctgcatt gaatggctgt acttacgggc cctcgccatt
    10141 gtcctgatca tttccagctt caccctcgtt gctgcaaagt agttagtgac tagtcaagga
    10201 ctagttgaaa tgggagaaga aactcacgaa ttctcgacac ccttagtatt gtggtccttg
    10261 gacttggtgc tgctatatat tagctaatac actagttaga ctcacagaaa cttacgcagc
    10321 tcgcttgcgc ttcttggtag gagtcggggt tgggagaaca gtgccttcaa acaagccttc
    10381 ataccatgct acttgactag tcagggacta gtcaccaagt aatctagata ggacttgcct
    10441 ttggcctcca tcagttcctt catagtggga ggtccattgt gcaatgtaaa ctccatgccg
    10501 tgggagttct tgtccttcaa gtgcttgacc aatatgtttc tgttggcaga gggaacctgt
    10561 caactagtta ataactagtc agaaactagt atagcagtag actcactgta cgcttgaggc
    10621 atcccttcac tcggcagtag acttcatatg gatggatatc aggcacgcca ttgtcgtcct
    10681 gtggactagt cagtaactag gcttaaagct agtcgggtcg gcttactatc ttgaaatccg
    10741 gcagcgtaag ctccccgtcc ttaactgcct cgagatagtg acagtactct ggggactttc
    10801 ggagatcgtt atcgcgaatg ctcggcatac taatcgttga ctagtcttgg actagtcccg
    10861 agcaaaaagg attggaggag gaggaggaag gtgagagtga gacaaagagc gaaataagag
    10921 cttcaaaggc tatctctaag cagtatgaag gttaagtatc tagttcttga ctagatttaa
    10981 aagagatttc gactagttat gtacctggag tttggatata ggaatgtgtt gtggtaacga
    11041 aatgtaaggg ggaggaaaga aaaagtcggt caagaggtaa ctctaagtcg gccattcctt
    11101 tttgggaggc gctaaccata aacggcatgg tcgacttaga gttagctcag ggaatttagg
    11161 gagttatctg cgaccaccga ggaacggcgg aatgccaaag aatcccgatg gagctctagc
    11221 tggcggttga caaccccacc ttttggcgtt tctgcggcgt tgcaggcggg actggatact
    11281 tcgtagaacc agaaaggcaa ggcagaacgc gctcagcaag agtgttggaa gtgatagcat
    11341 gatgtgcctt gttaactagg tcaaaatctg cagtatgctt gatgttatcc aaagtgtgag
    11401 agaggaaggt ccaaacatac acgattggga gagggcctag gtataagagt ttttgagtag
    11461 aacgcatgtg agcccagcca tctcgaggag attaaacacg ggccggcatt tgatggctat
    11521 gttagtaccc caatggaaag cctgagagtc cagtggcgct gtgagcaaaa ggccagcaaa
    11581 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
    11641 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
    11701 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
    11761 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
    11821 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
    11881 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
    11941 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
    12001 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
    12061 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
    12121 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
    12181 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtggagcg
    12241 caaaaaaccc cgcccctgac agggcggggt tttttcgcga tcggaggtac agtggccatg
    12301 aaatccaatc atttccttct ggccgccctc gggcaagaga tagtgccgca gaggactctc
    12361 acagcatcta catctgcgac cgcaacagcc accaagcgag gcgcacatga gcttgtcctc
    12421 ctcccatgcc aaagtttggc cctcttcgtt tctgtgatgc tgaaggaagt caaactcgtc
    12481 gatgatagga ccagatggtt tgtcaagggt caacgctttc catgccttct ggcaccggta
    12541 gtaatgctct tctgcaaggg agacttgacg tttcggatcc gcgggccccg ggacatgctg
    12601 gaagggattt tctggctcaa taccacgtct gtatttgacc ctttccagac agttaatccg
    12661 ctgcaggagg gcgaactgta gctcctcgtt ctccttgtag cgcttgatcc agtctttttg
    12721 gatgttgcac ttgcttggcc tatgcttctc atataatctt gccctgtcat agagacgacg
    12781 tctgagattg tagcgttcgt ctttgatcac ccggagccag ataggcctga gtatatctga
    12841 cattagatca aagggtctgt ggatagtctc cttcagcatc agcgacgcat gtgactcgca
    12901 tgtcggagag agcttgtggg tggtcatctt tgatggcgtc ctctgctttc ccttgatttt
    12961 cgttgattgt ttttcgaaag ttaagtctgg aagtcaagag aatccttctg ccagacatta
    13021 tatttacgta tactgacgta gtagaaacag cgtcaggatg aggacatggt gtgtgctgga
    13081 ccacggaatc atagttcatc agtatattgg gttggacaaa taacgctgag catgtatatg
    13141 tctttacaca ctataaaagc cagcgaacgc caataaaata gggcatattg atgtgaaaat
    13201 atgacaccag ttaaaagcag tgtattgatt ttatctctct tcacctcgga cctatactac
    13261 cgtatacaag actcaactta cttccagata tagtaatata caccctatgg acgaaccagc
    13321 acaataatta cagccaaaca acaccaccca aatggcatat tcctaatcag cactaagcac
    13381 aaataccact gtcatcacag cataatcaat aagaatccca gacaaccgac tcactctgac
    13441 tcaccttaca caaaccccca agcaaagcgc agcccagaac ctcagccaac aatcgggcaa
    13501 cgtacgggga aagattggcc gatccatgat gtcagcagcc ctaacccaaa gcggactagc
    13561 gcataccgcc cctctgactc cgccatccca gggctcgaga agcttccgtg gcgtcgatat
    13621 aaattcagcg ggccttgaac atccctcctt acgacacacc tcacgcgatc gattttgaca
    13681 ctcacacacc gccaccctca catcctccac ccacaccaca ccccttaatc aacccaccat
    13741 caccgctaga acgtctatct catcaccgac ttctcatcca tcttcaaaca tgtgggatct
    13801 gatgagtccg tgaggacgaa acgagtaagc tcgtcatccc aatgctctgg cgaaggtttt
    13861 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
    13921 cgagtcggtg cttttggccg gcatggtccc agcctcctcg ctggcgccgg ctgggcaaca
    13981 tgcttcggca tggcgaatgg gacagctttg gactgcttga tccacttaac gttactgaaa
    14041 tcatcaaaca gcttgacgaa tctggatata agatcgttgg tgtcgatgtc agctccggag
    14101 ttgagacaaa tggtgttcag gatctcgata agatacgttc atttgtccaa gcagcaaaga
    14161 gtgccttcta gtgatttaat agctccatgt caacaagaat aaaacgcgtt tcgggtttac
    14221 ctcttccaga tacagctcat ctgcaatgca ttaatgcatt ggacctcgca accctagtac
    14281 gcccttcagg ctccggcgaa gcagaagaat agcttagcag agtctatttt cattttcggg
    14341 agacgagatc aagcagatca acggtcgtca agagtcctac gagactgagg aatccgctct
    14401 tggctccacg cgactatata tttgtctcta attgtacttt gacatgctcc tcttctttac
    14461 tctgatagct tgactatgaa aattccgtca ccagcccctg ggttcgcaaa gataattgca
    14521 ctgtttcttc cttgaactct caagcctaca ggacacacat tcatcgtagg tataaacctc
    14581 gaaaatcatt cctactaaga tgggtataca atagtaacca tgcatggttg cctagtgaat
    14641 gctccgtaac acccaatacg ccggccgaaa cttttttaca actctcctat gagtcgttta
    14701 cccagaatgc acaggtacac ttgtttagag gtaatccttc tttctagaag tcctcgtgta
//
