LOCUS       sequence-181691-       14566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BB3_L 23_gRNAempty_cas9_BbsI; CRISPR plasmid with empty gRNA spacer 
            to incorporate gRNA, Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14566)
  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 14566)
  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: "Bioresour 
            Technol. 2017 May 4. pii: S0960-8524(17)30643-0. doi: 
            10.1016/j.biortech.2017.05.004."
COMMENT     Sequence Label: pMST665_BB3_L 23_gRNAempty_cas9_BbsI
FEATURES             Location/Qualifiers
     source          1..14566
                     /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"
     terminator      2269..3042
                     /gene="trpC"
                     /label=trpC terminator
                     /note="transcription terminator from the Aspergillus 
                     nidulans trpC gene (Punt et al., 1987)"
     CDS             4357..8460
                     /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             4357..8460
                     /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             4357..8460
                     /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             8473..8493
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      8501..9274
                     /gene="trpC"
                     /label=trpC terminator
                     /note="transcription terminator from the Aspergillus 
                     nidulans trpC gene (Punt et al., 1987)"
     regulatory      9430..9439
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             10023..11048
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             10023..11048
                     /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"
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 gtacagtggc catgaaatcc aatcatttcc ttctggccgc cctcgggcaa
      781 gagatagtgc cgcagaggac tctcacagca tctacatctg cgaccgcaac agccaccaag
      841 cgaggcgcac atgagcttgt cctcctccca tgccaaagtt tggccctctt cgtttctgtg
      901 atgctgaagg aagtcaaact cgtcgatgat aggaccagat ggtttgtcaa gggtcaacgc
      961 tttccatgcc ttctggcacc ggtagtaatg ctcttctgca agggagactt gacgtttcgg
     1021 atccgcgggc cccgggacat gctggaaggg attttctggc tcaataccac gtctgtattt
     1081 gaccctttcc agacagttaa tccgctgcag gagggcgaac tgtagctcct cgttctcctt
     1141 gtagcgcttg atccagtctt tttggatgtt gcacttgctt ggcctatgct tctcatataa
     1201 tcttgccctg tcatagagac gacgtctgag attgtagcgt tcgtctttga tcacccggag
     1261 ccagataggc ctgagtatat ctgacattag atcaaagggt ctgtggatag tctccttcag
     1321 catcagcgac gcatgtgact cgcatgtcgg agagagcttg tgggtggtca tctttgatgg
     1381 cgtcctctgc tttcccttga ttttcgttga ttgtttttcg aaagttaagt ctggaagtca
     1441 agagaatcct tctgccagac attatattta cgtatactga cgtagtagaa acagcgtcag
     1501 gatgaggaca tggtgtgtgc tggaccacgg aatcatagtt catcagtata ttgggttgga
     1561 caaataacgc tgagcatgta tatgtcttta cacactataa aagccagcga acgccaataa
     1621 aatagggcat attgatgtga aaatatgaca ccagttaaaa gcagtgtatt gattttatct
     1681 ctcttcacct cggacctata ctaccgtata caagactcaa cttacttcca gatatagtaa
     1741 tatacaccct atggacgaac cagcacaata attacagcca aacaacacca cccaaatggc
     1801 atattcctaa tcagcactaa gcacaaatac cactgtcatc acagcataat caataagaat
     1861 cccagacaac cgactcactc tgactcacct tacacaaacc cccaagcaaa gcgcagccca
     1921 gaacctcagc caacaatcgg gcaacgtacg gggaaagatt ggccgatcca tgatgtcagc
     1981 agccctaacc caaagcggac tagcgcatac cgcccctctg actccgccat cccagggctc
     2041 gagaagcttc cgtggcgtcg atataaattc agcgggcctt gaacatccct ccttacgaca
     2101 cacctcacgc gatcgatttt gacactcaca caccgccacc ctcacatcct ccacccacac
     2161 cacacccctt aatcaaccca ccatcaccgc tagaacgtct atctcatcac cgacttctca
     2221 tccatcttca aacatggcgt cttcggatcc ctcgaggaag acgcgcttga tccacttaac
     2281 gttactgaaa tcatcaaaca gcttgacgaa tctggatata agatcgttgg tgtcgatgtc
     2341 agctccggag ttgagacaaa tggtgttcag gatctcgata agatacgttc atttgtccaa
     2401 gcagcaaaga gtgccttcta gtgatttaat agctccatgt caacaagaat aaaacgcgtt
     2461 tcgggtttac ctcttccaga tacagctcat ctgcaatgca ttaatgcatt ggacctcgca
     2521 accctagtac gcccttcagg ctccggcgaa gcagaagaat agcttagcag agtctatttt
     2581 cattttcggg agacgagatc aagcagatca acggtcgtca agagtcctac gagactgagg
     2641 aatccgctct tggctccacg cgactatata tttgtctcta attgtacttt gacatgctcc
     2701 tcttctttac tctgatagct tgactatgaa aattccgtca ccagcccctg ggttcgcaaa
     2761 gataattgca ctgtttcttc cttgaactct caagcctaca ggacacacat tcatcgtagg
     2821 tataaacctc gaaaatcatt cctactaaga tgggtataca atagtaacca tgcatggttg
     2881 cctagtgaat gctccgtaac acccaatacg ccggccgaaa cttttttaca actctcctat
     2941 gagtcgttta cccagaatgc acaggtacac ttgtttagag gtaatccttc tttctagaag
     3001 tcctcgtgta ctgtgtaagc gcccactcca catctccact cgctccgggg agaaccgaag
     3061 aggatgggaa ccccgaccat ggcctgcggc cgcagctatc agtcctctca gctgacgccc
     3121 ctcacgaacc ctcgcactct cccgaaatgc tcttcggtgt ccatggcgca tcatctcagc
     3181 agcatcccat gtcaaaccat ggctttgggc ccacggactc tgtggccctg ccgcaacatc
     3241 accatcacca tcgactcccg ccccatgcag cgctgcgcgc cccagggcgt catggagtga
     3301 atgtggaatc tcctcctttg ccctcgggcc cgccgagtgt ggtgggccag cctggaatgc
     3361 ctgatccagc ccccaggcct cgaggaccaa aactgaagtt tactcccgaa gaggacgctc
     3421 tactggtgga gttgaaggaa aacaagaact tgacgtggaa gcaaattgca gacttcttcc
     3481 cgggccgaac gagcggtacc ttgcaagtcc gatactgcac caagctgaag gctaaggatg
     3541 tagcttggag tgacgaaatg gttcgatttg ctcctgatgt atttctacgc ctgtctcaca
     3601 catgctaatg aagggaatag gtacaaaggc tgcagcgggc aatgcacgag tacgagaacg
     3661 atcggtggcg catcattgca gggaaggttg gaaatggctt caccccagct gcttgccgcg
     3721 agaaagccat gcagctccat gagtaaaagc gttggggaat tttcatattt atatctactg
     3781 tcgccagatt cggccctgct tggaccctct gatctcctta ctctccatat tggttcaaat
     3841 gtcgggtcac cgatagggct ggtggtgcag gcttgttgta ggcacgggag gatgatcagc
     3901 ataactctga gtcactatag ggacgggttg atgtaaggta ttaagtgatg tatgataatt
     3961 cattttagcc cgggggaaca tatggcgccg gcatttgttc gttcgcaatg aaccgacact
     4021 agcgtccgct ctcgcagttt agcaccggct gatcccgggc tgaacgcggc cattgctcgg
     4081 ccggggcatg tgttccttat ctacggcaga ccgcagatga ccactggagc agattataga
     4141 ccctaagccc taagccggac acccaatcga gtaggtctgc ggaccaggtc actgcgggca
     4201 gccggagaag ctccgcaacc aatcaatccc cggcgctgac taagggcagg cgaccacggg
     4261 ccgaagcggc ttcaaactca cctcaacctc caaactccct catctccaaa cgtccttgcc
     4321 ttgtctgccg tcattgcaac ccacccacca ggacacatgg acaagaagta ctccattggg
     4381 ctcgatatcg gcacaaacag cgtcggttgg gccgtcatta cggacgagta caaggtgccg
     4441 agcaaaaaat tcaaagttct gggcaatacc gatcgccaca gcataaagaa gaacctcatt
     4501 ggcgccctcc tgttcgactc cggggagacg gccgaagcca cgcggctcaa aagaacagca
     4561 cggcgcagat atacccgcag aaagaatcgg atctgctacc tgcaggagat ctttagtaat
     4621 gagatggcta aggtggatga ctctttcttc cataggctgg aggagtcctt tttggtggag
     4681 gaggataaaa agcacgagcg ccacccaatc tttggcaata tcgtggacga ggtggcgtac
     4741 catgaaaagt acccaaccat atatcatctg aggaagaagc ttgtagacag tactgataag
     4801 gctgacttgc ggttgatcta tctcgcgctg gcgcatatga tcaaatttcg gggacacttc
     4861 ctcatcgagg gggacctgaa cccagacaac agcgatgtcg acaaactctt tatccaactg
     4921 gttcagactt acaatcagct tttcgaagag aacccgatca acgcatccgg agttgacgcc
     4981 aaagcaatcc tgagcgctag gctgtccaaa tcccggcggc tcgaaaacct catcgcacag
     5041 ctccctgggg agaagaagaa cggcctgttt ggtaatctta tcgccctgtc actcgggctg
     5101 acccccaact ttaaatctaa cttcgacctg gccgaagatg ccaagcttca actgagcaaa
     5161 gacacctacg atgatgatct cgacaatctg ctggcccaga tcggcgacca gtacgcagac
     5221 ctttttttgg cggcaaagaa cctgtcagac gccattctgc tgagtgatat tctgcgagtg
     5281 aacacggaga tcaccaaagc tccgctgagc gctagtatga tcaagcgcta tgatgagcac
     5341 caccaagact tgactttgct gaaggccctt gtcagacagc aactgcctga gaagtacaag
     5401 gaaattttct tcgatcagtc taaaaatggc tacgccggat acattgacgg cggagcaagc
     5461 caggaggaat tttacaaatt tattaagccc atcttggaaa aaatggacgg caccgaggag
     5521 ctgctggtaa agcttaacag agaagatctg ttgcgcaaac agcgcacttt cgacaatgga
     5581 agcatccccc accagattca cctgggcgaa ctgcacgcta tcctcaggcg gcaagaggat
     5641 ttctacccct ttttgaaaga taacagggaa aagattgaga aaatcctcac atttcggata
     5701 ccctactatg taggccccct cgcccgggga aattccagat tcgcgtggat gactcgcaaa
     5761 tcagaagaga ctatcactcc ctggaacttc gaggaagtcg tggataaggg ggcctctgcc
     5821 cagtccttca tcgaaaggat gactaacttt gataaaaatc tgcctaacga aaaggtgctt
     5881 cctaaacact ctctgctgta cgagtacttc acagtttata acgagctcac caaggtcaaa
     5941 tacgtcacag aagggatgag aaagccagca ttcctgtctg gagagcagaa gaaagctatc
     6001 gtggacctcc tcttcaagac gaaccggaaa gttaccgtga aacagctcaa agaggactat
     6061 ttcaaaaaga ttgaatgttt cgactctgtt gaaatcagcg gagtggagga tcgcttcaac
     6121 gcatccctgg gaacgtatca cgatctcctg aaaatcatta aagacaagga cttcctggac
     6181 aatgaggaga acgaggacat tcttgaggac attgtcctca cccttacgtt gtttgaagat
     6241 agggagatga ttgaagaacg cttgaaaact tacgctcatc tcttcgacga caaagtcatg
     6301 aaacagctca agaggcgccg atatacagga tgggggcggc tgtcaagaaa actgatcaat
     6361 gggatccgag acaagcagag tggaaagaca atcctggatt ttcttaagtc cgatggattt
     6421 gccaaccgga acttcatgca gttgatccat gatgactctc tcacctttaa ggaggacatc
     6481 cagaaagcac aagtttctgg ccagggggac agccttcacg agcacatcgc taatcttgca
     6541 ggtagcccag ctatcaaaaa gggaatactg cagaccgtta aggtcgtgga tgaactcgtc
     6601 aaagtaatgg gaaggcataa gcccgagaat atcgttatcg agatggcccg agagaaccaa
     6661 actacccaga agggacagaa gaacagtagg gaaaggatga agaggattga agagggtata
     6721 aaagaactgg ggtcccaaat ccttaaggaa cacccagttg aaaacaccca gcttcagaat
     6781 gagaagctct acctgtacta cctgcagaac ggcagggaca tgtacgtgga tcaggaactg
     6841 gacatcaatc ggctctccga ctacgacgtg gatcatatcg tgccccagtc ttttctcaaa
     6901 gatgattcta ttgataataa agtgttgaca agatccgata aaaatagagg gaagagtgat
     6961 aacgtcccct cagaagaagt tgtcaagaaa atgaaaaatt attggcggca gctgctgaac
     7021 gccaaactga tcacacaacg gaagttcgat aatctgacta aggctgaacg aggtggcctg
     7081 tctgagttgg ataaagccgg cttcatcaaa aggcagcttg ttgagacacg ccagatcacc
     7141 aagcacgtgg cccaaattct cgattcacgc atgaacacca agtacgatga aaatgacaaa
     7201 ctgattcgag aggtgaaagt tattactctg aagtctaagc tcgtctcaga tttcagaaag
     7261 gactttcagt tttataaggt gagagagatc aacaattacc accatgcgca tgatgcctac
     7321 ctgaatgcag tggtaggcac tgcacttatc aaaaaatatc ccaagcttga atctgaattt
     7381 gtttacggag actataaagt gtacgatgtt aggaaaatga tcgcaaagtc tgagcaggaa
     7441 ataggcaagg ccaccgctaa gtacttcttt tacagcaata ttatgaattt tttcaagacc
     7501 gagattacac tggccaatgg agagattcgg aagcgaccac ttatcgaaac aaacggagaa
     7561 acaggagaaa tcgtgtggga caagggtagg gatttcgcga cagtccggaa ggtcctgtcc
     7621 atgccgcagg tgaacatcgt taaaaagacc gaagtacaga ccggaggctt ctccaaggaa
     7681 agtatcctcc cgaaaaggaa cagcgacaag ctgatcgcac gcaaaaaaga ttgggacccc
     7741 aagaaatacg gcggattcga ttctcctaca gtcgcttaca gtgtactggt tgtggccaaa
     7801 gtggagaaag ggaagtctaa aaaactcaaa agcgtcaagg aactgctggg catcacaatc
     7861 atggagcgat caagcttcga aaaaaacccc atcgactttc tcgaggcgaa aggatataaa
     7921 gaggtcaaaa aagacctcat cattaagctt cccaagtact ctctctttga gcttgaaaac
     7981 ggccggaaac gaatgctcgc tagtgcgggc gagctgcaga aaggtaacga gctggcactg
     8041 ccctctaaat acgttaattt cttgtatctg gccagccact atgaaaagct caaagggtcc
     8101 cccgaagata atgagcagaa gcagctgttc gtggaacaac acaaacacta ccttgatgag
     8161 atcatcgagc aaataagcga attctccaaa agagtgatcc tcgccgacgc taacctcgat
     8221 aaggtgcttt ctgcttacaa taagcacagg gataagccca tcagggagca ggcagaaaac
     8281 attatccact tgtttactct gaccaacttg ggcgcgcctg cagccttcaa gtacttcgac
     8341 accaccatag acagaaagcg gtacacctct acaaaggagg tcctggacgc cacactgatt
     8401 catcagtcaa ttacggggct ctatgaaaca agaatcgacc tctctcagct cggtggagac
     8461 agcagggctg accccaagaa gaagaggaag gtgtgagctt gatccactta acgttactga
     8521 aatcatcaaa cagcttgacg aatctggata taagatcgtt ggtgtcgatg tcagctccgg
     8581 agttgagaca aatggtgttc aggatctcga taagatacgt tcatttgtcc aagcagcaaa
     8641 gagtgccttc tagtgattta atagctccat gtcaacaaga ataaaacgcg tttcgggttt
     8701 acctcttcca gatacagctc atctgcaatg cattaatgca ttggacctcg caaccctagt
     8761 acgcccttca ggctccggcg aagcagaaga atagcttagc agagtctatt ttcattttcg
     8821 ggagacgaga tcaagcagat caacggtcgt caagagtcct acgagactga ggaatccgct
     8881 cttggctcca cgcgactata tatttgtctc taattgtact ttgacatgct cctcttcttt
     8941 actctgatag cttgactatg aaaattccgt caccagcccc tgggttcgca aagataattg
     9001 cactgtttct tccttgaact ctcaagccta caggacacac attcatcgta ggtataaacc
     9061 tcgaaaatca ttcctactaa gatgggtata caatagtaac catgcatggt tgcctagtga
     9121 atgctccgta acacccaata cgccggccga aactttttta caactctcct atgagtcgtt
     9181 tacccagaat gcacaggtac acttgtttag aggtaatcct tctttctaga agtcctcgtg
     9241 tactgtgtaa gcgcccactc cacatctcca ctcgctaatt catgaataac ggtgagacta
     9301 gcggccggtc cccttatccc agctgttcca cgttggcctg cccctcagtt agcgctcaac
     9361 tcaatgcccc tcactggcga ggcgagggca aggatggagg ggcagcatcg cctgagttgg
     9421 agcaaagcgg ccgccatggg agcagcgaac caacggaggg atgccgtgct ttgtcgtggc
     9481 tgctgtggcc aatccgggcc cttggttggc tcacagagcg ttgctgtgag tccatgagct
     9541 attattgcta ggtacagtat agagagagga gagagagaga gagagagaga ggggaaaaaa
     9601 ggtgaggttg aagtgagaaa aaaaaaaaaa aaaatccaac cactgacggc tgccggctct
     9661 gccacccccc tccctccacc ccagaccacc tgcacactca gcgcgcagca tcacctaatc
     9721 ttggctcgcc ttcccgcagc tcaggttgtt ttttttttct ctctccctcg tcgaagccgc
     9781 ccttgttccc ttatttattt ccctctccat ccttgtctgc ctttggtcca tctgcccctt
     9841 tgtctgcatc tcttttgcac gcatcgcctt atcgtcgtct cttttttcac tcacgggagc
     9901 ttgacgatga cctgactcgt gagcctcacc tgctgatttc tctccccccc tcccgaccgg
     9961 cttgactttt gtttctcctc cagtacctta tcgcgaagcc ggaagaacct cttaacctct
    10021 agatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa gttcctgatc gaaaagttcg
    10081 acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg
    10141 atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag
    10201 atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca
    10261 ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt
    10321 tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg
    10381 atgcgatcgc tgcggccgat ctcagccaga cgagcgggtt cggcccattc ggaccgcaag
    10441 gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt
    10501 atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg
    10561 agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg
    10621 gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg
    10681 cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg
    10741 cttgtatgga gcagcagacg cgctacttcg agcggaggca cccggagctt gcaggatcgc
    10801 cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg
    10861 acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg
    10921 gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg
    10981 gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa
    11041 aggaatagat gcatggcttt cgtgaccggg cttcaaacaa tgatgtgcga tggtgtggtt
    11101 cccggttggc ggagtctttg tctactttgg ttgtctgtcg caggtcggta gaccgcaaat
    11161 gagcaactga tggattgttg ccagcgatac tataattcac atggatggtc tttgtcgatc
    11221 agtagctagt gagagagaga gaacatctat ccacaatgtc gagtgtctat tagacatact
    11281 ccgagaataa agtcaactgt gtctgtgatc taaagatcga ttcggcagtc gagtagcgta
    11341 taacaactcc gagtaccagc gaaagcacgt cgtgacagga gcagggcttt gccaactgcg
    11401 caaccttgct tgaatgagga tacacggggt gcaacatggc tgtactgatc catcgcaacc
    11461 aaaatttctg tttatagatc aagctggtag attccaatta ctccacctct tgcgcttctc
    11521 catgacatgt aagtgcacgt ggaaaccata cccaaattgc ctacagctgc ggagcatgag
    11581 cctatggcga tcagtctggt catgttaacc agcctgtgct ctgacgttaa tgcagaatag
    11641 cttatgctgc agacgcgtta cggcggagat gctgcagacg cgttacgcgg agatgctgca
    11701 gacgcgttac gtatcggatc cagaattcgt gatatctgaa ttcgtcgaca agcttatttt
    11761 ttgtatactg ttttgtgata gcacgaagtt tttccacggt atcttgttaa aaatatatat
    11821 ttgtggcggg cttacctaca tcaaattaat aagagactaa ttataaacta aacacacaag
    11881 caagctactt tagggtaaaa gtttataaat gcttttgacg tataaacgtt gcttgtattt
    11941 attattacaa ttaaaggtgg atagaaaacc tagagactag ttagaaacta atctcaggtt
    12001 tgcgttaaac taaatcagag cccgagaggt taacagaacc tagaagggga ctagatatcc
    12061 gggtagggaa acaaaaaaaa aaaacaagac agccacatat tagggagact agttagaagc
    12121 tagttccagg actaggaaaa taaaagacaa tgataccaca gtctagttga caactagata
    12181 gattctagat tgaggccaaa gtctctgaga tccaggttag ttgcaactaa tactagttag
    12241 tatctagtct cctataactc tgaagctaga ataacttact actattatcc tcaccactgt
    12301 tcagctgcgc aaacggagtg attgcaaggt gttcagagac tagttattga ctagtcagtg
    12361 actagcaata actaacaagg tattaaccta ccatgtctgc catcaccctg cacttcctcg
    12421 ggctcagcag ccttttcctc ctcattttca tgctcatttt ccttgtttaa gactgtgact
    12481 agtcaaagac tagtccagaa ccacaaagga gaaatgtctt accactttct tcattgcttg
    12541 tctcttttgc attatccatg tctgcaacta gttagagtct agttagtgac tagtccgacg
    12601 aggacttgct tgtctccgga ttgttggagg aactctccag ggcctcaaga tccacaacag
    12661 agccttctag atgactggtc aataactagt tggtctttgt ctgagtctga cttacgaggt
    12721 tgcatactcg ctccctttgc ctcgtcaatc gatgagaaaa agcgccaaaa ctcgcaatat
    12781 ggctttgaac cacacggtgc tgagactagt tagaatctag tcccaaacta gcttggatag
    12841 cttacctttg ccctttgcgt tgcgacaggt cttgcagggt atggttcctt tctcaccagc
    12901 tgatttagct gccttgctac cctcacggcg gatctgcata aagagtggct agaggttata
    12961 aattagcact gatcctaggt acggggctga atgtaacttg cctttccttt ctcatcgcgc
    13021 ggcaagacag gcttgctcaa attcctacca gtcacagggg tatgcacggc gtacggacca
    13081 cttgaactag tcacagatta gttagcaact agtctgcatt gaatggctgt acttacgggc
    13141 cctcgccatt gtcctgatca tttccagctt caccctcgtt gctgcaaagt agttagtgac
    13201 tagtcaagga ctagttgaaa tgggagaaga aactcacgaa ttctcgacac ccttagtatt
    13261 gtggtccttg gacttggtgc tgctatatat tagctaatac actagttaga ctcacagaaa
    13321 cttacgcagc tcgcttgcgc ttcttggtag gagtcggggt tgggagaaca gtgccttcaa
    13381 acaagccttc ataccatgct acttgactag tcagggacta gtcaccaagt aatctagata
    13441 ggacttgcct ttggcctcca tcagttcctt catagtggga ggtccattgt gcaatgtaaa
    13501 ctccatgccg tgggagttct tgtccttcaa gtgcttgacc aatatgtttc tgttggcaga
    13561 gggaacctgt caactagtta ataactagtc agaaactagt atagcagtag actcactgta
    13621 cgcttgaggc atcccttcac tcggcagtag acttcatatg gatggatatc aggcacgcca
    13681 ttgtcgtcct gtggactagt cagtaactag gcttaaagct agtcgggtcg gcttactatc
    13741 ttgaaatccg gcagcgtaag ctccccgtcc ttaactgcct cgagatagtg acagtactct
    13801 ggggactttc ggagatcgtt atcgcgaatg ctcggcatac taatcgttga ctagtcttgg
    13861 actagtcccg agcaaaaagg attggaggag gaggaggaag gtgagagtga gacaaagagc
    13921 gaaataagag cttcaaaggc tatctctaag cagtatgaag gttaagtatc tagttcttga
    13981 ctagatttaa aagagatttc gactagttat gtacctggag tttggatata ggaatgtgtt
    14041 gtggtaacga aatgtaaggg ggaggaaaga aaaagtcggt caagaggtaa ctctaagtcg
    14101 gccattcctt tttgggaggc gctaaccata aacggcatgg tcgacttaga gttagctcag
    14161 ggaatttagg gagttatctg cgaccaccga ggaacggcgg aatgccaaag aatcccgatg
    14221 gagctctagc tggcggttga caaccccacc ttttggcgtt tctgcggcgt tgcaggcggg
    14281 actggatact tcgtagaacc agaaaggcaa ggcagaacgc gctcagcaag agtgttggaa
    14341 gtgatagcat gatgtgcctt gttaactagg tcaaaatctg cagtatgctt gatgttatcc
    14401 aaagtgtgag agaggaaggt ccaaacatac acgattggga gagggcctag gtataagagt
    14461 ttttgagtag aacgcatgtg agcccagcca tctcgaggag attaaacacg ggccggcatt
    14521 tgatggctat gttagtaccc caatggaaag cctgagagtc cagtgg
//
