LOCUS       sequence-240614-        4107 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Gene knockout/in for actinomycetes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4107)
  AUTHORS   Tong Y, Charusanti P, Zhang L, Weber T, Lee SY
  TITLE     CRISPR-Cas9 Based Engineering of Actinomycetal Genomes.
  JOURNAL   ACS Synth Biol. 2015 Sep 18;4(9):1020-9. doi: 
            10.1021/acssynbio.5b00038. Epub 2015 Apr 7.
  PUBMED    25806970
REFERENCE   2  (bases 1 to 4107)
  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"; doi: 
            "10.1021/acssynbio.5b00038"; journalName: "ACS Synth Biol"; date: 
            "2015-09-18- 18"; volume: "4"; issue: "9"; pages: "1020-9"
COMMENT     Sequence Label: pCRISPR-Cas9
FEATURES             Location/Qualifiers
     source          1..4107
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..4107
                     /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             1..4104
                     /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"
ORIGIN
        1 atggacaaga agtactccat cggcctcgac atcggcacca actccgtggg ctgggcggtc
       61 atcaccgacg agtacaaggt cccctccaag aagttcaagg tcctgggcaa caccgaccgg
      121 cactcgatca agaagaacct gatcggcgcc ctgctcttcg acagcggcga gaccgccgag
      181 gcgacccgcc tgaagcggac cgcgcgtcgc cgctacaccc ggcgcaagaa ccgcatctgc
      241 tacctgcagg aaatcttctc caacgagatg gccaaggtgg acgactcgtt cttccaccgc
      301 ctggaggaga gcttcctggt ggaggaggac aagaagcacg agcgccaccc gatcttcggc
      361 aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctccgcaag
      421 aagctggtgg actcgaccga caaggcggac ctgcggctca tctacctggc cctcgcgcac
      481 atgatcaagt tccgcggcca cttcctcatc gagggcgacc tgaacccgga caactccgac
      541 gtggacaagc tcttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc
      601 atcaacgcca gcggcgtgga cgccaaggcg atcctctccg cgcgcctgag caagtcccgg
      661 cgcctggaga acctcatcgc ccagctgccg ggcgagaaga agaacggcct cttcggcaac
      721 ctgatcgcgc tgtcgctcgg cctgaccccc aacttcaaga gcaacttcga cctggccgag
      781 gacgcgaagc tccagctgtc caaggacacc tacgacgacg acctggacaa cctgctcgcc
      841 cagatcggcg accagtacgc ggacctcttc ctggccgcga agaacctctc ggacgccatc
      901 ctgctcagcg acatcctgcg ggtcaacacc gagatcacca aggccccgct gtcggcgagc
      961 atgatcaagc ggtacgacga gcaccaccag gacctgaccc tgctcaaggc cctcgtgcgc
     1021 cagcagctgc ccgagaagta caaggaaatc ttcttcgacc agtccaagaa cggctacgcc
     1081 ggctacatcg acggcggcgc gtcgcaggag gagttctaca agttcatcaa gccgatcctg
     1141 gagaagatgg acggcaccga ggagctgctc gtcaagctga accgcgagga cctgctccgc
     1201 aagcagcgga ccttcgacaa cggctccatc ccgcaccaga tccacctggg cgagctccac
     1261 gccatcctcc ggcgccagga ggacttctac cccttcctga aggacaaccg cgagaagatc
     1321 gagaagatcc tgaccttccg catcccgtac tacgtcggcc ccctggcccg cggcaactcc
     1381 cggttcgcgt ggatgacccg gaagtcggag gagaccatca ccccgtggaa cttcgaggag
     1441 gtcgtggaca agggcgcgtc cgcgcagtcg ttcatcgagc gcatgaccaa cttcgacaag
     1501 aacctcccga acgagaaggt cctgcccaag cactccctgc tctacgagta cttcaccgtg
     1561 tacaacgagc tgaccaaggt caagtacgtg accgagggca tgcggaagcc ggccttcctg
     1621 tcgggcgagc agaagaaggc gatcgtggac ctgctcttca agaccaaccg caaggtcacc
     1681 gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgactc cgtcgagatc
     1741 agcggcgtgg aggaccgctt caacgcctcc ctgggcacct accacgacct gctcaagatc
     1801 atcaaggaca aggacttcct cgacaacgag gagaacgagg acatcctgga ggacatcgtc
     1861 ctcaccctga ccctcttcga ggaccgcgag atgatcgagg agcggctcaa gacctacgcc
     1921 cacctgttcg acgacaaggt gatgaagcag ctgaagcgtc gccgctacac cggctggggc
     1981 cgcctctccc ggaagctgat caacggcatc cgggacaagc agagcggcaa gaccatcctg
     2041 gacttcctca agtccgacgg cttcgccaac cgcaacttca tgcagctcat ccacgacgac
     2101 agcctgacct tcaaggagga catccagaag gcccaggtct cgggccaggg cgacagcctc
     2161 cacgagcaca tcgccaacct ggcgggctcc ccggcgatca agaagggcat cctccagacc
     2221 gtcaaggtcg tggacgagct ggtcaaggtg atgggccgcc acaagcccga gaacatcgtg
     2281 atcgagatgg cccgggagaa ccagaccacc cagaagggcc agaagaactc gcgcgagcgg
     2341 atgaagcgga tcgaggaggg catcaaggag ctcggcagcc agatcctgaa ggagcacccg
     2401 gtcgagaaca cccagctgca gaacgagaag ctgtacctct actacctgca gaacggccgc
     2461 gacatgtacg tggaccagga gctcgacatc aaccggctgt ccgactacga cgtggaccac
     2521 atcgtgccgc agtccttcct gaaggacgac tcgatcgaca acaaggtcct gacccgctcg
     2581 gacaagaacc ggggcaagtc cgacaacgtg ccctcggagg aggtcgtgaa gaagatgaag
     2641 aactactggc gccagctgct caacgccaag ctcatcaccc agcgcaagtt cgacaacctg
     2701 accaaggccg agcggggcgg cctgagcgag ctcgacaagg cgggcttcat caagcgccag
     2761 ctggtcgaga cccggcagat caccaagcac gtggcccaga tcctggactc ccggatgaac
     2821 accaagtacg acgagaacga caagctgatc cgcgaggtca aggtgatcac cctcaagagc
     2881 aagctggtct ccgacttccg caaggacttc cagttctaca aggtccggga gatcaacaac
     2941 taccaccacg cccacgacgc gtacctgaac gccgtcgtgg gcaccgcgct gatcaagaag
     3001 tacccgaagc tggagtccga gttcgtctac ggcgactaca aggtctacga cgtgcgcaag
     3061 atgatcgcca agagcgagca ggagatcggc aaggccaccg cgaagtactt cttctactcc
     3121 aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat ccgcaagcgg
     3181 cccctgatcg agaccaacgg cgagaccggc gagatcgtct gggacaaggg ccgcgacttc
     3241 gccaccgtcc ggaaggtgct gtcgatgccg caggtcaaca tcgtgaagaa gaccgaggtg
     3301 cagaccggcg gcttcagcaa ggagtccatc ctccccaagc gcaacagcga caagctgatc
     3361 gcccggaaga aggactggga cccgaagaag tacggcggct tcgacagccc caccgtcgcc
     3421 tactccgtgc tggtcgtggc gaaggtcgag aagggcaaga gcaagaagct gaagtccgtg
     3481 aaggagctgc tcggcatcac catcatggag cgctcctcgt tcgagaagaa cccgatcgac
     3541 ttcctggagg ccaagggcta caaggaggtc aagaaggacc tcatcatcaa gctgcccaag
     3601 tacagcctgt tcgagctgga gaacggccgc aagcggatgc tcgcctccgc gggcgagctg
     3661 cagaagggca acgagctggc cctcccgtcg aagtacgtca acttcctgta cctcgcgtcc
     3721 cactacgaga agctgaaggg ctcgcccgag gacaacgagc agaagcagct cttcgtggag
     3781 cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttcag caagcgcgtc
     3841 atcctggccg acgcgaacct cgacaaggtg ctgtccgcct acaacaagca ccgcgacaag
     3901 ccgatccggg agcaggcgga gaacatcatc cacctgttca ccctcaccaa cctgggcgcc
     3961 cccgccgcgt tcaagtactt cgacaccacc atcgaccgca agcggtacac ctccaccaag
     4021 gaggtcctcg acgcgaccct gatccaccag agcatcaccg gcctgtacga gacccgcatc
     4081 gacctgtccc agctcggcgg cgactga
//
