LOCUS       sequence-212156-        4299 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression and purification of NLSH2BCas9 for fungal gene editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4299)
  AUTHORS   Wang Q, Cobine PA, Coleman JJ
  TITLE     Efficient genome editing in Fusarium oxysporum based on CRISPR/Cas9 
            ribonucleoprotein complexes.
  JOURNAL   Fungal Genet Biol. 2018 May 12. pii: S1087-1845(18)30081-1. doi: 
            10.1016/j.fgb.2018.05.003.
  PUBMED    29763675
REFERENCE   2  (bases 1 to 4299)
  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: "Fungal 
            Genet Biol. 2018 May 12. pii: S1087-1845(18)30081-1. doi: 
            10.1016/j.fgb.2018.05.003."
COMMENT     Sequence Label: pHis-parallel1-NLSH2BCas9
FEATURES             Location/Qualifiers
     source          1..4299
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             169..4272
                     /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             169..4272
                     /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             4276..4296
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 atggctccca aggctgctga caagaagccc gcctccaagg ctcccgctac tgcctccaag
       61 gctcctgaga agaaggatgc tggcaagaag actgccgcct ctggtgacaa gaagaagcgc
      121 tccaagaccc gcaaggagac ctactcttct tacatctaca aggtcgacat ggacaagaag
      181 tatagcatcg ggctggacat tggaacgaac tcggttggtt gggctgtgat tacggacgaa
      241 tacaaggtgc catccaagaa gtttaaggtc ctgggaaaca ccgaccgtca ctcaatcaag
      301 aagaatctca ttggagccct gctcttcgat agtggggaga ccgccgaagc tactcgactg
      361 aagcgaacgg ctcgccggcg ttatacacga cgcaagaatc gcatctgcta cctccaggag
      421 attttcagca acgaaatggc taaggttgat gactcattct ttcatcgact cgaagaaagt
      481 ttcttggtcg aggaggataa gaagcacgag cgccatccga tctttggtaa cattgtggat
      541 gaggttgcct atcacgaaaa gtacccaact atctatcatc ttcgtaagaa gctggtcgat
      601 agcacggaca aggctgattt gcgacttatc tacctggcac tcgcgcacat gattaagttc
      661 cgcggccatt ttcttatcga gggtgacctg aaccccgata attctgacgt tgataagctc
      721 ttcatccagt tggtccaaac ctacaatcag ctgtttgagg aaaaccctat taatgcatct
      781 ggcgtggacg ccaaggctat cctttcggcg cgcctgtcta agtcgcggcg tttggagaac
      841 cttatcgcac aactccccgg cgaaaagaag aacggcctct tcggtaattt gattgcgttg
      901 tcacttggtc tgactcctaa cttcaagagt aattttgacc tggcagagga tgcgaagctc
      961 cagttgtcta aggatacgta tgatgacgat ctcgacaact tgcttgccca aatcggtgac
     1021 cagtacgctg atcttttcct ggccgctaag aatctctcag atgcaatcct gctcagtgac
     1081 attttgcggg tcaacaccga gattactaag gcccccctgt cagctagtat gatcaagcgg
     1141 tatgatgagc accatcagga cctcaccttg cttaaggccc tcgtgcgtca gcaattgcct
     1201 gagaagtaca aggaaatctt ctttgaccaa tccaagaacg gatacgcagg gtatattgat
     1261 ggcggtgcga gccaggagga attctacaag tttatcaagc cgattttgga gaagatggac
     1321 ggcactgagg aactgctcgt caagctgaat cgcgaagatt tgcttcgtaa gcaacgaacg
     1381 ttcgacaacg gctccatccc gcaccagatt catctgggcg agctccacgc catccttcga
     1441 cgccaggaag atttctaccc atttctgaag gacaaccgtg agaagatcga aaagattctt
     1501 acattccgaa tcccctacta tgtgggacct ttggcccgtg ggaattcccg atttgcttgg
     1561 atgacccgaa agagcgagga aaccatcact ccgtggaact tcgaggaagt cgtggacaag
     1621 ggtgcatccg cgcagagctt cattgagcgg atgaccaatt ttgataagaa ccttccgaat
     1681 gaaaaggtcc tgccaaagca ttcgctgctc tacgagtatt tcaccgtgta taacgaactg
     1741 actaaggtca agtacgtgac ggagggaatg cggaagccag ccttcctctc aggggaacaa
     1801 aagaaggcta tcgtcgattt gctttttaag accaatcgta aagtgactgt taagcagctg
     1861 aaggaggatt atttcaagaa gattgaatgt ttcgactccg tcgagatcag cggcgtggaa
     1921 gatcgcttta acgcttccct cggtacctac cacgacctgc tcaagatcat taaggacaag
     1981 gatttcctcg ataacgagga aaatgaggac atcttggaag atattgtcct cacgttgaca
     2041 ctttttgagg accgcgaaat gatcgaggaa cggctcaaga catatgccca tttgttcgac
     2101 gataaggtga tgaagcagct gaagcggcgt cgatacaccg gatggggtcg ccttagccgg
     2161 aagctgatca acggcattcg agataagcaa tctggtaaga ctatcttgga tttccttaag
     2221 tcggacggct tcgccaaccg caattttatg cagcttattc acgacgattc cctgacgttc
     2281 aaggaggaca tccagaaggc acaagtctca ggacaagggg attccctgca cgagcatatc
     2341 gccaacctgg ctggatcccc ggcgatcaag aaggggattc ttcagaccgt caaggttgtc
     2401 gacgagctgg tcaaggtgat gggccgtcat aagccagaaa acatcgtgat tgagatggcc
     2461 cgagaaaatc agaccactca aaagggtcag aagaacagcc gcgagcggat gaagcggatc
     2521 gaggaaggca ttaaggaact tggttctcag atcctgaagg agcaccctgt tgaaaacaca
     2581 cagctccaaa atgagaagct gtatctctac tatttgcaaa atggacgcga catgtacgtc
     2641 gatcaggagc tcgacattaa ccggttgtcg gactacgatg ttgaccatat cgtcccgcaa
     2701 tccttcctta aggacgatag cattgataac aaggtgctga ctcgctcaga taagaaccgg
     2761 ggcaagtccg acaatgttcc aagcgaggaa gtggttaaga agatgaagaa ctactggcgc
     2821 caattgctta atgccaagct catcacacag cgcaagtttg acaacttgac caaggccgag
     2881 cggggagggc tgagtgaact cgataaggct ggcttcatca agcgtcaact cgtggagacg
     2941 cgacagatca caaagcacgt tgctcagatt ctggactccc ggatgaacac aaagtacgac
     3001 gagaatgata agctcatccg tgaagttaag gtcattaccc tcaagtctaa gttggtgtcg
     3061 gatttccgca aggacttcca attttataag gttcgggaga tcaacaatta tcaccatgca
     3121 catgatgcgt acctcaacgc agtcgtggga actgcgctca tcaagaagta tcccaagttg
     3181 gagtccgaat tcgtctacgg ggattataag gtttacgacg tccgcaagat gatcgccaag
     3241 agtgagcagg aaattggcaa ggccacggct aagtatttct tttactccaa catcatgaat
     3301 ttctttaaga cggagatcac actcgccaat ggagaaatcc gtaagcgacc tttgattgag
     3361 accaacggcg agactggtga aatcgtttgg gataaggggc gcgacttcgc taccgtgcgg
     3421 aaggttctga gcatgccgca agtcaatatc gtcaagaaaa ccgaggtgca gacaggcggt
     3481 ttctctaagg aatcgattct tccaaagcgt aactctgaca agctgatcgc tcgaaagaag
     3541 gattgggacc ccaagaagta tggagggttc gattctccta cagtggcata ctcggttctc
     3601 gttgtcgcga aggttgagaa gggaaagtct aagaagctga agtcggtcaa ggaactgctc
     3661 gggatcacca ttatggagcg ctccagcttc gaaaagaatc ccatcgactt tctcgaggcc
     3721 aagggctata aggaagtcaa gaaggatctt atcattaagc tgcctaagta ctctttgttc
     3781 gagcttgaaa acggtcgaaa gcgaatgctc gcatcggcag gagagttgca gaaggggaat
     3841 gaattggcac ttccctcaaa gtacgtgaac ttcctgtatc tcgcgtccca ctacgagaag
     3901 ctgaagggta gccctgagga caacgaacag aagcaacttt ttgttgagca acacaagcat
     3961 tatctggatg agatcattga acagatttca gagttcagta agcgcgtcat cctcgccgat
     4021 gctaatctcg acaaggtgtt gtcggcctac aacaagcacc gtgacaagcc gatccgagag
     4081 caggctgaaa atatcattca tctgttcacc ctcactaact tgggagcacc agcagcgttc
     4141 aagtattttg atacgacaat cgaccgtaag cgatacacgt ccacaaagga ggtgcttgat
     4201 gcgaccctga ttcatcaatc catcactggg ctctatgaaa cccgtatcga ccttagtcaa
     4261 ctggggggcg acccccccaa gaagaagcgc aaggtctga
//
