LOCUS       sequence-177090-        4272 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression plasmid for human codon-optimized high-fidelity 
            SpCas9-HF1, px330-like backbone (without U6-sgRNA coding sequence).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4272)
  AUTHORS   Kulcsar PI, Talas A, Huszar K, Ligeti Z, Toth E, Weinhardt N, Fodor 
            E, Welker E
  TITLE     Crossing enhanced and high fidelity SpCas9 nucleases to optimize 
            specificity and cleavage.
  JOURNAL   Genome Biol. 2017 Oct 6;18(1):190. doi: 10.1186/s13059-017-1318-8.
  PUBMED    28985763
REFERENCE   2  (bases 1 to 4272)
  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: "Genome 
            Biol."; date: "2017-10-6"; volume: "18(1):190. doi"; pages: " 
            10.1186/s13059-017-1318-8"
COMMENT     Sequence Label: pX330-Flag-SpCas9-HF1  (without sgRNA)
FEATURES             Location/Qualifiers
     source          1..4272
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             4..69
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             76..96
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4222..4269
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 atggactata aggaccacga cggagactac aaggatcatg atattgatta caaagacgat
       61 gacgataaga tggccccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc
      121 gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc
      181 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
      241 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
      301 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
      361 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
      421 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
      481 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
      541 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
      601 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
      661 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
      721 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
      781 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg
      841 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
      901 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
      961 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
     1021 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
     1081 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
     1141 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
     1201 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
     1261 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
     1321 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
     1381 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
     1441 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
     1501 ttcgcctgga tgacaagaaa gtccgaagaa acgattactc cctggaattt tgaggaagtt
     1561 gtcgataaag gtgcgtcagc tcaatcgttc atcgagagga tgaccgcctt tgacaagaat
     1621 ttaccgaacg aaaaagtatt gcctaagcac agtttacttt acgagtattt cacagtgtac
     1681 aatgaactca cgaaagttaa gtatgtcact gagggcatgc gtaaacccgc ctttctaagc
     1741 ggagaacaga agaaagcaat agtagatctg ttattcaaga ccaaccgcaa agtgacagtt
     1801 aagcaattga aagaggacta ctttaagaaa attgaatgct tcgattctgt cgagatctcc
     1861 ggggtagaag atcgatttaa tgcgtcactt ggtacgtatc atgacctcct aaagataatt
     1921 aaagataagg acttcctgga taacgaagag aatgaagata tcttagaaga tatagtgttg
     1981 actcttaccc tctttgaaga tcgggaaatg attgaggaaa gactaaaaac atacgctcac
     2041 ctgttcgacg ataaggttat gaaacagtta aagaggcgtc gctatacggg ctggggagcc
     2101 ttgtcgcgga aacttatcaa cgggataaga gacaagcaaa gtggtaaaac tattctcgat
     2161 tttctaaaga gcgacggctt cgccaatagg aactttatgg ccctgatcca tgatgactct
     2221 ttaaccttca aagaggatat acaaaaggca caggtttccg gacaagggga ctcattgcac
     2281 gaacatattg cgaatcttgc tggttcgcca gccatcaaaa agggcatact ccagacagtc
     2341 aaagtagtgg atgagctagt taaggtcatg ggacgtcaca aaccggaaaa cattgtaatc
     2401 gagatggcac gcgaaaatca aacgactcag aaggggcaaa aaaacagtcg agagcggatg
     2461 aagagaatag aagagggtat taaagaactg ggcagccaga tcttaaagga gcatcctgtg
     2521 gaaaataccc aattgcagaa cgagaaactt tacctctatt acctacaaaa tggaagggac
     2581 atgtatgttg atcaggaact ggacataaac cgtttatctg attacgacgt cgatcacatt
     2641 gtaccccaat cctttttgaa ggacgattca atcgacaata aagtgcttac acgctcggat
     2701 aagaaccgag ggaaaagtga caatgttcca agcgaggaag tcgtaaagaa aatgaagaac
     2761 tattggcggc agctcctaaa tgcgaaactg ataacgcaaa gaaagttcga taacttaact
     2821 aaagctgaga ggggtggctt gtctgaactt gacaaggccg gatttattaa acgtcagctc
     2881 gtggaaaccc gcgccatcac aaagcatgtt gccgagatac tagattcccg aatgaatacg
     2941 aaatacgacg agaacgataa gctgattcgg gaagtcaaag taatcacttt aaagtcaaaa
     3001 ttggtgtcgg acttcagaaa ggattttcaa ttctataaag ttagggagat aaataactac
     3061 caccatgcgc acgacgctta tcttaatgcc gtcgtaggga ccgcactcat taagaaatac
     3121 ccgaagctag aaagtgagtt tgtgtatggt gattacaaag tttatgacgt ccgtaagatg
     3181 atcgcgaaaa gcgaacagga gataggcaag gctacagcca aatacttctt ttattctaac
     3241 attatgaatt tctttaagac ggaaatcact ctggcaaacg gagagatacg caaacgacct
     3301 ttaattgaaa ccaatgggga gacaggtgaa atcgtatggg ataagggccg ggacttcgcg
     3361 acggtgagaa aagttttgtc catgccccaa gtcaacatag taaagaaaac tgaggtgcag
     3421 accggagggt tttcaaagga atcgattctt ccaaaaagga atagtgataa gctcatcgct
     3481 cgtaaaaagg actgggaccc gaaaaagtac ggtggcttcg atagccctac agttgcctat
     3541 tctgtcctag tagtggcaaa agttgagaag ggaaaatcca agaaactgaa gtcagtcaaa
     3601 gaattattgg ggataacgat tatggagcgc tcgtcttttg aaaagaaccc catcgacttc
     3661 cttgaggcga aaggttacaa ggaagtaaaa aaggatctca taattaaact accaaagtat
     3721 agtctgtttg agttagaaaa tggccgaaaa cggatgttgg ctagcgccgg agagcttcaa
     3781 aaggggaacg aactcgcact accgtctaaa tacgtgaatt tcctgtattt agcgtcccat
     3841 tacgagaagt tgaaaggttc acctgaagat aacgaacaga agcaactttt tgttgagcag
     3901 cacaaacatt atctcgacga aatcatagag caaatttcgg aattcagtaa gagagtcatc
     3961 ctagctgatg ccaatctgga caaagtatta agcgcataca acaagcacag ggataaaccc
     4021 atacgtgagc aggcggaaaa tattatccat ttgtttactc ttaccaacct cggcgctcca
     4081 gccgcattca agtattttga cacaacgata gatcgcaaac gatacacttc taccaaggag
     4141 gtgctagacg cgacactgat tcaccaatcc atcacgggat tatatgaaac tcggatagat
     4201 ttgtcacagc ttgggggtga caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa
     4261 aagaaaaagt aa
//
