LOCUS       sequence-133470-         837 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Human expression plasmid for SpCas9-HF4 variant: 
            CMV-T7-humanSpCas9-HF4(Y450A, N497A, R661A, Q695A, 
            Q926A)-NLS-3xFLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 837)
  AUTHORS   Kleinstiver BP, Pattanayak V, Prew MS, Tsai SQ, Nguyen NT, Zheng Z, 
            Keith Joung J
  TITLE     High-fidelity CRISPR-Cas9 nucleases with no detectable genome-wide 
            off-target effects.
  JOURNAL   Nature. 2016 Jan 6. doi: 10.1038/nature16526.
  PUBMED    26735016
REFERENCE   2  (bases 1 to 837)
  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: "Nature. 
            2016 Jan 6. doi: 10.1038/nature16526."
COMMENT     Sequence Label: MSP2133
FEATURES             Location/Qualifiers
     source          1..837
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(34..99)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(106..126)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ggtgatggtg atgatgaccg gtcatcctgc agccttgtca tcgtcatcct tgtaatcgat
       61 gtcatgatct ttataatcac cgtcatggtc tttgtagtcg ctcgagactt tcctcttctt
      121 cttgggggat ccgtcacccc caagctgtga caaatctatc cgagtttcat ataatcccgt
      181 gatggattgg tgaatcagtg tcgcgtctag cacctccttg gtagaagtgt atcgtttgcg
      241 atctatcgtt gtgtcaaaat acttgaatgc ggctggagcg ccgaggttgg taagagtaaa
      301 caaatggata atattttccg cctgctcacg tatgggttta tccctgtgct tgttgtatgc
      361 gcttaatact ttgtccagat tggcatcagc taggatgact ctcttactga attccgaaat
      421 ttgctctatg atttcgtcga gataatgttt gtgctgctca acaaaaagtt gcttctgttc
      481 gttatcttca ggtgaacctt tcaacttctc gtaatgggac gctaaataca ggaaattcac
      541 gtatttagac ggtagtgcga gttcgttccc cttttgaagc tctccggcgc tagccaacat
      601 ccgttttcgg ccattttcta actcaaacag actatacttt ggtagtttaa ttatgagatc
      661 cttttttact tccttgtaac ctttcgcctc aaggaagtcg atggggttct tttcaaaaga
      721 cgagcgctcc ataatcgtta tccccaataa ttctttgact gacttcagtt tcttggattt
      781 tcccttctca acttttgcca ctactaggac agaataggca actgtagggc tatcgaa
//
