LOCUS       sequence-133477-         996 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Human expression plasmid for SpCas9-VRQR-HF1 variant: 
            CMV-T7-humanSpCas9-VRQR-HF1(N497A, R661A, Q695A, Q926A, D1135V, 
            G1218R, R1335Q, T1337R)-NLS-3xFLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 996)
  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 996)
  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: MSP2443
FEATURES             Location/Qualifiers
     source          1..996
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(1..18)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(36..101)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(108..128)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 atggtgatgg tgatgatgac cggtcatcct gcagccttgt catcgtcatc cttgtaatcg
       61 atgtcatgat ctttataatc accgtcatgg tctttgtagt cgctcgagac tttcctcttc
      121 ttcttggggg atccgtcacc cccaagctgt gacaaatcta tccgagtttc atataatccc
      181 gtgatggatt ggtgaatcag tgtcgcgtct agcacctcct tggtagatct gtactgtttg
      241 cgatctatcg ttgtgtcaaa atacttgaat gcggctggag cgccgaggtt ggtaagagta
      301 aacaaatgga taatattttc cgcctgctca cgtatgggtt tatccctgtg cttgttgtat
      361 gcgcttaata ctttgtccag attggcatca gctaggatga ctctcttact gaattccgaa
      421 atttgctcta tgatttcgtc gagataatgt ttgtgctgct caacaaaaag ttgcttctgt
      481 tcgttatctt caggtgaacc tttcaacttc tcgtaatggg acgctaaata caggaaattc
      541 acgtatttag acggtagtgc gagttcgttc cccttttgaa gctctctggc gctagccaac
      601 atccgttttc ggccattttc taactcaaac agactatact ttggtagttt aattatgaga
      661 tcctttttta cttccttgta acctttcgcc tcaaggaagt cgatggggtt cttttcaaaa
      721 gacgagcgct ccataatcgt tatccccaat aattctttga ctgacttcag tttcttggat
      781 tttcccttct caacttttgc cactactagg acagaatagg caactgtagg gctcacgaag
      841 ccaccgtact ttttcgggtc ccagtccttt ttacgagcga tgagcttatc actattcctt
      901 tttggaagaa tcgattcctt tgaaaaccct ccggtctgca cctcagtttt ctttactatg
      961 ttgacttggg gcatggacaa aacttttctc accgtc
//
