LOCUS       sequence-116548-         865 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression plasmid for Sp-dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 865)
  AUTHORS   Kleinstiver BP, Prew MS, Tsai SQ, Topkar VV, Nguyen NT, Zheng Z, 
            Gonzales AP, Li Z, Peterson RT, Yeh JJ, Aryee MJ, Joung JK
  TITLE     Engineered CRISPR-Cas9 nucleases with altered PAM specificities.
  JOURNAL   Nature. 2015 Jun 22. doi: 10.1038/nature14592.
  PUBMED    26098369
REFERENCE   2  (bases 1 to 865)
  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. 
            2015 Jun 22. doi: 10.1038/nature14592."
COMMENT     Sequence Label: MSP712
FEATURES             Location/Qualifiers
     source          1..865
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      complement(242..289)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     285..303
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     misc_RNA        complement(305..380)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(448..467)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(449..467)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(537..602)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(609..629)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggt
       61 tgctacgcct gaataagtga taataagcgg atgaatggca gaaattcgaa agcaaattcg
      121 acccggtcgt cggttcaggg cagggtcgtt aaatagccgc ttatgtctat tgctggttta
      181 ccggtttatt gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc tgaggtttca
      241 gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt tattgctcag
      301 cggagcaccg actcggtgcc actttttcaa gttgataacg gactagcctt attttaactt
      361 gctatttcta gctctaaaac gagaccacta ggtgttggtg aatctgttca aaaattcata
      421 ttccgaacaa cgctatggca tgggtctccc tatagtgagt cgtattaatt tcgattatgc
      481 ggccgtgtac aatacgatta ctttctgttc gacttaagca ttatgcggcc gcttcacttg
      541 tcatcgtcat ccttgtaatc gatgtcatga tctttataat caccgtcatg gtctttgtag
      601 tcgctcgaga ctttcctctt cttcttgggg gatccgtcac ccccaagctg tgacaaatct
      661 atccgagttt catataatcc cgtgatggat tggtgaatca gtgtcgcgtc tagcacctcc
      721 ttggtagaag tgtatcgttt gcgatctatc gttgtgtcaa aatacttgaa tgcggctgga
      781 gcgccgaggt tggtaagagt aaacaaatgg ataatatttt ccgcctgctc acgtatgggt
      841 ttatccctgt gcttgttgta tgcgc
//
