LOCUS       sequence-137956-         776 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  bacterial pMCSG7 expression vector expressing Nme cas9 nickase with 
            a D16A mutation (RuvC domain) with T7 promoter, N-terminal His tag 
            and TEV site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 776)
  AUTHORS   Zhang Y, Rajan R, Seifert HS, Mondragon A, Sontheimer EJ
  TITLE     DNase H Activity of Neisseria meningitidis Cas9.
  JOURNAL   Mol Cell. 2015 Oct 15;60(2):242-55. doi: 
            10.1016/j.molcel.2015.09.020.
  PUBMED    26474066
REFERENCE   2  (bases 1 to 776)
  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"; doi: 
            "10.1016/j.molcel.2015.09"; journalName: "Mol Cell"; date: 
            "2015-10-15- 15"; volume: "60"; issue: "2"; pages: "242-55"
COMMENT     Sequence Label: pMCSG7-D16A-NmeCas9
FEATURES             Location/Qualifiers
     source          1..776
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     57..76
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        57..75
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    76..100
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             115..137
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             148..165
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             190..210
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
ORIGIN
        1 gccggccacg atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg cgaaattaat
       61 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt
      121 taactttaag aaggagatat acatatgcac catcatcatc atcattcttc tggtgtagat
      181 ctgggtaccg agaacctgta cttccaatcc aatgccattg ctgccttcaa acctaattca
      241 atcaactaca tcctcggcct cgctatcggc atcgcatccg tcggctgggc gatggtagaa
      301 attgacgaag aagaaaaccc catccgcctg attgatttgg gcgtgcgcgt atttgagcgt
      361 gccgaagtac cgaaaacagg cgactccctt gccatggcaa ggcgtttggc gcgcagtgtt
      421 cgccgcctga cccgccgtcg cgcccaccgc ctgcttcgga cccgccgcct attgaaacgc
      481 gaaggcgtat tacaagccgc caattttgac gaaaacggct tgattaaatc cttaccgaat
      541 acaccatggc aacttcgcgc agccgcatta gaccgcaaac tgacgccttt agagtggtcg
      601 gcagtcttgt tgcatttaat caaacatcgc ggctatttat cgcaacggaa aaacgagggc
      661 gaaactgccg ataaggagct tggcgctttg cttaaaggcg tagccggcaa tgcccatgcc
      721 ttacagacag gcgatttccg cacaccggcc gaattggctt taaataaatt tgagaa
//
