LOCUS       sequence-83911-e        1083 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses N-terminal His tag fused human codon-optimized Cas9 
            nuclease having C-terminal Cysteine .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1083)
  AUTHORS   Ramakrishna S, Kwaku Dad AB, Beloor J, Gopalappa R, Lee SK, Kim H
  TITLE     Gene disruption by cell-penetrating peptide-mediated delivery of 
            Cas9 protein and guide RNA.
  JOURNAL   Genome Res. 2014 Apr 2.
  PUBMED    24696462
REFERENCE   2  (bases 1 to 1083)
  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 Res.
            2014 Apr 2."
COMMENT     Sequence Label: pET28a/Cas9-Cys
FEATURES             Location/Qualifiers
     source          1..1083
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     68..87
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        68..86
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    87..111
                     /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             126..148
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             167..184
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             194..211
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             215..247
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
ORIGIN
        1 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc
       61 gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa
      121 ataattttgt ttaactttaa gaaggagata taccatgggc agcagccatc atcatcatca
      181 tcacagcagc ggcctggtgc cgcgcggcag ccatatggct agcatgactg gtggacagca
      241 aatgggtcgc ggatccgaat tcgagctccg tcgacaagct tgcggccgca tggacaagaa
      301 gtacagcatc ggcctggaca tcggtaccaa cagcgtgggc tgggccgtga tcaccgacga
      361 gtacaaggtg cccagcaaga agttcaaggt gctgggcaac accgaccgcc acagcatcaa
      421 gaagaacctg atcggcgccc tgctgttcga cagcggcgag accgccgagg ccacccgcct
      481 gaagcgcacc gcccgccgcc gctacacccg ccgcaagaac cgcatctgct acctgcagga
      541 gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccaccgcc tggaggagag
      601 cttcctggtg gaggaggaca agaagcacga gcgccacccc atcttcggca acatcgtgga
      661 cgaggtggcc taccacgaga agtaccccac catctaccac ctgcgcaaga agctggtgga
      721 cagcaccgac aaggccgacc tgcgcctgat ctacctggcc ctggcccaca tgatcaagtt
      781 ccgcggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct
      841 gttcatccag ctggtgcaga cctacaacca gctgttcgag gagaacccca tcaacgccag
      901 cggcgtggac gccaaggcca tcctgagcgc ccgcctgagc aagagccgcc gcctggagaa
      961 cctgatcgcc cagctgcccg gcgagaagaa gaacggcctg ttcggcaacc tgatcgccct
     1021 gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg acgccaagct
     1081 gca
//
