LOCUS       sequence-82983-e        1128 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 1128)
  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 1128)
  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..1128
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             27..49
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             68..85
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             95..112
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             116..148
                     /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 ggggggaaaa ttccctctag aataattttg tttaacttta agaaggagat ataccatggg
       61 cagcagccat catcatcatc atcacagcag cggcctggtg ccgcgcggca gccatatggc
      121 tagcatgact ggtggacagc aaatgggtcg cggatccgaa ttcgagctcc gtcgacaagc
      181 ttgcggccgc atggacaaga agtacagcat cggcctggac atcggtacca acagcgtggg
      241 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aagttcaagg tgctgggcaa
      301 caccgaccgc cacagcatca agaagaacct gatcggcgcc ctgctgttcg acagcggcga
      361 gaccgccgag gccacccgcc tgaagcgcac cgcccgccgc cgctacaccc gccgcaagaa
      421 ccgcatctgc tacctgcagg agatcttcag caacgagatg gccaaggtgg acgacagctt
      481 cttccaccgc ctggaggaga gcttcctggt ggaggaggac aagaagcacg agcgccaccc
      541 catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca
      601 cctgcgcaag aagctggtgg acagcaccga caaggccgac ctgcgcctga tctacctggc
      661 cctggcccac atgatcaagt tccgcggcca cttcctgatc gagggcgacc tgaaccccga
      721 caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga
      781 ggagaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgagcg cccgcctgag
      841 caagagccgc cgcctggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct
      901 gttcggcaac ctgatcgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga
      961 cctggccgag gacgccaagc tgcagctgag caaggacacc tacgacgacg acctggacaa
     1021 cctgctggcc cagatcggcg accagtacgc gactgttctg tcgcaagaac tgagcgacgc
     1081 atctgctgag cgacatcctg gcggcggtga acaccgagat caccaagc
//
