LOCUS       sequence-131906-         765 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression plasmid for KKH SaCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 765)
  AUTHORS   Kleinstiver BP, Prew MS, Tsai SQ, Nguyen NT, Topkar VV, Zheng Z, 
            Joung JK
  TITLE     Broadening the targeting range of Staphylococcus aureus CRISPR-Cas9 
            by modifying PAM recognition.
  JOURNAL   Nat Biotechnol. 2015 Nov 2. doi: 10.1038/nbt.3404.
  PUBMED    26524662
REFERENCE   2  (bases 1 to 765)
  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: "Nat 
            Biotechnol. 2015 Nov 2. doi: 10.1038/nbt.3404."
COMMENT     Sequence Label: MSP2253
FEATURES             Location/Qualifiers
     source          1..765
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      complement(249..296)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     292..310
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     primer_bind     complement(421..440)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(422..440)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(510..575)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(582..602)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac
       61 gcctggttgc tacgcctgaa taagtgataa taagcggatg aatggcagaa attcgaaagc
      121 aaattcgacc cggtcgtcgg ttcagggcag ggtcgttaaa tagccgctta tgtctattgc
      181 tggtttaccg gtttattgac taccggaagc agtgtgaccg tgtgcttctc aaatgcctga
      241 ggtttcagca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctagttat
      301 tgctcagcgg aaaaaaaatc tcgccaacaa gttgacgaga taaacacggc attttgcctt
      361 gttttagtag attctgtaat tttcattaca gagtactaaa acccggcaag ctgcccgtgc
      421 ccctatagtg agtcgtatta atttcgatta tgcggccgtg tacaatacga ttactttctg
      481 ttcgacttaa gcattatgcg gccgctttac ttgtcatcgt catccttgta atcgatgtca
      541 tgatctttat aatcaccgtc atggtctttg tagtcgctcg agactttcct cttcttcttg
      601 ggggatccac cttttttaat gatttgtggg tgcttctttg acttcacttc atagagattc
      661 cccaggatat cggtagagta cttttttatt gactgagttt tagatgcgat tgtcttgatg
      721 atgtgaggcg gccgtttatc attcatgttt tcgagatact cccgg
//
