LOCUS       sequence-116546-         665 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression plasmid for SpCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 665)
  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 665)
  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: BPK764
FEATURES             Location/Qualifiers
     source          1..665
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      complement(230..277)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     273..291
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     misc_RNA        complement(293..368)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(423..442)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(424..442)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(512..577)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(584..604)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ggggcgtaat ttttttaagg cagttattgg tgcccttaaa cgcctggttg ctacgcctga
       61 ataagtgata ataagcggat gaatggcaga aattcgaaag caaattcgac ccggtcgtcg
      121 gttcagggca gggtcgttaa atagccgctt atgtctattg ctggtttacc ggtttattga
      181 ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg aggtttcagc aaaaaacccc
      241 tcaagacccg tttagaggcc ccaaggggtt atgctagtta ttgctcagcg gagcaccgac
      301 tcggtgccac tttttcaagt tgataacgga ctagccttat tttaacttgc tatttctagc
      361 tctaaaacgg agaccactag gtgttggtga atctgttccg aacaacgcta tggcatgggt
      421 ctccctatag tgagtcgtat taatttcgat tatgcggccg tgtacaatac gattactttc
      481 tgttcgactt aagcattatg cggccgcttc acttgtcatc gtcatccttg taatcgatgt
      541 catgatcttt ataatcaccg tcatggtctt tgtagtcgct cgagactttc ctcttcttct
      601 tgggggatcc gtcaccccca agctgtgaca aatctatccg agtttcatat aatcccgtga
      661 tggat
//
