LOCUS       sequence-349574-         720 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Double cutting Cas9 plasmid with puromycin resistance and a single 
            guide RNA targeting tdTomato fluorescent protein sites 90 and 816..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 720)
  AUTHORS   Madireddy I, Pierson Smela M
  TITLE     Stably Integrating an Inducible CRISPR-Cas9 to Protect Against Viral
            Infections in Vitro.
  JOURNAL   MicroPubl Biol. 2022 Jun 16;2022. doi: 
            10.17912/micropub.biology.000590. eCollection 2022.
  PUBMED    35789697
REFERENCE   2  (bases 1 to 720)
  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: "MicroPubl 
            Biol."; date: "2022-06-16"; volume: "2022. doi"; pages: " 
            10.17912/micropub.biology.000590. eCollection 2022"
COMMENT     Sequence Label: pX459-sg-tdTomato90/816
FEATURES             Location/Qualifiers
     source          1..720
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     57..76
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        155..230
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        327..612
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
       61 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
      121 aaggacgaaa caccggccac gagttcgaga tcgagtttta gagctagaaa tagcaagtta
      181 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttgttt
      241 tagagctaga aatagcaagt taaaataagg ctagtccgtt tttagcgcgt gcgccaattc
      301 tgcagacaaa tggctctaga ggtacccgtt acataactta cggtaaatgg cccgcctggc
      361 tgaccgccca acgacccccg cccattgacg tcaatagtaa cgccaatagg gactttccat
      421 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      481 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattgt
      541 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      601 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
      661 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg
//
