LOCUS       sequence-349576-         722 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 166 and 892..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 722)
  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 722)
  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-tdTomato166/892
FEATURES             Location/Qualifiers
     source          1..722
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     61..80
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        160..235
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        332..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg
       61 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg
      121 tggaaaggac gaaacaccga tgtcccaggc gaagggcagg ttttagagct agaaatagca
      181 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
      241 tgttttagag ctagaaatag caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc
      301 aattctgcag acaaatggct ctagaggtac ccgttacata acttacggta aatggcccgc
      361 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attgtgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
      661 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
      721 gg
//
