LOCUS       sequence-156555-         829 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses eGFP with ecDHFR along with an U6 promoter driven gRNA 
            which cleaves the plasmid in-cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 829)
  AUTHORS   Talas A, Kulcsar PI, Weinhardt N, Borsy A, Toth E, Szebenyi K, 
            Krausz SL, Huszar K, Vida I, Sturm A, Gordos B, Hoffmann OI, 
            Bencsura P, Nyeste A, Ligeti Z, Fodor E, Welker E
  TITLE     A convenient method to pre-screen candidate guide RNAs for 
            CRISPR/Cas9 gene editing by NHEJ-mediated integration of a 
            'self-cleaving' GFP-expression plasmid.
  JOURNAL   DNA Res. 2017 Jun 30. doi: 10.1093/dnares/dsx029.
  PUBMED    28679166
REFERENCE   2  (bases 1 to 829)
  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: "DNA Res. 
            2017 Jun 30. doi: 10.1093/dnares/dsx029."
COMMENT     Sequence Label: pSc1-DD
FEATURES             Location/Qualifiers
     source          1..829
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      42..51
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             54..530
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="MISLIAALAVDYVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRH
                     TWESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVIEQFL
                     PKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     primer_bind     complement(621..642)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 agcagagctg gtttagtgaa ccgtcagatc cgctagcggc cgccaccatg gccatgatca
       61 gtctgattgc ggcgttagcg gtagattacg ttatcggcat ggaaaacgcc atgccgtgga
      121 acctgcctgc cgatctcgcc tggtttaaac gcaacacctt aaataaaccc gtgattatgg
      181 gccgccatac ctgggaatca atcggtcgtc cgttgccagg acgcaaaaat attatcctca
      241 gcagtcaacc gagtacggac gatcgcgtaa cgtgggtgaa gtcggtggat gaagccatcg
      301 cggcgtgtgg tgacgtacca gaaatcatgg tgattggcgg cggtcgcgtt attgaacagt
      361 tcttgccaaa agcgcaaaaa ctgtatctga cgcatatcga cgcagaagtg gaaggcgaca
      421 cccatttccc ggattacgag ccggatgact gggaatcggt attcagcgaa ttccacgatg
      481 ctgatgcgca gaactctcac agctattgct ttgagattct ggagcggcga ggatcccttg
      541 cagaggcggc tgctaaagcg gcagcgggct cgagtgtggt gagcaagggc gaggagctgt
      601 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
      661 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
      721 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
      781 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttctt
//
