LOCUS       sequence-156544-         886 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses eGFP 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 886)
  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 886)
  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
FEATURES             Location/Qualifiers
     source          1..886
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      47..56
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(53..55,56..58,59..769)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(53..55,56..58,59..769)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(98..119)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(359..378)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     706..727
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    770..835
                     /label=MCS
                     /note="multiple cloning site"
ORIGIN
        1 gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggtcgcca ccatggtgag
       61 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
      121 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
      181 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
      241 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
      301 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
      361 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
      421 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
      481 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
      541 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
      601 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
      661 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
      721 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt ccggactcag
      781 atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg gatccaccgg
      841 atctagataa ctgatcataa tcagccatac cacatttgta gaggtt
//
