LOCUS       sequence-156552-         877 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses eGFP along with an U6 promoter driven gRNA which cleaves 
            the plasmid in-cell, and also confers resistance to puromycin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 877)
  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 877)
  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-puro
FEATURES             Location/Qualifiers
     source          1..877
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      48..57
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(54..56,57..59,60..770)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(54..56,57..59,60..770)
                     /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(99..120)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(360..379)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     707..728
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    771..836
                     /label=MCS
                     /note="multiple cloning site"
ORIGIN
        1 agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggtcgcc accatggtga
       61 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
      121 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
      181 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
      241 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
      301 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
      361 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
      421 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
      481 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
      541 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
      601 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
      661 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
      721 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag tccggactca
      781 gatctcgagc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccaccg
      841 gatctagata actgatcata atcagccata ccacatt
//
