LOCUS       sequence-158962-         818 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 818)
  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 818)
  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..818
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(276..295)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    320..367
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     583..602
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        681..756
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(782..801)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
       61 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
      121 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
      181 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
      241 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
      301 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
      361 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccacgccta
      421 tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga gataattgga
      481 attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa
      541 tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc
      601 gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc
      661 ggcgcaacgc gatcgcgtaa gttttagagc tagaaatagc aagttaaaat aaggctagtc
      721 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgcggccgc taagatacat
      781 tgatgagttt ggacaaacca caactagaat gcagtgaa
//
