LOCUS       sequence-86577-e         892 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Allows expression of shRNA from U6 promoter and a rescue cDNA from a
            doxycycline (Tet) inducible promoter in the same plasmid. Contains 
            puromycin resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 892)
  AUTHORS   Ma HT, On KF, Tsang YH, Poon RY
  TITLE     An inducible system for expression and validation of the specificity
            of short hairpin RNA in mammalian cells.
  JOURNAL   Nucleic Acids Res. 2007  . 35(4):e22.
  PUBMED    17234679
REFERENCE   2  (bases 1 to 892)
  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: "Nucleic 
            Acids Res. 2007  . 35(4):e22."
COMMENT     Sequence Label: pKAR1/Pur
FEATURES             Location/Qualifiers
     source          1..892
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    41..59
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    83..101
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    125..143
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    167..185
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    209..227
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    251..269
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        302..340
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     337..361
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             443..466
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             473..496
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    528..662
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(552..571)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     606..625
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa
       61 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta
      121 ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat
      181 agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg
      241 agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagctcggt acccgggtcg
      301 aggtaggcgt gtacggtggg aggcctatat aagcagagct cgtttagtga accgtcagat
      361 cgcctggaga cgccatccac gctgttttga cctccataga agagaccggg accgatccag
      421 cctccgcggg ctagccacaa tggactacaa agacgatgac gataaagcca tgctggaagt
      481 tctgtttcag gggcccatgg ctcgagaatt cgagctcggt acccggggat ccagacatga
      541 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      601 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      661 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
      721 tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcct gcaagcctcg
      781 tcgtcctggc cggaccacgc tatctgtgca aggtccccgg ccccggacgc gcgctccatg
      841 agcagagcgc ccgccgccga ggcgaagctt ctaccgggta ggggaggcgc tt
//
