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    Plasmid map of pLVCT-tTR-KRAB

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    Gene/insert name: CAG promoters, GFP, tTR-KRAB, Tet-on
    Insert size (bp): Unknown
    Vector backbone: None (Search Vector Database)
    Type of vector: Mammalian expression,Lentiviral
    Backbone size (bp): 12877
    5' Sequencing primer: See map (List of Sequencing Primers)
    Bacteria resistance: Ampicillin
    High or low copy: High Copy
    Grow in standard E. coli @ 37C: No
    Bacterial strain for growth and growth condition: Use Stbl3 or HB101 to reduce chance of recombination. Grow at 37C
    Plasmid Provided In: Stbl3
    Principal Investigator: Patrick Aebischer
    Comments: Transgenes can be expressed from any RNA Pol II promoter as part of bicistronic unit comprising the KRAB-based
    repressor; tetO sequences are inserted into the vector LTR. Tet-on and Tet-off versions rely on repressors that bind in
    the absence or the presence of doxycycline, respectively. Addition of Pol III promoter-small hairpin RNA cassette allows
    for drug-controllable RNA interference (Tet-on shRNA).
    Cloning shRNA into pLVET, pLVCT, and pLVPT vectors: first clone shRNA into pLVTHM downstream of the tetO-H1 region.
    Then cut pLVTHM containing your shRNA with MscI-FspI and clone the insert containing the 3'LTR to target plasmid opened
    with MscI-FspI. FspI cuts into AmpR. This means for inverted clones AmpR will not be restored; after selection you will
    be left with clones with the shRNA cassette and functional AmpR.
    Article: A versatile tool for conditional gene expression and knockdown. Szulc J et al. (Nat Methods. 2006 Feb . 3(2):109-16. Pubmed)

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