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    Plasmid map of pLKO.1-TRC_cloning_vector

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    pLKO.1 - TRC cloning vector
    Gene/insert name: stuffer
    Insert size (bp): 1900
    Vector backbone: pLKO.1 (Search Vector Database)
    Type of vector: Mammalian expression,Lentiviral,RNAi
    Backbone size (bp): 8901
    Cloning site 5': AgeI
    Site destroyed during cloning: No
    Cloning site 3': EcoRI
    Site destroyed during cloning: No
    5' Sequencing primer: LKO.1 5' (List of Sequencing Primers)
    Bacteria resistance: Ampicillin
    High or low copy: High Copy
    Grow in standard E. coli @ 37C: Yes
    Selectable markers: Puromycin
    Plasmid Provided In: DH5a
    Principal Investigator: David Root
    Comments: This is the recommended vector for cloning and expressing new shRNA sequences. This plasmid is being used by The RNAi Consortium to produce their shRNA library (http://www.broad.mit.edu/genome_bio/trc/). The 1.9kb stuffer can be released with AgeI and EcoRI and replaced with your shRNA sequence of choice.
    The link to the author's map shows the key features of this vector before the 1.9kb stuffer was cloned in. The link to the sequence shows the entire sequence with the stuffer sequence in capital letters.
    Article: A lentiviral RNAi library for human and mouse genes applied to an arrayed viral high-content screen. Moffat J et al. (Cell. 2006 Mar 24. 124(6):1283-98. Pubmed)



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