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    Plasmid map of pDW1775

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    pDW1775
    Gene/insert name: none
    Insert size (bp): Unknown
    Fusion proteins or tags: AcV5
    Terminal: N terminal on backbone
    Vector backbone: pDW1775
    Type of vector: Plant expression
    Backbone size (bp): 5528
    Cloning site 5': XbaI
    Site destroyed during cloning: No
    Cloning site 3': BamHI
    Site destroyed during cloning: No
    5' Sequencing primer: T7
    3' Sequencing primer: T3
    Bacteria resistance: Ampicillin
    High or low copy: High Copy
    Grow in standard E. coli @ 37C: Yes
    Plasmid Provided In: DH5a
    Principal Investigator: Daniel Voytas
    Comments: Plant zinc finger nuclease cloning vector. Zinc fingers are cloned in between unique XbaI and BamHI sites in front of the FokI nuclease domain. pDW1775 consists of the following components in the given order: Hinc II, SexA I, EcoN I, lambda T0 terminator, T7 primer site, BsiW I, rrnB terminator, modified NOS promoter, Bgl II, Xho I, plant leader, MASS translation signal, SV40 nuclear localization signal, AcV5 epitope tag, Xba I, Xma I, Age I, BamH I, plant optimized Fok I nuclease domain, Sac I, NOS terminator, Apa I, T3 primer site, phage FD gene VIII terminator, Bpu 10 I, modified yeast His3 gene, Pac I, yeast CEN/ARS, Bsu36 I (not unique), modified beta lactamase gene (ampicillin resistance), E. coli Trp A terminator, BstE II, Blp I, PpuM I, Rsr II, PflF I, Bgl I, modified high copy ColEI replicon, and back to Hinc II. This vector can be maintained in E. coli and yeast (Saccharomyces cerevisiae) and is also able to express a zinc finger nuclease in yeast and plants using the modified NOS promoter to drive expression. Additionally, this vector was designed to have minimal expression of the zinc finger nuclease in E. coli to facilitate cloning and reduce toxicity effects.



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