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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.