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. 2010 Oct 8;5(10):e13244. doi: 10.1371/journal.pone.0013244

Figure 1. Construction of pBAV1K-T5-lux, a very broad host range expression vector.

Figure 1

The cryptic plasmid, pWV01, exhibits broad host range but is unstable in many species. The ORF D, and inverted repeats IV, V and VI were deleted from its plasmid origin; terminators t0 and T1 were inserted on opposite ends of the shortened origin (upper right). The selectable marker, the Enterococcus 3′,5″-aminoglycoside phosphotransferase type III, and a T5 promoter within a BioBrick multiple cloning site were cloned into the plasmid (top circle). The lux genes of Photorhabdus luminescens were individually PCR amplified, cloned, assembled with ribosome binding sites (middle) and cloned into the plasmid to create pBAV1k-T5-luxABCDE (bottom circle).