FIG. 2.

One-step construction of plasmids pZR80 and pZR79 from three independent restriction fragments, numbered I (white), II (grey), and III (black) for pZR80 and I, II, and IV (black) for pZR79. The individual fragments have been marked, as have the three relevant restriction sites used for their cloning. Acinetobacter DNA present on the ColE1 vector fragment I is crosshatched. Fragment II has been enlarged at the top of the figure to show the presence of the constitutive P_tet and the unique cloning sites present downstream of this promoter in the small polylinker. Plasmid pZR79 was used to generate transformation recipient ADP1200 and carries a truncated aphA3 gene (aphA3′; see the text).