FIG. 2.
Construction of a Tn5-based insertion delivery plasmid containing an inducible cell lysis system based on lysis gene E from bacteriophage φX174. Restriction sites relevant for the constructions are shown. Plasmid pUC18Not and the pUT-based plasmid pJMSB4 have been described elsewhere (9, 33). Abbreviations: geneE, lysis gene E of bacteriophage φX174; RBS, ribosome binding site from the E. coli expression plasmid pUHE24-1 (16); PA1-04/03, synthetic lactose promoter from plasmid pUHE24-1; ori, origin of replication; ori R6K, origin of replication dependent on the Pir protein; ori ColE1, origin of replication from plasmid ColE1; oriT RP4, origin of transfer; tnp*, transposase; ′lacI, gene encoding the repressor protein for the lac operon.