FIG. 2.

Representation of the 9,505-bp M13-attTn7 bacteriophage genome and the position of the TnsE-meditated mini-Tn7 insertion events. Numbering follows the previously established convention for M13mp18 (28). The M13mp18 sequence extends from bp 8529 to 9505/1 to 6266. The M13 origin (ori; bp 5487 to 5867), ′glmS (bp 6267 to 7371), pstS′ (bp 7708 to 8529), and the position of TnsD-mediated insertion events (attTn7) are indicated. Independent transposition events are indicated by arrows either inside (left-to-right insertion events) or outside (right-to-left insertion events) the circle. The positions of mini-Tn7 insertion events are indicated in parentheses along with the strain background according to the following letter code: A, pCW15 (TnsABC) pJP104 (TnsE^wt); B, pCW15 pJP103 (TnsE^A453V); C, pCW15 pJP103 (TnsE^D523N); D, pCW15 pJP103 (TnsE^A453V+D523N). The position and orientation of individual transposition events were determined by isolating the replicative (double-stranded) form of the bacteriophage and sequencing from the left end of the element using a primer complementary to this end. To confirm that the process of transposition was responsible for relocating the element into the bacteriophage, we sequenced a subset of the insertions from both ends and in all cases identified a 5-bp duplication that is indicative of Tn7 transposition. The asterisk indicates where identical insertions were found in this experiment that are likely siblings.