FIGURE 5.

Schematic of a Tn1721-mediated plasmid-to-plasmid translocation. (A) The genetic content of Tn1721 translocated to plasmid p4 and the original KPC locus carried by Tn1721 from plasmid p2 in strain KP20194a2. IRL, the left terminal inverted repeat of Tn1721; IRR, the right terminal inverted repeat of Tn1721. DR, Tn1721-mediated translocation resulted in the generation of a 5-bp direct repeat at the target site. Arrow with a dotted line indicates the site of the additional IRL2 in Tn1721-like transposon-mediated bla_KPC–2 translocation reported elsewhere (Tang et al., 2017), which was not seen in our strains or other highly similar plasmids from strains KP58 and L39_2. The arrows around Tn1721 indicate the relative location of the designed primers (but not the length). (B) PCR was performed with the indicated primers using genomic DNA extracted from KP20194a or KP20194a2 as a template to confirm the targeted insertion. The amplified fragment was verified by Sanger sequencing. Strains: a, KP20194a; a2, KP20194a2. Primer locations are indicated in (A) and sequences in Supplementary Table 1.