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During recent years, several Tn554-related transposons have been identified in Gram-positive bacteria.1,2 These transposons share a rather conserved transposition module comprising the genes tnpA, tnpB and tnpC, whereas their antimicrobial resistance gene content varies considerably, such as erm(A) and spc in Tn554, erm(A), spc and vga(E) in Tn6133, erm(A), spc, fexA and optrA in Tn6674, fexA and optrA in Tn6823, fexA in Tn558, lnu(G) in Tn6260, vga(A) in Tn5406 and dfrK in Tn559. Another transposon of the Tn554 family, Tn6188, harbours the biocide resistance gene qacH (Figure 1a). All these transposons move by a unique mechanism as identified in studies on serial transposition of Tn554. With each new transposition event, the 6 bp sequence representing the target site is located at the left junction of Tn554, while the 6 bp sequence representing the former left junction is now present at the right junction and the former 6 bp sequence at the right junction is lost.3–5 Tn554-like transposons often use the chromosomal radC gene as a preferential integration site and do not form target site duplications at their integration site.5,6 In the present study, we describe a new member of the Tn554 family, which is only distantly related to Tn554 and apparently prefers a different integration site.