Integrons are genetic elements which are capable of acquiring genes by site-specific recombination. The most common integron structure consists of two conserved segments flanking a variable region where many different antibiotic resistance genes have been found. The integrons In6 and In7, present in the plasmids pSa and pDGO100, respectively, are unusual in that they include a duplication of the sulI gene which is located within the integron 3′-conserved segment. To further investigate the structure of these integrons, the DNA sequence of the segment located between the two sulI genes was determined. In In7 this segment is 2822 bases long and includes a trimethoprim resistance gene, dhfrX, at one end. The corresponding region in In6 is 4.5 kb and is nearly identical to the In7 segment over the first 2105 bases. In the region unique to In6, a cat gene, conferring chloramphenicol resistance, has replaced the dhfrX gene of In7. This location thus represents a second variable region where different antibiotic resistance genes are found, but the way in which genes become associated with this second variable region is not known. The overall similarity of the structures of In6 and In7 suggests that the additional DNA segments found in these integrons have a common origin, and a possible mechanism for the origin of integrons with partial 3′-conserved segment duplications is presented.