FIG. 2.

Consensus map of the antibiotic resistance region of pSEM. Thin arrows show the orientation of genes. Δ5′CS and 3′CS are the 5′ and 3′ conserved region of the In-t3 integron. The short boldface arrows indicate the position and direction of primers used in PCR amplifications. The aacA4R-OS6 amplicon was obtained by the Expand 20 kb PCR system (Boehringer GmbH, Mannheim, Germany). The DNA sequence of the aacA4R primer is: 5′-TGTGACGGAATCGTTGC-3′. Thin bars below the map represent cloned inserts used for sequencing, with the plasmid designation given on the right of each construct. Dashed lines show the In-t3 and In-t3/bla_SHV-5 intervening sequences released under EMBL accession no. AJ009820 and AJ245670, respectively. Black thick bars represent pSEM regions homologous to pPAG-KE, pPa-1, pZMP1, and pACM1 plasmids; hatched thick bars are gaps between homologous regions; white thick bars are nonhomologous regions. The numbering is relative to positions of nucleotides in the released plasmid sequences.