Structure of Tn5062 and the osaAB gene region. (A) Tn5062 is comprised of the following elements (not drawn to scale): two Tn5 mosaic ends (ME) flanking a three-frame translational stop and a consensus streptomycete ribosome-binding site (rbs) preceding a promoterless copy of eGFP, the apramycin resistance gene, aac3(IV), itself flanked by two T4 transcriptional terminators, and oriT(RK2). The locations of the two sequencing primers, EZL2 and EZR1, are indicated. (B) Arrows above the osaAB gene region indicate the location of representative Tn5062 insertions (1–7) and the Tn5493 insertion (x). Tn5062 insertions 1–6 in osaAB are all oriented to place eGFP under transcriptional control of osaAB promoters. S. coelicolor strain AB201 contains an insertion at position 1 (position 6281495 in the chromsome), and strain AB001 contains an insertion at position 5 (position 6285442). The extent of the fragment and the restriction sites used for cloning osaB and its promoter for complementation are indicated beneath the gene. This gene was initially cloned linked to aac3(IV) of the Tn5062 insertion in SCO5751 (insertion 7).
