A promoter-probe shuttle plasmid (pGL7011) containing the promoterless aminoglycoside--acetyltransferase I gene () of Tn was used to isolate DNA fragments from phage 119 that possessed promoter activity in TK23. Analysis of gentamicin (Gm) resistance levels in and in TK23, and of mRNA levels in identified a fragment (F14) that exhibited a high level of promoter activity in both species. Subsequent analysis revealed that the promoter activity of SF14 (a subcloned fragment of F14) was about twice that of , one of the strongest characterized actinomycete promoters. SF14 contained two tandemly arranged promoters, 14-1p and p14-llp, with overlapping and adjacent -10 and -35 regions, respectively. Both promoters appear to be recognized with different efficiencies by the major RNA polymerase holoenzyme (Es) of A3(2).


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