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Significance To meet the increasing demands of drug discovery and biosynthetic studies, we established a precise quantitative method based on flow cytometry at single-cell (protoplast) resolution inStreptomyces for the identification of regulatory elements. A series of native or synthetic promoters and ribosomal binding sites has been characterized. Moreover, an insulator was demonstrated to eliminate element–element interference. As a proof of concept, a native silent gene cluster was activated by the synthetic modular regulatory elements in a predictable manner. The universality of these elements is of high value to the synthetic biology ofStreptomyces .

Exploiting a precise design of universal synthetic modular regulatory elements to unlock the microbial natural products in Streptomyces