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Yeast homologous recombination-based promoter engineering for the activation of silent natural product biosynthetic gene clusters To solve this problem, we added random 15-bp in- sulator sequences and unique natural Streptomyces RBSs (19) to 24 promoters identified by Seghezzi et al. (18). The spacer and RBS sequences were matched to give an overall GC content of ～65% for each promoter element. Pairs of promoter/spacer/RBS sequences were synthesized as primers containing 20-bp sequences homologous to the ends of yeast selectable marker genes. |
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