FIGURE 1.

Riboswitch-mediated reporter expression under the control of a potent promoter. Schematic diagram of a potent promoter dampening strategy with one (A) or two (B) riboswitches. (C) Key features of pVK-f-lux, a parts-swappable mini-CTX-lux derived backbone for convenient cloning of promoters and RNA regulatory parts. MCS1 contains CsiI-XmaJI-SacI restriction sites and MCS2 contains NcoI-SdaI-ScaI restriction sites. A type IIS restriction site is located at the beginning of luxC allowing digestion into the second codon for scar-free translational fusion. GA overlaps for double digestions with SacI and NcoI or ScaI and AarI have been designed for allowing interchangeability of parts as described (Supplementary Material: Quick User Manual for pVK-f-lux). These double digestions are buffer compatible and yield fragments visible on an agarose gel (approximately 300 and 400 bp in length, see Supplementary Figure S3 for more details). (D) Plasmid map of pVK-f-lux and pVK-f2-lux.