FIGURE 3.

SMARTT schematic. (A) Schematic of the strategy used for Sequencing-based Mutational Analysis of RNA Transcription Termination (SMARTT). Mutant libraries were produced through error-prone PCR (mutations are depicted by yellow boxes) and used as template DNA in transcription reactions containing varying amounts of ligand. The resulting RNA was prepared for high-throughput sequencing (Illumina adapters are shown in cyan). Each sequence was computationally analyzed to produce response profiles for all variants with zero or one mutations between positions 8 and 235 of the parent construct simultaneously. Representative response profiles are shown for mutants that alter the fit parameters Y_min (B), Y_max (C), amplitude (D), and K_1/2 (E). Error bars represent the standard deviation across two replicates at each data point. Figure and legend are adapted from Torgerson et al. (2018).