Source data for: Effect of seed length and binding motifs on Hfq-mediated sRNA-mRNA annealing analyzed using single-molecule FRET

This dataset contains raw and processed single-molecule FRET (smFRET) data used to quantify Hfq-mediated annealing between bacterial sRNAs and mRNAs. It includes fluorescence time traces, extracted dwell times, FRET distributions, and analysis scripts supporting measurements of how seed length, Hfq-binding motifs, and motif spacing affect interaction stability and kinetics. The data enable independent reproduction of the analyses and reuse for modeling RNA–RNA interaction dynamics and RNA chaperone function. Funding: This research was funded by the National Science Centre, Poland, grant no 2022/46/E/NZ1/00462 and an EMBO Installation Grant IG 5730-2024.