KEY RESOURCES TABLE

REAGENT or RESOURCE	SOURCE	IDENTIFIER
Bacterial and Virus Strains
Escherichia coli DH5α	Thermo Fisher	Cat# 18265017
Escherichia coli BL21 (DE3)	Thermo Fisher	Cat# EC0114
Chemicals, Peptides, and Recombinant Proteins
AlexaFluor488 (A488) NHS ester	Thermo Fisher	Cat# A20000
Cy3 maleimide mono-reactive dye	GE Healthcare	Cat# PA23031
Cy5 NHS ester mono-reactive dye	GE Healthcare	Cat# PA15101
RNaseOUT Recombinant ribonuclease inhibitor	Thermo Fisher	Cat# 10777019
Gibson Assembly Master mix	New England Biolabs	Cat# E2611S
Klenow Fragment	New England Biolabs	Cat# M0212
Biotin-11-dUTP	Thermo Fisher	Cat# R0081
Biotin-14-dCTP	Thermo Fisher	Cat# 19518018
Biotin-14-dATP	Thermo Fisher	Cat# 19524016
Trolox	Sigma-Aldrich	Cat# 238813
4-Nitrobenzyl alcohol (NBA)	Sigma-Aldrich	Cat# N12821
Cyclooctatetraene (COT)	Sigma-Aldrich	Cat# 138924
Protocatechuate 3,4-Dioxygenase from Pseudomonas sp. (PCD)	Sigma-Aldrich	Cat# P8279
3,4-Dihydroxybenzoic acid (PCA)	Sigma-Aldrich	Cat# 37580
Streptavidin-coated polystyrene particles (2.0–2.9 μm)	Spherotech	Cat# SVP-20-5
Rifampicin	Sigma-Aldrich	Cat# R3501
S6-RNAP	This paper	N/A
σ70	40	N/A
Sfp synthase	41	N/A
Recombinant DNA
pVS11-Sfp-rpoC (expressing S6-RNAP)	This paper	N/A
pLW83 (Template1)	This paper	N/A
pLW86 (Template2)	This paper	N/A
pLW87 (Template3)	This paper	N/A
pLW88 (Template4)	This paper	N/A
pLW89 (Template5)	This paper	N/A
pLW103 (Template6)	This paper	N/A
Software and Algorithms
Bluelake	LUMICKS	https://lumicks.github.io/bluelake-api/
Origin	OriginLab	https://www.originlab.com
Prism	GraphPad	https://www.graphpad.com/scientific-software/prism/
Python scripts for single-molecule data analysis	This paper	doi:10.5281/zenodo.7618698
Deposited data
Unprocessed single-molecule images	This paper	doi:10.17632/djgmvpxjj9.1
Unprocessed gels	This paper	doi:10.17632/yp3x9k53gf.1