KEY RESOURCES TABLE

REAGENT or RESOURCE	SOURCE	IDENTIFIER
Bacterial and virus strains
E. coli BL21-DE3	NEB	C2527H
Chemicals, peptides, and recombinant proteins
NEBNext High-Fidelity 2x PCR Master Mix	NEB	M0541
SYBR Green	Invitrogen	S7563
SYBR Gold	Invitrogen	S11494
PhiX Control V3	Illumina	110-3001
HiTrap SP HP	GE Healthcare	17115201
Proteinase K	NEB	P8107S
Alexa Fluor 647-aminohexylacrylamido-dUTP	Life Technologies	A32763
Alexa Fluor 555 NHS Ester	Life Technologies	A20009
Klenow fragment (3′-to-5′ exo‐minus)	NEB	M0212
Cover glasses (24 Å~ 60 mm)	Gold Seal	Cat. #3423
glass coverslips (25 Å~ 25 mm)	Gold Seal	Cat. #3307
NanoStrip	KMG Chemicals	210034
Streptavidin	NEB	N7021S
Heparin	Sigma	H4784
Protocatechuic acid	Aldrich	37580
Pseudomonas sp. protocatechuate 3,4-Dioxygenase	Sigma	P8279
trolox	Aldrich	238813
propyl gallate	Sigma	P3130
4-nitrobenzyl alcohol	Aldrich	N12821
Critical commercial assays
QIAquick PCR Purification Kit	QIAGEN	28106
MiSeq Reagent Kit v3 (150 cycle)	Illumina	MS-102-3001
Deposited data
Mendeley Data: Fit parameters for TtAgo binding and cleavage	This Paper	http://doi.org/10.17632/vrb6n8gm3x.1
Oligonucleotides
Table S2	This Paper
Recombinant DNA
pET-SUMO	Invitrogen	K30001
Software and algorithms
NUPACK	J. N. Zadeh, C. D. Steenberg, J. S. Bois, B. R. Wolfe, M. B. Pierce, A. R. Khan, R. M. Dirks, N. A. Pierce. NUPACK: analysis and design of nucleic acid systems. J Comput Chem, 32:170-173, 2011	http://www.nupack.org/
GelAnalyzer 19.1	GelAnalyzer 19.1 by Istvan Lazar Jr., PhD and Istvan Lazar Sr., PhD, CSc	www.gelanalyzer.com
CoSMoS pipeline	C. S. Smith, K. Jouravleva, M. Huisman, S.M. Jolly, P. D. Zamore, D. Grunwald. An automated Bayesian pipeline for rapid analysis of single-molecule binding data. Nat Commun. 2019 Jan 17;10(1):272.	https://github.com/quantitativenanoscopy/cosmos_pipeline
TtAgo curve fitting scripts	This Paper	https://github.com/GreenleafLab/TtAgo and Mendeley Data: http://doi.org/10.17632/vrb6n8gm3x.1
Other