REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
Mouse anti-CD69 conjugated to Alexa647 (FN50)	Thermo Fisher Scientific	Cat#: MA5-18150; RRID: AB_2539524
anti-phosphoERK (rabbit polyclonal)	Cell Signaling Technology	Cat#: 9101; RRID:AB_331646
Chemicals, Peptides, and Recombinant Proteins
His10-SNAPf-ybbr13	This study
His10-CLIPf-ybbr13	This study
Latrunculin A	Sigma	Cat#: L5163-100UG
BC-GLA-NHS	New England Biolabs	Cat#: S9237S
BG-GLA-NHS	New England Biolabs	Cat#: S9151S
Puromycin dihydrochloride	Sigma	Cat#: P8833-10MG
Experimental Models: Cell Lines
Human: JRT3	Art Weiss; Ohashi et al., 1985
Human: JRT3 expressing DNA-CARζ-GFP	This study
Human: JRT3 expressing DNA-CARζ-GFP/ZAP70-mCherry	This study
Human: JRT3 expressing DNA-CARζ-IRESpuro/ZAP70-GFP	This study
Human: JRT3 expressing DNA-CARζ-IRESpuro/ZAP70-GFP	This study
Human: JRT3 expressing DNA-CARTCR/ZAP70-GFP	This study
Human: P116	Art Weiss; Williams et al., 1998
Human: P116 expressing DNA-CARζ-IRESpuro/ZAP70-GFP	This study
Human: HEK293T	ATCC	Cat#: CRL-3216
Recombinant DNA
pHR-DNA-CARζ-GFP	This study
pHR-DNA-CARζ-IRESpuro	This study
pHR-SP-SNAPf:TCRβ-IRESpuro	This study
pHR-TCRα-E2A-SNAPf:TCRβ-P2A-CD3ε-P2A-CD3ζ	This study
pHR-ZAP70-GFP	James and Vale, 2012
pHR-ZAP70-mCherry	James and Vale, 2012
Oligonucleotides
A full list of oligonucleotides is presented in Table S1.	N/A	N/A
Software and Algorithms
GraphPad Prism v6	GraphPad	http://www.graphpad.com/scientific-software/prism/
MATLAB	Mathworks	https://www.mathworks.com/
vbFRET	Bronson et al., 2009	http://vbfret.sourceforge.net/
FIJI	NIH	https://fiji.sc/
Cell Profiler 2.1.1	The Broad Institute	http://cellprofiler.org/
μManager	Open Imaging	https://open-imaging.com/
C script to analyze stochastic signaling model	This study	https://github.com/kabirhusain/mjtayloretal_clustergillespie