REAGENT or RESOURCE	SOURCE	IDENTIFIER
Bacterial and virus strains
E.coli BL21 (DE3) pLysS	Thermo Fisher Scientific	Cat# 10604973
E.coli DH5α	Thermo Fisher Scientific	Cat# 11583117
E.coli DH10Bac	Thermo Fisher Scientific	Cat# 10361-012
Chemicals, peptides, and recombinant proteins
DSS-H12/D12	Creative Molecules Inc	Cat# 001S
Trypsin	Promega, UK	Cat# V5111
Glu-C protease	Promega, UK	Cat# V1651
2XFlag-peptide	Peptide synthetics, UK	Quote Q23477
L-Glutathione reduced	Thermo Fisher Scientific	Cat# 10628343
ANTI-FLAG® M2 Affinity Gel	Sigma-Aldrich	Cat# A2220
Complete, EDTA-free protease inhibitor tablets	Roche Diagnostics	Cat# 05056489001
PhosSTOP	Sigma-Aldrich	Cat# 4906845001
PreScission and TEV protease	University of Sussex	N/A
Phospho-4EBP1 (Thr37/46) polyclonal antibody	Cell Signaling Technology	Cat# 236B4
Critical commercial assays
PiColorLock Gold Phosphate Detection System	Innova Biosciences	Cat# 303-0030
Cellfectin II Reagent	Thermo Fisher Scientific	Cat# 10362100
Deposited data
R2TP-TTT cryoEM volume	this work	Electron Microcopy Databank (EMDB): EMD:12957
R2-TTT cryoEM volume	this work	Electron Microcopy Databank (EMDB): EMD:12979
Atomic Coordinates for R2-TTT	this work	Protein Databank (PDB): PDB:7OLE
Experimental models: Cell lines
Sf9 cells	Thermo Fisher Scientific	Cat# 11496015
Recombinant DNA
P3E plasmid	University of Sussex	N/A
pFBDM plasmid expressing human and yeast TTT and mTOR-KD proteins	This study	N/A
KmTOR-KmLst8	A Gift from Prof. Roger Williams	N/A
Software and algorithms
PRISM 8.0	GraphPad	https://www.graphpad.com/scientific-software/prism/
Relion-3.1	Zivanov et al., 2018	https://www2.mrc-lmb.cam.ac.uk/groups/scheres/impact.html
EPU software	Thermo Fisher Scientific	https://www.fei.com/software/epu-automated-single-particles-software-for-life-sciences/
MotionCor2	Zheng et al., 2017	https://msg.ucsf.edu/software
GCTF and CTFFIMND4	Zhang, 2016; Rohou and Grigorieff, 2015	https://www2.mrc-lmb.cam.ac.uk/research/locally-developed-software/zhang-software/#gctf and https://grigoriefflab.umassmed.edu/ctf_estimation_ctffind_ctftilt
Phenix	Adams et al., 2010	https://www.phenix-online.org/
Coot	Emsley and Cowtan, 2004	https://www2.mrc-lmb.cam.ac.uk/personal/pemsley/coot
Pymol	Schrödinger	https://pymol.org/2/
Chimera	Pettersen et al., 2004	http://www.rbvi.ucsf.edu/chimera
I-TASSER	Yang et al., 2015	https://zhanglab.ccmb.med.umich.edu/I-TASSER/
Phyre2	Kelley et al., 2015	http://www.sbg.bio.ic.ac.uk/∼phyre2/html/page.cgi?id=index
Other
Quantifoil 1.2/1.3, 300 mesh copper	Agar Scientific	Cat# AGS143-2
FEI-Vitrobot MAG IV	Thermo Fisher Scientific	N/A
Leica EM GP2	Leica microsystems	N/A
Titan Krios	Thermo Fisher Scientific	N/A
K2 direct detector camera	GATAN	N/A