A subcellular map of translational machinery composition and regulation at the single-molecule level.

Zhang Z, Xu A, Bai Y, Chen Y, Cates K, Kerr C, Bermudez A, Susanto TT, Wysong K, Garcia Marques FJ, Nolan GP, Pitteri S, Barna M
Science. 2025 Mar 7;387(6738):eadn2623. doi: 10.1126/science.adn2623. Epub 2025 Mar 7. PubMed Article

Plasmids from Article

ID	Plasmid	Purpose
235608	pCR4-TOPO_L31-FLAG-TEV-AviTag_repair_template	Repair template for inserting FLAG-TEV protease cleavage site-AviTag into the C terminus of mouse Rpl31 via CRISPR
235609	PX459_L31_CRISPR_upstream_sgRNA	Encodes CRISPR gRNA for cleaving upstream of insertion site at C terminus of mouse Rpl31
235610	PX459_L31_CRISPR_downstream_sgRNA	Encodes CRISPR gRNA for cleaving downstream of insertion site at C terminus of mouse Rpl31
235611	pCAGEN_GFP	Encodes GFP. For use in transfections as a negative control without epitope tags.
235612	pCAGEN_N65_eMagBF_V5_NES	Encodes ALIBi N-terminal fragment, residues 1-65, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235613	pCAGEN_NES_V5_eMagAF_C66	Encodes ALIBi C-terminal fragment, residues 66-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235614	pCAGEN_N73_eMagBF_V5_NES	Encodes ALIBi N-terminal fragment, residues 1-73, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235615	pCAGEN_NES_V5_eMagAF_C74	Encodes ALIBi C-terminal fragment, residues 74-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235616	pCAGEN_N77_eMagBF_V5_NES	Encodes ALIBi N-terminal fragment, residues 1-77, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235617	pCAGEN_NES_V5_eMagAF_C78	Encodes ALIBi C-terminal fragment, residues 78-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235618	pCAGEN_N99_eMagBF_V5_NES	Encodes ALIBi N-terminal fragment, residues 1-99, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235619	pCAGEN_NES_V5_eMagAF_C100	Encodes ALIBi C-terminal fragment, residues 100-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235620	pCAGEN_N140_eMagBF_V5_NES	Encodes ALIBi N-terminal fragment, residues 1-140, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235621	pCAGEN_NES_V5_eMagAF_C141	Encodes ALIBi C-terminal fragment, residues 141-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235622	pCAGEN_N167_eMagBF_V5_NES	Encodes ALIBi N-terminal fragment, residues 1-167, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235623	pCAGEN_NES_V5_eMagAF_C168	Encodes ALIBi C-terminal fragment, residues 168-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235624	pCAGEN_N195_eMagBF_V5_NES	Encodes ALIBi N-terminal fragment, residues 1-195, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235625	pCAGEN_NES_V5_eMagAF_C196	Encodes ALIBi C-terminal fragment, residues 196-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235626	pCAGEN_N221_eMagBF_V5_NES	Encodes ALIBi N-terminal fragment, residues 1-221, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235627	pCAGEN_NES_V5_eMagAF_C222	Encodes ALIBi C-terminal fragment, residues 222-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235628	pCAGEN_N232_eMagBF_V5_NES	Encodes ALIBi N-terminal fragment, residues 1-232, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235629	pCAGEN_NES_V5_eMagAF_C233	Encodes ALIBi C-terminal fragment, residues 233-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235630	pCAGEN_N256_eMagBF_V5_NES	Encodes ALIBi N-terminal fragment, residues 1-256, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235631	pCAGEN_NES_V5_eMagAF_C257	Encodes ALIBi C-terminal fragment, residues 257-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235632	pCAGEN_NES_V5_GFP	Encodes V5-GFP with an N-terminal nuclear export sequence. Used as a no-ALIBi negative control.
235633	pCAGEN_NES_V5_GFP_FTA	Encodes V5-GFP with an N-terminal nuclear export sequence and a C-terminal FLAG-TEV cleavage site-AviTag. Used to express a control substrate for ALIBi.
235634	pCAGEN_NES_V5_BirA	Encodes full-length BirA fused to a nuclear export sequence and V5. Used as a positive control in ALIBi experiments
235635	pCAGEN_NES_V5_eMagBF_N73	Encodes fusion protein of nuclear export sequence, V5, eMagBF (enhanced Magnet, basic, fast), ALIBi N-terminal fragment (residues 1-73) (order reversed)
235636	pCAGEN_C74_eMagAF_V5_NES	Encodes fusion protein of ALIBi C-terminal fragment (residues 74-321), eMagAF (enhanced Magnet, acidic, fast), V5, nuclear export sequence (order reversed)
235637	pCAGEN_NES_V5_eMagAF_C74_Sec61b	Encodes ALIBi C-terminal fragment, residues 74-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, nuclear export sequence, Sec61b for targeting ALIBi to endoplasmic reticulum
235638	pCAGEN_NES_V5_eMagAF_C74_MAVS	Encodes ALIBi C-terminal fragment, residues 74-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, nuclear export sequence, MAVS residues 500-540 for targeting to outer mitochondrial membrane
235639	pCAGEN_MyrPalm_NES_V5_eMagAF_C74	Encodes ALIBi C-terminal fragment, residues 74-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, nuclear export sequence, MyrPalm for targeting to plasma membrane
235640	pCAGEN_V5_eMagAF_C74_cMycNLS	Encodes ALIBi C-terminal fragment, residues 74-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, cMyc nuclear localization sequence for targeting to nucleus
235641	pCAGEN_V5_Fbl_eMagAF_C74	Encodes ALIBi C-terminal fragment, residues 74-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, Fbl for targeting to nucleolus
235642	pCAGEN_N73_eMagBF_V5	Encodes ALIBi N-terminal fragment, residues 1-73, fused to eMagBF (enhanced Magnet, basic, fast), V5. No nuclear export sequence to allow nuclear localization.
235643	pCAGEN_Fbl_N73_eMagBF_V5	Encodes ALIBi N-terminal fragment, residues 1-73, fused to eMagBF (enhanced Magnet, basic, fast), V5, Fbl for targeting to nucleolus.
236087	pBX_RPL7a_smRuby2_HA	Constitutively encodes RPL7a-smRuby2-HA (mRuby2-based spaghetti monster fluorescent protein embedding HA epitope tags), for targeting the 60S subunit.
236088	pBX_RPS2_smGFP_V5	Constitutively encodes RPS2-smGFP-V5 (sfGFP-based spaghetti monster fluorescent protein embedding V5 epitope tags), for targeting the 40S subunit.
236089	pBX_RPS8_smRuby2_HA	Constitutively encodes RPS8-smRuby2-HA (mRuby2-based spaghetti monster fluorescent protein embedding HA epitope tags), for targeting RPS8 in the 40S subunit.
236090	pBX_RPS25_smRuby2_HA	Constitutively encodes RPS25-smRuby2-HA (mRuby2-based spaghetti monster fluorescent protein embedding HA epitope tags), for targeting RPS25 in the 40S subunit.
236091	pBX_RPL29_smGFP_V5	Constitutively encodes RPL29-smGFP-V5 (sfGFP-based spaghetti monster fluorescent protein embedding V5 epitope tags), for targeting RPL29 in the 60S subunit.
236092	pBX_Sec61b_smGFP_Myc	Constitutively encodes Sec61b-smGFP-Myc (sfGFP-based spaghetti monster fluorescent protein embedding Myc epitope tags), for targeting the endoplasmic reticulum.
236093	pcDNA4TO_24xGCN4_MUT_24xPP7	Constitutively encodes 24xGCN4-MUT-24xPP7 stem loops, for visualization of the Mut mRNA.
236094	pcDNA4TO_24xGCN4_PCC_24xPP7	Constitutively encodes 24xGCN4-PCC-24xPP7 stem loops, for visualization of the Pcc mRNA.