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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. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
235609PX459_L31_CRISPR_upstream_sgRNAEncodes CRISPR gRNA for cleaving upstream of insertion site at C terminus of mouse Rpl31
235610PX459_L31_CRISPR_downstream_sgRNAEncodes CRISPR gRNA for cleaving downstream of insertion site at C terminus of mouse Rpl31
235611pCAGEN_GFPEncodes GFP. For use in transfections as a negative control without epitope tags.
235612pCAGEN_N65_eMagBF_V5_NESEncodes ALIBi N-terminal fragment, residues 1-65, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235613pCAGEN_NES_V5_eMagAF_C66Encodes ALIBi C-terminal fragment, residues 66-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235614pCAGEN_N73_eMagBF_V5_NESEncodes ALIBi N-terminal fragment, residues 1-73, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235615pCAGEN_NES_V5_eMagAF_C74Encodes ALIBi C-terminal fragment, residues 74-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235616pCAGEN_N77_eMagBF_V5_NESEncodes ALIBi N-terminal fragment, residues 1-77, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235617pCAGEN_NES_V5_eMagAF_C78Encodes ALIBi C-terminal fragment, residues 78-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235618pCAGEN_N99_eMagBF_V5_NESEncodes ALIBi N-terminal fragment, residues 1-99, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235619pCAGEN_NES_V5_eMagAF_C100Encodes ALIBi C-terminal fragment, residues 100-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235620pCAGEN_N140_eMagBF_V5_NESEncodes ALIBi N-terminal fragment, residues 1-140, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235621pCAGEN_NES_V5_eMagAF_C141Encodes ALIBi C-terminal fragment, residues 141-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235622pCAGEN_N167_eMagBF_V5_NESEncodes ALIBi N-terminal fragment, residues 1-167, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235623pCAGEN_NES_V5_eMagAF_C168Encodes ALIBi C-terminal fragment, residues 168-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235624pCAGEN_N195_eMagBF_V5_NESEncodes ALIBi N-terminal fragment, residues 1-195, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235625pCAGEN_NES_V5_eMagAF_C196Encodes ALIBi C-terminal fragment, residues 196-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235626pCAGEN_N221_eMagBF_V5_NESEncodes ALIBi N-terminal fragment, residues 1-221, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235627pCAGEN_NES_V5_eMagAF_C222Encodes ALIBi C-terminal fragment, residues 222-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235628pCAGEN_N232_eMagBF_V5_NESEncodes ALIBi N-terminal fragment, residues 1-232, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235629pCAGEN_NES_V5_eMagAF_C233Encodes ALIBi C-terminal fragment, residues 233-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235630pCAGEN_N256_eMagBF_V5_NESEncodes ALIBi N-terminal fragment, residues 1-256, fused to eMagBF (enhanced Magnet, basic, fast), V5, and nuclear export sequence.
235631pCAGEN_NES_V5_eMagAF_C257Encodes ALIBi C-terminal fragment, residues 257-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, and nuclear export sequence
235632pCAGEN_NES_V5_GFPEncodes V5-GFP with an N-terminal nuclear export sequence. Used as a no-ALIBi negative control.
235633pCAGEN_NES_V5_GFP_FTAEncodes 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.
235634pCAGEN_NES_V5_BirAEncodes full-length BirA fused to a nuclear export sequence and V5. Used as a positive control in ALIBi experiments
235635pCAGEN_NES_V5_eMagBF_N73Encodes fusion protein of nuclear export sequence, V5, eMagBF (enhanced Magnet, basic, fast), ALIBi N-terminal fragment (residues 1-73) (order reversed)
235636pCAGEN_C74_eMagAF_V5_NESEncodes fusion protein of ALIBi C-terminal fragment (residues 74-321), eMagAF (enhanced Magnet, acidic, fast), V5, nuclear export sequence (order reversed)
235637pCAGEN_NES_V5_eMagAF_C74_Sec61bEncodes 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
235638pCAGEN_NES_V5_eMagAF_C74_MAVSEncodes 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
235639pCAGEN_MyrPalm_NES_V5_eMagAF_C74Encodes ALIBi C-terminal fragment, residues 74-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, nuclear export sequence, MyrPalm for targeting to plasma membrane
235640pCAGEN_V5_eMagAF_C74_cMycNLSEncodes ALIBi C-terminal fragment, residues 74-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, cMyc nuclear localization sequence for targeting to nucleus
235641pCAGEN_V5_Fbl_eMagAF_C74Encodes ALIBi C-terminal fragment, residues 74-321, fused to eMagAF (enhanced Magnet, acidic, fast), V5, Fbl for targeting to nucleolus
235642pCAGEN_N73_eMagBF_V5Encodes ALIBi N-terminal fragment, residues 1-73, fused to eMagBF (enhanced Magnet, basic, fast), V5. No nuclear export sequence to allow nuclear localization.
235643pCAGEN_Fbl_N73_eMagBF_V5Encodes ALIBi N-terminal fragment, residues 1-73, fused to eMagBF (enhanced Magnet, basic, fast), V5, Fbl for targeting to nucleolus.

Antibodies from Article