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Spatiotemporal functional assembly of split protein pairs through a light-activated SpyLigation.

Ruskowitz ER, Munoz-Robles BG, Strange AC, Butcher CH, Kurniawan S, Filteau JR, DeForest CA
Nat Chem. 2023 May;15(5):694-704. doi: 10.1038/s41557-023-01152-x. Epub 2023 Apr 17. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
207629pSC-LPETG A plasmid encoding photocaged SpyCatcher (pSC) with a C-terminal Sortase recognition sequence.
207630nUnaG-ST A plasmid encoding the N-terminal UnaG fragment fused to SpyTag.
207631pSC-cUnaG A plasmid encoding photocaged SpyCatcher (pSC) fused to C-terminal UnaG fragment.
207632pSC-SmBiT A plasmid encoding photocaged SpyCatcher (pSC) fused to SmBiT
207633LgBiT-ST A plasmid encoding LgBiT fused to SpyTag
207634pSC-cUnaG-P2A-nUnaG-ST-P2A-mChA plasmid encoding photocaged SpyCatcher (pSC)-cUnaG, nUnaG-SpyTag (ST), and mCherry separated by P2A cleaving sequences for mammalian expression.
207635pSC-cUnaG-LPETGA plasmid encoding photocaged SpyCatcher (pSC) fused to C-terminal UnaG fragment and LPETG sortase recognition sequence.
207636pSC-mCh-P2A-ST-EGFP-CAAXA plasmid encoding photocaged SpyCatcher (pSC) fused to mCherry and a SpyTag-EGFP localized to the cell membrane via CAAX tag
207637EGFP-ST-P2A-pSC-CAAXA plasmid encoding EGFP fused to SpyTag (ST) and photocaged SpyCatcher (pSC) localized to the cell membrane via CAAX tag.
207638nCre-ST-NLS-P2A-NLS-pSC-cCreA plasmid encoding N-terminal Cre fused to SpyTag (ST) and photocaged SpyCatcher (pSC) fused to C-terminal Cre with a nuclear localization signal (NLS) sequence
207639pEVOL-NBK-1 A plasmid for the expression of M. mazei pyrrolysine aminoacyl-tRNA synthetase/tRNA pair (PylRS/tRNAPyl) in E. Coli.

Antibodies from Article