S2

MATERIAL AND METHODS

Plasmid assembly
MaPylRS coding sequence was synthesised as double stranded DNA fragment (Integrated DNA Technologies) and introduced into the previously reported plasmid encodes all of the 20 canonical amino acids, using the following codons:
ACA (2)), Ala (GCT (1), GCC (7)), Tyr (TAG (1)), His (CAT (4), CAC (12)), Gln (CAG (7)), Asn (AAT (1), ACC (11)), Lys (AAA (1), AAG (19)), Asp (GAT (4),
GAC (14)), Glu (GAA (7), GAG (9)), Cys (TGC (2)), Trp (TGG (1) 
Fluorescence microscopy
The cells were imaged after 24 hours post transfection using Leica DM IL LED inverted fluorescence microscope with QCapture Suite PLUS software.
LC-MS analysis
HEK293 cells were seeded into 75 ml cell culture flasks. After reaching ~ 80% confluency, the media was exchanged for fresh media and the cells were transfected M. alvus (6) CUA

{'paper_id': '52876158', 'title': 'An Evolved Methanomethylophilus alvus Pyrrolysyl-tRNA Synthetase/tRNA Pair Is Highly Active and Orthogonal in Mammalian Cells', 's2_url': 'https://api.semanticscholar.org/CorpusID:52876158', 'source': 'S2ORC'}