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Generating Efficient Methanomethylophilus alvus Pyrrolysyl-tRNA Synthetases for Structurally Diverse Non-Canonical Amino Acids.
Avila-Crump S, Hemshorn ML, Jones CM, Mbengi L, Meyer K, Griffis JA, Jana S, Petrina GE, Pagar VV, Karplus PA, Petersson EJ, Perona JJ, Mehl RA, Cooley RB
ACS Chem Biol. 2022 Dec 16;17(12):3458-3469. doi: 10.1021/acschembio.2c00639. Epub 2022 Nov 16.
PubMed Article

Plasmids from Article

ID Plasmid Purpose
197566pAcBac1-Ma-Acridone-RS1 (A7)Expression of Methanomethylophilus alvus (Ma) Acridone RS1 (A7) tRNA synthetase/Pyl-tRNA pair for the encoding of Acridone at TAG codons in HEK293 cells
197567pAcBac1-Ma-sfGFP WTExpression of sfGFP WT with Methanomethylophilus alvus (Ma) Pyl-tRNA for the encoding of non-canonical amino acids at TAG codons in HEK293 cells
197568pAcBac1-Ma-sfGFP 150TAGExpression of sfGFP-150TAG with Methanomethylophilus alvus (Ma) Pyl-tRNA for the encoding of non-canonical amino acids at TAG codons in HEK293 cells
197569pAcBac1-Mb-sfGFP WTExpression of sfGFP WT with Methanosarcina barkeri (Mb) Pyl-tRNA for the encoding of non-canonical amino acids at TAG codons in HEK293 cells
197570pAcBac1-Mb-sfGFP 150TAGExpression of sfGFP 150TAG with Methanosarcina barkeri (Mb) Pyl-tRNA for the encoding of non-canonical amino acids at TAG codons in HEK293 cells
197571Ma-pREPPositive selection plasmid for selecting ncAA-encoding Methanomethylophilus alvus Pyl-RS mutants. Expresses TAG-codon interrupted chloramphenicol resistance cassette and M. alvus Pyl-tRNA(6). p15a ori
197572Ma-BarnaseNegative selection plasmid for selecting ncAA-encoding Methanomethylophilus alvus Pyl-RS mutants. Expresses 2xTAG-codon interrupted barnase gene and M. alvus Pyl-tRNA(6). p15a origin of replication.
197573pBK- Ma PylRS WTExpresses wild-type Methanomethylophilus alvus pyrrolysine tRNA synthetase under GlnS promoter. Used for Pyl tRNA-synthetase selections.
197574pALS2-sfGFP 150TAGFluorescence selection plasmid for selecting ncAA-encoding Methanomethylophilus alvus Pyl-RS mutants. Expresses sfGFP-150TAG and M. alvus Pyl-tRNA(6). p15a origin of replication.
197575pALS2-sfGFP WTFluorescence control plasmid for selecting ncAA-encoding Methanomethylophilus alvus Pyl-RS mutants. Expresses sfGFP-WT and M. alvus Pyl-tRNA(6). p15a origin of replication

Antibodies from Article