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By overexpressing the C-terminal domain of the ribosomal protein L11 to decrease release factor 1-mediated termination of protein translation, enhanced amber suppression is achieved in E. coli. This enhanced amber suppression efficiency allows the genetic incorporation of three Nε-acetyl-L-lysines into one GFPUV protein in E. coli.

Graphical abstract: A convenient method for genetic incorporation of multiple noncanonical amino acids into one protein in Escherichia coli

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