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To create genetically programmed cell-based synthesis of non-natural depsipeptide macrocycles (or any non-α-l-amino acid linked macrocycle), they further utilized PylRS/tRNA pairs from different classes to facilitate the cotranslational integration of various α-hydroxy acids encompassing both aliphatic and aromatic side chains, and finally identified a total of 49 engineered, mutually orthogonal pairs capable of recognizing distinct ncAAs or alpha hydroxy acids and deciphering distinct codons
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