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Recoding a stop codon to an amino acid may afford orthogonal genetic systems for biosynthesizing new protein and organism properties.Although reassignment of stop codons has been found in extant organisms,they represent only the endpoint of code evolution;a model organism is lacking that enables real-time investigation of reassignment process and directed evolution with genetically encoded new amino acids.Release factors (RFs) recognize stop codons to terminate translation,preventing complete reassignment of a stop codon.