Compositions and methods of use thereof for making polypeptides with many instances of nonstandard amino acids

A composition comprising

• a plurality of a translated peptide or polypeptide comprising at least four instances of a non-standard amino acid,
• mRNA encoding the peptide or polypeptide, and
• a translation system comprising a heterologous aminoacyl tRNA synthetase (AARS) and a cognate tRNA that can be charged with the non-standard amino acid by the AARS express a superfolder green fluorescent protein (GFP) comprising three instances of the non-standard amino acid at more than 33% yield of the corresponding wildtype superfolder green fluorescent protein (GFP) with no non-standard amino acids,
• wherein the translated peptide or polypeptide is prepared by translation of the mRNA encoding the peptide or polypeptide in genomically recoded organism (GRO) E. coli cells expressing the translation system, and
• wherein the plurality of the peptide has at least 95% correct non-standard amino acid incorporated at the desired residue(s) relative to an undesired amino acid at the same residue.