Abstract: The invention relates to prokaryotic cells for the production of polymers containing non-canonical amino acids, and to methods for making said cells. The invention also relates to newly obtainable polymers as produced by the cells of the invention. In addition, the invention relates to new orthogonal aminoacyl-tRNA synthetases (aaRSs) and orthogonal tRNAs, which may be used in pairs and find utility in host cells such as, but not limited to, the prokaryotic cells of the invention.

Abstract: The current invention provides a synthetic prokaryotic genome comprising 5 or fewer occurrences of one or more sense codons; and/or a synthetic prokaryotic genome derived from a parent genome, wherein the synthetic prokaryotic genome comprises less than 10%, 5%, 2%, 1%, 0.5%, 0.1% of the occurrences of one or more sense codons, relative to the parent genome; and/or a synthetic prokaryotic genome comprising 100 or more, 200 or more, or 1000 or more genes with no occurrences of one or more sense codons.

Abstract: The invention relates to prokaryotic cells for the production of polymers containing non-canonical amino acids, and to methods for making said cells. The invention also relates to newly obtainable polymers as produced by the cells of the invention. In addition, the invention relates to new orthogonal aminoacyl-tRNA synthetases (aaRSs) and orthogonal tRNAs, which may be used in pairs and find utility in host cells such as, but not limited to, the prokaryotic cells of the invention.

Abstract: The current invention provides a synthetic prokaryotic genome comprising 5 or fewer occurrences of one or more sense codons; and/or a synthetic prokaryotic genome derived from a parent genome, wherein the synthetic prokaryotic genome comprises less than 10%, 5%, 2%, 1%, 0.5%, 0.1% of the occurrences of one or more sense codons, relative to the parent genome; and/or a synthetic prokaryotic genome comprising 100 or more, 200 or more, or 1000 or more genes with no occurrences of one or more sense codons.

Abstract: The invention relates to a method comprising a) providing a host cell said host cell comprising an episomal replicon, said episomal replicon comprising a donor nucleic acid sequence, said host cell further comprising a target nucleic acid, b) providing helper protein(s) capable of supporting nucleic acid recombination in said host cell c) providing helper protein(s) and/or RNAs capable of supporting nucleic acid excision in said host cell wherein said donor nucleic acid sequence comprises in order. 5?-homologous recombination sequence 1-sequence of interest-homologous recombination sequence 2-3? wherein said sequence of interest comprises a positive selectable marker d) inducing excision of said donor nucleic acid sequence e) incubating to allow recombination between the excised donor nucleic acid and said target nucleic acid f) selecting for recombinants having incorporated said donor nucleic acid into said target nucleic acid. Also described are nucleic acids and cells.

Abstract: The current invention provides a synthetic prokaryotic genome comprising 5 or fewer occurrences of one or more sense codons; and/or a synthetic prokaryotic genome derived from a parent genome, wherein the synthetic prokaryotic genome comprises less than 10%, 5%, 2%, 1%, 0.5%, 0.1% of the occurrences of one or more sense codons, relative to the parent genome; and/or a synthetic prokaryotic genome comprising 100 or more, 200 or more, or 1000 or more genes with no occurrences of one or more sense codons.

Abstract: The invention relates to a method comprising a) providing a host cell said host cell comprising an episomal replicon, said episomal replicon comprising a donor nucleic acid sequence, said host cell further comprising a target nucleic acid, b) providing helper protein(s) capable of supporting nucleic acid recombination in said host cell c) providing helper protein(s) and/or RNAs capable of supporting nucleic acid excision in said host cell wherein said donor nucleic acid sequence comprises in order: 5?-homologous recombination sequence 1-sequence of interest-homologous recombination sequence 2-3? wherein said sequence of interest comprises a positive selectable marker d) inducing excision of said donor nucleic acid sequence e) incubating to allow recombination between the excised donor nucleic acid and said target nucleic acid f) selecting for recombinants having incorporated said donor nucleic acid into said target nucleic acid. Also described are nucleic acids and cells.