Abstract: Disclosed are methods, components, compositions, and kits for preparing and identifying engineered E. coli ribosomes. The E. coli ribosomes may be prepared and identified under a set of defined conditions, such as in the presences of antibiotics, in order to obtain an engineered ribosome that is resistant to the antibiotic.

Abstract: An engineered ribosome that includes a tethered subunit arrangement is disclosed, in which the engineered ribosome supports translation of a sequence defined polymer. Methods for making and using the engineered ribosome are also disclosed, including a method for preparing a sequence defined polymer using the engineered ribosome and a method for preparing a sequence defined polymer using the engineered ribosome in a two-protein translation system. The engineered ribosomes may be utilized in methods for incorporating unnatural amino acids into a sequence defined polymer. Also disclosed are optimized polynucleotide sequences for use as tethers and Shine-Dalgarno/anti-Shine-Dalgarno sequences.

Abstract: Disclosed are cell-free systems for metabolic engineering, methods for cell-free metabolic engineering, kits for preparing the disclosed systems, and kits for performing the disclosed methods. The disclosed systems, methods, and kits may be utilized to prepare a chemical product and to optimize conditions for preparing a chemical product. The disclosed systems, methods, and kits also may be utilized for combinatorial cell-free metabolism engineering.

Abstract: Improved methods are provided in vitro synthesis of biological molecules, providing for improved yields, lowered costs, and enhanced utility. Improved yield and lowered cost is obtained by the use of a phosphate free energy source in the presence of exogenous phosphate, and optionally in the absence of exogenous nucleoside triphosphates.

Abstract: A platform for preparing a sequence defined biopolymer in vitro is disclosed. The platform includes a ribosome-depleted cellular extract ribosomal RNAs prepared by in vitro transcription and purified ribosomal proteins depleted of ribosomal RNAs. A method of synthesizing and assembling ribosomes in vitro for use in the platform is provided, as well as a method for preparing a sequence defined biopolymer in vitro using assembling ribosomes and the platform.

Abstract: Biological macromolecules are synthesized in vitro under conditions and in a reaction composition wherein oxidative phosphorylation is activated and protein folding is improved.

Abstract: Biological macromolecules are synthesized in vitro under conditions wherein oxidative phosphorylation is activated.

Abstract: Biological macromolecules are synthesized in vitro under conditions and in a reaction composition wherein oxidative phosphorylation is activated and protein folding is improved.

Abstract: Biological macromolecules are synthesized in vitro under conditions and in a reaction composition wherein oxidative phosphorylation is activated and protein folding is improved.