The invention provides, inter alia, a nucleic acid (e.g. expression vector) that comprises at least a first coding sequence and a second coding sequence. Each conding sequence is under the control of an inducible promoter of defined strength. Different promoters can have different strengths. Each promoter is responsive to the same inducer. The invention also provides: methods of expressing coding regions, methods of making a product of a multi-enzyme pathway, and methods of optimizing the yield of a product of a multi-enzyme metabolic pathway using the nucleic acids provided by the invention. Also disclosed is a method of non-enzymatic gene cloning useful for practicing the invention.

Provided herein are compositions and systems comprising an engineered release factor and methods for producing the same. Also provided herein are compositions, systems, and methods for producing a polypeptide comprising a non-canonical amino acid.

The invention provides compositions and methods for manufacturing pluripotent cells. In particular, the invention provides improved culture platforms for manufacturing pluripotent cells with ground state pluripotency.

Methods and compositions are provided for durably influencing microbiological ecosystems (microbiomes) in a subject in order to prevent infection and reduce recurrence of infection by microorganisms. In some embodiments, compositions and methods are provided for the creation and use of molecularly-modified bacterial strains with the potential to prevent a variety of microorganism infections.

Compositions and methods for production of monoclonal antibodies are provided according to aspects of the present invention including a synthetic gene circuit system, comprising: a first expression construct comprising a nucleic acid encoding tetracycline transactivator (tTA) operably linked to a ubiquitous promoter; a second expression construct comprising a nucleic acid encoding tetracycline transactivator (tTA) operably linked to a tetracycline responsive element (TRE); a third expression construct comprising a nucleic acid encoding a heavy chain component of an monoclonal antibody operably linked to a TRE; and a fourth expression construct comprising a nucleic acid encoding a light chain component of an monoclonal antibody operably linked to a TRE, functioning of the system includes a positive feedback loop which amplifies expression of the heavy chain component of the monoclonal antibody and the light chain component of the monoclonal antibody.

A method of detecting Polycomb Repressive Complex (PRC) activity in a cell providing a cell with a DNA having a protein binding site and at least one reporter gene expression site is operatively connected to the protein binding site, and with a DNA containing a recombinant gene of a binding protein, the binding protein being capable of binding to the protein binding site, wherein the binding protein is fused to a member of the PRC, the method including the step of expressing the recombinant gene, letting the fused binding protein bind to the protein binding site and detecting at least one reporter gene expression.

Methods of making synthetic yeast cells by mating together two diploid (or higher ploidy) yeast species or hybrids to generate multiploid yeast hybrids are provided herein. The synthetic yeast cells made by this process and kits for performing the process are also provided.

Disclosed herein are protein translation switches and conditional gene expression systems that are compatible with retroviral and lentiviral gene delivery. The linking of a protein translation switch to a 3 gene of interest suppresses translation of the gene of interest, and the alteration of the protein translation switch by DNA recombinase-mediated DNA recombination relieves the suppressed translation of the 3 gene of interest. Also disclosed herein are methods of mimicking clinical pharmacology in a pre-clinical setting.

The presently disclosed subject matter relates to targeted integration (TI) host cells suitable for the expression of recombinant proteins where the TI host cells are also subjected to transposon-mediated genomic integration of one or more exogenous nucleic acids, as well as methods of producing and using said combined transposon-mediated genomic integration and TI host cells.

The present invention provides methods for light-dependent gene regulation using a light-responsive DNA-binding protein. Also provided are related nucleic acid molecules, and protein molecules, such as those encoding or comprising the light-responsive DNA-binding protein or DNA-binding sites recognizing the light-responsive DNA-binding protein. Kits using the present light-dependent gene regulation system are further provided by the present invention.