Provided herein are regulatable expression systems and methods of using said regulatable expression systems to express proteins of interest. The regulatable expression systems comprise a unidirectional regulatable promoter operably linked to a single transcription unit encoding a protein of interest, a ribosome skip, and a transactivator protein.

Provided herein are regulatable expression systems and methods of using said regulatable expression systems to express proteins of interest. The regulatable expression systems comprise a unidirectional regulatable promoter operably linked to a single transcription unit encoding a protein of interest, a ribosome skip, and a transactivator protein.

Described herein are materials and methods for user-controlled adjustment of plasmid copy numbers.

Described herein are materials and methods for user-controlled adjustment of plasmid copy numbers.

Disclosed herein are methods for the production of low to medium expressing constitutive promoters in bacteria and promoters produced therewith.

Disclosed herein are methods for the production of low to medium expressing constitutive promoters in bacteria and promoters produced therewith.

A nucleic acid molecule comprising a variant inc coding strand is disclosed as a regulator of plasmid copy number. Also disclosed is a replicon comprising the nucleic acid molecule, a promoter, and an origin of replication. Also disclosed is a vector comprising the replicon. Also disclosed is a recombinant microorganism comprising the vector.

The present disclosure provides methods for producing recombinant adeno-associated virus (rAAV) virions. The present disclosure provides compositions for producing rAAV virions.

The present disclosure provides methods for producing recombinant adeno-associated virus (rAAV) virions. The present disclosure provides compositions for producing rAAV virions.

Nucleic acid molecule comprising a coding sequence and a region of increased folding energy upstream of a stop codon are provided. Expression vectors and cells comprising the nucleic acid molecule are also provided. Methods for optimizing a coding sequence comprising increasing folding energy in a region upstream of the stop codon are also provided.