ProteInProgress: a cellular assembly line for protein manufacturing

Solutions

Self-inducible promoters

Integrative standard vector for E. coli

The integration of the genetic circuits of interest into the microbial host genome can eliminate the need of expensive selection techniques, such as antibiotics or auxotrophic media, in cell cultures.

In order to simplify the engineering of the host genome, two standard and modular integrative vectors have been designed for Escherichia coli and Saccharomyces cerevisiae, two commonly used hosts for industrial protein production. Here, a detailed description of the integrative vector for E. coli is reported, while the following section deals with the integrative vector for yeast.

The structure of the designed vector, here named <partinfo>BBa_K300000</partinfo>, is shown in Fig.1. Most of its features have been inspired by <partinfo>BBa_I51020</partinfo> (BioBrick base vector) and <partinfo>BBa_J72007</partinfo> (BamHI methyltransferase encoding CRIM plasmid), described by [Shetty RP et al., 2008] and [Anderson JC et al., 2010] respectively.

BioBrick integrative base vector BBa_K300000.

Parts notation.

This vector can be considered as a base vector, which can be specialized to target the desired integration site in the host genome. The default version of this backbone has the bacteriophage Phi80 attP (<partinfo>BBa_K300991</partinfo>) as integration site.

This vector enables multiple integrations in different positions of the same genome.

Glossary

The passenger is the desired DNA part to be integrated into the genome.

The guide is the DNA sequence that is used to target the passenger into a specific locus in the genome.

Integrative standard vectors for yeast

Self-cleaving affinity tags to easily purify proteins