Chromosomal integration of biosynthetic pathways for the production of high‐value chemicals is a necessity to develop industrial strains with a high long‐term stability and a low production variability. Here we present SIRE, Serine Integrase Recombinational Engineering, a novel methodology which combines the ease of recombinase mediated cassette exchange (RMCE) with the selectivity of orthogonal att sites of the PhiC31 integrase. As a proof of concept this toolbox is developed for Escherichia coli.
Using SIRE we were able to introduce a 10.3 kb biosynthetic gene cluster on different locations throughout the genome with an efficiency of 100% for the integrating step and without the need for selection markers on the knock‐in cassette. If you want to know more about SIRE and its possibilities: read our new article in Biotechnology and Bioengineering!