Table 1 Genetic tools constructed during this study and made publicly available

pDOC-GG

Acceptor plasmid for Golden Gate-style cloning of gene fragments to generate any Gene Doctoring donor plasmid in a single reaction.

149377

mUAVbla

Derivative of the universal acceptor vector from the Mobius Assembly toolkit [12], conferring an ampicillin resistance phenotype rather than chloramphenicol resistance.

149378

mUAVbla:tetR 5′

Gene part with a tetracycline resistance selectable marker flanked by FRT sites for BsaI-guided Golden Gate assembly. Removal from the chromosome will leave an FRT scar between homologous region 1 and an inserted sequence.

149379

mUAVbla:tetR 3′

Gene part with a tetracycline resistance selectable marker flanked by FRT sites for BsaI-guided Golden Gate assembly. Removal from the chromosome will leave an FRT scar between an inserted sequence and homologous region 2.

149380