Cloning System : Golden Gate Assembly

As described on our engineering page, our assemblies have 8 parts in order: the plasmid backbone, the 5' HR region, terminator #1, the promoter, the Ribosome Binding Site (RBS), the gene itself, terminator #2 and the 3' HR region. As such, we are able to give each of these a specific prefix that is complementary to the suffix of the previous part.


Here we present the new parts for the 3' & 5' homology regions (HR), as well as the genes themselves. Each part has with it the set of primers that we used to extract the gene from the B. subtilis genome, to which overhangs can be added in the primers to add on the required prefixes and suffixes for golden gate assembly.

We would like to highlight part BBa_K4769420, a composite part of the 3' HR and 5' HR of spoIIE, that together allow for the knockout of sporulation. You can read more about the characterisation on the iGEM parts page, or in our Results page.

3' & 5' Homology Regions

Coding Sequences