Proof of Concept
We constructed a set of RFP/GFP reporters (BBa_K2145000-BBa_K2145003) with a large spacer between the two reporter genes. The reporter set includes all of the combinations of orientations of the two genes, and will demonstrate orientation-dependent expression interference between genes on the same plasmid. This set of constructs demonstrates the need for new techniques for insulating the components of multi-gene circuits on the same piece of DNA.