In Fig 4, we wanted to expand the host range even further by incorporating the broad-host range ori, RSF1010. This enabled successful CCP expression in A. baumannii. We look forward to seeing the pKEK-Chrom plasmids expand the use of CCPs to other non-model bacteria!

In Fig 3, we demonstrate how promoter swapping can be used to incorporate alternate promoter elements in the pKEK-Chrom plasmids. This enables CCP expression in species with unique transcriptional requirements, such as Francisella! #GoldenGateCloning

In Fig 2, we evaluated expression of each of the 8 CCPs in several gram negative bacterium our lab routinely works with. You can also see the unique color offered by each of CCPs!

Fig 1 gives an overview of the 32 pKEK-Chrom plasmids. We carefully designed the plasmid set to readily overcome many of the challenges our lab faces working with non-model bacteria. All plasmids are available at Addgene or upon request!

Thank you Sebastian, was great working with you to expand the host range of CCPs!