Fig. 4.

Deletion series of the conserved backbone from pLA6_12. (A) Module #01 with the complete pLA6_12 backbone and modules #02 to #08, representing stepwise deletions of each of the 6 backbone genes mobACX, repL, and the toxin/antitoxin system genes (here designated “Tox” and “a,” respectively), were amplified from pLA6_12 via specific PCRs. The size of each module and the primers used for its respective creation are listed to the Right. The respective primer sequences are given in SI Appendix, Table S4. (B) Each module was individually cloned into the commercially available E. coli vector pCR2.1 encoding kanamycin resistance as a universal selective marker.