Fig 5. Vector map of plasmid pMTL-YZ14.
Expression of the hyperactive mariner transposase gene Himar1 C9 was driven by the C. difficile toxin B promoter, PtcdB. The plasmid backbone consisted of the conditional replicon between restriction sites AscI and FseI, the macrolide-lincosamide-streptogramin B antibiotic resistance gene ermB, and the Gram-negative replicon, ColE1. The whole mariner element (i.e., transposase gene and catP mini-transposon) can be excised as a SbfI fragment. The control plasmid pMTL-YZ13 was identical, except that the Gram-positive replicon is the pCB102 replicon from C. butyricum. This plasmid conforms to the pMTL80000 modular system for Clostridium shuttle plasmids [32].