FIG 1.

Map of the pRAM18dSGA[RmRickA] shuttle vector. The shuttle vector pRAM18dSGA[RmRIckA] was constructed using the pGEM vector backbone (Promega, Madison, WI), which is selectable by ampicillin (Amp) and thus enables production of pRAM18dSGA[RmRickA] shuttle vectors in Escherichia coli. It includes the region containing the parA and dnaA genes of the R. amblyommii plasmid pRAM18 and the intervening sequence (23), and it carries the genes for spectinomycin and streptomycin resistance (labeled Spec) through insertion of the aadA gene and a GFPuv reporter, both driven by ompA promoters. The rickA gene from R. monacensis, along with the region 5′ of the gene, were PCR amplified and inserted into the multiple-cloning site at the Nhel restriction site.