FIG 3.

Construction of L. interrogans gfp reporter plasmid. The thick arrows denote important open reading frames. The restriction sites shown are unique except for the XmaI sites of pGreenTIR. The numbers next to restriction enzyme names refer to the nucleotide positions after the cleavage site. The transcription terminators and a multicloning site were inserted as double-stranded oligonucleotides with overhangs compatible with the ends of the restriction-digested parent plasmid DNA. aadA, spectinomycin resistance; parA and parB, partition loci; rep, replication initiator; ENH, T7 gene 10 translational enhancer; SD, Shine-Dalgarno sequence; T7t, early T7 transcription terminator; rrnDt, E. coli rrnD transcription terminator; RP4 oriT, RP4 origin of transfer; R6K ori, R6K origin of replication; MCS, multicloning site; *, restriction site subject to dam methylation.