FIG 1.
Restriction map of plasmids pSIP409 and pSIPSYN. (A) pSIP409. pSIP409 is a Gram-positive/Gram-negative shuttle vector with the Gram-negative replication origin pUC(GEM)ori, Gram-positive origin of replication 256rep, erythromycin selection (ermL), and expression of the reporter gene encoding β-glucuronidase (gusA) inducible by addition of a 19-amino-acid peptide pheromone (SppIP, MAGNSSNFIHKIKQIFTHR). (B) pSIPSYN. pSIPSYN is a derivative of pSIP409 with removal of the inducible expression system (sppK, sppR) and replacement of the PorfX inducible promoter with a synthetic promoter from O. oeni strain ATCC BAA-1163 upstream of the gene gusA between the SalI and NcoI restriction sites.