Enterococcus faecalis N06-0364, exhibiting a vancomycin MIC of 8 mu g/ml, was found to harbor a novel D-Ala-D-Ser gene cluster, designated vanL. The vanL gene cluster was similar in organization to the vanC operon, but the VanT serine racemase was encoded by two separate genes, vanTm(L) (membrane binding) and vanTr(L) (racemase).