TABLE 4.

Primers used in this study

Method and primera	Sequence (5′–3′)	Size of PCR product (bp)	Noteb
Construction of the vraSR deletion mutant
vra-DS-F	GGGGACAAGTTTGTACAAAAAAGCAGGATTATTAGCACTTTTCCCAATG	972	attB1
vra-DS-R	CCGGAATTCTAATTCAATAAATTATTAAAGGCG		EcoRI
vra-US-S	CCGGAATTCTAGTGATTCATCTGTAAATC	920	EcoRI
vra-US-R	GGGGACCACTTTGTACAAGAAAGCTGGGTTAGATTTAGTAAACAATCAA		attB2
Verification of the vraSR deletion mutant
vra-con-F	GGGTCCTTTGCATTCGGTTAC	2,200
vra-con-R	ACGGCACTGCTTATGTGAACG
Cotranscription analysis of vraSR operon
V-12F	ATCAGTATTTGAACCAGAAG	519
V-12R	ATTATTCACCAAAGAAAAGT
V-23F	ACAAAAGAAAGTACCAATGA	534
V-23R	TATCGTCCATAAGTAAATCC
V-34F	TAAGGAAACAAATACCATAG	641
V-34R	ATATTGTAGTTTTCATTAGTTA
V-45F	AATCTATTGTGCTGAGGAAA	435
V-45R	AAATAGAATAATAATCGTGT
vraSR complementation
pRAB11-vra-F	CGGGGTACCAAGATTGAGAATAACCATG		KpnI
pRAB11-vra-R	CCGGAATTCTTATTGAATTAAATTATGCTGG		EcoRI
VraR expression
pET28a-vraR-F	CGGGATCCGTGGCGATTAAAGTTTTATTTG	630	BamHI
pET28a-vraR-R	CCGCTCGAGTTGAATTAAATTATGCTGGAAC		XhoI
Amplification of promoter fragments
P-vra-F	TGAAACTAAAGATAAAAG	281
P-vra-R	TTGTTGTAAATATAAATCT
P-ica-F	ATTCTAAAATCTCCCCCT
P-ica1-R	CTTGTTGATGTTCAGATT	301
P-ica2-R	GGTAGGTGAAAAAATGCA	170
P-ica3-R	GTTGTTATACTGAAACAGT	92
P-ica4-R	GTAATTTTAACTTAATTTTTC	66
P-pbp2-F	CAAGTTTGTTCCTATTTT	329
P-pbp2-R	TTAGATGCCTCCTACTTA
P-sgtB-F	ATCTGCACTCATTATTTT	302
P-sgtB-R	ATGGGTTTTCTCCTTTT
P-murAA-F	CATTACAAGTTCAAGTT	252
P-murAA-R	TCCACCATTTATTACTA
P-rpsJ-F	AAGATTCTCGTGAACAATTC	119
P-rpsJ-R	GATGTCTACACCTGATGG

^aThe primers were designed according to the genomic sequence of S. epidermidis RP62A (GenBank accession number NC_002976). F, forward primer; R, reverse primer.

^bUnderlined sequences represent the BP Clonase reaction sites or restriction enzyme sites.