TABLE 1.

Strains, plasmids, and primers used in this study

Strain, plasmid, or oligonucleotide	Description	Source or reference
E. coli Novablue	General E. coli cloning strain	Novagen, Madison, Wis.
S. aureus strains
EBII42	SA178RI, RN4220 derivative containing T7 RNA polymerase	10
EBII59	SA178RI yjeQ::erm	This study
EBII61	Wild-type Newman strain	11
EBII74	Newman yjeQ::erm	This study
EBII83	Newman yjeQ::erm/pLI50-yjeQ	This study
Plasmids
pDG1664	Source for a erythromycin resistance cassette, used for gene replacement	17
pSAKO	E. coli replicating vector containing sacB[BamP]W29 and kanamycin resistance cassette	10
pLI50	E. coli-S. aureus shuttle plasmid; Ap (E. coli), Cm (S. aureus)	22
Oligonucleotides
SA1234-A	5′-GGGAAACTCGAGCAATGGCAATGTTTGGTAATAAATACG-3′
SA1234-B	5′-GATACTGCACTATCAACACACTCTTAAGTTTGCTTCTAAGGGCACCTCTCGATTAATTTTACTGC-3′
SA1234-C	5′-AGCTTCCAAGGAGCTAAAGAGGTCCCTAGACTCTAGACCCATCGATATGGTGAAACATGTAAGTTTAGGAATTGTAATC-3′
SA1234-D	5′-GGGAAACTCGAGGGTATAAATGAAATGTACGGATAAC-3′
SA1234-F	5′-GATAAAGATAATGACGGTTC-3′
SA1234-R	5′-GGGAAACTCGAGGATAATAATGATGGATATAGTTTTG-3′
Erm-F	5′-CTTAGAAGCAAACTTAAGAGTG-3′
Erm-R	5′-GGGTCTAGAGTCTAGGGACC-3′