TABLE 1.

Strains and plasmids used in this study

Strain or plasmid	Relevant characteristic(s)a	Reference(s) or source
Strains
E. faecium
TX82	Endocarditis isolate; Vanr Ampr	7
TX6051	TX82 Δacm::cat; acm allelic replacement mutant with an incidental ccpA premature stop codon	7; this study
TX6086	TX82 Δacm; nonpolar markerless acm deletion mutant	This study
TX6130	TX6051::ccpA nucleotide mutation corrected	This study
TX6127	TX82 ΔccpA; ccpA deletion mutant	This study
TX6140	TX6127 complemented with ccpA (in situ in the chromosome)	This study
TX6145	TX6127 complemented with 300 bp of the ccpA gene	This study
E. coli
DH5α	E. coli cloning host	Invitrogen
EC1000	E. coli host strain, provides RepA	45
E. faecalis CK111	Conjugative donor for genetic manipulations	18
Plasmids
pHOU1	Plasmid for mutagenesis; Genr	19
TX6143	Plasmid for ccpA gene deletion; 820 bp upstream and 912 bp downstream of the ccpA gene cloned into pHOU1	This study
TX6144	Plasmid for initial 300-bp complementation of the ccpA gene; 452 bp upstream of the start codon of ccpA along with the initial 300 bp of ccpA and 507 bp downstream of the stop codon of the ccpA gene cloned into pHOU1	This study
TX6146	Plasmid for correcting the ccpA gene mutation of TX6051; fragment containing 863 bp upstream and 572 bp downstream of the ccpA stop codon cloned into pHOU1	This study
TX6147	Plasmid for acm gene deletion; flanking regions of the acm gene cloned into pHOU1	This study
TX6148	Plasmid for second-step complementation (restoration) of the ccpA gene; fragment containing bp 21–1020 of the ccpA gene along with 507 bp of downstream sequence cloned into pHOU1	This study

^aAmp^r, ampicillin resistance; Gen^r, gentamicin resistance; Van^r, vancomycin resistance.