TABLE 2.

Oligonucleotides used in plasmid construction

Oligonucleotidea	Restriction enzyme	Comment
F, 5′-GCTTCCGGAATGGTGAGCAAGGGCGAGG	BspEI	YFP cloning (PCR)
R, 5′-GCTGGATCCTTACTTGTACAGCTCGTCCATG	BamHI
5′-CTTTGCCCGTTATCCCGATCATATGAAACGG		YFP A206K mutantb
5′-CCGTTTCATATGATCGGGATAACGGGCAAAG
F, 5′-GCTACTAGTTGACAATTAATCATCGGCTCG	SpeI	p15A/Ptac cloning (PCR), cMyc-YFP and RepA-WH1 WT, A31V, or ΔN37 variant with YFP tag
R, 5′-GCTGGATCCTTACTTGTACAGCTCGTCCATG	BamHI
F, 5′-GCTACTAGTTGACAATTAATCATCGGCTCG	SpeI	pRK2/Ptac cloning (PCR), His6-RepA-WH1 WT, A31V, or ΔN37 variant with mCherry tag
R, 5′-AGTGAATTCGAGCTCGGTAC	EcoRI
F, 5′-GCTCGGTACCTCGATCCTCTACGCCGGAC	KpnI	lacIq cloning (PCR), pRK2/p15A
R, 5′-CGAGGAATTCTCATGCCCCGCGCCCAC	EcoRI	lacIq cloning (PCR), pRK2
R, 5′-CGAGGAGCTCTCATGCCCCGCGCCCAC	SacI	lacIq cloning (PCR), p15A

^aRestriction sites are underlined; mutations are in boldface. F, forward; R, reverse.

^bQuikChange kit (Stratagene).