TABLE 2.

Oligonucleotides used

Oligonucleotide	Sequencea	Product
D70ori F	5′-GATCCCTTTTTCTGTAATCTG-3′	pD70ori (1,160 bp)
D70ori R	5′-GATCATAGGCTCAATTCTCGC-3′
CT109 F	5′-CCTACTACCAAAAAACCATTCAA-3′	TS plasmid, pCT109PCR
CT109 R	5′-AACGCTTCGCAATCTCTTG-3′
GA301 F	5′-ATAAGCGAGATTTAAGGATAACAG-3′	TS plasmid, pGA301PCR
GA301 R	5′-TAGGTGTGCTTGTATTTCTTG-3′
5′Δ0ori F	5′-GAACGCACGAAACAGATTACAGA-3′	pD70-5′Δ0oriKanR
5′Δ0ori R	5′-TTTGCTAAGATACAGACCCTAG-3′	0 bases deleted
5′Δ0ori F	5′-GAACGCACGAAACAGATTACAGA-3′	pD70-5′Δ76oriKanR
5′Δ76ori R	5′-CTACTACCAAAAAACCATTCAAC-3′	34-110 bases deleted
3′Δ0ori R	5′-GAAAAAAATCTCAAAAAATTGCGAG-3′	pD70-3′Δ0oriKanR
3′Δ0ori F	5′-AATTTCTCGCAAGGTCGCC-3′	0 bases deleted
3′Δ0ori R	5′-GAAAAAAATCTCAAAAAATTGCGAG-3′	pD70-3′Δ100oriKanR
3′Δ100ori F	5′-CAGCTCGAACAGTAACG-3′	1,021-1,119 bases deleted
3′Δ0ori R	5′-GAAAAAAATCTCAAAAAATTGCGAG-3′	pD70-3′Δ200oriKanR
3′Δ200ori F	5′-TTTCTCTCTAACAAGGCTTG-3′	921-1,119 bases deleted
3′Δ0ori R	5′-GAAAAAAATCTCAAAAAATTGCGAG-3′	pD70-3′Δ300oriKanR
3′Δ300ori F	5′-TGACCGCTGAAAGCG-3′	831-1,119 bases deleted
3′Δ0ori R	5′-GAAAAAAATCTCAAAAAATTGCGAG-3′	pD70-3′Δ400oriKanR
3′Δ400ori F	5′-ACACCAAAAGCCCCG-3′	721-1,119 bases deleted
3′Δ0ori R	5′-GAAAAAAATCTCAAAAAATTGCGAG-3′	pD70-3′Δ500oriKanR
3′Δ500ori F	5′-TTATTATTAGGGAAGTGGTTATTG-3′	611-1,119 bases deleted
3′Δ0ori R	5′-GAAAAAAATCTCAAAAAATTGCGAG-3′	pD70-3′Δ600oriKanR
3′Δ600ori F	5′-CATCAGCTTTTCAATGGCT-3′	523-1,119 bases deleted
3′Δ0ori R	5′-GAAAAAAATCTCAAAAAATTGCGAG-3′	pD70-3′Δ700oriKanR
3′Δ700ori F	5′-TTTCTATTCGGTTCTCAAATGTT-3′	421-1,119 bases deleted

^aBoldface indicates single-nucleotide substitutions for reverse engineering of temperature-sensitive origins of replication.