Materials used in the mutagenesis process for creating plasmids with two Lac repressor binding sites.
Primer sequences(5′→3′):
Mut0: ctaactcacattaattgcgttgAgctcGAGgTTcgctttccagtc.
Mut1: catacgagccggaa (G) cataaagtgtaaagc.
Mut2: ctcggaaagaaca AATTGTGAGCGCTCACAATT aaggccaggaacc.
Mut3: ctcggaaagaacat AATTGTGAGCGCTCACAATT aggccaggaaccg.
Mut4: cggaaagaacatg AATTGTGAGCGCTCACAATT ggccaggaaccgt.
Mut5: ggaaagaacatgt AATTGTGAGCGCTCACAATT gccaggaaccgta.
Mut6: gaaagaacatgtg AATTGTGAGCGCTCACAATT ccaggaaccgtaa.
Mut7: cggaaagaacatgtga AATTGTGAGCGCTCACAATT caggaaccgtaaaaag.
Mut8: ggaaagaacatgtgag AATTGTGAGCGCTCACAATT aggaaccgtaaaaagg.
Mut9: gaaagaacatgtgagc AATTGTGAGCGCTCACAATT ggaaccgtaaaaaggc.
Mut10: catacgagccggaag [C] cataaagtgtaaagc.
Mut11: catacgagccggaag [CG] cataaagtgtaaagc.
The capital letters in the primer sequences indicate the mutations. ‘()’ indicates bp deletion and ‘[ ]’ indicates bp addition. The inter-operator distance indicated here is the distance between two inner edges of the operators instead of center to center distance that is commonly used in in vivo experiments [11]–[13], [86], [93].