Table 1. Primers used for generating promoter-luciferase constructs and PCR site-directed mutagenesis.
| Name | Sequence 5’ to 3’ | Orientation |
|---|---|---|
| Promoter-luciferase constructs | ||
| ek1-1966-5’ | CTA GCT AGC TAC ATC CTG GTA GGG TTG GTC C | Forward |
| ek1-154-5’ | CTA GCT AGC GTT CCC AGG GAT GGG TGT G | Forward |
| ek1-100-5’ | CTA GCT AGC GAG GTC CCA TTG TGA CCG GAG | Forward |
| ek1-69-5’ | CTA GCT AGC CCG CCT CGG CAC CCT GAC G | Forward |
| ek1-3’ | GGA AGA TCT TGC CGG GGC TGG CCT GAC G | Reverse |
| Site-directed mutagenesis | ||
| ek1-mut[Sp(-40/-31)] | CTA GCT AGC CCG CCT CGG CAC CCT GAC GCA GCG CAG GAC CtG CtC CGC GCG TGA CGC CAG | Forward |
| ek1-mut[Sp(-69/-60)] | CCT GAG CTC GCT AGC CCttCT CGG CAC CCT GAC GCA GCG CAG | Forward |
| ek1-3’ | GGA AGA TCT TGC CGG GGC TGG CCT GAC G | Reverse |
| Real-time PCR | ||
| ek1-F | AAAGGTTCCTAAGTGATATCCC | Forward |
| ek1-R | GCCAGGTAGTTGTATCCAGA | Reverse |
| ChIP assay | ||
| ek1-154-5’ | CTA GCT AGC GTT CCC AGG GAT GGG TGT G | Forward |
| ek1-3’ | GGA AGA TCT TGC CGG GGC TGG CCT GAC G | Reverse |
The mutations introduced into the binding sites are in lower cases and italicized. Underlined nucleotides are the restriction enzyme recognition sites for cloning.