Table 1.
Primers used for preparation of constructs
| Primers | Amplified fragment (bp) | Sequence (5′–3′) |
|---|---|---|
| 1F (−1455 to −1428) | 1122 | cttttctCtagaggtctttgaggatagg |
| 2R (−332 to −353) | attacgcGtctgccttgcctgt | |
| 3F (−340 to −318) | 1585 | cagacgcGtgtctggtcagtatt |
| 4R (+1244 to +1220) | gaaacacactcGAgaaagaagaagaa |
| Mutagenic oligonucleotides | pGL3 construct generated | |
|---|---|---|
| 5F (−157/−127) | (−138/+1244) | gccaccaatcgggacgCgTggagcgacagcc |
| 6R | ggctgtcgctccAcGcgtcccgattggtggc | |
| 7F (−97 to −66) | (−78/+1244) | cccgtggccctatgggaCgCgTgtcctgcggc |
| 8R | gccgcaggacAcGcGtcccatagggccacggg | |
| 9F (−63 to −29) | (−47/+1244) | gggggcgggacgacgCGTggcgctgcccggcgc |
| 10R | gcgccgggcagcgccACGcgtcgtcccgccccc | |
| 11F (−30 to +1) | (−15/+1244) | gcactagcggctCgAgggcgctgccagtctc |
| 12R | gagactggcagcgcccTcGagccgctagtgc |
Primers are shown along with their binding sites relevant to the TSS and the length of the amplified fragment. Capitalized are depicted the nucleotides that were changed from the original sequence to generate restriction sites (underlined). F, Forward; R, reverse. For more details see Materials and Methods.