Adapted fnr binding site
- 10Illegal XbaI site found at 76
- 12COMPATIBLE WITH RFC
- 21Illegal XhoI site found at 97
- 23Illegal XbaI site found at 76
- 25Illegal XbaI site found at 76
- 1000COMPATIBLE WITH RFC
The size of the sequence and number of included binding sites had to be greatly reduced in order to minimize repeats and facilitate synthesis.
The DNA sequence recognized by FNR consists of an inverted repeat with a consensus sequence of TTGATnnnnATCAA. To create an appropriately sized and spaced eukaryotic promoter that would be recognized by FNR, we adopted the GAL4 Upstream Activating Sequences (UAS) from the Invitrogen pGene/V5-His A plasmid. The 6 GAL4 binding sites were replaced with 3 FNR binding sites (to minimize repeats and, therefore, simplify DNA production), and the sequence was truncated about 30 bp after the included TATA box.