Part:BBa_M36100:Design
Promoter, HY5 regulated (3 HY5 binding sites)
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The Gemini regulatory promoter is comprised of the -35 and -10 sites from J23100 and 3 HY5 binding sites. Two of these HY5 binding sites lie between the -35 and -10 regions, and an additional site exists downstream of the promoter. When present, HY5 bZIP protein will inhibit translation by binding to one or all of these sites, preventing RNA polymerase from moving downstream.
Location of the binding sites was chosen to maximize the probability of inhibitory function and minimization of HY5 self-obstruction. HY5 has a size of 168 aa, and will best inhibit RNA polymerase when located closest to the -35 and -10 regions. Additionally, HY5 binding sites should be placed sufficiently far apart from each other such that the binding of one HY5 protein is unlikely to prevent an additional HY5 from binding. As previously mentioned, HY5 is a bZIP protein, and thus binding to two adjacent 3-bp sequences.
Source
Modified J23100 constitutive promoter.
HY5 DNA binding region was found using: http://www.plantphysiol.org/content/146/4/1862.full
HY5 bZIP binding information: http://en.wikipedia.org/wiki/File:CREB.png