Regulatory

Part:BBa_M36707:Design

Designed by: Katie Lund, Wyatt Woodson   Group: Stanford BIOE44 - S11   (2011-04-26)

IclR binding sites on aceBAK promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

In order to measure pyruvate levels, we want to utilize the aceBAK promoter region to which IclR binds and inhibits transcription. IclR is an inhibitory transcription factor that efficiently inhibits the aceBAK promoter, as well as it's own promoter, when bound to pyruvate. For our sensor, we wanted to eliminate as many variables as possible, thus we only selected binding sites for IclR that were upstream of the promoter. We found that there were many binding sites upstream of the aceBAK coding domain, and we incorporated all of the detailed sites in our promoter as we hope this should allow for variable expression as opposed to binary expression. There were a few other binding sites we were unable to remove due to proximity to the IclR binding sites.

Source

Ecocyc.org provided a detailed picture of the region upstream of the promoter to which IclR binds. Specific link: http://tinyurl.com/aceBAK-Depiction

Genomic sequence found at genome.jp (http://tinyurl.com/aceBAK-Sequence). The promoter region is written in black, while the aceBAK coding sequence is written in blue.

References