Composite
Part:BBa_K4387978:Design
Designed by: Nathalie Weibel Group: iGEM22_UZurich (2022-09-30)
Nitric Oxide Sensing Genetic Circuit
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 708
Illegal BglII site found at 1287 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
An important part of our engineering cycle was the characterization of GFP expression when changing the amount of ribosome binding sites after the NO sensor. We found that the best solution with the highest GFP yields contains two different RBSs inserted right after the promoter pNorVß. For further information about the promoter, please go to the part BBa_K4387000.
Source
The sequence of the nitric oxide sensor was take from the paper by Xiaoyu J. Chen et al. [1] Nanobody sequence was obtained from the patent mentioned in the respective nanobody part registry (BBa_K4387996). [2]
References
- [1] Xiaoyu J. Chen et al., 2021, Rational Design and Characterization of Nitric Oxide Biosensors in E. coli Nissle 1917 and Mini SimCells
- [2] Silence, Karen, Lauwereys, Marc, De Haard, Hans, et al. "Single domain antibodies directed against tumour necrosis factor-alpha and uses therefor", Int. Publication Number: WO 2004/041862 A2, 21 May 2004