Part:BBa_K3945009:Design
Lucifer: A luminescence-based measurement system for rare earth elements
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 586
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
When designing the Lucifer construct, it was extremely important to note that the two luciferase subunits, Large BiT and Small BiT, were as close to each other as possible. As mentioned previously, unless these subunits come into very close proximity, the likelihood of signal output is low [1]. As such, the best linker sequence selected was done through protein modelling experiments, where the linkers were tested through homology modelling to determine the atomic distance between the NanoBiT subunits in the desired fusion protein. To further test this, molecular docking simulations were performed to ensure LanM still retains functionality even with the added protein.
Source
The sequence was obtained from the genome of Methylobacterium extorquens and the NanoBIT sequence was obtained from Promega