Part:BBa_K4932002:Design
LuCage
- 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
We used this concept to create a diagnosing tool for E. coli. We decided to focus our project on a naturally occurring family of proteins, bacteriocins. Bacteriocins are produced by many types of bacteria and they bind to the same species in order to compete against them. They have co-evolved meaning they target extremely conserved regions. Specifically we used colicins. These are specific to E. coli and can be used to test for E. coli Note that we had to reverse the sequences because colicins are encoded in reverse. The sequence published here NOT reversed. This is because the normal sequences are more useful. If you are designing your own protein, currently (2023) it only really works well with small, alpha-helical targets. Fuse this to the C-terminal part of your biosensor.
Source
https://www.ipd.uw.edu/wp-content/uploads/2021/01/Rubio_et_al_Nature_COVID_LOCKR_sensors.pdf
For the whole labelled plasmid, please see https://benchling.com/s/seq-eNnthiZccI1twOVzJvKy?m=slm-HN7zXQYQc78f914MWcZ5
The sequence provided is just the coding sequence for the LucCage protein but in our guides we will be using our whole plasmid. We could not get the igem system to accept the labels for this.
References
Alfredo Quiiano-Rubio et al 2021 https://www.ipd.uw.edu/wp-content/uploads/2021/01/Rubio_et_al_Nature_COVID_LOCKR_sensors.pdf