Coding
mleR
Part:BBa_K4137002:Design
Designed by: Jen-Hao Yao Group: iGEM22_GEMS_Taiwan (2022-08-08)
Malate-binding Transcriptional Activator + 6xHis
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 18
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The 6xHis tag was added to the mleR coding sequence during part design, as we wanted to characterize the toxin-antitoxin system and thus had to derive a parameter for its quantity; we thus needed to perform protein quantification, and believed adding a 6xHis tag to be the most straightforward method of achieving this end. The sequence of mleR and 6xHis was then reversed, as we would go on to use it in the composite part BBa_K4130, which uses a T7 promoter to overexpress mleR; the T7 promoter is optimal when reversed, so we adjusted this part's direction in response to this fact.
Source
https://journals.asm.org/doi/10.1128/jb.171.6.3108-3114.1989