Coding
mleR

Part:BBa_K4137002:Design

Designed by: Jen-Hao Yao   Group: iGEM22_GEMS_Taiwan   (2022-08-08)
Revision as of 07:41, 5 October 2022 by Stuberosum ryan (Talk | contribs) (→‎References)


Malate-binding Transcriptional Activator + 6xHis


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 18
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The 6xHis tag was added to the mleR coding sequence during part design, as we needed to purify mleR and then quantify its production to characterize the toxin-antitoxin system through the derivation of kinetic constants.

The sequence of mleR and 6xHis was then reversed alongside the entirety of BBa_K4137008 such that the transcription of CcdA would not interfere with mleR, and vice versa; this was taken into our design as our constructs only have a single terminator.

Unfortunately, the sequence features view tool does not reflect the fact that this part is reversed; attached below is a benchling screenshot of how the sequence features should look.

Figure 1. Benchling Linear Map View of the BBa_K4137002 Basic Part.

Source

https://journals.asm.org/doi/10.1128/jb.171.6.3108-3114.1989

References

https://journals.asm.org/doi/10.1128/jb.171.6.3108-3114.1989