Part:BBa_K4137002:Design

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. This sequence is codon optimized for E. coli.

However, for registry documentation purposes, the sequenced attached here is in the forward direction; attached below is a benchling diagram of the sequence as it is used in our project experimentation

Figure 1. Benchling Linear Map View of the BBa_K4137000 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