Part:BBa_K5127027:Design

IPTG-inducible promoter with IAA degradation enzyme (T7-iadCDE）

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 2374
  Illegal PstI site found at 2191
  Illegal PstI site found at 2769
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 2374
  Illegal NheI site found at 696
  Illegal PstI site found at 2191
  Illegal PstI site found at 2769
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 2374
  Illegal BglII site found at 2383
  Illegal BglII site found at 2435
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 2374
  Illegal PstI site found at 2191
  Illegal PstI site found at 2769
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 2374
  Illegal PstI site found at 2191
  Illegal PstI site found at 2769
  Illegal NgoMIV site found at 642
  Illegal NgoMIV site found at 1633
  Illegal AgeI site found at 406
  Illegal AgeI site found at 1435
  Illegal AgeI site found at 1876
  Illegal AgeI site found at 2299
  Illegal AgeI site found at 2733
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

The iadC, iadD, and iadE were synthesized by Genscript. To verify the successful expression of iadC, iadD, and iadE, we performed PAGE to the expressed protein of E. coli transformed with PiadCDE with or without IPTG added. The bands at indicated lengths showed the successful expression of iadCDE (Figure 2).

Source

In the IAA degradation module, we used IPTG to induce the expression of iadC, iadD, and iadE. They function together to degrade IAA (Figure 1).

References

Gordon, S. A., & Weber, R. P. (1951). Colorimetric Estimation of indoleacetic Acid. Plant Physiology, 26(1), 192–195. https://doi.org/10.1104/pp.26.1.192