Part:BBa_K4197014

ihfB800 promoter

Constitutive promoter for E. coli.

Introduction

This part is composed of the gene coding for the 800 first base pairs of the ihfB promoter. This promoter has been identified as a constitutive E. coli promoter (Weglenska et al., 1996). It was used for constitutive expression of fluorescent proteins in E. coli (Barthe et al., 2020) and appears as strong enough to permit sufficient expression without inclusion bodies.

Construction

The objective of the INSA-UPS 2022 team was to use the ihfB800 promoter to express mTagBFP (BBa_K4197013), mRFP1 (BBa_K4197012), and mScarlet-I (BBa_K4197022) into E. coli Tuner (DE3) cells. The functionality of the promoter was confirmed as the mTagBFP and mScarlet-I were successfully expressed (see BBa_K4197001 and BBa_K4197021 respectively).

Conclusion

We found that this promoter is very efficient for constitutive expression in E. coli. Do not hesitate to use it for your experimentations.

References

More information about the project for which the part was created: DAISY (INSA-UPS 2022)

1. Wȩgleńska, A., Jacob, B., & Sirko, A. (1996). Transcriptional pattern of Escherichia coli ihfB (himD) gene expression. Gene, 181(1-2), 85–88. https://doi.org/10.1016/s0378-1119(96)00468-4
2. Barthe, M., Tchouanti, J., Gomes, P. H., Bideaux, C., Lestrade, D., Graham, C., Steyer, J.-P., Meleard, S., Harmand, J., Gorret, N., Cocaign-Bousquet, M., & Enjalbert, B. (2020). Availability of the Molecular Switch XylR Controls Phenotypic Heterogeneity and Lag Duration during Escherichia coli Adaptation from Glucose to Xylose. mBio, 11(6), Article e02938-20. https://doi.org/10.1128/mbio.02938-20

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 213
  Illegal PstI site found at 224
  Illegal PstI site found at 342
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 213
  Illegal PstI site found at 224
  Illegal PstI site found at 342
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 213
  Illegal PstI site found at 224
  Illegal PstI site found at 342
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 213
  Illegal PstI site found at 224
  Illegal PstI site found at 342
  Illegal AgeI site found at 329
  
• 1000
  COMPATIBLE WITH RFC[1000]