Part:BBa_K4197014

ihfB800 promoter

Constitutive promoter of E. coli.

Introduction

This part is composed of the gene coding for the 800 first bp of the ihfB promoter. This promoter has been identified as a constitutive E. coli promoter (Weglenska et al., 1996). It is often used by researchers of the Toulouse Biotechnology Institute to express recombinant fluorescent proteins in E. coli (Barthe et al., 2020), as it is strong enough to allow correct expression and weak enough to avoid inclusion bodies.

Construction

The objective of the INSA-UPS 2022 team was to use 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.

References

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]