Part:BBa_K4197022

mSCARLET-I under control of ihfB800 promoter

Red fuorescent protein mScarlet-I expressed with a constitutive promoter of E. coli.

Introduction

This part is composed of the gene coding for the mScarlet-I (BBa_K2333414), a red fluorescent protein. It also contains the gene coding for the 800 first bp of the ihfB promoter, which was used to express mScarlet-I. 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 mScarlet-I into E. coli Tuner (DE3) cells. The functionality of the promoter and protein were confirmed as the mScarlet was successfully expressed (see BBa_K4197020 and BBa_K4197021).

References

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

Other parts of fluorescent proteins with ihfB800:
- mRFP1
- mTagBFP

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 EcoRI site found at 1520
  Illegal XbaI site found at 1505
  Illegal PstI site found at 213
  Illegal PstI site found at 224
  Illegal PstI site found at 342
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1520
  Illegal PstI site found at 213
  Illegal PstI site found at 224
  Illegal PstI site found at 342
  Illegal NotI site found at 1512
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1520
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 1520
  Illegal XbaI site found at 1505
  Illegal PstI site found at 213
  Illegal PstI site found at 224
  Illegal PstI site found at 342
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 1520
  Illegal XbaI site found at 1505
  Illegal PstI site found at 213
  Illegal PstI site found at 224
  Illegal PstI site found at 342
  Illegal AgeI site found at 329
  Illegal AgeI site found at 1475
  
• 1000
  COMPATIBLE WITH RFC[1000]