Regulatory

Part:BBa_K4207000

Designed by: Jesper Mickos   Group: iGEM22_Aboa   (2022-09-26)
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T7max promoter

This promoter is an optimized version of the standard T7 promoter for usage in cell-free transcription systems.This promoter recruits the standard T7 RNA-polymerase, so it can be used in standard applications, which take advantage of the T7 transcription reactions.


1. Usage and Biology

Figure. 1 T7max promoter performance compared to classic T7 promoter in different cell-free expression systems by Deich et al. (2021).

The T7max promoter, introduced by Deich et al. in 2021, is a T7 promoter optimized for in vitro transcription. The promoter recruits the standard T7 RNA-polymerase so it can be integrated into existing systems to reap the benefits of more efficient transcription. The sequence of this promoter has been established before, but the recent thorough characterization deemed it more efficient for in vitro transcription compared to the classic T7 promoter.

Using T7max in a T7 system requires only changing the promoter to T7max promoter since T7max is compatible with all other existing elements in a T7 system. The benefits of T7max promoter include higher protein synthesis yields from linear plasmids compared to the regular T7 promoter due to an elevated transcription rate that compensates for linear DNA degradation in in vitro systems. Deich et al. compared T7max promoter with the classic T7 promoter in several different cell-free expression systems, e.g. PURE system, wheat germ extract, and rabbit reticulocyte extract, and concluded that T7max was more efficient with each of them (Figure. 1).


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

2. References

Deich, C., Cash, B., Sato, W., Sharon, J., Aufdembrink, L., Gaut, N., Heili, J., Stokes, K., Engelhart, A., Adamala, K. (2021) T7Max transcription system. https://doi.org/10.1101/2021.10.17.464727


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