Composite

Part:BBa_K3790225

Designed by: Chongwen Cao   Group: iGEM21_Fudan   (2021-10-17)


GFP controlled by σS promoter csiD

2021 Fudan

RNA polymerase in E. coli. relies on combined with σ factors to recognize promoter sequences. Although σ70 factor is responsible for the transcription of most of the genes in the genome. E. coli has other six kinds of alternative σ factors that are triggered in stressful conditions. By switching σ factors, bacteria can dramatically change the whole transcription pattern in order to express specific proteins to help themself survive in a bad day.

Among the six alternative σ factors, σS factor recognizes DNA sequences that are very similar to the typical promoter sequences recognized by σ70. Therefore, it is thought that σS promoters rely on upstream elements[1], which are protein-binding DNA sequences distributed upstream, to show their dependence on σS factors.

We noticed that there’s no standard nor well-characterized σS promoter in the registry. Therefore, we tested the core regions of several σS promoters and examined where they can be recognized by σS factors independently.This is one of the devices used to test σS promoters.

Usage and Biology

It can be used to generate GFP during plateau stage in E. coli.

Characterization

T--Fudan--sigmaSpromoters.png


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 20
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 20
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 20
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 707



References

  1. What makes an Escherichia coli promoter sigma(S) dependent? Role of the -13/-14 nucleotide promoter positions and region 2.5 of sigma(S). Becker G,  Hengge-Aronis R. Mol Microbiol, 2001 Mar;39(5):1153-65. PMID:11251833
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