Regulatory

Part:BBa_K3606014

Designed by: Yuanyuan Liu   Group: iGEM20_Fudan   (2020-10-11)


P5 promoter

Usage and Biology:

This promoter comes from a kit which contains combinations of specific constitutive bacterial promoters that vary in strength.

Design:

We have chosen a series of constitutive promoters with different strength which include P5. They are used to reliably drive expression of CaAP and McbABCEFG in E. coli in a controlled, reliable manner. We have tested these different, unique combinations to find the best expression level of CaAP and McbABCEFG.

Characterization:

We characterized the promotor's express efficiency by measuring the fluorescent intensity of the RFP following the promotor. The bacteria are incubated under two different conditions. Different promotors had varied features of expression.

Figure 1. Measurement of bacteria incubated in 3mL medium for 12h
Figure 2. Measurement of bacteria incubated in 3mL medium for 24h

Further Application:

This part comes from a collection of precisely quantitated bacterial transcription and translation initiation elements. You can directly choose a promoter with suitable strength to drive expression of a gene of interest in E. coli from those we have tested. You can also combine them with other elements to achieve your goal.

References:

[1] Mutalik VK, Guimaraes JC, Cambray G, Lam C, Christoffersen MJ, Mai QA, Tran AB, Paull M, Keasling JD, Arkin AP, Endy D. Precise and reliable gene expression via standard transcription and translation initiation elements. Nat Methods. 2013 Apr;10(4):354-60. doi: 10.1038/nmeth.2404. Epub 2013 Mar 10. PMID: 23474465.


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]


[edit]
Categories
Parameters
None