Composite

Part:BBa_K5111004

Designed by: Thiago Espurio Moraes, Angus Lusty   Group: iGEM24_City-of-London-UK   (2024-09-25)


GFP reporter regulated by ComR

Uses a strong constitutive promoter (BBa_J23119), regulatory sequence (BBa_K5111000), strong ribosome binding site (BBa_B0030), a GFP (BBa_E0040), and terminator (BBa_B0010).

This sequence is designed to report ComR concentration via the downregulation of GFP synthesis. The regulatory sequence is the promoter region for the periplasmic protein ComC, which can be bonded to by ComR that downregulates it (M. Mermod et al., 2017). The strong constitutive promoter was added to provide a baseline expression to compare luminescence.

Sources Mermod M, Magnani D, Solioz M, Stoyanov JV. The copper-inducible ComR (YcfQ) repressor regulates expression of ComC (YcfR), which affects copper permeability of the outer membrane of Escherichia coli. Biometals. 2012 Feb;25(1):33-43. doi: 10.1007/s10534-011-9510-x. Epub 2011 Nov 17. PMID: 22089859.

Mermod M, Magnani D, Solioz M, Stoyanov JV. The copper-inducible ComR (YcfQ) repressor regulates expression of ComC (YcfR), which affects copper permeability of the outer membrane of Escherichia coli. Biometals. 2012 Feb;25(1):33-43. doi: 10.1007/s10534-011-9510-x. Epub 2011 Nov 17. PMID: 22089859.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 738


[edit]
Categories
//classic/reporter/constitutive
Parameters
None