Composite

Part:BBa_K4390042

Designed by: Maarten van den Ancker   Group: iGEM22_Edinburgh-UHAS_Ghana   (2022-08-17)


pbrR regulated mCherry expression

This part is not compatible with BioBrick RFC10 assembly but is compatible with the iGEM Type IIS Part standard which is also accepted by iGEM.

This is a level 1 part formed by assembly of the following level 0 parts:

Promoter I721001
RBS B0034
N-O-C part K180008
Terminator K4390001

Usage and Biology

mCherry is a red fluorescent protein which derived from DsRed of Discosoma (Shaner, N. C. et al., 2004). This protein will generate bright red colour when expressed which is always been used as reporter in research. PbrR is a repressor for lead resistance operons in bacteria. In gram-negative bacteria, pbrR can bind to the promoter of lead resistance operons which repress the expression. This repression activity is controlled by lead apperance. When lead is appeared in the environment, the pbrR will bind to lead ions which will no longer be able to bind to the promoter and release the lead resistance operons to be expressed (Borremans, B. et al., 2001).

In this designed part, mCherry will be expressed under regulation of pbrR, thus mCherry will only be expressed when there are lead present in the environment. In iGEM22_Edinburgh-UHAS_Ghana SELIS design, the expressed mCherry will be used to detect the present of heavy metals, the expression of mCherry will perform a red colour when lead is present. For further information please refer to SELIS pbrR evolution construct.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 470
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 470
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 470
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 470
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 84

References

D'Oelsnitz, S. et al., (2022) Using fungible biosensors to evolve improved alkaloid biosyntheses. Nature chemical biology. 18 (9), 981–989.

Johnson, A. D. et al. (1979) Interactions between DNA-Bound Repressors Govern Regulation by the $\lambda $ Phage Repressor. Proceedings of the National Academy of Sciences - PNAS. 76 (10), 5061–5065.

Shaner, N. C. et al. (2004) Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nature biotechnology. 22 (12), 1567–1572.


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