Composite

Part:BBa_K4390039

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


pbrR controlled lambda cI 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 K4390037
Terminator K4390001

Usage and Biology

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). Lambda cI is a transcriptional repressor which allows Lambda phage to establish and maintain latency after infect E. coli. It regulates the entry of lytic cycle by repressing the lytic promoters (Johnson, A. D. et al., 1979). This Lambda cI sequence was codon optimised for expression in E. coli K12, and was be used in Seamless Enrichment of Ligand-Inducible Sensors (SELIS) as the repressor (d’Oelsnitz, S. et al., 2022).

In this designed part, Lambda cI will be expressed under regulation of pbrR, thus Lambda cI will only be expressed when there are lead present in the environment. In iGEM22_Edinburgh-UHAS_Ghana SELIS design, the expressed Lambda cI will be used to repress CmR gene which generates chloramphenicol resistance in bacteria. For further information please refer to SELIS pbrR evolution construct.


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 84

References

Borremans, B. et al. (2001) Cloning and functional analysis of the pbr lead resistance determinant of Ralstonia metallidurans CH34. Journal of bacteriology. 183 (19), 5651–5658.

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.


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