Regulatory

Part:BBa_K4849002

Designed by: Edvardas Eigminas and Devansh Kumar   Group: iGEM23_Edinburgh   (2023-10-10)

PcopM195-BCD zinc-inducible Synechocystis promoter

 

Description

In Synechocystis, PcopM promoter regulates the copMRS operon, which is involved in copper response (Giner-Lamia et al., 2015). Under native conditions, PcopM responds robustly to Cu2+. As little as 1 µM Cu2+ triggers about 26-fold increase in CopM protein levels, and 3 µM Cu2+ increases copM mRNA levels 75-fold (Giner-Lamia et al., 2015, 2012). Other metal ions (Zn2+, Cd2+, and Ni2+) trigger a poorer PcopM response (Giner-Lamia et al., 2012).

Čelešnik et al. (2016) constructed a modified version of the PcopM promoter, called PcopM195-BCD, in order to increase the efficiency of their kill switch. PcopM195-BCD includes a 195 nt-long sequence segment preceding the copMRS operon, and the 26 nucleotides of the native sequence (from the copM transcriptional start site to the start codon) are replaced by bicistronic design (BCD). BCD is a genetic element that standardizes the mRNA secondary structure over the RBS–CDS (ribosome binding site – coding sequence) junction, thereby diminishing variation in ribosome binding and expression when combined with different genes of interest. It allows reliable initiation of translation of random genes.

 

References

Čelešnik, H., Tanšek, A., Tahirović, A., Vižintin, A., Mustar, J., Vidmar, V. and Dolinar, M., 2016. Biosafety of biotechnologically important microalgae: intrinsic suicide switch implementation in cyanobacterium Synechocystis sp. PCC 6803. Biology Open5(4), pp.519-528.

Giner-Lamia, J., López-Maury, L., Reyes, J.C. and Florencio, F.J., 2012. The CopRS two-component system is responsible for resistance to copper in the cyanobacterium Synechocystis sp. PCC 6803. Plant physiology159(4), pp.1806-1818.

GinerLamia, J., LópezMaury, L. and Florencio, F.J., 2015. CopM is a novel copperbinding protein involved in copper resistance in S ynechocystis sp. PCC 6803. MicrobiologyOpen4(1), pp.167-185.


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]
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Categories
Parameters
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