Composite

Part:BBa_K4217005:Design

Designed by: Madeline Gruenberg   Group: iGEM22_WrightState   (2022-09-30)


pTrc-(6xHis)MntR


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 156


Design Notes

The pTRC plasmid backbone for this construct came from Addgene #50603, pTrc-trGPPS(CO)-LS, a plasmid used by the 2021 Wright State iGEM team that had a truncated codon optimized geranylpyrophosphate synthase and truncated limonene synthase. The backbone, isolated from an NcoI / Hind III digest, was used as a target vector for the 6x-(HIS)-MntR (BBa_K4217004) geneblock resulting in an IPTG-inducible mntR plasmid with an ampicillin selection marker.

Source

This part is a composite of the 6X-HIS-mntR (part # BBa_K4217004) and the plasmid backbone E. coli DH10B isolated as an NcoI/HindIII digest of Addgene #50603, pTrc-trGPPS(CO)-LS.

References

Que, Q., & Helmann, J. D. (2000). Manganese homeostasis in Bacillus subtilis is regulated by MntR, a bifunctional regulator related to the diphtheria toxin repressor family of proteins. Molecular microbiology, 35(6), 1454–1468. https://doi.org/10.1046/j.1365-2958.2000.01811.x

Patzer, S. I., & Hantke, K. (2001). Dual repression by Fe(2+)-Fur and Mn(2+)-MntR of the mntH gene, encoding an NRAMP-like Mn(2+) transporter in Escherichia coli. Journal of bacteriology, 183(16), 4806–4813. https://doi.org/10.1128/JB.183.16.4806-4813.2001