Regulatory
pLacUV5e

Part:BBa_K4451000:Design

Designed by: Brooks J Rady   Group: iGEM22_Sheffield   (2022-09-30)


pLacUV5e (lacO1-minus35cons-minus10cons-lacOsym)


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]


Design Notes

Team Sheffield 2022 assembled BBa_K4451000 (IDT-synthesised gBlock) into pSB1C3-GFP (BBa_I20270) in place of the constitutive promoter BBa_J23151 via NEB HiFi assembly, to create BBa_K4451017. Though preliminary experiments found that IPTG-induced cultures expressing BBa_K4451017 gave off noticeably more fluorescence than the control plasmid (BBa_K4451016) at the same optical density, we were unable to collect any quantitative data before the project deadline.

Source

https://doi.org/10.1038/s41467-020-20094-3

References

Spronk, C. A. E. M. et al. Hinge-helix formation and DNA bending in various lac repressor–operator complexes. EMBO J. 18, 6472–6480 (1999).

Yu, T. C. et al. Multiplexed characterization of rationally designed promoter architectures deconstructs combinatorial logic for IPTG-inducible systems. Nat. Commun. 12, 325 (2021).