Regulatory

Part:BBa_K4721004:Design

Designed by: Celeste Bos   Group: iGEM23_Leiden   (2023-10-11)


PL/O4/A1


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 4
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 4
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 4
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The upstream region was included to facilitate primer hybridization.

Source

PL/O4/A1 is a synthetic promoter designed by Carrillo et al. (2019)[1]. In order to function as an inducible promoter, the lacI gene with its respective lacIq promoter is expressed, which expresses the lac repressor, LacI. This repressor binds the _lacO _sequence, present in the PL/04/A1 in the interspacer region between the -10 and -33 sites. When Isopropyl β-d-1-thiogalactopyranoside (IPTG) is present, this repressor is released, and the respective gene (here mCherry) is expressed.

References

[1] Carrillo M, Wagner M, Petit F, Dransfeld A, Becker A, Erb TJ. Design and Control of Extrachromosomal Elements in Methylorubrum extorquens AM1. ACS Synth Biol. 2019;8(11):2451-2456. doi:10.1021/acssynbio.9b00220