Regulatory

Part:BBa_K4673024:Design

Designed by: Chieh Yu Lee   Group: iGEM23_Taipei-KCISLK-V1   (2023-10-02)


The repressor binds to the operator


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

The lac operator was engineered into two vectors using different restriction enzymes. On the commercial pGEM-T EASY vector, it originally exists between M13_pUC_rev_primer and lac primer. In the second vector pET-22b-LK, the lac operator is cleaved by the two restriction enzymes BglII and XbaI and inserted between the BglII site and XbaI site.



Source

GSL Biotech LLC. (2023). pET-22b(+) Sequence and Map. SnapGene. https://www.snapgene.com/plasmids/pet_and_duet_vectors_(novagen)/pET-22b(%2B)

In addition to the pET-22b-LK vector, the lac operator was also inserted into the pGEM-T EASY vector purchased from a company called Promega. https://worldwide.promega.com/products/pcr/pcr-cloning/pgem-t-easy-vector-systems/?catNum=A1360

References

Nishizaki, S. S., McDonald, T. L., Farnum, G. A., Holmes, M. J., Drexel, M. L., Switzenberg, J. A., & Boyle, A. P. (2021, June 18). The Inducible lac Operator-Repressor System Is Functional in Zebrafish Cells. Frontiers in genetics. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8249864/