Composite

Part:BBa_K3285000:Design

Designed by: PRANAV S R   Group: iGEM19_IISER-Pune-India   (2019-10-11)


Blue Light Repressible Promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 605
    Illegal NgoMIV site found at 677
    Illegal NgoMIV site found at 767
    Illegal NgoMIV site found at 785
    Illegal NgoMIV site found at 1297
    Illegal NgoMIV site found at 1590
    Illegal NgoMIV site found at 1684
    Illegal AgeI site found at 319
    Illegal AgeI site found at 1465
    Illegal AgeI site found at 2815
    Illegal AgeI site found at 2927
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1354
    Illegal BsaI.rc site found at 218


Design Notes

We had to consider the insertion of RBS in between pFixK2 and mrfp. The RBS used in cloning is BBa_B0034. We also had to make sure to add 6-8 bps on either side of these RBSs as that greatly determines RBS efficiency and downstream gene expression (2)


Source

All the individual basic parts of this promoter were obtained from the iGEM 2019 distribution kit plates. Originally, the fixL, fixJ and pFixK2 promoter are part of the fix operon of B.japonicum. The YF1 fusion protein was developed by Andreas Moglich and Keith Moffat (1).

References

(1) Moglich A, Ayers RA and Moffat K. (2009) Design and Signaling Mechanism of Light-Regulated Histidine Kinases. J. Mol. Bio. 385, 5, 1433-1444.

(2)