DNA

Part:BBa_K1316017:Design

Designed by: Joan Cortada Garcia   Group: iGEM14_TU_Delft-Leiden   (2014-09-29)

T7 lacO + mtrCAB with His tag


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 BsaI site found at 359


Design Notes

The big gene complex is supposed to be very toxic for the cell, thus, having a low leakage promoter such as the T7 lacO is crucial to ensure the viability of the cells.

In order to eliminate the illegal restriction sites, the entire construct was PCRed in different parts and reassembled via the Golden Gate Assembly method

A His tag was placed at the C-terminus of the MtrB protein in order to easily be able to detect and purify this protein.


Source

Caroline M. Ajo-Franklin laboratory (Goldbeck et al. ACS Synth. Biol. 2013, 2, 150−159)

References