DNA

Part:BBa_K3791020:Design

Designed by: Laura Sánchez Ruiz, Auba Fuster Palŕ   Group: iGEM21_UPF_Barcelona   (2021-10-12)


Efficient gRNA Ampicillin construct


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 81
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The gRNA architecture includes two DR separated by the spacer with 4 additional base pairs downstream. So the final structure is: repeat + spacer + 4 nucleotides + repeat. Adding the promoter and the terminator right before and after the gRNA, the final architecture would be: T7 promoter + repeat + spacer + 4 nucleotides + repeat + L2S2P21 terminator.


Source

The repeat sequence was extracted from the parts registry (BBa_K2927006), whereas the spacer one was designed with the constraint that it had to be complementary to the sequence of the ampicillin-resistance gene aiming to be detected. (as explained in its own part page: BBa_K3791000).
The promoter and the terminator were also taken from Parts registry: BBa_K1614000, BBa_K2675031.

References