Composite

Part:BBa_K1638030:Design

Designed by: Jens Sivkær Pettersen   Group: iGEM15_SDU-Denmark   (2015-08-18)


Leucine zipper fused to T25 domain of cyaA with cAMP-induced RFP generator


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1851
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 712
    Illegal AgeI site found at 824
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part was first constructed by amplifying the leucine zipper region from BBa_C2001 using the primers:

- Forward: 5’-CGCTTCTAGAGGGATCCGAAAATTTGTATTTTCAATCTGGTATGAAACAGCTGGAAGACAAAGTTGA-3’ (BamHI res. site included) - Reverse: 5’-ATATCTGCAGCGGCCGCTACTAGTAACGTTCACCAACCAGTTTTTTCAGA-3’

By digesting the amplified leucine zipper product and the backbone containing either the T25 or T18 domain of the adenylate cyclase cyaA from Bordetella pertussis with BamHI and PstI, we were able to ligate the two bricks together without creating a scar-site.

Source

pKT25, BBa_C2001 and BBa_K861173

References

[1]: Karimova G, Pidoux J, Ullmann A, Ladant D. A bacterial two-hybrid system based on a reconstituted signal transduction pathway. Proceedings of the National Academy of Sciences of the United States of America. 1998;95(10):5752-6.