Composite

Part:BBa_K1638011:Design

Designed by: Jens Sivkr Pettersen   Group: iGEM15_SDU-Denmark   (2015-05-22)
Revision as of 09:17, 18 September 2015 by Jpettersen (Talk | contribs) (→‎Source)


Leucine zipper fused to T18 domain of cyaA from Bordetella pertussis


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 711
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 252
    Illegal NgoMIV site found at 662
    Illegal AgeI site found at 468
  • 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

pUT18C and BBa_C2001

References