Device

Part:BBa_K1638032

Designed by: Jens Sivkćr Pettersen   Group: iGEM15_SDU-Denmark   (2015-09-09)
Revision as of 10:46, 11 September 2015 by Jpettersen (Talk | contribs)

T18 domain of cyaA with cAMP-induced RFP generator

The T18 domain of CyaA to be used in the bacterial two-hybrid system. The expression of the gene is under control of a lac operator and is induced by Isopropyl β-D-1-thiogalactopyranoside (IPTG). When protein-coding genes is suffixed to the T25 and T18 domains of the bacterial two-hybrid system, the interaction of these two proteins can be examined. If the conjugated proteins associates, T25 and T18 associates too. This leads to the catalysation of the conversion of ATP to cAMP. The rise in cyclic AMP can trigger the expression of genes by using a cAMP-induced promotor. For this construct a reporter system based on RFP controlled by the promoter PcstA is used. The presence of red-fluorescent cells can be used to verify protein-protein interactions.

See BBa_K1638033 for the T25 domain.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1719
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1260
    Illegal NgoMIV site found at 1670
    Illegal AgeI site found at 712
    Illegal AgeI site found at 824
    Illegal AgeI site found at 1476
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None