Coding

Part:BBa_K1962004

Designed by: Frank Sargent   Group: iGEM16_Dundee   (2016-10-11)
Revision as of 23:36, 22 October 2016 by Frulhuq (Talk | contribs)


Immunity Protein Im-E3

This biobrick encodes for the Immunity protein specific for Colicin E3. This Immunity Protein inhibits the 16S RNA hydrolyzing activity of Colicin E3 by binding with high affinity to the C-terminal catalytic domain of E3. The sequence was obtained from Uniprot and then codon optimised for E. coli K-12 and synthesised by IDT as a gBlock gene fragment.


Usage and Biology

Our idea was to generate a device that would express Colicin E3 in the presence of low pH and bile salts. In order to protect the host cells from the toxic activity of Colicin E3 we began by cloning the E3-Immunity protein downstream of a pH sensitive promoter Pasr (BBa_K1231000) and a bile salt sensitive promoter PacrRA (BBa_K1231001) to generate the composite parts (BBa_K1962016) and (BBa_K1962012), respectively. Unfortunately we were unable to clone Colicin E3 downstream of the E3-Immunity proteins.

Sequence and Features


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
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None