Coding

Part:BBa_K1855003

Designed by: Iain Galpin   Group: iGEM15_Warwick   (2015-09-17)

Lpp-OmpA sZF14

A fusion protein of a transmembrane domain (Lpp-OmpA) and a zinc-finger domain (sZF14),under pL_lac promotion and containing a C-terminal FLAG tag. It is used to exhibit a zinc-finger dsDNA binding domain complimentary to TCCTCACTC on the cell surface of E.coli.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 558
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 519
  • 1000
    COMPATIBLE WITH RFC[1000]


sZF14

WarwickZif14.png

Cells transformed with the Lpp-OmpA-sZF14 construct; wild-type, uninduced and induced in order from left to right. The cells were washed first with an Anti-FLAG antibody, followed by an Anti-mouse fluorescent antibody. The cells were then observed under a fluorescence microscope. The results were as expected, with the wild-type and uninduced showing no surface expression, and the induced showing high surface expression.

[edit]
Categories
//chassis/prokaryote/ecoli
//proteindomain/dnabinding
//proteindomain/transmembrane
Parameters
None