Coding

Part:BBa_K2022012

Designed by: Jessica Mazalo, Daniel Winter   Group: iGEM16_UNSW_Australia   (2016-10-12)

INPNC-GFP, i.e. membrane-anchored GFP

A commonly used reporter is Green Fluorescent Protein (GFP) (BBa_E0040). Here, we have fused it to the C-terminus of the Ice Nucleation Protein, N- and C-termini (BBa_K811005), which is a well-characterised membrane anchor. As such, this part is intended to function like GFP, but simply localised to the membrane of cells

Usage and Biology

Info coming!

Clone this part into an expression vector or construct, cells should become fluorescence but to a large extent only at plasma membranes. GFP has a excitation and emission maxima at 488nm and 509nm, respectively (note: this is enhanced-GFP, not wild-type GFP)

Sequence and Feature Information


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 952
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 72
    Illegal NgoMIV site found at 405
    Illegal AgeI site found at 823
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1604


This part is simply an ORF containing INPNC fused with GFP. To express it, it will need to be put into an expression vector or assembled with a promoter, RBS, and terminator.

Characterisation by UNSW Australia (2016)

Coming soon!

References

Coming soon!

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Categories
Parameters
None