Coding

Part:BBa_K523008

Designed by: Mun Ching Lee, Sylvia Ispasanie   Group: iGEM11_Edinburgh   (2011-08-04)

INP for BioSandwich

This is a modified version of BBa_K265008 for use with the BioSandwich protocol (RFC 81) of Edinburgh 2011.

When used to make fusion proteins, BioSandwich assembly requires an in-frame BglII site (formed by the last 2 bases of the RFC10 prefix and the first 4 bases at the start of this BioBrick) and an in-frame SpeI site (present in the RFC10 suffix). Because the SpeI site is out of frame, two extra "G" bases have been added at the end of the BioBrick. When combined with the T at the start of the RFC10 suffix, they code for a glycine residue and correct the frameshift.

Incidentally, although this part has been made in RFC10 format, the two extra bases might help in some fusion applications using other protocols.

Usage and Biology

The DNA is a codon-optimised version of the N and C terminal domains of Pseudomonas syringae Ice Nucleation Protein. It has been shown that, when INPNC is fused to another protein (the "passenger"), it causes the passenger to be displayed on the outer membrane of E. coli.


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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 409
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None