Composite

Part:BBa_K2622028

Designed by: Auksė Gaižauskaitė   Group: iGEM18_Vilnius-Lithuania   (2018-10-10)
Revision as of 13:39, 17 October 2018 by GAMBIT (Talk | contribs)

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Mstx-Lpp-OmpA-GFPN

Mstx-Lpp-OmpA-GFPN has a GFP nanobody (BBa_K2622005) attached to Lpp-OmpA's (BBa_K1991004) C-terminus via glycine/serine linker, so that it could be displayed and detected in the outer surface of the membrane by binding to GFP. On the N-terminus it has a fused Mstx protein (BBa_K2622006) that improves recombinant membrane protein integration. There is no signal sequence as this part was designed to be used in vitro systems (liposomes, nanodiscs). This part should be expressed in IVTT's or bacteria lysates based on a T7 RNR polymerase as it has T7 promoter and terminator.

This composite part has a RFC10 prefix and suffix, though it has an incompatible XbaI site between T7 promoter and RBS that must be avoided.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 39
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 792
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 39
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 39
    Illegal NgoMIV site found at 747
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 20


[edit]
Categories
Parameters
None