Composite

Part:BBa_K2622034

Designed by: Auksė Gaižauskaitė   Group: iGEM18_Vilnius-Lithuania   (2018-10-10)


GFPN-IgA-Mstx

GFPN-IgA-Mstx has a GFP nanobody (BBa_K2622005) fused to translocator domain's (BBa_K2622003) N-terminus, so that it could be displayed and detected in the outer surface of the membrane by binding GFP. On the C-terminus it has a 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 incompatible XbaI and SpeI sites that must be avoided when cloning.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 39
    Illegal SpeI site found at 432
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 432
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 39
    Illegal SpeI site found at 432
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 39
    Illegal SpeI site found at 432
    Illegal NgoMIV site found at 1335
    Illegal AgeI site found at 1110
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 20


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Categories
Parameters
None