Composite

Part:BBa_K2387029:Design

Designed by: Bart Scholten   Group: iGEM17_Wageningen_UR   (2017-10-18)


CpxR-mVenus[1-157] and CpxR-mVenus[158-238] + araC/pBAD promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
    Illegal XhoI site found at 1283
    Illegal XhoI site found at 2498
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
    Illegal AgeI site found at 1694
    Illegal AgeI site found at 2909
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


Design Notes

A long flexible linker was used to fuse CpxR and mVenusc to maximize the availability of mVenusc and thus increase the chances to reassemble with mVenusn, and vice versa. Both fusions were put under control of one promoter and the same RBS to ensure similar levels of protein expression.


Source

mVenus is a mutated version of eYFP, which originates from the jellyfish Aequorea victoria It was amplified fromPlasmid #27794 mVenus C1 from Addgene.

araC/pBAD is an endogenous E. coli promoter

These original parts are taken from the iGEM repository; [http://parts.igem.com/BBa_E0030 eYFP] [http://parts.igem.com/BBa_I0030 araC/pBAD promoter] [http://parts.igem.com/BBa_B0034 RBS BBa_B0034] [http://parts.igem.com/Part:BBa_K1486004 Flexible Linker] [http://parts.igem.com/Part:BBa_K2387002 CpxR]

N-terminus of mVenus and [https://parts.igem.org/Part:BBa_K2387039

C-terminus of mVenus] were created in during [http://2017.igem.org/Team:Wageningen_UR Wageningen_URs 2017 iGEM project].


References