Composite

Part:BBa_K3338023:Design

Designed by: Jonas Scholz   Group: iGEM20_Hannover   (2020-10-23)


IL-6 Pmut-MagA-P2A-hGLuc


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1347
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 220
    Illegal BamHI site found at 395
    Illegal XhoI site found at 1581
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 340
    Illegal BsaI site found at 2803
    Illegal BsaI.rc site found at 387
    Illegal BsaI.rc site found at 696
    Illegal BsaI.rc site found at 2135
    Illegal BsaI.rc site found at 2684
    Illegal SapI site found at 1825


Design Notes

The coding sequences of MagA, P2A and hGLuc have to be in one reading frame. Due to the fact, that the reporter hGLuc has a much lower detection limit compared to MagA, it was cloned behind the P2A peptide. The IL-6 promoter is mutagenized to be compatible with the RFC10 assembly standard.

Source

The human Interleukin-6 promoter exhibits a point mutation in one XbaI restriction site making it RFC10 compatible. MagA originates from Magnetospirillum magneticum, P2A from the porcine teschovirus 2A and hGLuc is a human codon optimized form of Gaussia luciferase from Gaussia princeps.

References