Composite
Part:BBa_K3338023:Design
Designed by: Jonas Scholz Group: iGEM20_Hannover (2020-10-23)
IL-6 Pmut-MagA-P2A-hGLuc
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1347
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 220
Illegal BamHI site found at 395
Illegal XhoI site found at 1581 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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.
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.