Composite

Part:BBa_K2958012:Design

Designed by: Angelica Sabandal   Group: iGEM19_ULaVerne_Collab   (2019-07-25)


Fast Acting Single Chain Insulin Analog Gene Block (with tags for purification)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 56
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 56
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 56
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 56
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The various tags were added to this sequence for protein folding and purification purposes. The 6GGS-6 His tag-6GGS linker is included to allow for the His tag to be exposed because instead of being at the C or N terminus of the protein, the His tag is in the middle of it. This is because the ecotin tag, which is essential for the formation of disulfide bonds in the insulin, must be at the beginning of the sequence. The TEV cleavage site is included in order to isolate the insulin from the tags once extracted from E. coli. The linker used for the insulin in lieu of the C chain is BBa_K2958004.


Source

This part comes from a genomic sequence designed on IDT. The A and b chain of this single chain native proinsulin sequence is from BBa_K2958006. The C chain has been replaced with a linker.

References