Composite
Part:BBa_K2958009:Design
Designed by: Angelica Sabandal Group: iGEM19_ULaVerne_Collab (2019-07-24)
Native Proinsulin Gene Block (with Tags for purification)
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 56
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 56
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 56
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 56
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The various tags were added to this sequence for protein folding and purification purposes. The 6GGS-6His tag-6GGS linker is included in order to allow for the His tag to be exposed because instead of being at the C or N terminus of the protein, it is in the middle of it. This is because the Ecotin tag, which is essential for the formation of disulfide bonds, 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 the E. coli.
Source
This part comes from a genomic sequence designed on IDT. The proinsulin is from BBa_K2417006 (iGEM17_Sydney_Australia).