Part:BBa_K3237020
His-mRFP1-Proinsulin-CPP with furin cut sites codon optimized for E.coli
This part was designed for targeted delivery of insulin into the blood stream from the intestinal wall. The Cell penetrating peptide (a protein transduction domain) has been found to increase the intestinal absorption of insulin [1] and is therefore able to increase insulin uptake from the GI tract and into the blood stream. This CPP was fused onto the C-terminus of proinsulin with a furin cut site in between. The proinsulin itself was modified by the removal of original protease sites and replaced by furin cut sites as proposed originally by Tsinghua 2014. The proinsulin-CPP was attached to a his-mRFP1 coding sequence with linkers and furin cut sites [2] to remove the RFP after expression. In summary, this part expresses the his-mRFP1-prinsulin-CPP construct. After expression the CPP, and his-mRFP1 peptides are removed by the action of furin protease which also processes the insulin to an active form. This will only occur in vitro or after the construct passes through the intestinal wall.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 649
Illegal AgeI site found at 761 - 1000COMPATIBLE WITH RFC[1000]
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