Composite

Part:BBa_K3237021

Designed by: Kristi Turton   Group: iGEM19_Lethbridge   (2019-10-14)


His-mRFP1-Proinsulin-CPP with furin cut sites codon optimized for C. reinhardtii

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 iteslt was modified by the removal of original protease sites and replaced by furin cut sites as proposed originally by Tsingua2014. The proinsulin-CPP are 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. This part is codon optimized for chlamydomonas reinhardtii and has a complementary part codon optimized for E.coli BBa_K3237009.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1829
    Illegal XbaI site found at 1817
    Illegal XbaI site found at 1859
    Illegal SpeI site found at 1853
    Illegal PstI site found at 1367
    Illegal PstI site found at 1835
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1829
    Illegal NheI site found at 1381
    Illegal SpeI site found at 1853
    Illegal PstI site found at 1367
    Illegal PstI site found at 1835
    Illegal NotI site found at 1865
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1829
    Illegal BamHI site found at 1847
    Illegal XhoI site found at 656
    Illegal XhoI site found at 1197
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1829
    Illegal XbaI site found at 1817
    Illegal XbaI site found at 1859
    Illegal SpeI site found at 1853
    Illegal PstI site found at 1367
    Illegal PstI site found at 1835
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1829
    Illegal XbaI site found at 1817
    Illegal XbaI site found at 1859
    Illegal SpeI site found at 1853
    Illegal PstI site found at 1367
    Illegal PstI site found at 1835
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None