Composite

Part:BBa_K3237023

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


His-mRFP1-SCI57-CPP with furin cut sites codon optimized for C. reinhardtii

This part is an expression construct for SCI57, a super-stable single chain insulin variant [1]. This part is fused to a hexahistidine tagged mRFP1 on the N-terminus and a protein transduction domain (that acts as a cell penetrating peptide) on the C-terminus. The mRPF1 is based on the registry part BBa_E1010. The protein transduction domain was added as it has been shown to improve insulin uptake into the intestinal wall, allowing for increased introduction into the bloodstream from the GI tract [2]. Furin cut sites have been added between the mRFP1-SCI57 fusion and the SCI57-Protein transduction domain fusion to allow for removal as the protein enters the blood stream as furin is a human specific protein [3]and during in vitro studies. this part is codon optimized for Chlamydomonas reinhardtii and has a counterpart construct codon optimized for E.coli, BBa_K3237007.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1745
    Illegal XbaI site found at 1733
    Illegal XbaI site found at 1775
    Illegal SpeI site found at 1769
    Illegal PstI site found at 1283
    Illegal PstI site found at 1751
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1745
    Illegal NheI site found at 1297
    Illegal SpeI site found at 1769
    Illegal PstI site found at 1283
    Illegal PstI site found at 1751
    Illegal NotI site found at 1781
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1745
    Illegal BamHI site found at 1763
    Illegal XhoI site found at 656
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1745
    Illegal XbaI site found at 1733
    Illegal XbaI site found at 1775
    Illegal SpeI site found at 1769
    Illegal PstI site found at 1283
    Illegal PstI site found at 1751
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1745
    Illegal XbaI site found at 1733
    Illegal XbaI site found at 1775
    Illegal SpeI site found at 1769
    Illegal PstI site found at 1283
    Illegal PstI site found at 1751
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None