Composite

Part:BBa_K3237023:Design

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


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]


Design Notes

The part was codon optimized for Chlamydomonas reinhardtii .


Source

The mRFP1 sequence is based off of part BBa_E1010 and the SCI57 sequence was found in the literature [1]. The construct itself was synthesized by TWIST.


References

1. Hua, Q., Nakagawa, S., Jia, W., Huang, K., Philips, N., Hu, S., and Wiess, M. (2008) Design of an active ultrastable single-chain insulin analog: synthesis, structure, and therapuetic implications. The Journal of Biological Chemistry. 21, 1473-14716.

2.Liang, J., and Yang, V. (2005) Insulin-cell penetrating peptide hybrids with improved intestinal absorption efficiency. Communications. 335, 734-738.

3. Hay, C., and Docherty, K. (2003) Enhanced expression of a furin-cleavable proinsulin. Journal of molecular endocrinology. 3, 597-607.