Composite

Part:BBa_K3237018:Design

Designed by: Kristi Turton   Group: iGEM19_Lethbridge   (2019-10-11)


His-mRFP1-SCI57 with furin cut sites codon optimized for E.coli


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 649
    Illegal AgeI site found at 761
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The linkers were added after the furin cut sites to allow for space for the protease to be able to cut the amino acid sequence and to prevent steric hinderance.



Source

The SCI57 coding sequence was found from the literature [3] and the RFP sequence was taken from the registry (BBa_E1010). The part was synthesized by IDT.

References

1. Hua,Q. (2010) Insulin: a small protein with a long journey. Protein Cell. 6, 537-551.


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


3. 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.