Reporter

Part:BBa_K2605000

Designed by: Kaitlin Schaaf   Group: iGEM18_Calgary   (2018-10-09)


mCherry + BGH

mCherry red fluorescent protein (RFP) codon-optimized for expression in human/mammalian cells with a bovine growth hormone (BGH) terminator attached.

This is an improvement of Part BBa_J176005.


Usage and Biology

This mCherry protein is derived from mushroom coral, Discoma sp. (Shaner et al. 2004). The BGH polyadenylation signal is one of the most common terminator sequences used for trasngenic protein expression in eukaryotic cells (Petitclerc et al. 1995)

A BmtI restriction site (sequence GCTAGC) was added to the 3' end of the BGH terminator so that this can be used as a reporter gene within our Multiple-Cloning Site flanked by FRT sites Part BBa_K2605002 system.

References

Shaner N.C., Campbell R.E., Steinbach P.A., Giepmans B.N., Palmer A.E., Tsien R.Y. (2004).Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nature Biotechnology, , 1567-1572

Petitclerc,D., Attal, J., Theron, M.C., Bearzotti, M., Bolifraud, P., Kann, G., Stinnarke, M.G., Pointu, H., Puissant, C., Houdebine, L.M. (1995). The effect of various introns and transcription terminators on the efficiency of expression vectors in various cultured cell lines and in the mammary gland of transgenic mice. Journal of Biotechnology, 40(3), 169-178.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 979
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None