Coding

Part:BBa_K3755007:Design

Designed by: Kaijun Wang   Group: iGEM21_ShanghaiTech_China   (2021-09-29)


GCaMP6m


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 34
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 175
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1016
    Illegal SapI.rc site found at 922


Design Notes

We used PCR to obtain GCaMP6m from pAAV.Syn.Flex.GCaMP6m.WPRE.SV40(Part:BBa_K3755032). Then we inserted it behind CMV promoter using pEGFP as the backbone.

Source

It's a fusion protein which consists of a circularly permuted GFP (cpGFP), calmodulin (CaM) and a calmodulin-binding peptide(CBP). We get pAAV.Syn.Flex.GCaMP6m.WPRE.SV40 containing it from Wei Shen' lab, Shanghaitech university.

References

[1]Chen TW, Wardill TJ, Sun Y, Pulver SR, Renninger SL, Baohan A, Schreiter ER, Kerr RA, Orger MB, Jayaraman V, Looger LL, Svoboda K, Kim DS. Ultrasensitive fluorescent proteins for imaging neuronal activity. Nature. 2013 Jul 18;499(7458):295-300. doi: 10.1038/nature12354. PMID: 23868258; PMCID: PMC3777791.