Composite

Part:BBa_K1634004:Design

Designed by: Xinhong Chen   Group: iGEM15_China_Tongji   (2015-09-14)


pmyo3-ChR2-YFP ( C.elegans )


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1068
    Illegal XhoI site found at 1106
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1747
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2556
    Illegal SapI site found at 1471
    Illegal SapI.rc site found at 259


Design Notes

Since we use the restriction enzyme site to do the ligation, we should pay attention to the sequence of Pmyo3 and ChR2-YFP, avoiding using the restriction site exits in it.


Source

We get Pmyo3 from the C.elegans' genomic DNA by PCR. ChR2 is a channelrhodopsin in green algae, but we didn't get it directly from the algae. Instead, we get ChR2 from some plamids by PCR. Then we ligated the Pmyo3 into PPD95.75, a vector commonly used in C.elegans. Finally, we construted the fused protein, ChR2-YFP,and ligated it into PPD95.75_Pmyo3.

References