Composite
pmyo2-ChR2

Part:BBa_K1634007:Design

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


pmyo2-ChR2-YFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 948
    Illegal XhoI site found at 986
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 288
    Illegal AgeI site found at 1627
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2436
    Illegal SapI site found at 1351
    Illegal SapI.rc site found at 198


Design Notes

Since we use the restriction enzyme site to do the ligation, we should pay attention to the sequence of Pmyo2 and ChR2-YFP, avoiding using the restriction site exits in it.


Source

We get Pmyo2 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 Pmyo2 into PPD95.75, a vector commonly used in C.elegans. Finally, we construted the fused protein, ChR2-YFP,and ligated it into PPD95.75_Pmyo2.

References