Reporter

Part:BBa_K1943011:Design

Designed by: Shixin Lu, Shiqiang Tang   Group: iGEM16_SUSTech_Shenzhen   (2016-09-04)


msfGFP+B0015, green fluorescence protein reporter system


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
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 74


Design Notes

In our lab, we usually use sfGFP as a report for molecular experiment. In the plasmid, there is a sfGFP coding device which contains promoter BBa_J23116 and RBS BBa_B0034. Some of our backbones contain KpnI site, and we need a single KpnI site, while sfGFP contains a KpnI site. So we do site-directed mutagenesis of it. And then we design primer and do PCR of this msfGFP coding device and ligate it to BBa_B0015 (adding a terminator) that we do double enzyme digestion, EcoRI & XbaI.

Source

sfGFP coding sequence was from our instructor, Huangwei's lab.
The following list is the Biobricks we’ve used in construction.
1.BBa_B0015

References

  1. Parts Registry Assembly Help
  2. Parts Registry Transformation Guideline