Composite

Part:BBa_K3365054:Design

Designed by: Zou Aixuan   Group: iGEM20_SJTU-BioX-Shanghai   (2020-10-22)


enhanced GFP with double terminator


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 721
    Illegal XhoI site found at 730
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Due to the complex secondary structure of double terminator, it is quite hard to add double terminator to eGFP fragment only by performing PCR twice.


Source

We get double terminator from plasmid pE1a-RFP offered by team Tongji_China and get eGFP from plasmid HGT. We first obtain terminator by PCR on pE1a-RFP and then link it with eGFP through overlap PCR. Success is proven by both gel electrophoresis of PCR product and DNA sequencing.

References