Composite
Part:BBa_K3365054:Design
Designed by: Zou Aixuan Group: iGEM20_SJTU-BioX-Shanghai (2020-10-22)
enhanced GFP with double terminator
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 721
Illegal XhoI site found at 730 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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.