Part:BBa_K4654000:Design

superfolder GFP

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

With sfGFP, we have chosen an optimized version of GFP that is better suited for our project. It is crucial for our testing system to obtain results as quickly as possible. SuperfolderGFP is a particularly fast-folding variant that allows us to achieve this and is already widely used. For all our reporters, we have removed the start codons since we already have a start codon upstream in the nhaA gene. Additionally, we have added stop codons downstream. Furthermore, we have adjusted the sfGFP sequence upstream and downstream to enable Golden Gate Cloning. Here, we utilize the ability of Type IIIS restriction enzymes to cut outside their recognition sequence, allowing for a seamless transition between two components of our plasmid. The sequence has been extended on both sides with the recognition sequence.

Source

sfGFP belongs to the family of green fluorescent proteins. The protein originates from Aequorea victoria, a species of jellyfish found in the Pacific Ocean, and was published in 2005.

References

Pédelacq J-D, Cabantous S, Tran T, Terwilliger Tc, Waldo Gs (2005). Nature Biotechnology, 24(1) , 79-88. doi: 10.1038/nbt1172.

fpbase