Coding
fuGFP-CBDc

Part:BBa_K4488007:Design

Designed by: Donovan Wu, Jackie Yau, Oliver Nicholls, Jasmin Li   Group: iGEM22_Sydney_Australia   (2022-09-30)
Revision as of 08:48, 30 September 2022 by Jali2097 (Talk | contribs)

Fusion of free-use GFP with CBDclos (cellulose-binding domain) at the C-terminal end


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 163
  • 1000
    COMPATIBLE WITH RFC[1000]

Design Notes

The construct has a constitutive RBS (BBa_B0034). 6bp cloning scar (“accgcc”) was also inserted between the fuGFP and CBDclos, similar to BBa_K1321341, to act as a small linker. NdeI recognition site was removed from CBDclos sequence by changing the codon for threonine (“aca” to “acT”). A few base pairs were removed to avoid an inverted repeat and the codon usage of ≈200bp at a GC rich region was changed.

Source

The design of the part was based on the sfGFP-CBD fusion protein (BBa_K1321341) developed by the 2014 Imperial team Aqualose. The fuGFP sequence can be found here (BBa_K3814004).

References