Coding
fuGFP-CBDc

Part:BBa_K4488008:Design

Designed by: Donovan Wu, Jackie Yau, Oliver Nicholls, Jasmin Li   Group: iGEM22_Sydney_Australia   (2022-09-30)
Revision as of 05:00, 9 October 2022 by Jali2097 (Talk | contribs) (→‎Source)

Fusion of free-use GFP with CBDcex (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). A 6bp cloning scar (“accgcc”) was also inserted between the fuGFP and CBDclos, similar to BBa_K1321342, to act as a small linker. NdeI recognition site was removed from CBDclos sequence by changing the codon for threonine (“aca” to “acT”). Many base pairs were removed to avoid an inverted repeat and the codon usage at a GC rich region was changed.

Source

The design of the part was based on the sfGFP-CBD fusion protein (BBa_K1321342) developed by the 2014 Imperial team Aqualose with the type of GFP replaced. The fuGFP sequence can be found here (BBa_K3814004).

References