Composite
aGFP-SpAB

Part:BBa_K4071000:Design

Designed by: María Calvo Noriega   Group: iGEM21_Stockholm   (2021-08-03)

B domain of Staphylococcal protein A labelled with amilGFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 860
    Illegal BamHI site found at 778
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Two STOP codons (TAA) at the end of amilGFP had to be engineered. Using Quikchange site-directed mutagenesis, the TAA Stop codons were mutated to TCA Serine codons. Serine is a small amino acid with an uncharged side chain, ideal to create a flexible linker region between amilGFP and protein A that allows both proteins to acquire their native folding state.

Detailed information on how the BBa_K4071000 was engineered can be found here: https://2021.igem.org/Team:Stockholm/Experiments#BioBrickEngineering

Source

Both BBa_K1073024 and BBa_K103003 subparts were obtained from the 2021 DNA Distribution Kit Plates


References