Composite

Part:BBa_K4733011:Design

Designed by: Diego Perez Hidalgo   Group: iGEM23_UBC-Okanagan   (2023-10-05)


MWPp-RBS-VP3-HisTag-ThrClLinker-CotB-Term


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 286
    Illegal EcoRI site found at 1259
    Illegal SpeI site found at 307
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 286
    Illegal EcoRI site found at 1259
    Illegal SpeI site found at 307
    Illegal NotI site found at 2239
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 286
    Illegal EcoRI site found at 1259
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 286
    Illegal EcoRI site found at 1259
    Illegal SpeI site found at 307
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 286
    Illegal EcoRI site found at 1259
    Illegal SpeI site found at 307
    Illegal NgoMIV site found at 1150
    Illegal AgeI site found at 1063
    Illegal AgeI site found at 1156
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1526


Design Notes

Given that MoClo produces a four-nucleotide scar between each CDS, the team had to insert two extra amino acids per scar to avoid a codon shift. To avoid having protein structure disruption the added nucleotides would produce Glycine amino acid, which is known to not affect protein structure. If you were to use this composite part you must also make this change.


Source

The UBC-Okanagan 2023 team used Modular Cloning (MoClo) standardization for this part: added Type II restriction sites and standard flanks to this part.

References