Composite

Part:BBa_K4225006:Design

Designed by: Leon Ritchie Salim   Group: iGEM22_HKUST   (2022-10-07)


katGp-RFP-Pc-OxyR


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 160
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 160
    Illegal NheI site found at 1305
    Illegal NheI site found at 1328
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 160
    Illegal BglII site found at 231
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 160
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 160
    Illegal AgeI site found at 979
    Illegal AgeI site found at 1091
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

It is important to note that katGp has an EcoRI cut site. Thus, to ligate Pc-OxyR (BBa_K4225000) and katGp-RFP(BBa_K4225005) composite parts using RFC[10], we used SpeI and PstI enzymes to linearize Pc-OxyR, and XbaI and PstI enzymes to digest the katGp-RFP part out. They are then ligated together to for this composite part that is used for our experiment. Therefore, a scar is formed between the 2 composite parts (BBa_K4225000 and BBa_K4225005)


Source

Source for katGp sequence can refer to BBa_K4225004, while the other parts are taken from biobricks.

References