Composite

Part:BBa_K3634016:Design

Designed by: Laurence Seeley   Group: iGEM20_St_Andrews   (2020-08-08)
Revision as of 12:32, 8 August 2020 by Ls268 (Talk | contribs) (→‎Design Notes)


PlacIq + LacI Repressor (+ mf-Lon deg. tag)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The sequence for lacI was taken from BBa_K1695000 with the 3' tga stop codon removed and the initiation start codon atg replaced with the native gtg start codon to improve protein expression efficiency. BBa_K2333001, the strong mf-Lon degradation tag taken from Mesoplasma florum, was then directly attached to the 3' end of the lacI gene to give the in silico fusion product.

Source

LacI alongside its promoter is found natively in E.coli. The initial sequence used for lacI was taken from BBa_K1695000 (see design notes) and the mutant promoter taken from BBa_K091111. The strong mf-Lon degradation tag is native to Mesoplasma florum and the sequence taken from BBa_K2333001.

References