Part:BBa_K3634015:Design

LacI + mf-Lon Degradation 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 is found natively in E.coli and the initial sequence used here was taken from BBa_K1695000 (see design notes). The strong mf-Lon degradation tag is native to Mesoplasma florum and the sequence taken from BBa_K2333001.

References

Jacob F., Monod J. 1961. Genetic Regulatory Mechanisms in the Synthesis of Proteins. J. Mol. Biol. 3: p818-356.

Oehler S., Eismann E.R., Krämer H., Müller-Hill B. 1990. The three operators of the lac operon cooperate in repression. EMBO J. 9(v): p973-979.

City University of Hong Kong iGEM 2015 - https://parts.igem.org/Part:BBa_K1695000

William and Mary iGEM 2017 - https://parts.igem.org/Part:BBa_K2333001