Part:BBa_K3634016:Design

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

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

Calos M.P. 1978. DNA sequence for a low-level promoter of the lac repressor gene and an 'up' promoter mutation. Nature. 274: p762-765.

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.

Glascock C.B., Weickert M.J. 1998. Using chromosomal lacIQ1 to control expression of genes on high-copy-number plasmids in Escherichia coli. Gene. 223(1-2): p221-231.

Missouri Western State University iGEM 2008 - https://parts.igem.org/Part:BBa_K091111

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