Part:BBa_K2680257

3G LacIAM

Optimized LacI repressor LacI-AM flanked by prefix sticky end C and suffix sticky end D in a 3G-compatible backbone. For more information on 3G hybrid DNA assembly and 3G parts, see the [http://2018.igem.org/Team:William_and_Mary/Part_Collection William and Mary 3G Parts Page].

LacI regulates the lac operon by repressing it in the absence of lactose. Building on earlier studies, Adam Meyer "partially mutagenized key amino acid residues" of LacI and designed an optimized version, LacI-AM.¹. Meyer and his colleagues aimed to simultaneously select for "less background, high dynamic range, increased sensitivity, and low crosstalk" among sensors. ¹

In the image above: "Each genetically encoded sensor is shown, with coding (red) and non-coding (grey/black) mutations noted. Mutations in grey were also applied to the parental sensor. The corresponding response functions comparing the evolved (blue) and parental (grey) sensors. The fit of Equation 1 to the mean of three replicates performed on different days is shown (see Supplementary Figure 5 for data). The response function parameters for evolved sensors are provided in Table 1."¹

References

¹Meyer, A. J., Segall-Shapiro, T. H., & Voigt, C. A. (2018). Marionette: E. coli containing 12 highly-optimized small molecule sensors. BioRxiv, 285866. doi:https://doi.org/10.1101/285866

Sequence and Features

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
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 47
  Illegal BsaI.rc site found at 1142