Regulatory
lacI+pL

Part:BBa_R0011:Design

Designed by: Neelaksh Varshney, Grace Kenney, Daniel Shen, Samantha Sutton   Group: Antiquity   (2003-01-31)

Promoter (lacI regulated, lambda pL hybrid)


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]

Use

To induce this promoter region, generally add 1mM of IPTG to a plate (amount will vary by cell strain). That is, total volume of stuff in plate (usually around 0.025 liters) * 0.001 = X amount of liters that you will add to the solution.

Design Notes

hybrid promoter design to create strong promoter that is, at the same time, highly repressible. note that the upstream operator installed in this hybrid is slightly different than the one in the original source (Lutz and Bujard, 1997). the most upstream operator region is slightly truncated in the represillator version, so that both operators in the hybrid are the same sequence. see references for details. also, the sequence has been truncated after the transcriptional start site.

LacI binds to this regulator. This part is incompatible with species containing active LacI coding regions. Lactose and IPTG disable the operation of LacI and increase transcription. This part is incompatible with environments containing lactose or lactose analogs.

Source

represillator of Elowitz and Leibler (2000)

References

  • Lutz R, Bujard H. Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucleic Acids Res. 1997 Mar 15;25(6):1203-10.