Composite

Part:BBa_K2680116:Design

Designed by: Ethan M Jones   Group: iGEM18_William_and_Mary   (2018-10-06)


3G pLac/ara-1


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 476
    Illegal XhoI site found at 443
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 41
    Illegal BsaI.rc site found at 556


Design Notes

Used PCR to add William and Mary Pad overhangs, which were used to clone the part into William and Mary Pad backbone. Forbidden restriction sites were checked for and removed if necessary.

Conserved upstream region due to comments made in Stricker paper suggesting its necessity.

Source

Sequence originally from Lutz and Bujard 1997; later used for the creation of oscillator from Stricker et al 2008.1,2

References

1Lutz, R., & Bujard, H. (1997). 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 Research, 25(6), 1203–1210.

2Stricker, J., Cookson, S., Bennett, M. R., Mather, W. H., Tsimring, L. S., & Hasty, J. (2008). A fast, robust and tunable synthetic gene oscillator. Nature, 456(7221), 516–519. https://doi.org/10.1038/nature07389