Regulatory
pBAD

Part:BBa_K845001:Design

Designed by: Grégory Hansen   Group: iGEM12_Grenoble   (2012-09-26)
Revision as of 13:39, 30 September 2012 by Greghansen (Talk | contribs) (→‎References)

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New paraBAD part


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 344
    Illegal XhoI site found at 494
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 179
    Illegal AgeI site found at 484
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 161


Design Notes

Our pBAD promoter has a longer sequence compared to the ones already registered and is composed of six regulation sites.( http://www.univ-orleans.fr/sciences/BIOCHIMIE/L/Illustrations%20cours/SLO-5BC03%20Regulation%20expression%20genome/Procaryotes/SLO-5BC03-COURS3.pdf )

417 bp from the promoter to the ATG of araB plus the 82 first bp of the araB gene.

Source

pBAD is a natural promoter in E. Coli and is part of its genomic sequence. It regulates the production of three proteins (AraA, AraB and AraD) which form the arabinose operon. Their production enables the use of arabinose as a carbon source. It has six regulation sites, five of them are devoted to AraC fixation (three are repressing, one has a dual activity and one is an activator); the last one is a CRP binding site (positive activity). This promoter is activated when both activated CRP and AraC are bind to it.

References

Zaslaver, A., Bren, A., Ronen, M., Itzkovitz, S., Kikoin, I., Shavit, S., Liebermeister, W., et al (2006). A comprehensive library of fluorescent transcriptional reporters for Escherichia coli. Nature Methods, 3(8), 623-628. doi:10.1038/nmeth895 ( http://www.ncbi.nlm.nih.gov/pubmed/16862137 )

http://ecocyc.org/ECOLI/NEW-IMAGE?type=OPERON&object=TU00214