Regulatory

Part:BBa_K540001:Design

Designed by: Mathilde Dumond   Group: iGEM11_Lyon-INSA-ENS   (2011-09-15)

rcn, cobalt-sensitive promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 47
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

An internal iGEM restriction site had to be mutated by QuickChange mutagenesis.


Source

PCR from E.coli MC4100 genome with the following primers (5'-3') : FW:GTTTCTTCGAATTCGCGGCCGCTTCTAGAGTTATTATTATTTGATATATGAATCCAGCACCTTCAG REV:CTGCAGCGGCCGCTACTAGTACTGAAGAAGAGTTGTAAATTCGGTCATG

This PCR was followed by a Quickchange mutagenesis with the following oligos (5'-3') : CTGTTGTAAAACTGCtGCGCATTCGTGCGGC GCCGCACGAATGCGCaGCAGTTTTACAACAG

References

Biochimie. 201193(3):434-9. The Escherichia coli metallo-regulator RcnR represses rcnA and rcnR transcription through binding on a shared operator site: Insights into regulatory specificity towards nickel and cobalt Didier Blaha, Safia Arous, Camille Blériot, Corinne Dorel, Marie-Andrée Mandrand-Berthelot, Agnès Rodrigue