Composite

Part:BBa_K551000:Design

Designed by: Laurence Van Melderen   Group: iGEM11_ULB-Brussels   (2011-09-19)

FRT-CM-FRT


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]


Design Notes

The initial part (BBa_J61025) we wanted to use don't confer the chloramphenicol resistance; so we remade it from the CmR biobrick (BBa_P1004). This Cm resistance gene was amplified by PCR using primers containing a mutated FRT sequence (FRT'). As the wild-type FRT sequence contains a Xba1 restriction site, a mutation was introduced to mutate the Xba1 site in order to be able to use this biobrick in iGEM standard 10. This FRT' is the same as BBa_J61020. Since there is no biobrick scar between the FRT' and the CmR gene we encoded it as a new biobrick part compared to BBa_J61025.

Source

PCR from BBa_J61024.

References

"One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products" Datsenko KA, Wanner BL. PNAS 2000.