Composite
Part:BBa_K551000:Design
Designed by: Laurence Van Melderen Group: iGEM11_ULB-Brussels (2011-09-19)
FRT-CM-FRT
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The initial part (BBa_J61025) wasn't giving the resistance, so we remade it from the intermediate CmR-FRT (BBa_J61024). For that we made a PCR with a primer containing the FRT sequence and the RFC10 prefix; this means there is no scar between the first FRT and CmR. This is the reason we encoded it as a new biobrick to differentiate it from 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.