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Part:BBa_K1076001:Design
Designed by: Luna Lacerda, Adolfo Mota, Carlos Gustavo Nunes Group: iGEM13_Manaus_Amazonas-Brazil (2013-09-13)
Flanking genes for FadR deletion
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 768
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
To design the upstream gene we used primers with Apa1(5') and NdeI(3') and to downsteam gene we used primers with NdeI(5') and BamH1(3')ends, this parts have Nde1 complementary site and they were ligate together. This method garantee generation of a deleted copy of the target gene using a two step assymmetric/crossover PCR amplification. The target in this case is the FadR.
Source
It comes from amplified genomic sequence