Composite
Part:BBa_K1433005:Design
Designed by: Hanqing Liu Group: iGEM14_ZJU-China (2014-09-28)
RBS-gamma protein-beta protein-exonuclease
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1422
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
No
Source
Derive from pKD46 by PCR.
Mutate by PCR.
Mutation Primers:
Mutant1-F: atactgcatacacGgcagaacg
Mutant1-R: cgttctgcCgtgtatgcagtat
Mutant2-F: CGGACATTATCCTGCAACGTA
Mutant2-R: TACGTTGCAGGATAATGTCCG
Mutant3-F: tgtttgaGttcacttccggc
Mutant3-R: gccggaagtgaaCtcaaaca
References
- Baba T, Ara T, Hasegawa M, et al. Construction of Escherichia coli K‐12 in‐frame, single‐gene knockout mutants: the Keio collection[J]. Molecular systems biology, 2006, 2(1).
- Mosberg J A, Lajoie M J, Church G M. Lambda red recombineering in Escherichia coli occurs through a fully single-stranded intermediate[J]. Genetics, 2010, 186(3): 791-799.
- Datsenko K A, Wanner B L. One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products[J]. Proceedings of the National Academy of Sciences, 2000, 97(12): 6640-6645.
- Sharan S K, Thomason L C, Kuznetsov S G, et al. Recombineering: a homologous recombination-based method of genetic engineering[J]. Nature protocols, 2009, 4(2): 206-223.