Composite

Part:BBa_K1433005:Design

Designed by: Hanqing Liu   Group: iGEM14_ZJU-China   (2014-09-28)
Revision as of 17:06, 17 October 2014 by Lilina (Talk | contribs) (→‎Source)

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RBS-gamma protein-beta protein-exonuclease


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1422
  • 1000
    COMPATIBLE 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

  1. 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).
  2. 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.
  3. 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.
  4. 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.