Difference between revisions of "Part:BBa K1433005:Design"

(→‎References)
(→‎Source)
 
Line 12: Line 12:
  
 
===Source===
 
===Source===
 +
<p>Derive from pKD46 by PCR.</p>
 +
<p>Mutate by PCR.</p>
  
−
No
+
 
 +
<b><big>Mutation Primers:</big></b><br />
 +
<p>Mutant1-F: atactgcatacacGgcagaacg</p>
 +
<p>Mutant1-R: cgttctgcCgtgtatgcagtat</p>
 +
<p>Mutant2-F: CGGACATTATCCTGCAACGTA</p>
 +
<p>Mutant2-R: TACGTTGCAGGATAATGTCCG</p>
 +
<p>Mutant3-F: tgtttgaGttcacttccggc</p>
 +
<p>Mutant3-R: gccggaagtgaaCtcaaaca</p>
  
 
===References===
 
===References===

Latest revision as of 17:06, 17 October 2014


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.