Difference between revisions of "Part:BBa K777100:Design"

(→‎Source)
(→‎Design Notes)
 
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*The PstI restriction site was mutated to CTGCGG using the reverse primer.
 
*The PstI restriction site was mutated to CTGCGG using the reverse primer.
 
*''flhDC'' was amplified from genomic DNA of ''E. coli'' DH10B via PCR using the following primers:
 
*''flhDC'' was amplified from genomic DNA of ''E. coli'' DH10B via PCR using the following primers:
−
**Fwd: ACTGAATTCGCGGCCGCTTCTAGATGCATACCTCCGAGTTGCTG <br>
+
**Fwd: 5'-actgaattcgcggccgcttctagatgcatacctccgagttgctg-3'<br>
−
**Rev: TCCTGCAGCGGCCGCTACTAGTTACTGCCCGGGATGGCGGTTGACATAAGC'''c'''GCAGGCAAAGCTGCCAACAG <br>(the uncapitalized "c" induces the mutation for removal of the PstI site)
+
**Rev: 5'-tcctgcagcggccgctactagttactgcccgggatggcggttgacataagcCgcaggcaaagctgccaacag-3' <br>(the capitalized "C" induces the mutation for removal of the PstI site)
  
 
===Source===
 
===Source===

Latest revision as of 19:18, 18 September 2012

flhDC


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 609
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

  • The PstI restriction site was mutated to CTGCGG using the reverse primer.
  • flhDC was amplified from genomic DNA of E. coli DH10B via PCR using the following primers:
    • Fwd: 5'-actgaattcgcggccgcttctagatgcatacctccgagttgctg-3'
    • Rev: 5'-tcctgcagcggccgctactagttactgcccgggatggcggttgacataagcCgcaggcaaagctgccaacag-3'
      (the capitalized "C" induces the mutation for removal of the PstI site)

Source

  • The part was amplified from genomic DNA of E. coli str. K-12 substr. DH10B, complete genome (CP000948.1).

References

  • Chevance F.F., Hughes K.T. Coordinating assembly of a bacterial macromolecular machine. Nat Rev Microbiol. 2008. 6:455–465.
  • Ling H., Kang A., Tan M.H., Qi X., Chang M.W. The absence of the luxS gene Increases swimming motility and flagella synthesis in Escherichia coli K12. Biochem Biophys Res Commun. 2010. 401: 521-526.