Difference between revisions of "Part:BBa K524000:Design"

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===Design Notes===
 
===Design Notes===
  
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To make this part compatible with Biobrick standard assembly protocols, overlapping PCR was used to mutate and render non-functional a SpeI cut site originally present inside the construct.
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To make this part compatible with Biobrick standard assembly protocols, overlapping PCR was used to mutate and render non-functional a SpeI cut site originally present inside the construct, resulting in a base substitute of C to G at nucleotide residue 1391.
  
 
===Source===
 
===Source===

Latest revision as of 02:39, 6 October 2011

Heat sensitive origin of replication (oriR101 & repA101-ts)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 1709
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

To make this part compatible with Biobrick standard assembly protocols, overlapping PCR was used to mutate and render non-functional a SpeI cut site originally present inside the construct, resulting in a base substitute of C to G at nucleotide residue 1391.

Source

From the plasmid pKD46 maintained inside the E. coli strain BW25113, courtesy of E.coli Genetic Resources at Yale CGSC, The Coli Genetic Stock Center.

References

Datsenko KA, Wanner BL.(2000).One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products, Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6640-5.

D. Manen, L. Caro.(1991).The replication of plasmid pSC101, Mol Microbiol. 1991 Feb;5(2):233-7.

Phillips GJ.(1995). New Cloning Vectors with Temperature-Sensitive Replication, Plasmid. 1999 Jan;41(1):78-81.