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 a SpeI cut site originally present inside the construct to render it nonfunctional.
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To make this part compatible with Biobrick standard assembly protocols, overlapping PCR was used to mutate a SpeI cut site originally present inside the construct to render it non-functional.
  
 
===Source===
 
===Source===

Revision as of 22:07, 5 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 a SpeI cut site originally present inside the construct to render it non-functional.

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.