Difference between revisions of "Part:BBa K524000:Design"
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===Design Notes=== | ===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 | + | 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)
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 1709
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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.