Difference between revisions of "Part:BBa K524005:Design"
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===References=== | ===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. | + | Datsenko KA, Wanner BL.(2000).One-step inactivation of chromosomal genes in <i>Escherichia coli</i> K-12 using PCR products, Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6640-5. |
| − | F Wu, I Goldberg, and M Filutowicz.(1992).Roles of a 106-bp origin enhancer and Escherichia coli DnaA protein in replication of plasmid R6K, Nucleic Acids Res. 1992 February 25; 20(4): 811–817. | + | F Wu, I Goldberg, and M Filutowicz.(1992).Roles of a 106-bp origin enhancer and <i>Escherichia coli</i> DnaA protein in replication of plasmid R6K, Nucleic Acids Res. 1992 February 25; 20(4): 811–817. |
F Wu, I Goldberg, and M Filutowicz.(1994).Binding of DnaA protein to a replication enhancer counteracts the inhibition of plasmid R6K gamma origin replication mediated by elevated levels of R6K pi protein, J Bacteriol. 1994 November; 176(22): 6795–6801. | F Wu, I Goldberg, and M Filutowicz.(1994).Binding of DnaA protein to a replication enhancer counteracts the inhibition of plasmid R6K gamma origin replication mediated by elevated levels of R6K pi protein, J Bacteriol. 1994 November; 176(22): 6795–6801. | ||
Latest revision as of 14:14, 5 October 2011
pToolkit
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1215
Illegal EcoRI site found at 2724
Illegal PstI site found at 2219
Illegal PstI site found at 2466 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1215
Illegal EcoRI site found at 2724
Illegal PstI site found at 2219
Illegal PstI site found at 2466 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1215
Illegal EcoRI site found at 2724
Illegal BamHI site found at 1154 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1215
Illegal EcoRI site found at 2724
Illegal PstI site found at 2219
Illegal PstI site found at 2466 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1215
Illegal EcoRI site found at 2724
Illegal PstI site found at 2219
Illegal PstI site found at 2466
Illegal AgeI site found at 989
Illegal AgeI site found at 2646 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 4422
Illegal SapI site found at 971
Design Notes
The heat sensitive replication origin oriR101 & repA101-ts from the pKD46 backbone was replaced by the ori-gamma of the R6K origin of replication. Since the function of the adjacent orf60a to oriR101 & repA101ts was unknown, it was retained in pToolkit.
Source
The backbone of the pToolkit, including the arabinose inducible lambda RED recombination system and the ampicillin resistance gene (bla) comes from the RED recombinase plasmid pKD46, 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.
F Wu, I Goldberg, and M Filutowicz.(1992).Roles of a 106-bp origin enhancer and Escherichia coli DnaA protein in replication of plasmid R6K, Nucleic Acids Res. 1992 February 25; 20(4): 811–817.
F Wu, I Goldberg, and M Filutowicz.(1994).Binding of DnaA protein to a replication enhancer counteracts the inhibition of plasmid R6K gamma origin replication mediated by elevated levels of R6K pi protein, J Bacteriol. 1994 November; 176(22): 6795–6801.