Difference between revisions of "Part:BBa K389010"
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Latest revision as of 16:17, 10 May 2013
VirA receptor (expressed)
This BioBrick contains a VirA receptor from Agrobacterium tumefaciens C58 under the control of a medium strong constitutive promoter.
Usage and Biology
We used this BioBrick to mutate virA by error-prone PCR so the mutated VirA receptor is expressed and ready to be screened for several substances.
The screening system for a mutated virA gene contains this BioBrick in a plasmid with ColE1 ori and the BioBrick BBa_K389011 in a plasmid with R6K ori. Both plasmids will be transformed to and screened in E. coli EC100D. Plasmids with R6K ori are only replicated in pir+ or pir116 E. coli strains. Once a receptor with high sensitivity for a screened substance is found, the plasmids are isolated and transformed to e.g. E. coli TOP10. Because the R6K ori does not work in this strain because it is not a pir+ / pir116 strain, it is easy to separate the mutated virA BioBrick from the screening plasmid.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 647 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1632
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 1768