Difference between revisions of "Part:BBa K3128016:Design"
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Euromedex BACTH kit contains the pKNT25 plasmid with a MCS cassette followed by the T25 subpart of the Bordetella pertussis adenylate cyclase. It allows the cloning of the protein of interest in fusion with the T25 subpart. To respect RFC rules, the plasmid has been amplificated by PCR using primers that bind from either site of the MCS cassette to eliminate it. Those primers contain also restriction sites enabling the cloning of the protein in the system. The plasmid was digested by the corresponding enzymes and the protein gene was inserted. | Euromedex BACTH kit contains the pKNT25 plasmid with a MCS cassette followed by the T25 subpart of the Bordetella pertussis adenylate cyclase. It allows the cloning of the protein of interest in fusion with the T25 subpart. To respect RFC rules, the plasmid has been amplificated by PCR using primers that bind from either site of the MCS cassette to eliminate it. Those primers contain also restriction sites enabling the cloning of the protein in the system. The plasmid was digested by the corresponding enzymes and the protein gene was inserted. | ||
| + | [[File:BBa K3128016 design1.png|900px|thumb|left]] | ||
| + | [[File:BBa K3128016 design2.png|900px|thumb|left]] | ||
| + | <partinfo>BBa_K3128016 SequenceAndFeatures</partinfo> | ||
===Source=== | ===Source=== | ||
Revision as of 15:24, 30 September 2019
COMP fused with T25 subpart of Bordetella Pertussis adenylate cyclase under lactose promoter
Euromedex BACTH kit contains the pKNT25 plasmid with a MCS cassette followed by the T25 subpart of the Bordetella pertussis adenylate cyclase. It allows the cloning of the protein of interest in fusion with the T25 subpart. To respect RFC rules, the plasmid has been amplificated by PCR using primers that bind from either site of the MCS cassette to eliminate it. Those primers contain also restriction sites enabling the cloning of the protein in the system. The plasmid was digested by the corresponding enzymes and the protein gene was inserted.
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1597
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Source
pKNT25 plasmid from Euromedex BACTH kit was used. OmpX gene and GGS linker were synthesized by IDT because they were unavailable on iGEM plates.