Difference between revisions of "Part:BBa K4606000"
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In this part, we designed a plasmid for fusion protein recombination. With this device, we can insert the coding sequence of a novel protein before Poly A. When transfected into E. coli or other recipient cells, it amplifies and expresses the corresponding gene product. | In this part, we designed a plasmid for fusion protein recombination. With this device, we can insert the coding sequence of a novel protein before Poly A. When transfected into E. coli or other recipient cells, it amplifies and expresses the corresponding gene product. | ||
Due to the addition of the AmpR gene to the plasmid, we could easily confirm whether the recombination was successful or not, as only the recombinants can grow into colonies on the medium supplemented with Ampicillin. | Due to the addition of the AmpR gene to the plasmid, we could easily confirm whether the recombination was successful or not, as only the recombinants can grow into colonies on the medium supplemented with Ampicillin. |
Latest revision as of 07:24, 16 September 2023
Vector-AmpR
In this part, we designed a plasmid for fusion protein recombination. With this device, we can insert the coding sequence of a novel protein before Poly A. When transfected into E. coli or other recipient cells, it amplifies and expresses the corresponding gene product. Due to the addition of the AmpR gene to the plasmid, we could easily confirm whether the recombination was successful or not, as only the recombinants can grow into colonies on the medium supplemented with Ampicillin. To use this device, all you need to do is cut the plasmid at Xba1 and insert your target fragment into this position. We have proved its feasibility by inserting our destination fragment (Firefly Luciferase) to the plasmid. The recombinants show resistance to Ampicillin.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 1163
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 12
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 1163
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 1163
Illegal NgoMIV site found at 487 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 279
Illegal BsaI.rc site found at 2637
Illegal SapI site found at 1557