Difference between revisions of "Part:BBa K5034227:Design"
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===Design Notes=== | ===Design Notes=== | ||
− | |||
+ | PPK2 gene was PCR-amplified and inserted into the plasmid pBBR1MCS-terminator(after EcoRI and XhoI digestion) to get the expression vector. It then got transformed into E. coli DH5α to amplify, and was purified by picking monoclonal culture and sequencing. | ||
+ | The terminator(BBa_B0015) is on the plasmid backbone(BBa_K5034201). | ||
===Source=== | ===Source=== | ||
− | + | Polyphosphate kinase 2(PPK2) from <i>Pseudomonas paraeruginosa</i>. NCBI reference sequence: NZ_CP020560.1:c164262-163366 | |
===References=== | ===References=== | ||
+ | |||
+ | Zhang, H., Ishige, K., & Kornberg, A. (2002). A polyphosphate kinase (PPK2) widely conserved in bacteria. Proceedings of the National Academy of Sciences, 99(26), 16678-16683. doi:10.1073/pnas.262655199 | ||
+ | |||
+ | Neville, N., Roberge, N., & Jia, Z. (2022). Polyphosphate Kinase 2 (PPK2) Enzymes: Structure, Function, and Roles in Bacterial Physiology and Virulence. International Journal of Molecular Sciences, 23(2), 670. doi:10.3390/ijms23020670 |
Revision as of 11:25, 30 September 2024
Pi <-> PolyP
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 4981
Illegal suffix found in sequence at 1 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 4981
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NotI site found at 9
Illegal NotI site found at 2834
Illegal NotI site found at 4987 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 4981
Illegal BglII site found at 3580 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 4981
Illegal suffix found in sequence at 2 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 4981
Illegal XbaI site found at 4996
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NgoMIV site found at 562
Illegal NgoMIV site found at 4244
Illegal NgoMIV site found at 4527
Illegal AgeI site found at 402 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
PPK2 gene was PCR-amplified and inserted into the plasmid pBBR1MCS-terminator(after EcoRI and XhoI digestion) to get the expression vector. It then got transformed into E. coli DH5α to amplify, and was purified by picking monoclonal culture and sequencing.
The terminator(BBa_B0015) is on the plasmid backbone(BBa_K5034201).
Source
Polyphosphate kinase 2(PPK2) from Pseudomonas paraeruginosa. NCBI reference sequence: NZ_CP020560.1:c164262-163366
References
Zhang, H., Ishige, K., & Kornberg, A. (2002). A polyphosphate kinase (PPK2) widely conserved in bacteria. Proceedings of the National Academy of Sciences, 99(26), 16678-16683. doi:10.1073/pnas.262655199
Neville, N., Roberge, N., & Jia, Z. (2022). Polyphosphate Kinase 2 (PPK2) Enzymes: Structure, Function, and Roles in Bacterial Physiology and Virulence. International Journal of Molecular Sciences, 23(2), 670. doi:10.3390/ijms23020670