Part:BBa_K5034223:Design
Pi <-> Poly P Poly P -> NADP
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 11
Illegal PstI site found at 3787 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 11
Illegal PstI site found at 3787
Illegal NotI site found at 2828 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 11
Illegal BglII site found at 3574
Illegal XhoI site found at 4985 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 11
Illegal PstI site found at 3787 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 11
Illegal PstI site found at 3787
Illegal NgoMIV site found at 556
Illegal NgoMIV site found at 4238
Illegal NgoMIV site found at 4521
Illegal AgeI site found at 396 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 3150
Illegal SapI.rc site found at 4087
Illegal SapI.rc site found at 4297
Design Notes
PPK2 and NADK genes were PCR-amplified and inserted into the plasmid pYYDT(after NdeI 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
BBa_K5034205: Polyphosphate kinase 2(PPK2) from Pseudomonas paraeruginosa. NCBI reference sequence: NZ_CP020560.1:c164262-163366
BBa_K5034206: Inorganic polyphosphate/ATP-NAD kinase(PPNK) from Mycobacterium tuberculosis H37Rv. NCBI reference sequence: NC_000962.3:1918746-1919669
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
Itoh, H., & Shiba, T. (2004). Polyphosphate synthetic activity of polyphosphate:AMP phosphotransferase in Acinetobacter johnsonii 210A. Journal of Bacteriology, 186(15), 5178-5181. doi:10.1128/jb.186.15.5178-5181.2004