Difference between revisions of "Part:BBa K5034221:Design"
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===Design Notes=== | ===Design Notes=== | ||
− | PPK1 gene was PCR-amplified and inserted into the plasmid | + | PPK1 gene was PCR-amplified and inserted into the plasmid pBBR1MCS-terminator to get the expression vector. It then got transformed into <i>E. coli</i> 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 1(PPK1) from Citrobacter freundii. NBCI reference sequence: NZ_LR890181.1:c1367205-1365139 | + | Polyphosphate kinase 1(PPK1) from <i>Citrobacter freundii</i>. NBCI reference sequence: NZ_LR890181.1:c1367205-1365139 |
===References=== | ===References=== | ||
Wang, X., Wang, X., Hui, K., Wei, W., Zhang, W., Miao, A., . . . Yang, L. (2018). Highly Effective Polyphosphate Synthesis, Phosphate Removal, and Concentration Using Engineered Environmental Bacteria Based on a Simple Solo Medium-Copy Plasmid Strategy. Environmental Science & Technology, 52(1), 214-222. doi:10.1021/acs.est.7b04532 | Wang, X., Wang, X., Hui, K., Wei, W., Zhang, W., Miao, A., . . . Yang, L. (2018). Highly Effective Polyphosphate Synthesis, Phosphate Removal, and Concentration Using Engineered Environmental Bacteria Based on a Simple Solo Medium-Copy Plasmid Strategy. Environmental Science & Technology, 52(1), 214-222. doi:10.1021/acs.est.7b04532 |
Latest revision as of 11:38, 1 October 2024
Pi <-> Poly P
- 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
PPK1 gene was PCR-amplified and inserted into the plasmid pBBR1MCS-terminator 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 1(PPK1) from Citrobacter freundii. NBCI reference sequence: NZ_LR890181.1:c1367205-1365139
References
Wang, X., Wang, X., Hui, K., Wei, W., Zhang, W., Miao, A., . . . Yang, L. (2018). Highly Effective Polyphosphate Synthesis, Phosphate Removal, and Concentration Using Engineered Environmental Bacteria Based on a Simple Solo Medium-Copy Plasmid Strategy. Environmental Science & Technology, 52(1), 214-222. doi:10.1021/acs.est.7b04532