Difference between revisions of "Part:BBa K5034231:Design"
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===Design Notes=== | ===Design Notes=== | ||
− | PPN1 gene was PCR-amplified and inserted into the plasmid pBBR1MCS-terminator | + | ''PPN1'' 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). | The terminator(BBa_B0015) is on the plasmid backbone(BBa_K5034201). |
Latest revision as of 11:40, 1 October 2024
Poly P -> Poly P(smaller) or Pi
Assembly Compatibility:
- 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
PPN1 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
Endopolyphosphatase(PPN1) from Saccharomyces cerevisiae. NCBI reference sequence: NC_001136.10.
References
Lichko, L. P., Kulakovskaya, T. V., & Kulaev, I. S. (2006). Inorganic polyphosphate and exopolyphosphatase in the nuclei of Saccharomyces cerevisiae: dependence on the growth phase and inactivation of the PPX1 and PPN1 genes. Yeast, 23(10), 735-740. doi:10.1002/yea.1391