Difference between revisions of "Part:BBa K5034219:Design"
 
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===Design Notes===
 
===Design Notes===
  
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PAP gene was PCR-amplified and inserted into the plasmid pYYDT(after NdeI and XhoI digestion) to get the expression vector. It then got transformed into <i>E. coli</i> DH5&#945; to amplify, and was purified by picking monoclonal culture and sequencing.
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''PAP'' 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> DH5&#945; to amplify, and was purified by picking monoclonal culture and sequencing.
  
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The Terminator(BBa_B0015) is on the plasmid backbone(BBa_K5034201).
+
The terminator(BBa_B0015) is on the plasmid backbone(BBa_K5034201).
  
 
===Source===
 
===Source===

Latest revision as of 11:38, 1 October 2024


Poly P + AMP-> ADP


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 4981
    Illegal suffix found in sequence at 1
  • 12
    INCOMPATIBLE 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
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4981
    Illegal BglII site found at 3580
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 4981
    Illegal suffix found in sequence at 2
  • 25
    INCOMPATIBLE 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
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

PAP 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

Phosphate-AMP phosphotransferase(PAP) from Acinetobacter johnsonii. NCBI reference sequence: AB092983.1

References

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