Coding

Part:BBa_K3866004

Designed by: Venetios Michelioudakis   Group: iGEM21_Thessaly   (2021-09-23)


pta- Phosphate acetyltransferase GB compatible with B3-B5

Figure 1. The level 0 module : pupd2- pta(illustration from SnapGene)

Usage and Biology

Involved in acetate metabolism. Pta catalyzes the interconversion of acetyl-CoA and acetyl phosphate. The direction of the reaction changes depending the conditions. On minimal medium acetyl-CoA is produced. In rich mediums acetyl-CoA is converted to acetate. In our System is one of the two genes for Acetate production.

Design Notes

The coding sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is presents in pUPD2 and has overhangs compatible for Golden Braid cloning. The CDS has position B3-B5.


Figure 2.The overhangs of this part in the GoldenBraid Grammar.


Verification of Cloning

Fig.3:(U=Uncut C=Cut) Positive clone. 4 Expected bands 3027,1174, 50

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1028
    Illegal PstI site found at 1574
    Illegal PstI site found at 1874
    Illegal PstI site found at 2039
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1028
    Illegal PstI site found at 1574
    Illegal PstI site found at 1874
    Illegal PstI site found at 2039
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1678
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1028
    Illegal PstI site found at 1574
    Illegal PstI site found at 1874
    Illegal PstI site found at 2039
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1028
    Illegal PstI site found at 1574
    Illegal PstI site found at 1874
    Illegal PstI site found at 2039
    Illegal AgeI site found at 134
    Illegal AgeI site found at 532
    Illegal AgeI site found at 1778
    Illegal AgeI site found at 1976
  • 1000
    COMPATIBLE WITH RFC[1000]


Source

Synthesized by Twist Biosciences.

References

  • Dittrich, C. R., Bennett, G. N., & San, K. Y. (2005). Characterization of the acetate-producing pathways in Escherichia coli. Biotechnology progress, 21(4), 1062–1067. https://doi.org/10.1021/bp050073s

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