Part:BBa_K3866009

Acetate production construct

Usage and Biology

This TU includes the Pta gene placed under the control of the arabinosed-induced promoter. The inducible promoter is used because cloning wasn't done. After the constitutive expression, the metabolism went out of balance so no colonies survied to be chosen for screening.

Design Notes

The coding sequence was domesticated . We removed BsmBI and BsaI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in seva a2 vector and has overhangs compatible for GoldenBraid cloning.

File:T--Thessaly--grandaAC.png

Figure 1. The final level omega module of the Actetate Production

Verification of Cloning

File:T--Thessaly--grandACgelgel.png

Fig.2:: (U=Uncut , C= Cut)Positive Clone 1 Expected Bands 6345, 3544

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 2853
  Illegal PstI site found at 3399
  Illegal PstI site found at 3699
  Illegal PstI site found at 3864
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 2853
  Illegal PstI site found at 3399
  Illegal PstI site found at 3699
  Illegal PstI site found at 3864
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 3503
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 2853
  Illegal PstI site found at 3399
  Illegal PstI site found at 3699
  Illegal PstI site found at 3864
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 2853
  Illegal PstI site found at 3399
  Illegal PstI site found at 3699
  Illegal PstI site found at 3864
  Illegal AgeI site found at 1959
  Illegal AgeI site found at 2357
  Illegal AgeI site found at 3603
  Illegal AgeI site found at 3801
  
• 1000
  COMPATIBLE WITH RFC[1000]

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