Part:BBa_K3866029

ParaBAD:sbm:Terminator - stuffer

Usage and Biology

This composite part consists of the sbm Trancription Unit BBa_K3866026, and the stuffer BBa_K3866019

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 ω1 vector and has overhangs compatible for GoldenBraid cloning.

Figure 1. The level ω module of the Propionate Production: ω1:ParaBAD:RBS-sbm-Double terminator:stuffer

Verification of Cloning

Figure 2.: (C=Cut, U=Uncut) Restriction digestion of ω1-sbm-stuffer (C6-C7-C8) with: EcoRI + EcoRV, Expected bands: 7439bp, 1509bp, 1132bp, 302bp Positive result: C6-C7-C8

Experimental Use and Experience

This part showed functionality at part BBa_K3866031

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 2948
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 2948
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 2948
  Illegal BamHI site found at 1148
  Illegal BamHI site found at 2236
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 2948
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 2948
  Illegal NgoMIV site found at 3263
  Illegal AgeI site found at 983
  Illegal AgeI site found at 1400
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 965
  

References

Akawi L, Srirangan K, Liu X, Moo-Young M, Perry Chou C. Engineering Escherichia coli for high-level production of propionate. J Ind Microbiol Biotechnol. 2015 Jul;42(7):1057-72. https://doi.org/10.1007/s10295-015-1627-4