Part:BBa_K3866026

ParaBAD:sbm:Terminator

Usage and Biology

This TU includes the AckA 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 α1 vector and has overhangs compatible for GoldenBraid cloning.

Figure 1. The first level α module of the Propionate Production: α1:ParaBAD:RBS-smb-Double terminator

Verification of Cloning

Fig.2:: (U=Uncut , C= Cut)Positive clone: 6,Seva a2 arac ACK. Expected bands 4601, 853

Experimental Use and Experience

This part showed functionality at this part BBa_K3866009

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