Difference between revisions of "Part:BBa K3866026"
(→‎Design Notes)
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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.
 
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.
  
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[[Image:T--Thessaly--arac-ack-term.png|900px|thumb|none|<I><b>Figure 1.</b> The level a module of the Actetate Production : a2:ParaBAD:RBS-AckA-Double terminator </i>]]
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[[Image:T--Thessaly--arac-sbm-term.png|900px|thumb|none|<I><b>Figure 1.</b> The first level α module of the Propionate Production: α1:ParaBAD:RBS-smb-Double terminator </i>]]
  
 
===Verification of Cloning===
 
===Verification of Cloning===

Revision as of 22:07, 25 September 2021


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 a2 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


References