Difference between revisions of "Part:BBa K3866006"

 
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===Usage and Biology===
 
===Usage and Biology===
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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.
  
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===Design Notes===
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<span class='h3bb'>Sequence and Features</span>
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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.
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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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===Verification of Cloning===
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[[File:T--Thessaly--arac-ackgel.png|700px|thumb|none|<i><b>Fig.2:</b>: (U=Uncut , C= Cut)Positive clone: 6,Seva a2 arac ACK. Expected bands 4601, 853</i>]]
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===Experimental Use and Experience===
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This part showed functionality at this part <bbpart>BBa_K3866009</bbpart>
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===Sequence and Features===
 
<partinfo>BBa_K3866006 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3866006 SequenceAndFeatures</partinfo>
  
  
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===References===
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===Functional Parameters===
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*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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<partinfo>BBa_K3866006 parameters</partinfo>
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Latest revision as of 15:07, 25 September 2021


ParaBAD-ackA-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 level a module of the Actetate Production : a2:ParaBAD:RBS-AckA-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
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1148
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 983
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 965


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