Difference between revisions of "Part:BBa K3866007"

 
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===Usage and Biology===
 
===Usage and Biology===
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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.
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===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.
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[[Image:T--Thessaly--arac-pta-term.png|900px|thumb|none|<I><b>Figure 1.</b> The level a module of the Actetate Production : a2:ParaBAD:RBS-Pta-Double terminator </i>]]
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===Verification of Cloning===
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[[File:T--Thessaly--arac-ptagel.png|700px|thumb|none|<i><b>Fig.2:</b>: (U=Uncut , C= Cut)Positive Clone 1  Expected Bands 6345, 3544</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===
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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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Revision as of 15:15, 25 September 2021


ParaBAD-pta-Terminator

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.

Figure 1. The level a module of the Actetate Production : a2:ParaBAD:RBS-Pta-Double terminator

Verification of Cloning

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


Experimental Use and Experience

This part showed functionality at this part BBa_K3866009

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 2279
    Illegal PstI site found at 2825
    Illegal PstI site found at 3125
    Illegal PstI site found at 3290
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 2279
    Illegal PstI site found at 2825
    Illegal PstI site found at 3125
    Illegal PstI site found at 3290
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1148
    Illegal BamHI site found at 2929
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 2279
    Illegal PstI site found at 2825
    Illegal PstI site found at 3125
    Illegal PstI site found at 3290
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 2279
    Illegal PstI site found at 2825
    Illegal PstI site found at 3125
    Illegal PstI site found at 3290
    Illegal AgeI site found at 983
    Illegal AgeI site found at 1385
    Illegal AgeI site found at 1783
    Illegal AgeI site found at 3029
    Illegal AgeI site found at 3227
  • 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