Difference between revisions of "Part:BBa K3866031"

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===Verification of Cloning===
 
===Verification of Cloning===
 
[[File:T--Thessaly--grand1gel.png|800px|thumb|none|<i><b>Figure 2.</b>: (C=Cut, U=Uncut) Restriction digestion of α1-sbm-ygfG-ygfH (C5-C6-C7-C8-C9) with: EcoRV, Expected bands: 5727bp, 2167bp, 1509bp, 1434bp, 745bp, Positive result: C5-C6-C7-C8-C9</i>]]
 
[[File:T--Thessaly--grand1gel.png|800px|thumb|none|<i><b>Figure 2.</b>: (C=Cut, U=Uncut) Restriction digestion of α1-sbm-ygfG-ygfH (C5-C6-C7-C8-C9) with: EcoRV, Expected bands: 5727bp, 2167bp, 1509bp, 1434bp, 745bp, Positive result: C5-C6-C7-C8-C9</i>]]
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[[File:T--Thessaly--grand2gel.png|800px|thumb|none|<i><b>Figure 3.</b>: (C=Cut, U=Uncut) Restriction digestion of α1-sbm-ygfG-ygfH with: BamHI, Expected bands: 5727bp, 2612bp, 1903bp, 1076bp, 264bp, Positive result: C1-C2-C3-C4-C5</i>]]
  
 
===Experimental Use and Experience===
 
===Experimental Use and Experience===

Revision as of 23:47, 25 September 2021


Propionate Production Construct

Usage and Biology

This composite part consists of the following two Composite Parts:

sbm:stuffer: BBa_K3866029 & ygfG:ygfH: BBa_K3866030

Design Notes

The coding sequences were domesticated. We removed BsmBI and BsaI sites in order to be compatible with GoldenBraid and MoClo. The sequences are cloned in seva ω2 vector and have overhangs compatible for GoldenBraid cloning.

Figure 1. The ω module of the Propionate Production: α1:ParaBAD:RBS-sbm-Double terminator:ParaBAD:RBS-ygfG-Double terminator:ParaBAD:RBS-ygfH-Double terminator

Verification of Cloning

Figure 2.: (C=Cut, U=Uncut) Restriction digestion of α1-sbm-ygfG-ygfH (C5-C6-C7-C8-C9) with: EcoRV, Expected bands: 5727bp, 2167bp, 1509bp, 1434bp, 745bp, Positive result: C5-C6-C7-C8-C9
Figure 3.: (C=Cut, U=Uncut) Restriction digestion of α1-sbm-ygfG-ygfH with: BamHI, Expected bands: 5727bp, 2612bp, 1903bp, 1076bp, 264bp, Positive result: C1-C2-C3-C4-C5

Experimental Use and Experience

This part showed functionality at part BBa_K3866031

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Unknown
  • 21
    INCOMPATIBLE WITH RFC[21]
    Unknown
  • 23
    INCOMPATIBLE WITH RFC[23]
    Unknown
  • 25
    INCOMPATIBLE WITH RFC[25]
    Unknown
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 965
    Illegal SapI site found at 3160
    Illegal SapI.rc site found at 1883


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