Difference between revisions of "Part:BBa K3866029"
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===Usage and Biology=== | ===Usage and Biology=== | ||
− | This composite part consists of the <i>sbm</i> Trancription Unit <bbpart> | + | This composite part consists of the <i>sbm</i> Trancription Unit <bbpart>BBa_K3866026</bbpart>, and the stuffer <bbpart>BBa_K3866019</bbpart> |
===Design Notes=== | ===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 | + | 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 ω1 vector and has overhangs compatible for GoldenBraid cloning. |
− | [[Image:T--Thessaly- | + | [[Image:T--Thessaly--sbm-stuffer-term.png|800px|thumb|none|<I><b>Figure 1.</b> The level ω module of the Propionate Production: ω1:ParaBAD:RBS-sbm-Double terminator:stuffer </i>]] |
===Verification of Cloning=== | ===Verification of Cloning=== | ||
− | [[File:T--Thessaly-- | + | [[File:T--Thessaly--sbm-stuffergel.png|800px|thumb|none|<i><b>Figure 2.</b>: (C=Cut, U=Uncut) Restriction digestion of ω1-sbm-stuffer (C6-C7-C8) with: EcoRI + EcoRV, Expected bands: 7439bp, 1509bp, 1132bp, 302bp Positive result: C6-C7-C8</i>]] |
===Experimental Use and Experience=== | ===Experimental Use and Experience=== | ||
− | This part showed functionality at | + | This part showed functionality at part <bbpart>BBa_K3866031</bbpart> |
===Sequence and Features=== | ===Sequence and Features=== |
Latest revision as of 10:55, 12 October 2021
ParaBAD:sbm:Terminator - stuffer
Usage and Biology
This composite part consists of the sbm Trancription Unit BBa_K3866026, and the stuffer BBa_K3866019
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 ω1 vector and has overhangs compatible for GoldenBraid cloning.
Verification of Cloning
Experimental Use and Experience
This part showed functionality at part BBa_K3866031
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 2948
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 2948
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 2948
Illegal BamHI site found at 1148
Illegal BamHI site found at 2236 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 2948
- 25INCOMPATIBLE 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 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 965
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