Part:BBa_K3866026
ParaBAD:sbm:Terminator
Usage and Biology
This TU includes the sbm gene placed under the control of the arabinosed-induced promoter BBa_K3866000. Initially, the level 1 cloning was performed using the trc promoter BBa_K3866001, though we weren't able to isolate a colony with the desired construct, as there is a possibility that the bacteria's metabolism went out of balance so no colonies survived to be chosen for screening. Instead we used the arabinose-indused promoter, which seemed to be working as expected.
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 a PDGB3_α1 vector and has overhangs compatible for GoldenBraid cloning.
Verification of Cloning
Experimental Use and Experience
This part showed functionality at the following parts: BBa_K3866029, BBa_K3866031
Sequence and Features
- 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
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