Difference between revisions of "Part:BBa K3265026"
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This is an improved version of part BBa_K2213012 from team Manchester 2017. We down-regulated the base expression by exchanging the RBS to a the very weak RBS J61100. EDIT | This is an improved version of part BBa_K2213012 from team Manchester 2017. We down-regulated the base expression by exchanging the RBS to a the very weak RBS J61100. EDIT | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
Revision as of 07:11, 17 October 2019
EutSMN bacterial microcompartment
This is an improved version of part BBa_K2213012 from team Manchester 2017. We down-regulated the base expression by exchanging the RBS to a the very weak RBS J61100. EDIT
Usage and Biology
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Characterization
To test if our part could now produce the EutM and EutN proteins at levels low enough so that it would not be toxic to the cells anymore, we redid the OD measurement with the original EutSMN part.
100mL LB cultures were incoluated with a single colony from an overnight transformation of pSEVA441 vector carrying our improved EutSMN part. E. coli strain DH5alpha was used.
Cultures were grown for 4 hours at 37° 300 rpm and then induced with 4.5 ng/mL Anhydrotetracylcine (ATC) or 45 ng/mL Tetracycline (Tetc) and 1mM Arabinose ((+)araB) each. One culture was not induced (control).
Measurements were taken each hour for 9 hours and a final measurement was taken after 20 hours.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1293
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1232
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1067
Illegal AgeI site found at 3824 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 3212