Difference between revisions of "Part:BBa K1403019:Design"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K1403019 short</partinfo> | <partinfo>BBa_K1403019 short</partinfo> | ||
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===Design Notes=== | ===Design Notes=== | ||
− | + | The purpose of this part is to overexpress <i>ilvB/N</i> in <i>E. coli</i>. | |
+ | The <i>ilvB/N</i> genes were isolated from <i>E. coli</i> MG1655 through PCR, digested with NcoI and EcoRI, and cloned into the pETDUET-1 vector downstream a T7 promoter and RBS. | ||
+ | This part is not strictly necessary for production of acetoin in <i>E. coli</i> but improves the production thereof. | ||
+ | Oligos: | ||
+ | Fwd: 5'- GTTCAAGCCTTGAGCGGTTACTG -3' | ||
+ | |||
+ | Rev: 5'- GAACTTGCCATGCTCCAGTCCT -3' | ||
===Source=== | ===Source=== | ||
− | + | <i>ilvB/N</i> coding sequences from <i>E. coli</i> MG1655 ([http://www.ncbi.nlm.nih.gov/nuccore/545778205?from=3850802&to=3851092&sat=4&sat_key=117098803&report=gbwithparts U00096.3]) | |
− | E. coli MG1655 | + | |
===References=== | ===References=== | ||
+ | |||
+ | David R. Nielsen, Sang-Hwal Yoon, Clara J. Yuan and Kristala L. J. Prather, Metabolic engineering of acetoin and meso-2,3-butanediol biosynthesis in E. coli, January 2010. |
Latest revision as of 07:41, 31 October 2014
Acetohydroxybutanoate synthase /acetolactate synthase (ilvB/N) expression cassette
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 47
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 47
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 47
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The purpose of this part is to overexpress ilvB/N in E. coli.
The ilvB/N genes were isolated from E. coli MG1655 through PCR, digested with NcoI and EcoRI, and cloned into the pETDUET-1 vector downstream a T7 promoter and RBS. This part is not strictly necessary for production of acetoin in E. coli but improves the production thereof.
Oligos:
Fwd: 5'- GTTCAAGCCTTGAGCGGTTACTG -3'
Rev: 5'- GAACTTGCCATGCTCCAGTCCT -3'
Source
ilvB/N coding sequences from E. coli MG1655 ([http://www.ncbi.nlm.nih.gov/nuccore/545778205?from=3850802&to=3851092&sat=4&sat_key=117098803&report=gbwithparts U00096.3])
References
David R. Nielsen, Sang-Hwal Yoon, Clara J. Yuan and Kristala L. J. Prather, Metabolic engineering of acetoin and meso-2,3-butanediol biosynthesis in E. coli, January 2010.