Difference between revisions of "Part:BBa K1403019:Design"
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===Design Notes=== | ===Design Notes=== | ||
− | This sequence was submitted downstream a | + | The purpose of this part is to overexpress <i>ilvB/N</i> in <i>E. coli</i>. |
+ | This sequence was submitted downstream a T7 constitutive promoter inside a pETDUET vector. | ||
− | The | + | The <i>ilvB/N</i> sequence was isolated from <i>E. coli</i> MG1655 through PCR, digested with NcoI and EcoRI and cloned into the pETDUET-1 vector. |
+ | This part is not strictly necessary for production of acetoin in <i>E. coli</i>, but it improves its production. | ||
− | + | Oligos: | |
+ | Fwd: 5'- GTTCAAGCCTTGAGCGGTTACTG -3' | ||
− | + | Rev: 5'- GAACTTGCCATGCTCCAGTCCT -3' | |
− | E. coli MG1655 | + | ===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]) | ||
===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. | 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. |
Revision as of 04:01, 19 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. This sequence was submitted downstream a T7 constitutive promoter inside a pETDUET vector.
The ilvB/N sequence was isolated from E. coli MG1655 through PCR, digested with NcoI and EcoRI and cloned into the pETDUET-1 vector. This part is not strictly necessary for production of acetoin in E. coli, but it improves its production.
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.