Coding

Part:BBa_K1403019:Design

Designed by: Paris Bettencourt 2014   Group: iGEM14_aris_Bettencourt   (2014-10-06)
Revision as of 04:01, 19 October 2014 by Tovila (Talk | contribs)

Acetohydroxybutanoate synthase /acetolactate synthase (ilvB/N) expression cassette


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 47
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 47
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 47
  • 1000
    COMPATIBLE 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.