Device
Rubber Fuc

Part:BBa_K1088024

Designed by: Andreas Kjær   Group: iGEM13_SDU-Denmark   (2013-09-18)
Revision as of 21:11, 25 September 2013 by P.R.A (Talk | contribs)

HRT2 prenyltransferase from Hevea Brasilianis (ara promoter without araC: arabinose inducible)

This part encodes the rubberproducing prenyltransferase from the rubber tree Hevea brasiliensis. It uses isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) as substrates to produce cis-1,4 polyisoprene natural rubber.

The part is designed to express the enzyme when induced with arabinose under glucose scarce conditions. Arabinose binds the ara promoter regulator, AraC, and arabinose bound AraC then activates the transcription of the ara promoter.

Using Northern blot technique it was proved that the construct in E. coli K-12 MG1655 doesn't express the prenyltransferase when grown without arabinose. Upon addition of arabinose to the media prenyltranferase mRNA was detected. See experince for more details.

A 3xFLAG tag is behind the HRT2 prenyltransferase, but ins't part of the protein (HRT2 has it's natural stop-codon).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 125
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 65
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None