Generator

Part:BBa_K215000

Designed by: Chris Eiben   Group: iGEM09_Washington   (2009-09-30)
Revision as of 22:04, 18 October 2009 by Siegeljb (Talk | contribs)

R0011+B0034, strong IPTG-inducible promoter with strong RBS.

This part contains the strong, IPTG-inducible promoter R0011, combined with the strong RBS B0034. This part can be used as a strong protein expression system when combined with a protein coding sequence. This part is in pSB1A3, which has a built in transcriptional terminator located 3' of the insertion site for coding sequences. This allows for a protein coding sequence to be inserted into this vector and be immediately ready for expression.

BBa K215000 Induction.png E0040 (GFP) was inserted into BBa_K215000 between S and P using standard BioBrick assembly. The resulting constructs were grow in BL21(lacIq) cells overnight at 298K in either the presence or absence of 0.5mM IPTG. The cells were harvested, washed in PBS, and then fluorescence for the whole cells was measured on a fluorometer. Cells that did not contain GFP were used as a blank. As observed, the addition of IPTG significantly increased the observed fluorescence of the cells.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//plasmidbackbone/expression/inducible
Parameters
n/aR0011+B0034, strong IPTG-inducible promoter with strong RBS.