Device

Part:BBa_K886001

Designed by: edgar andres ochoa cruz   Group: iGEM12_USP-UNESP-Brazil   (2012-09-24)
Revision as of 22:14, 24 September 2012 by Andresochoa (Talk | contribs)

Recombination Device composed by lox71-Cre recombinase

This device was created to express the Cre recombinase enzyme and a target gene under the control of the same promoter (promoter not included in this part). The target gene needs to be amplified by PCR, using primers that will insert a loxP and a lox66 sites flanking the sequence, in order to integrate it in the lox71 recombination site. For this part the lox71 is located upstream the Cre recombinase gene. An alternative construction is submitted in the part BBa_K886002.

Once this device is located under the control of the desired promoter, it will express the Cre recombinase that recognizes the lox sites and inserts the target gene under the control of the same promoter.

The Cre expression levels are critical for the recombination, we created two different devices: BBa_K886002 (Cre-Lox71) and BBa_K886001 (Lox71-Cre) for testing if inserting the target gene either upstream or downstream the Cre gene will affect the expression efficiency of the target gene.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 396
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 82
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//chassis/prokaryote/ecoli
//classic/generator/plasmids
//function/recombination/cre
Parameters
chassisE. coli
functionRecombination/expression device