Part:BBa_K886001

Recombination Device composed by lox71-Cre recombinase

This device was created to express the Cre recombinase enzyme and any target gene, both under the control of the same promoter (promoter not included in this part). The target gene needs to be amplified by PCR with primers capable of inserting a loxP and a lox66 sites on both sides of the PCR product, "floxing" (flanking) it. This construction allows the target gene to be inserted into the lox71 recombination site (BBa K886000) of the device. The process occurs via Cre recombinase recognition and interaction with the lox sites.

However, in order to have this recombination a promoter is needed (there isn't a specific promoter for this purpose). The promoter (chosen by the user) will express the Cre recombinase and the target gene expression.

Knowing that Cre expression levels are critical for the recombination, we've created two different devices: BBa_K886001 (Lox71-Cre) and BBa_K886003 (Cre-Lox71) for testing if the insertion of the target gene either upstream or downstream the Cre gene will affect its expression efficiency.

As described above, in this part the lox71 is located upstream the Cre recombinase gene. An alternative construction can be found in the part BBa_K886003.

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]