From 2006.igem.org

Cloning

We have been able to successfully clone the following antibiotic pancakes

• Hin
• Hin with LVA degredation tag
• Cmr- forward
• Kan-forward
• Tet-forward
• Tet-backwards
• Cmr-backwards

KanB

• Kan backwards is the only gene that we are completely positive has not been cloned into a plasmid. Further testing is needed to see if it has indeed been cloned.

RE and Hix

We have been able to clone RE into cells. The RE that we are using has two point mutations but they should not compromise the activity of the Fis-binding sites please see the page for part Part BBa_J3101 [http://partsregistry.org/Part:BBa_J3101] for further details.

• As per the [http://2006.igem.org/Davidson_Assembly_Plan| assembly plan] we will be attaching the RE downstream of the double terminator BBa_B0015 [http://partsregistry.org/Part:BBa_B0015]

• We also have two cultures with the correct sequences for Hix C.
• We have sent a "Master Plate" with cultures of all of the things that we have cloned to the Mammoths.

Cloning Parts of the Constructs

The underlined portion of each construct represents parts that have been successfully ligated and cloned:

pAra-RBS-Hix- TetF-Hix-DFT-RE

pAra-Hix-RBS-TetF-Hix-DFT-RE

pAra-RBS-Hix-TetB-Hix-DFT-RE

pAra-Hix- TetB- SBR-Hix-DFT-RE

• SBR= backwards RBS