Landing Pads: aid in the insertion of biological constructs onto the chromosome of E. coli

Landing Pad Goals:

• BioBrick compatibility
• Easy phenotypic screening
• Easy sequencing
• Limit noise

Polylinker Features:

• BioBrick compatible restriction sites - these allow for standard BioBrick construction in forward and reverse directions
• Additional building restriction sites - these are for building the landing pad, as well as changing the homologous regions and drug resistance gene (can also be used for building if needed)
• Bar code sequence - a 20 bp segment that allows for easy sequencing
• Drug resistance gene - this allows for easy phenotypic screening
• Homologous regions - these specify where on the chromosome you wish to put your constructs.

Operon

Activator Operon

Repressor Operon

Topology

Landing Pad

2nd Operon in Arabinose Landing Pad

This is what our repressor operon would look like in our Arabinose BioBrick landing pad.

Notice that the landing pad plasmids are Ampicilin resistant, to allow for additional screeining. There is also a ScaI restriction site located in the AmpR gene, that is found nowhere else on the plasmid, that can be used for linearizing the plasmid. It is necessary to linearize the plasmid in order to cross it onto the chromosome.

For testing and results on these landing pads, please visit last year's iGEM page on Landing Pads [http://synthbio.engin.umich.edu/wiki/index.php/BioBrick_Landing_Pad HERE].