Team:Michigan/Project/LandingPads

From 2008.igem.org


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BioBrick Landing Pads

We will be using landing pads to insert our clock onto the chromosome of E. coli. Noisy behavior has proven detrimental to clock studies throughout the past, and we hope to reduce the noise in our system using landing pads.


BioBrick Landing Pad Polylinker

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 - our landing pads use Chloramphenicol resistance genes
  • Homologous regions - these specify where on the chromosome you wish to put your constructs.
Landing Pad Polylinker - gold.png


Landing Pads used for the Sequestilator

Operon



Activator Operon




Repressor Operon

Topology

New 1st operon - gold.PNG


New 2nd operon - gold.PNG

Landing Pad

Leucine Landing Pad

constructed by former Ninfa lab member, Don Eun Chang


Arabinose Landing Pad

iGEM 2007 project by Alyssa Delke & Khalid Miri


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].

Landing pad plasmid - gold.png





Learn more about Landing Pads [http://synthbio.engin.umich.edu/wiki/index.php/BioBrick_Landing_Pad Here!]