Translational_Unit

Part:BBa_K497004:Design

Designed by: Joanna Hare   Group: iGEM10_Macquarie_Australia   (2010-10-24)

Here, we have further improved part BBa_K497004 to facilate the expression of bacteriophytochrome gene. Please see our design below:

BBa K497004.jpg

Also we have particularly designed this part to be used as a BioBrick by coupling our upstream part (bacteriophytochrome gene and ribosome-binding-site) and our downstream part (double terminator available in iGEM2010 kit well 23L) in standard iGEM plasmid (pSB1C3). We have successfully proved that our biobrick is functional (please see the experience page).Please see our design below:


Biobrick Design.jpg

Note: Although we have designed our parts as a BioBrick. 5' of our BioBrick is missing Xpa1 site. It is because our bacteriophytochrome has an internal restriction site so when we cut with Xpa we end up cutting our bacteriophytochrome gene as well. However 3' of our Biobrick is still editable and allows us to add any other parts. Moreover we can add an Xpa site to the 5' by site-directed mutagenesis. Also we can still use site-directed mutagenesis to remove that internal site from our bacteriophytochrome gene. (We couldn't use site-directed mutagenesis because we discovered that our gene has an internal site just a week before the deadline after we sequenced it. And repeating the ligation/cloning/sub-cloning/sequencing took a week to design our new and intact Biobrick so we didn't have time for adding a restriction site to our gene. We couldn't think it during summer break either because as you know we don't have such a "summer break" in Australia and we had to work during the semester instead)