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− | <div style = 'padding-right: 190px; padding-left: 190px; text-indent: 50px;line-height: 25px;' ><a href='https://2017.igem.org/Team:William_and_Mary/Speed_Control'>Click through </a> + graphs</div> | + | <div style = 'padding-right: 190px; padding-left: 190px; text-indent: 50px;line-height: 25px;' ><a href='https://2017.igem.org/Team:William_and_Mary/Speed_Control' style='text-decoration: underline>Click through </a> + graphs</div> |
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<div style = 'padding-right: 190px; padding-left: 190px; text-indent: 50px;line-height: 25px;' > | <div style = 'padding-right: 190px; padding-left: 190px; text-indent: 50px;line-height: 25px;' > | ||
− | Graphs,<a href='https://2017.igem.org/Team:William_and_Mary/Readjustment'> click through</a>, modeling | + | Graphs,<a href='https://2017.igem.org/Team:William_and_Mary/Readjustment' style='text-decoration: underline> click through</a>, modeling |
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− | <div style = 'padding-right: 190px; padding-left: 190px; text-indent: 50px;line-height: 25px;' >Once we felt that we could understand and control gene expression speed using our system, we next wanted to make it accessible to future iGEM teams. While our system is inherently easy to clone and implement, as it only consists of only a 27 amino acid residue pdt and the associated mf-Lon protease, we wanted to make it even easier to implement. So we created a suite of ready to clone pdt constructs and placed them on the registry. Each part contains one of our six different strength E. coli optimized protein degradation tags, as well as a double stop codon and a double terminator. Combining all these parts together into one construct prevents extra cloning steps, saving time, money and aggravation. In addition to the functional elements above, each construct also contains two BsaI restriction sites for Golden Gate Assembly, two Universal Nucleotide Sequences for Gibson Assembly, as well as a number of well-tested primer sequences that can be used for any other type of cloning. We also made it easy to swap and design large libraries of constructs with different speeds, by making sure that the only difference between each ready cloning construct was a small unique region in the pdt. That means there is no need to switch primers to use a different strength pdt. Alongside our well-characterized construct <a href = "http://parts.igem.org/wiki/index.php?title=Part:BBa_K2333434">K2333434 </a> (pLac mf-Lon), these ready to clone parts should make it cheap and easy for future teams to test their constructs with a wide variety of different gene expression speeds, either by changing the pdt or the concentration of mf-Lon. | + | <div style = 'padding-right: 190px; padding-left: 190px; text-indent: 50px;line-height: 25px;' >Once we felt that we could understand and control gene expression speed using our system, we next wanted to make it accessible to future iGEM teams. While our system is inherently easy to clone and implement, as it only consists of only a 27 amino acid residue pdt and the associated mf-Lon protease, we wanted to make it even easier to implement. So we created a suite of ready to clone pdt constructs and placed them on the registry. Each part contains one of our six different strength E. coli optimized protein degradation tags, as well as a double stop codon and a double terminator. Combining all these parts together into one construct prevents extra cloning steps, saving time, money and aggravation. In addition to the functional elements above, each construct also contains two BsaI restriction sites for Golden Gate Assembly, two Universal Nucleotide Sequences for Gibson Assembly, as well as a number of well-tested primer sequences that can be used for any other type of cloning. We also made it easy to swap and design large libraries of constructs with different speeds, by making sure that the only difference between each ready cloning construct was a small unique region in the pdt. That means there is no need to switch primers to use a different strength pdt. Alongside our well-characterized construct <a href = "http://parts.igem.org/wiki/index.php?title=Part:BBa_K2333434" style='text-decoration: underline>K2333434 </a> (pLac mf-Lon), these ready to clone parts should make it cheap and easy for future teams to test their constructs with a wide variety of different gene expression speeds, either by changing the pdt or the concentration of mf-Lon. |
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<div style = 'padding-right: 190px; padding-left: 190px; text-indent: 0px;line-height: 25px;' > | <div style = 'padding-right: 190px; padding-left: 190px; text-indent: 0px;line-height: 25px;' > | ||
− | For more information please see our nominees for <a href="https://2017.igem.org/Team:William_and_Mary/Part_Collection">Best Part Collection</a> and <a href ="https://2017.igem.org/Team:William_and_Mary/Composite_Part">Best Composite Part.</a> | + | For more information please see our nominees for <a href="https://2017.igem.org/Team:William_and_Mary/Part_Collection">Best Part Collection style='text-decoration: underline</a> and <a href ="https://2017.igem.org/Team:William_and_Mary/Composite_Part" style='text-decoration: underline>Best Composite Part.</a> |
</div> | </div> | ||
Revision as of 22:57, 30 October 2017