Team:UiOslo Norway/Parts
Biological parts
<groupparts>iGEM17 UiOslo_Norway</groupparts>
We used CYC1 (BBa_K2110000) as the terminator for our composite part.
As few previous teams have done any work on Schizosaccharomyces Pombe, there are few parts in the registry that have been documented as useful for this yeast species. We wanted to design and document a system to express GFP in S. Pombe, which meant that we had to design and test parts specifically for this.
NMT1 was chosen as the promoter of choice as it's the most commonly used expression system in S. Pombe. One of our supervisors has a lot of experience with this system, which we also considered to be beneficial as it would make it easier to troubleshoot problems. This is the background for why we elected to use this particular sequence.
We elected to use sfGFP as the fluorescent protein of choice as this is a more stably folded form of GFP, and we wanted to ensure that the protein we used wouldn't be damaged by high light intensity. None of the sfGFP-sequences we found in the parts registry had been optimized for use in S. Pombe, so we decided that making a version of sfGFP specifically for our organism would be a good choice.
Like with NMT1, CYC1 was chosen as terminator because one of our supervisors had experience with using it in expression systems, and thus a natural sequence of choice.
Combining all of these, we created our composite part, which was made with the intention of cloning it into the pJK148 yeast expression vectoras well as into the pSB1C3 submission vector.
While doing the parts design and comparing our planned parts to existing sequences, we made a script that allowed us to search through the annual parts distributions for parts with particular names or properties.This sped up the process of figuring out if our parts were unique or had already been submitted greatly, and we are providing this as we believe this may be useful for future teams as well.
NMT1 was chosen as the promoter of choice as it's the most commonly used expression system in S. Pombe. One of our supervisors has a lot of experience with this system, which we also considered to be beneficial as it would make it easier to troubleshoot problems. This is the background for why we elected to use this particular sequence.
We elected to use sfGFP as the fluorescent protein of choice as this is a more stably folded form of GFP, and we wanted to ensure that the protein we used wouldn't be damaged by high light intensity. None of the sfGFP-sequences we found in the parts registry had been optimized for use in S. Pombe, so we decided that making a version of sfGFP specifically for our organism would be a good choice.
Like with NMT1, CYC1 was chosen as terminator because one of our supervisors had experience with using it in expression systems, and thus a natural sequence of choice.
Combining all of these, we created our composite part, which was made with the intention of cloning it into the pJK148 yeast expression vectoras well as into the pSB1C3 submission vector.
While doing the parts design and comparing our planned parts to existing sequences, we made a script that allowed us to search through the annual parts distributions for parts with particular names or properties.This sped up the process of figuring out if our parts were unique or had already been submitted greatly, and we are providing this as we believe this may be useful for future teams as well.
