We are working to develop a set of instructions for our future team members. With this set of instructions, they will see when they should be completing things by to succeed and meet the deadlines. We also keep track of any problems we encountered, whether that be with Wiki coding, team management, paperwork, or lab work.
We also hope to increase the size of our team, getting more and more members so that we can do even bigger, better things.
Overall, our goal is to make our iGEM team a “well-oiled machine” with clear roles, goals, and way to achieve these goals. We want to educate more and more Westview High School students on synthetic biology, and give them real-world experiences in STEM.
If you are making a contribution by adding information to an existing Part or creating a new Part, you must document your contribution on the Part's Main Page on the Registry for your team to be eligible for this criteria. You can use this page to link to that part and include additional information about your contribution.
FastPETase Full Transcriptional Unit (Composite Part)
RegistryThis transcriptional unit is made of a J23101 promoter (BBa_J23101), BBa_B0034 RBS, FastPETase (BBa_K5338000), 3x stop codon, and a rrNB (BBa_B0010) terminator. This transcriptional unit was successfully transformed into E.Coli when implemented on a pCA-odd1 backbone.
This transcriptional unit may also be used with other plasmid backbones. We are currently working on implementing it into a pRF backbone to directly conjugate into Alteromonas. If in pCA, you can combine with other L1s (e.g. MHETase) to create a L2 plasmid that covers the full PET enzymatic pathway. L2 Reactions have failed so far, but we will continue working on this in the next upcoming weeks.
FastPETase Gene Sequence (Basic Part)
RegistryDescription: PETase is a protein isolated from Ideonella Sakeinsis, that is found to degrade PET plastics into PET and MHET monomers. FastPETase is a synthetically modified version of PETase that was designed to be more efficient and heat-tolerant.
The original amino acid sequence was published by Nazirov,M.M., Qobilov,F.B. and Khalilov,I.M, but we performed codon optimization and implemented it into E.Coli via a L1 plasmid. The codon optimized sequence is found above. This protein has a signal peptide already added onto it.
Rhamnose Inducible Promoter (Basic Part)
RegistryDescription: This is the sequence for a rhamnose inducible promoter that was used in our attempted L2 FastPETase + MHETase plasmid. Genes of interest will be suppressed in the presence of glucose, but expression will occur with rhamnose. This rhamnose inducible promoter was used with the L2 plasmid to reduce MHETase production stress on cells, but L2 reactions failed nevertheless. We also used it with RFP to demonstrate fluorescence/expression of our plasmid, but this was not successfully implemented (Gel Electrophoresis revealed that reactions failed).
pCA Odd-1 backbone (Basic part)
RegistryDescription: This pCA backbone contains a kanamycin resistance gene (to determine successful plasmid transformations) and a pBR322 Ori. Sourced from JCVI.
Full L1 Plasmid (FastPETase Trancriptional Unit + pCA Backbone)
RegistryDescription: This is the full Level 1 Plasmid, which contains the L1 PETase Transcriptional Unit in a pCA backbone (K3338004, sourced from JCVI). SAP1 sites are located in the backbone for Loop Assembly. This plasmid can be directly transformed into E.Coli, or possibly combined with MHETase in a Golden-Gate Reaction to create a L2 plasmid with the PETase-MHETase pathway. Once again, the L2 reaction was attempted but did not succeed for Westview iGEM.