Team:MADRID UCM/Results

Cloning design - 4C_FUELS

iGEM is not an easy journey and often all the planification does not ends in the expected results, but that's how science work and every unexpected result is actually an opportunity to understand further and discover something new.

In 4C_Fuels we have found multiple difficulties in our road towards our final goal. However after an entire year working in the project we have made important achievements, which we aim they will inspire and ease the work of future iGEM teams. In this page you will find the most relevant outcomes of 4C_Fuels project.

4C_Fuels Project: The Results

After thoughtfully designing all the project contents and workplan, our team started working in all of its aspects. The results of all this work are furtherly detailed in the Results page, as well as in ever page of this wiki. However, a brief summary is explained below


As we have deeply explained within our sustainability page . 4C_Fuels was born to achieve a more sustaibable society, and this could only be achieved considering not only the technoeconomic aspect of sustainability, but also the social one.

With this idea in mind, we have developed various contents with the aim to promote social awareness about sustainability and raise the interest of general society for the potential of science and technology for solving our most pressing challenges. However, we also aimes to transmit the idea that technology alone is useless, unless people is aware about how its opinion, behavior and ideas are crucial for the definition of how technology is developed, and how it can be applied to transform our world.

In addition, we truly believe in the potential of iGEM to fuel this sustainable transition our world needs. Because of this we have taken an active role within iGEM community (know more about in the Collaborations page ), as well as promoted the iGEM concept within our region. Besides huge collaborative projects, like the participation in the iGEM PHototrophs Community, we have also offered various lectures in our region and created divulgative contents explaining what iGEM is. Our aim with all of these activities is not only to settle the foundations for the emergence of future iGEM teams in our region, but also to raise interest about synthetic biology in the society. We want people to understand how synthetic biology is a powerful tool that goe beyond the GMO debate and can actually offer solutions for many problems nowadays still unsolved.


After trial and error a few times without clear and promising results, it is easy to give up but we did try until the end. The work at the lab has been conducted iteratively, solving small problems till reaching the next one. Even though the results expected were not a reality yet, we have validated part of our theorethical design and documented all the issues we have found. We believe that our theorethical design can still be validated by future iGEM teams, which will take all of our work one step further to reality. Because of this, all the theorethical design has been extensively explained in this wiki, to ease the work of future scientist in the development of sustainable biomanufacturing technologies.

Nevertheless all our wet lab work has yileded some interesting outcomes.

  • Cell Encapsulation.On the one hand we have managed to study multiple procedures for cyanobacteria encapsulation in biohybrid materials. Our aim was the intial screening for the future development of cell-encapsulated photobiocatalyst. As a result of this research we have identified a promising silica-based nanomaterial where cyanobacterial cells can be kept alive for more than 30 days without losing its metabolic ability. All of this work has been extensively document in the encapsulation page .

  • Cyanobacteria Synthetic Biology We believe in the need to expand the available toolbox for cyanobacteria synthetic biology applications. In this aim we have proposed the theoretical design of an efficient Unmarking System for cyanobacteria, as well as created extensive documentation about cyanobacterial metabolic engineering. In addition, we have also documented our experience accessing novel fast-growin strains, which may pose important advantages to speed up the development of cyanobacteria metabolic engineering field. All of this contents can be found within our contribution to the Phototrophs Handbook as well as in thePhototrophs Synthetic Biology and Engineering pages.

  • Part Collection and Cloning design Despite we have been unable to succesfully test our expression constructs within a cyanobacteria. We managed to succesfully assemble and sequence-verify a set of useful regulatory and coding parts, which are furtherly detailed within the Contribution Page . In addition, because of our specific need to express a large number of enzymes we have adapted specific cloning procedures which are compatible with the utilization of the Marburg Collection MoClo Toolbox . All of this strategies are furtherly explained at the Cloning Design page


Besides woring in the wet lab, we have also employed all tools in our hands to adress the needs of our project. In this regard, we have worked in three main ambits.

  • Hardware Inspired by the work of previous iGEM teams we have commited ourselves to the task of optimizing and adapting for our needs a previous open-source photobioreactor system. In this regard, we have worked in the construction of a device which will help us to cultivate our cyanobacteria in tightly controlled conditions, as well as automatically record the relevant growth parameters. All this hardware work lead to the succesful construction of an improved version of the OpenPBR system designed by Humboldt_Berlin 2019 iGEM team . To know more about our findings and improvements you can visit our Hardware page

  • Software Bioinformatics is gaining more and more importance for the development of any kind of work within synthetic and molecular biology. In fact we found ourselves in the need of identifying new integration sites for the novel fast-growing cyanobacterial strain we were planning to transform. To achieve this task we have developed a python script: NSfinder. This software piece is capable of identifying putative neutral integration sites in any prokaryotik genome, requiring only genomic sequence information. Our aim with the development of this tool, was to either fulfill one of our project needs as well as to provide synthetic biology community with an additional resource for easing the utilization of non-conventional strains during their research. To now more about this you can visit our Software page

  • Modelling Eventually we have also used computational tools in order to model and simulate the behavior of our design. First, we have colaborated with MiamiU_OH 2019 iGEM Team to model our metabolic design and study how our synthetic pathways will perform in the cyanobacteria.

    Secondly, we have deveoped an intiial engineering design to model and simulate a complete photosynthetic n-butanol production bioprocess, considering current performance of n-butanol producing engineered cyanobacteria. The results of this simulation validated the techno-economic viability of our proposed technology. Further information about all of this results can be found at the Modelling page and Proof Of Concept Page .