Team:BostonU/MoClo

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MoClo Kit


One of our major contributions to the iGEM community is our MoClo Kit. This kit contains the 87 Level 0 MoClo parts that we built this summer, including a resubmission of the sequence verified parts made by the 2012 BostonU team. The kit also includes all of the destination vectors required for making new Level 0 parts, as well as Level 1 and four Level 2 vectors for stringing together 4-part transcriptional units and 2-to-4-transcriptional unit devices, respectively.



Our MoClo Protocol


As shown above in colored blocks, we have submitted a total of 49 promoters, 17 ribosomal binding sites and bicistronic design elements, 17 coding sequences, and 4 terminators. We have also submitted 10 Level 0 destination vectors and 4 each of the Level 1 and Level 2 destination vectors.

Primary selection markers are antibiotic resistance cassettes found in the backbones of each destination vector. Different antibiotic resistances are denoted by the three colors light green (chloramphenicol), orange (kanamycin), and light blue (ampicillin). Level 0 vectors were built using the pSB1C3 backbone while Level 1 vectors were built using the pSB1K3 backbone and Level 2 were built with the pSB1A2 backbone. The secondary marker of the alpha fragment of the lacZ gene allows the user to utilize blue/white screening when X-Gal and IPTG are added to the transformation plates.



This MoClo Kit contains 87 Level 0 basic DNA parts for creating genetic circuits following the Modular Cloning assembly method. The diagram above shows the variations of the Level 0 Parts we have in our Kit, shown in green. The Level 1 Parts are shown in orange and are comprised of green Level 0 Parts and the same is true for the Level 2 Parts in blue made up of orange Level 1 Parts.