Team:Bulgaria/Contribution


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Contribution


Bronze medal contribution


We assembled a test construct consisting of INPUT TetR constitutive generator (Part: BBa_K145201), Ptet promoter and a GFP generator. We made this construct since we wanted to test and optimize the induction of the pTet promoter. In course of our work we noticed that GFP is produced in high amounts even without an induction. The explanation we found most logical was that the TetR protein has a degradation tag (we found one that is not described in part BBa_K145201 but exists in the sequence) and it is not stable enough. So we decided to modify and improve this part. To do so we PCR amplify the part in a way that the degradation tag is removed and the TetR constitutive generator is in the opposite direction (limiting the chance for incidental transcription events). Our new part was characterized and submitted to the Registry.

Other contributions


1) We designed a BioBrick compatible gRNA vector and submitted it to the registry.


2) We tested and optimized the gRNA cloning process.