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Revision as of 01:35, 22 October 2021

Untitled Document i

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Overview

The parts we created

We designed several biobricks that helped us reach our objectives. All parts we developed were created using Snapgene while the synthesis of the DNA blocks were obtained from our sponsor IDT.

We created two main sets of parts: The Biosensor Parts and the Selection Marker Parts.

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T--TecCEM--lab4_edited.jpg

Biosensor Parts:



To complete the design of the Biosensor at a molecular level, we created two basic parts that code for two proteins and two composite parts. The basic parts correspond to the coding sequences for our biosensor while the composite parts were designed for the expression of the proteins.

To read more information about these parts, please check the information here. 

Part name Type Short Description Length (bp)
BBa_K3809001 Coding ESR1 with Linker, His Tag and Signal Peptide 1884
BBa_k3809011 Reporter mRFP with Linker, His Tag and Signal Peptide 774
BBa_k3809012 Composite Modified ESR1 protein under strong constitutive promoter and RBS 1948
BBa_k3809013 Composite Modified mRFP protein under strong constitutive promoter and RBS 838

Selection Marker Parts:

In order to design the new selection marker we proposed, we had to produce several pieces of DNA. The purpose of these pieces was to create a new strain of E. coli with the following characteristics: Knockout of genes oxyR, gldA and glpK and its substitution with genes PuDHT, mRFP and aldO. Besides, we designed a new plasmid based on some elements from the pUC plasmid. However, this new construct is Biobrick Assembly Friendly and has our new Selection Cassette.

To read more information about these parts, please check the information here. 

Part name Type Short Description Length (bp)
BBa_K3809001 Coding Novel Selection Cassette katE. 2562
BBa_K3809004 Coding aldO 1257
BBa_K3809006 Coding PuDHT 1728
BBa_K3809014 Reporter mRFP + oxyR homology regions + FLP recognition sequence 1040
BBa_K3809003 Composite katE regulated by BBa_J23100 2462
BBa_K3809005 Composite aldO + glpK homology regions 1487
BBa_K3809007 Composite PuDHT + gldA homology regions 1958