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Team:Hannover/Parts

Parts

Our aim is to circularize the TAL-effector so the proteins are solid during the purification. To do this we use the vector construct of the iGem team of Heidelberg in 2014 that has an Intein mechanism (more information at http://2014.igem.org/Team:Heidelberg). However, we changed a part of the Intein sequence because we wanted to optimize and stabilize the circulation. Another change was made on the repeat side of the TALE- we did into the vector of the team in Heidelberg. If you want to read more about our project concept click here Project.

We designed four circularized vectors with the help of the Heidelberg team’s mechanism and our own linker. All of them contain the TAL-effector sequence, the Strep Tag, the TEV side and the Intein sequence, a special TAL-effector sequence. This is so that they can bind to another DNA sequence. Additionally, two of them contain a GFP sequence.

Part NumberTypePart NameShort descriptionLength
BBa_K2075000TagStrep XT TagStrep Tag for protein purification and detection90 bp
BBa_K2075001CodingeGFPenhanced green fluorescence protein, used as reporter protein717 bp
BBa_K2075003Protein domainTEV Cleavage Siteencodes a amino acis sequencen, which is a specific cleavage site for the TEV protease21 bp
BBa_K2075004PlasmidVector iGem_02_Ax7L-DSCombination of TAL effector Ax7L-DS with TEV Site, Intein and Strep tag5841 bp
BBa_K2075005PlasmidVector iGem_02_Hax3-2xNGCombination of TAL effector Hax3-2xNG with TEV Site, Intein, eGFP and Strep Tag5396 bp
BBa_K2075007PlasmidVector iGem_02_Ax7R-RRCombination of TAL effector Ax7R-RR with TEV Site, Intein and Strep tag5739 bp
BBa_K2075009CodingHax3-2xNNTAL-effector protein, binds the DNA sequence: A C A C C C G/A G/A N C A1777 bp
BBa_K2075010CodingIntein and linker (N-Term)useful by the circularization of a protein by ligating the N- and C-terminal ends524 bp
BBa_K2075020CodingAx7L-DSTAL-effector protein, binds the DNA sequence: A G/A C A C T A T A T A A A C C C C C3210 bp
BBa_K2075021CodingAx7R-RRTAL-effector protein, binds the DNA sequence: T G/A A A G/A G/A C T T G/A A T G/A A G/A C T2946 bp
BBa_K2075022PlasmidVector iGem_02_Hax3-2xNNCombination of TAL effector Hax3-2xNN with TEV Site, Intein, eGFP and Strep Tag5396 bp
BBa_K2075023CodingHax3-2xNGTAL-effector protein, binds the DNA sequence: A C A C C C T T N C A T1803 bp
BB_aK2075024CodingIntein and linker (C-Term)useful by the circularization of a protein by ligating the N- and C-terminal ends336 bp
Sponsors

Our project would not have been possible without financial support from multiple sponsors and supporters.
Carl Roth IDT Leibniz University Hannover Leibniz Universitätsgesellschaft e.V. New England Biolabs Promega Sartorius SnapGene

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