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Find stop codons snapgene12/21/2023 ![]() 3: Vector map of N-terminal dsup with both BioBricks and His tag in N-terminus optimized for E. ![]() 2: Vector map of wildtype dsup with both BioBricks optimized for E. 1: Vector map of empty vector pBAD18 shown by Snapgene.įig. The dsup that was optimized for HeLa has a His-tag in the N-terminus (BBa_K3780019).Īll sequences for the fragments and primers can be found in the Wet-Lab section.įig. In BBa_K3780013 (C-terminal His-tag Dsup) the His-tag can be found in front of the stop codon. BBa_K3780017 (N-terminal His-tag Dsup) contains the same parts but the His-tag is located behind the ATG-Codon. The part BBa_K3780011 (wild-type Dsup) contains the dsup, the prefix BioBrick and the suffix BioBrick as well as the arabinose promoter and the promoter region. coli (N-His C-His and wt) and one for HeLa. Four dsup constructs were designed, three for E. Snapgene was used to insert the gene into a suitable plasmid and determine primers for the Gibson assembly. This has already been taken into account in the cause of the optimization. According to the rules of iGEM, the fragments should not contain any restriction sites that are part of the BioBricks. The dsup sequence can be found on database ( retrieved on 15.06.21) and codon optimized for the work with Escherichia Coli and HeLa-Cells by using the "Codon Optimization Tool" from IDT. We worked with a optimized sequence for E.coli to reduce the risks of unwanted influences.įor the HeLa plasmid, we decided to construct only one fragment with a His-tag on the N-Terminus.Ī vector with dsup and His-Tag and without the addition of the His-tag was designed As a result the His-tag was inserted at the C-Terminus and the N-Terminus. The addition of a tag could disturb the interaction of Dsup with the DNA. The Plasmid should contain the Dsup sequence and a His-tag.
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