Difference between revisions of "Part:BBa K5387001:Design"
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The sequence contains the amino acid sequence "QPPP", which is very difficult for the ribosomes in E. coli to translate KÄLLA. We did not do it, but for future projects we would recommend altering this part of the sequence, either by mutation or when ordering the sequence. | The sequence contains the amino acid sequence "QPPP", which is very difficult for the ribosomes in E. coli to translate KÄLLA. We did not do it, but for future projects we would recommend altering this part of the sequence, either by mutation or when ordering the sequence. | ||
| − | Primers for cloning into pET28a(+) between restriction sites NdeI and EcoRI: | + | Primers for cloning into pET28a(+) between restriction sites NdeI and EcoRI: |
| − | fwd: 5'-GCGCCATATGGCTGTTTATAGGCTATGTGTAACTACAGGACCG-3' | + | : ""fwd"": 5'-GCGCCATATGGCTGTTTATAGGCTATGTGTAACTACAGGACCG-3' |
| − | rev: 5'-GCGCGAATTCTTAGATGCTAACGCTGTTCTCAATCAGTGGTGGG-3' | + | : ""rev"": 5'-GCGCGAATTCTTAGATGCTAACGCTGTTCTCAATCAGTGGTGGG-3' |
===Source=== | ===Source=== | ||
Revision as of 08:36, 20 August 2024
Design Notes
The sequence has been optimized for expression in E. coli.
The sequence contains the amino acid sequence "QPPP", which is very difficult for the ribosomes in E. coli to translate KÄLLA. We did not do it, but for future projects we would recommend altering this part of the sequence, either by mutation or when ordering the sequence.
Primers for cloning into pET28a(+) between restriction sites NdeI and EcoRI:
- ""fwd"": 5'-GCGCCATATGGCTGTTTATAGGCTATGTGTAACTACAGGACCG-3'
- ""rev"": 5'-GCGCGAATTCTTAGATGCTAACGCTGTTCTCAATCAGTGGTGGG-3'
Source
This gene is expressed in humans KÄLLA