Difference between revisions of "Part:BBa K4477012:Design"
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===Design Notes=== | ===Design Notes=== | ||
Amino acid sequences were reverse transcribed and codon optimized for expression in E. coli B (the parent strain of SHuffle). In addition, further codon optimization was used to remove illegal restriction sites from the construct. | Amino acid sequences were reverse transcribed and codon optimized for expression in E. coli B (the parent strain of SHuffle). In addition, further codon optimization was used to remove illegal restriction sites from the construct. | ||
| + | |||
| + | A two-way terminator was used so that the KanR gene in our intended vector -- which is encoded on the strand opposite our insert -- would not be read backward into the insert and interfere with transcription of our antibody of interest. | ||
===Source=== | ===Source=== | ||
Revision as of 11:30, 12 October 2022
IK17 (anti-oxLDL) scFv - complete expression cassette
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 576
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Amino acid sequences were reverse transcribed and codon optimized for expression in E. coli B (the parent strain of SHuffle). In addition, further codon optimization was used to remove illegal restriction sites from the construct.
A two-way terminator was used so that the KanR gene in our intended vector -- which is encoded on the strand opposite our insert -- would not be read backward into the insert and interfere with transcription of our antibody of interest.
Source
1. https://www.jacc.org/doi/full/10.1016/j.jacc.2011.07.017
2. https://pubmed.ncbi.nlm.nih.gov/14644097/