Coding
Part:BBa_K4724072:Design
Designed by: Xinyao Li Group: iGEM23_NJTech-China-A (2023-10-07)
DsbA
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
In the initial design, because the signal peptide is a small protein molecule with a structural length of 20-30 amino acids, its coding sequence is approximately 60-90 bp. Therefore, we wanted to design primers to add the coding sequence of the signal peptide in front of the coding sequence of the ISPET enzyme through PCR. We used the previously designed reverse PCR primers between the RBS and ISPET enzyme sequences and added the signal peptide sequence to the primers. Subsequently, plasmid construction was carried out through homologous recombination. After inserting the signal peptide sequence, we ensured that the forward primer and reverse primer had a 20 bp homologous region with the signal peptide sequence.
Source
from E.coli