This part codes for CBD (cellulose binding domain)- PBP5 protein.
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12INCOMPATIBLE WITH RFCIllegal NheI site found at 7
Illegal NheI site found at 135
- 21INCOMPATIBLE WITH RFCIllegal BglII site found at 1322
Illegal BamHI site found at 168
Illegal XhoI site found at 1530
- 23COMPATIBLE WITH RFC
- 25INCOMPATIBLE WITH RFCIllegal AgeI site found at 636
Illegal AgeI site found at 1238
- 1000INCOMPATIBLE WITH RFCIllegal SapI site found at 184
The CBD sequence is retrieved from the GenBank. We modified its codon usage preference from eukaryotic to prokaryotic expression. We designed a plasmid with restriction site XbaI, promoter BBa_J23101, rbs, and restriction site BamHI, sent it for synthesis at Beijing Zi Xi Biotech, and the fragment we got was digested by enzymes. The result DNA and our PBP5 generator (BBa_K1979003) are digested by XbaI and BamHI, after which they are purified and ligated, and become our new construct.
Figure2. M: Marker; 1-3: deposit of E.coli after centrifugation
The CBD-PBP5 protein (red box) is expressed, with a molecular weight of 53kDa. E.coli in channel 3 is the blank comparison.