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Part:BBa_K1979005
CBD-PBP5
This part codes for CBD (cellulose binding domain)- PBP5 protein.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 135 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1322
Illegal BamHI site found at 168
Illegal XhoI site found at 1530 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 636
Illegal AgeI site found at 1238 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 184
Construct
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.
![](https://static.igem.org/mediawiki/parts/4/43/T--SDSZ_China--cbd-pbp_PCR.jpg)
![](https://static.igem.org/mediawiki/parts/6/60/T--SDSZ_China--cbd-pbp5_sds_.jpg)
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.
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