Generator

Part:BBa_K1979005

Designed by: MA Xinyi   Group: iGEM16_SDSZ_China   (2016-09-17)
Revision as of 03:50, 30 October 2016 by Xinyiloowhoo (Talk | contribs)


CBD-PBP5

This part codes for CBD (cellulose binding domain)- PBP5 protein.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 135
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1322
    Illegal BamHI site found at 168
    Illegal XhoI site found at 1530
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 636
    Illegal AgeI site found at 1238
  • 1000
    INCOMPATIBLE 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.





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.


[edit]
Categories
Parameters
None