Difference between revisions of "Part:BBa K1979005"
 
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<p class="title" style="font-size:20px;">Construct</p><br/>
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<p>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. </p><br/><br/>
 
<img src="https://static.igem.org/mediawiki/parts/4/43/T--SDSZ_China--cbd-pbp_PCR.jpg" style="width:60%;"/><br/><br/>
 
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<p class="text">Figure2. M: Marker; 1-3: deposit of E.coli after centrifugation<br/>The CBD-PBP5 protein (red box) is expressed, with a molecular weight of 53kDa. E.coli in channel 3 is the blank comparison.</p>
 
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Latest revision as of 03:51, 30 October 2016


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.