Difference between revisions of "Part:BBa K3076300:Design"
Line 7: Line 7:
  
 
===Source===
 
===Source===
−
The genomic sequence of CopA gene was  extracted from NCBI (ACCESSION: NC_000913 REGION: complement (508875..511379)). The kanamycin resistance gene was extracted from the sequence of pET28a plasmid sequence.  
+
The genomic sequence of <i>CopA</i> gene was  extracted from NCBI (ACCESSION: NC_000913 REGION: complement (508875..511379)). The kanamycin resistance gene was extracted from the sequence of pET28a plasmid sequence from addgene.org. Double terminator was extracted from iGEM part registry <html>(<a href="https://parts.igem.org/Part:BBa_B0015">BBa B0015</a>)</html> which is the most commonly used double terminator with high efficiency.  
  
 
===Design Notes===
 
===Design Notes===
−
The recombination site was chosen to be near the C terminal of the gene to increase the chance of successful knockout.
+
The recombination site was chosen to be near the 5' end of the coding strand to increase the chance of successful knockout.
  
 
===References===
 
===References===

Revision as of 13:14, 13 October 2019


dsDNA substrate with KanR gene for CopA knockout in E. coli by Lambda Red Recombineering


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Source

The genomic sequence of CopA gene was extracted from NCBI (ACCESSION: NC_000913 REGION: complement (508875..511379)). The kanamycin resistance gene was extracted from the sequence of pET28a plasmid sequence from addgene.org. Double terminator was extracted from iGEM part registry (BBa B0015) which is the most commonly used double terminator with high efficiency.

Design Notes

The recombination site was chosen to be near the 5' end of the coding strand to increase the chance of successful knockout.

References