DNA
amyE 5 IS

Part:BBa_K143001:Design

Designed by: Chris Hirst   Group: iGEM08_Imperial_College   (2008-08-27)

5' Integration sequence for the amyE locus of B. subtilis


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]

Design Notes

The amyE integration sequence was taken from the vector after comparison by BLAST to the B. subtilis chromosome to identify the homologous sequences. The sequence present in both the host chromosome and the plasmid at the 5' end of the gene is the 5' sequence required for integration

Template

B. subtilis integration plasmid pDR111#1

Forward Primer

5'- GCT CTA GAG ATG TTT GCA AAA CGA TTC AAA AC - 3'

Reverse Primer

5'- GGA CTA GTA TCG ATC AGA CCA GTT TTT AAT TTG -3'

Source

The 5’ integration sequence was taken from the shuttle vector pDR111 which has been used in many studies on B. subtilis, in particular in the studies of transcriptional control#2,#3,#4

References

<biblio>

  1. 1 pmid=12562810
  2. 2 pmid=14597697
  3. 3 pmid=15937167
  4. 4 pmid=12169614

</biblio>