Coding
LacI

Part:BBa_K143033:Design

Designed by: Chris Hirst   Group: iGEM08_Imperial_College   (2008-09-16)

LacI (Lva-, N-terminal deletion) regulatory protein



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

LacI was located in the sequence of the B. subtilis shuttle vector pDR111. This version of LacI lacks an Lva degradation sequence and has a small N-terminal deletion that is observed in many LacI used in studies on B.subtilis. In particular, this LacI protein is used in pDR111 to regulate expression of the inducible Phyper-spank protein (BBa_K143015) (also used in the pDR111 vector). The BioBrick prefix and suffix were applied to the gene.

Template

B. subtilis integration plasmid pDR111#1

Forward Primer

5'- GCT CTA GAT GAA ACC AGT AAC GTT ATA CGA TG - 3'

Reverse Primer

5'- GGA CTA GTA TTA TTA CTG CCC GCT TTC CAG TC -3'

Source

The LacI gene was cloned from the B. subtilis shuttle vector pDR111 using Pfu DNA polymerase PCR.

References

<biblio>

  1. 1 pmid=12562810

</biblio>