Part:BBa_K1321113

Phytochelatin (PC) EC20 + linker-CBDcipA-linker with LacI promoter

A LacI-promoter expression construct of Synthetic phytochelatin EC20 (metal binding peptide) fused N-terminally to CBDcipA (a cellulose-binding domain), which contains an endogenous N and C-terminal linker sequence.

At present the site-directed mutagenesis for this construct is in progress to correct an illegal EcoRI site which was identified in the CBDcipA.

This construct is part of a library of fusions with cellulose binding domains which we designed to bind to cellulose and enable capture of heavy metals ([http://2014.igem.org/Team:Imperial/Functionalisation project page]). Other fusion parts with this metal binding protein can be seen in the table below:

Note that the stop codon plus 6 bp at the end of the sequence are included the RFC25 suffix which is not shown. The prefix to this part is RFC10 format.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 512
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 512
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 512
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 512
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 512
  Illegal NgoMIV site found at 230
  
• 1000
  COMPATIBLE WITH RFC[1000]