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.

For reference the cellulose binding domain binding capability to bacterial cellulose was measured relative to other cellulose binding domains when fused to sfGFP, the data for which can be seen here - K1321371.

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]