Part:BBa_K1321126

linker-CBDcipA-linker + metallothionein SmtA

Metallothionein (metal binding peptide) SmtA 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 start and stop codon, plus 6 bp either side of the sequence, are included the RFC25 prefix and suffix which is not shown.

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 148
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 148
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 148
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 148
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 148
  
• 1000
  COMPATIBLE WITH RFC[1000]