Assembly standard 25/Overview

The 2007 Freiburg iGEM team proposed an extension of the original BioBrick assembly standard that enables in-frame assembly of protein domains. Two restriction sites are added within the original BioBrick prefix and suffix sites. These additional sites provide compatible ends, can be employed using the same cloning strategy as for the standard restriction sites, and code for amino acids suitable for linkers:

Prefix and suffix

               Prefix                                      Suffix
5' GAATTC GCGGCCGC T TCTAGA TG GCCGGC [part] ACCGGT TAAT ACTAGT A GCGGCCG CTGCAG 3'
   EcoRI    NotI      XbaI Met NgoMIV         AgeI   *    SpeI     NotI    PstI
                              (=NgoMI)

"N-part" prefix

5' GAATTC GCGGCCGC T TCTAG [ATG.part]
   EcoRI    NotI     XbaI 

Scar

Assembling two parts based on the new Freiburg standard restriction sites leaves the following scar:

5' [part A] ACCGGC [part B] 3'
             T  G

See [http://openwetware.org/wiki/The_BioBricks_Foundation:Standards/Technical/Formats The BioBricks Foundation wiki] for a discussion and comparison of different technical standards.