Synthetic Biology:Vectors/Modular construction scheme

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Summary

In order to both save on construction costs and facilitate generation of a larger library of vectors in the future, an extensible vector scaffold was designed. This scaffold serves as the basis for the pSB**5 series of plasmids.

The scaffold is designed such that

  1. It can be propagated and selected via ampicillin resistance by itself.
  2. Replication origins and antibiotic resistance markers specified in BioBricks format can be assembled and cloned into the vector scaffold thereby constructing a new pSB**5 plasmid.
  3. After insertion of the replication origin and antibiotic resistance marker, all BioBricks sites and BioBricks compatible sites are destroyed enabling their future reuse for other purposes.

Design overview

The vector scaffold contains two NheI sites flanking an ampicillin resistance marker. A replication origin.resistance marker composite BioBrick part can be cut with XbaI and SpeI and cloned into the vector scaffold at the NheI sites thereby replacing the ampicillin resistance marker. All XbaI/SpeI/NheI sites are destroyed in the process.

Implementation

Sample assembly scheme for vectors.  BBa_I50040 is a replication origin and BBa_P1000 is an antibiotic resistance marker.  BBa_I51000 is the vector scaffold.  The same scheme can be used for all combinations of replication origins and antibiotic resistance markers.  Click to view larger version.
Sample assembly scheme for vectors. BBa_I50040 is a replication origin and BBa_P1000 is an antibiotic resistance marker. BBa_I51000 is the vector scaffold. The same scheme can be used for all combinations of replication origins and antibiotic resistance markers. Click to view larger version.


Also see pSB**5 design notes for more information.

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