Talk:Synthetic Biology:Vectors

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Vector design review meeting

We held a meeting to review the proposed design for some new BioBricks standard vectors.

Meeting notes

  • Are there any other features that we want in these vectors?
    • All BioBricks compatible restriction sites should be removed from the vectors once the resistance marker and replication origin have been inserted.
    • Only include a single antibiotic resistance marker (rather than two).
    • Include a unique restriction site in the backbone for situations where the insert is equal in size to the backbone (such that they can be distinguished).
    • Since all components will be synthesized, remove unnecessary restriction sites. Also do codon optimization? See Codon pairs.
  • Should the flanking terminators be inside or outside the verification primer binding sites?
    • Inside.
  • Plasmid barcode
    • Use a 3-4 bp tag that can be read during sequencing. Also place this tag immediately 3' to VF2 and VR so that vector specific primers can be used in single colony PCR?
  • How can we "verify" the design?
    • Caitlin, Barry and TK have volunteered to inspect the design.
  • How should the vectors be fabricated?
    • Direct synthesis of useful vector components, scaffold and intact vectors (budget of $20,000). Prepare a flow chart of synthesis and cloning steps so that we can obtain useful intermediates and test whether they work or not.

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