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