Abstract
The Registry of Standard Biological Parts only accepts genetic parts compatible with the RFC 10 BioBrick format. This combined assembly and submission standard requires that four unique restriction enzyme sites must not occur in the DNA sequence encoding a part. We present evidence that this requirement places a nontrivial burden on iGEM teams developing large and novel parts. We further argue that the emergence of inexpensive DNA synthesis and versatile assembly methods reduces the utility of coupling submission and assembly standards and propose a submission standard that is compatible with current quality control strategies while nearly eliminating sequence constraints on submitted parts.
Highlights
The Registry of Standard Biological Parts only accepts genetic parts compatible with the RFC 10 BioBrick format
At the time BioBricks were introduced, restriction enzyme cloning was the dominant method for assembling multiple DNA sequences into a single construct, and E. coli was the host for most synthetic biology devices
As genes and gene clusters with new activities are discovered and iGEM teams seek to add these parts to the Registry, greater incidences of illegal restriction enzyme site sequences are expected to be found within the DNA sequences of prospective parts
Summary
The Registry of Standard Biological Parts only accepts genetic parts compatible with the RFC 10 BioBrick format. BioBrick parts are typically submitted on the Escherichia coli plasmid pSB1C3, and they must be flanked by defined prefix and suffix sequences containing restriction enzyme sites for 3A assembly [2] (Figure 1A).
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