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ACS Synth Biol. 2013 Feb 15;2(2):111-20. doi: 10.1021/sb300114d. Epub 2013 Jan 11.

Promoter element arising from the fusion of standard BioBrick parts.

Author information

1
Department of Biomedical Engineering, University of California Davis, One Shields Avenue, Davis, CA 95616, USA.

Erratum in

  • ACS Synth Biol. 2013 Jun 21;2(6):351.

Abstract

We characterize the appearance of a constitutive promoter element in the commonly used cI repressor-encoding BioBrick BBa_C0051. We have termed this promoter element pKAT. Full pKAT activity is created by the ordered assembly of sequences in BBa_C0051 downstream of the cI gene encoding the 11 amino acid LVA proteolytic degradation tag, a BioBrick standard double-TAA stop codon, a genetic barcode, and part of the RFC10 SpeI-XbaI BioBrick scar. Placing BBa_C0051 or other pKAT containing parts upstream of other functional RNA coding elements in a polycistronic context may therefore lead to the unintended transcription of the downstream elements. The frequent reuse of pKAT or pKAT-like containing basic parts in the Registry of Biological Parts has resulted in approximately 5% of registry parts encoding at least one instance of a predicted pKAT promoter located directly upstream of a ribosome binding site and ATG start codon. This example highlights that even seemingly simple modifications of a part's sequence (in this case addition of degradation tags and barcodes) may be sufficient to unexpectedly change the contextual behavior of a part and reaffirms the inherent challenge in carefully characterizing the behavior of standardized biological parts across a broad range of reasonable use scenarios.

PMID:
23656374
DOI:
10.1021/sb300114d
[Indexed for MEDLINE]

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