Promoter Element Arising from the Fusion of Standard BioBrick Parts

作者：Yao Andrew I; Fenton Timothy A; Owsley Keegan; Seitzer Philip; Larsen David J; Sit Holly; Lau Jennifer; Nair Arjun; Tantiongloc Justin; Tagkopoulos Ilias; Facciotti Marc T^*

来源：ACS Synthetic Biology, 2013, 2(2): 111-120.

DOI：10.1021/sb300114d

• synthetic biology
• standard assembly
• promoter
• transcription
• part characterization
• iGEM
• BioBrick

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.

• 出版日期2013-2