pEcoli-6xHN-GFPuv

pEcoli-6xHN-GFPuv is a control vector that encodes a GFPuv fusion protein containing an N-terminal 6xHN tag (composed of 6 repeating His-Asn subunits) and an enterokinase cleavage (EK) site. High-level, IPTG-inducible expression of the fusion is driven by the T7 lac hybrid promoter (PT7 lac), which consists of the strong T7 promoter (1–3) and the lac operator (4). Basal expression of the fusion is repressed by the Lac repressor (lacI), which binds to the lac operator, preventing expression from the promoter in the absence of IPTG. High-level, IPTG-inducible expression of 6xHN-tagged GFPuv is driven by the T7 promoter in the presence of T7 RNA polymerase.

The vector also contains an ampicillin resistance gene (Ampr) and a pBR322 origin of replication, which maintains the vector at a low-copy-number to further reduce basal levels of the fusion.