pGL3 Luciferase Reporter Vectors Technical Manual #TM033

III.B. pGL3-Enhancer VectorThe pGL3-Enhancer Vector contains an SV40 enhancer located downstream ofluc+ and the poly(A) signal. This aids in the verification of functional promoterelements because the presence of an enhancer will often result in transcriptionof luc+ at higher levels.Amp rSynthetic poly(A)signal / transcriptionalpause site(for backgroundreduction)2256 SalI2250 BamHIoripGL3-EnhancerVector(5064bp)f1 oriluc+KpnISacIMluINheISmaIXhoIBgIIIHindIIINcoI 86NarI 121511152128323653SV40 EnhancerSV40 latepoly(A) signal(for luc+ reporter)HpaI 1902XbaI 17420745VA08_4AFigure 2. The pGL3-Enhancer Vector circle map. Additional description: luc+,cDNA encoding the modified firefly luciferase; Amp r , gene conferring ampicillinresistance in E. coli; f1 ori, origin of replication derived from filamentous phage; ori,origin of plasmid replication in E. coli. Arrows within luc+ and the Amp r geneindicate the direction of transcription; the arrow in f1 ori indicates the direction ofssDNA strand synthesis.pGL3-Enhancer Vector Sequence Reference Points:Promoter(none)Multiple cloning region 1–58Luciferase gene (luc+) 88–1740GLprimer2 binding site 89–111SV40 late poly(A) signal 1772–1993Enhancer 2013–2249RVprimer4 binding site 2307–2326ColE1-derived plasmid replication origin 2564β-lactamase gene (Amp r ) 3329–4186f1 origin 4318–4773upstream poly(A) signal 4904–5057RVprimer3 binding site 5006–5025Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USAToll Free in USA 800-356-9526 · Phone 608-274-4330 · Fax 608-277-2516 · www.promega.comPart# TM033Printed in USA.Page 4 Revised 9/07