pGL3 Luciferase Reporter Vectors Technical Manual #TM033

III.A. pGL3-Basic VectorThe pGL3-Basic Vector lacks eukaryotic promoter and enhancer sequences,allowing maximum flexibility in cloning putative regulatory sequences.Expression of luciferase activity in cells transfected with this plasmid dependson insertion and proper orientation of a functional promoter upstream fromluc+. Potential enhancer elements can also be inserted upstream of thepromoter or in the BamHI or SalI sites downstream of the luc+ gene.Amp rSynthetic poly(A)signal / transcriptionalpause site(for backgroundreduction)oripGL3-BasicVector(4818bp)f1 oriKpnISacIMluINheISmaIXhoIBgIIIHindIIINcoI 862010 SalI2004 BamHIluc+NarI 121511152128323653SV40 latepoly(A) signal(for luc+ reporter)HpaI 1902XbaI 17420746VA08_4AFigure 1. pGL3-Basic Vector circle map. Additional description: luc+, cDNAencoding the modified firefly luciferase; Amp r , gene conferring ampicillin resistancein E. coli; f1 ori, origin of replication derived from filamentous phage; ori, origin ofreplication in E. coli. Arrows within luc+ and the Amp r gene indicate the direction oftranscription; the arrow in the f1 ori indicates the direction of ssDNA strandsynthesis.pGL3-Basic Vector Sequence Reference Points:Promoter(none)Enhancer(none)Multiple cloning region 1–58Luciferase gene (luc+) 88–1740GLprimer2 binding site 89–111SV40 late poly(A) signal 1772–1993RVprimer4 binding site 2080–2061ColE1-derived plasmid replication origin 2318β-lactamase gene (Amp r ) 3080–3940f1 origin 4072–4527upstream poly(A) signal 4658–4811RVprimer3 binding site 4760–4779Promega 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.comPrinted in USA.Part# TM033Revised 9/07 Page 3