Science Catalog 2012 Life Cell Signaling pGL2 Luciferase Reporter Vectors Product pGL2-Basic Vector pGL2-Control Vector pGL2-Enhancer Vector pGL2-Promoter Vector For Research Use Only. Not for Use in Diagnostic Procedures. Description: The pGL2 Luciferase Reporter Vectors provide a basis for the quantitative analysis of factors that potentially regulate mammalian gene expression. These factors may be cis-acting, such as promoters and enhancers, or trans-acting, such as various DNA-binding factors. The pGL2 Vectors carry the coding region for firefly (Photinus pyralis) luciferase, which is used to monitor transcriptional activity in transfected eukaryotic cells. The assay of this genetic reporter is rapid, sensitive and quantitative. In addition, the pGL2 Vectors contain numerous features that aid in the characterization and mutagenesis of the putative regulatory sequences. Features: • Versatile: Deletions and site-directed mutations can be made directly to inserted DNAs without subcloning. • Convenient: All vectors contain the firefly luciferase reporter gene, which enables sensitive and rapid quantitation of reporter activity. • Low Background: Upstream polyadenylation signal minimizes spurious transcription of the reporter gene. Storage Conditions: Store vector at –20°C. Store bacterial strain at –70°C. Protocol pGL2 Luciferase Reporter Vectors Technical Manual Part# TM003 ScaI 3552 XmnI 3671 3194 3188 SalI SV40 Ampr BamHI Enhancer poly(A) signal (for luc reporter) 2236 PflMI f1 ori pGL2-Control Vector (6047bp) SV40 luc poly(A) signal (for background reduction) SmaI KpnI SacI MluI NheI XhoI BglII 3 12 18 22 28 33 37 HindIII 239 SV40 Promoter start 1803 1790 1788 1777 1769 1767 1757 SP6 HindIII NsiI NotI (XbaI) ApaI (XbaI) 1702 BamHI KasI/NarI 1717/1718 (EcoRI) 1162 Ampr f1 ori T7 ori pGEM®-luc Vector (4931bp) lacZ′ luc Bsu36I 1137 BstEII 1141 SfiI SacI (EcoRI) SalI XhoI StuI ClaI 386 EcoRV 415 PpuMI 571 BspMI 351 17 27 29 35 41 47 EcoNI 135 1 start AatII 3994 NdeI 4243 NaeI 4423 DraIII 4529 Size Cat.# Price (Fr) 20 µg E1641 592.00 20 µg E1611 592.00 20 µg E1621 592.00 20 µg E1631 592.00 pGEM®-luc DNA Product pGEM®-luc DNA For Research Use Only. Not for Use in Diagnostic Procedures. Description: The pGEM®-luc Vector is a cassette vector designed to be a source of the luc gene encoding firefly luciferase, which is found in the pGL2 Vectors. The plasmid is not intended for the expression of luciferase in eukaryotic or prokaryotic cells. The pGEM®-luc Vector was constructed by positioning the luciferase gene (luc) in the center of the multiple cloning region of the pGEM®-11Zf(–) Vector, providing a number of unique restriction sites at both ends of the gene. Sites that are surrounded by parentheses are not unique, as additional sites for each also exist in the luciferase gene. Note also that using HindIII or NsiI to clone the luciferase gene will include upstream ATG codons, which may reduce the efficiency of expression in eukaryotes. The luciferase cassette does not contain the prokaryotic Shine-Delgarno sequence for bacterial expression. The pGEM®-luc Vector is supplied with a glycerol stock of bacterial strain JM109. Features: • Flexibility: Provides a luciferase cassette with several unique cloning sites at both ends for analysis of transcriptional activity, mRNA processing, protein structure/function, or labeling of cells and viruses. Storage Conditions: Store at –20°C. Store bacterial strain at –70°C. Protocol pGEM®-luc Vector Technical Bulletin Part# TB104 Size Cat.# Price (Fr) 20 µg E1541 175.00 276 For complete and up-to-date product information visit: www.promega.com/catalog   0317VA03_3A 0392VA05_3A Pagina 279

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