pVITRO1-Lucia/SEAP

Dual reporter plasmids - EF-1α promoters - luciferase & SEAP

Expression plasmids pVITRO1-Lucia/SEAP are developed mainly for in vitro studies.

pVITRO1-Lucia/SEAP allow the ubiquitous and constitutive co-expression of two secreted reporter genes: a secreted luciferase (Lucia) and SEAP (secreted embryonic alkaline phosphatase).

pVITRO1-Lucia/SEAP plasmids can be stably transfected in mammalian cells and the reporter genes are expressed at high levels.

pVITRO1-Lucia/SEAP plasmids carry two elongation factor 1 alpha (EF-1α) promoters, from rat and mouse origins combined to the CMV and SV40 enhancers respectively. 

pVITRO1-Lucia/SEAP plasmids are available with different selectable markers that are active both in E. coli and mammalian cells.

Specifications

Promoters: CMV enhancer / rat EF-1α & SV40 enhancer / mouse EF-1α.

Selection markers active both in E. coli and mammalian cells:

•   bsr (blasticidin resistance)
•   hph (hygromycin resistance)
•   neo (kanamycin/G418 resistance).

Reporter Genes:

•   Lucia secreted luciferase
•   SEAP (secreted embryonic alkaline phosphatase)

These products are covered by a Limited Use License (See Terms and Conditions).

Contents

pVITRO1-neo-Lucia/SEAP

• 20 µg of lyophilized DNA

pVITRO1-hygro-Lucia/SEAP

• 20 µg of lyophilized DNA
• 1 ml of Hygromycin B Gold (hygromycin) at 100 mg/ml

pVITRO1-blasti-Lucia/SEAP

• 20 µg of lyophilized DNA
• 2 x 1 ml blasticidin at 10 mg/ml

Product is shipped at room temperature

Description

Gene Combinations in a Single Plasmid

Co-expression of two or more genes from a single vector is more     efficient and convenient than using two separate vectors.   pVITRO1-Lucia/SEAP are multigenic plasmids that contain two distinct   transcription units   (TU).

Strong and Constitutive Expression of Two Transgenes

Each pVITRO1-Lucia/SEAP  plasmid features two constitutive promoters   that drive   high levels of expression from two separate TU in a large   number of   mammalian cell lines. Transcriptional interference is   minimized by using   promoters of different origins, and strong   polyadenylation signals   (polyA).
pVITRO1-Lucia/SEAP  plasmids carry   two elongation factor 1 alpha   (EF-1α) promoters, from rat and mouse   origins combined to the CMV and   SV40 enhancers respectively.

Single Selection Marker for E. coli and Mammalian Cells

pVITRO1-Lucia/SEAP  plasmids are available with a selectable marker that is active both in E. coli and mammalian cells: bsr (blasticidin resistance), hph (hygromycin resistance), neo (kanamycin/G418 resistance). In bacteria, the resistance gene is expressed from the E. coli EM7 promoter. In mammalian cells, it is transcribed from the promoter     located 3’ of the Ori, as a polycistronic mRNA and translated through     the IRES of the Foot and Mouth Disease Virus.

Two Reporter Genes

pVITRO1-Lucia/SEAP express two reporter genes: Lucia luciferase and SEAP
Lucia secreted luciferase and SEAP (secreted embryonic alkaline phosphatase) are commonly used reporter proteins.
Lucia is a secreted  luciferase with strong bioluminescent activity. The Lucia luciferase reporter gene  is ideal for promoter activity and gene expression studies. Lucia luciferase activity can be detected and quantified directly in the  culture medium of transfected cells using QUANTI-Luc™ detection reagent.

pVITRO1-Lucia/SEAP plasmids can be used as control plasmids for pVITRO1-MCS or cloning   vectors. The reporter genes are flanked by unique   restriction sites and   can be easily replaced by open reading frames   chosen from InvivoGen’s Gene A-List.

Details

•  rEF1 and mEF1 prom: pVITRO1-GFP/LacZ plasmids carry two
elongation factor 1 alpha (EF-1α) promoters, from rat and mouse origins.
Both   promoters display a strong activity that yield similar levels of   expression. EF-1α promoters are expressed at high levels in all cell   cycles and lower levels during G0 phase.
EF-1α promoters are also non-tissue specific; they are highly expressed in all cell types.

• SV40 enhancer which is comprised of a 72-base-pair   repeat allows the enhancement of gene expression in a large host  range.  The enhancement varies from 2-fold in non-permissive cells to  20-fold  in permissive cells. Furthermore, the SV40 enhancer is able to  direct  nuclear localization of plasmids.

• cMV enhancer: The major immediate early enhancer   of the human cytomegalovirus (HCMV) is composed of unique and repeated   sequence motifs. The HCMV enhancer can substitute for the 72-bp repeats   of SV40 and is several-fold more active than the SV40 enhancer.

• SV40 pAn: The Simian Virus 40 late polyadenylation   signal enables efficient cleavage and polyadenylation reactions   resulting in high levels of steady-state mRNA.

• pMB1 Ori is a minimal E. coli origin of replication to limit vector size, but with the same activity as the longer Ori.

• FMDV IRES: The internal ribosome entry site of the   Foot and Mouth Disease Virus enables the translation of two open   reading frames from one mRNA with high levels of expression.

• EM7 is a bacterial promoter that enables the constitutive expression of the antibiotic resistance gene in E. coli.

•  Antibiotic resistance gene :
- Resistance to Blasticidin S is conferred by the bsr gene from Bacillus cereus.
- hph gene confers resistance to Hygromycin B both in E. coli and mammalin cells.
- The neo gene from Tn5 confers resistance to Kanamycin in E.coli and G418 in mammalian cells.
In bacteria, the resistance gene is expressed from the constitutive E.    coli EM7  promoter. In mammalian cells, the resistance gene is    transcribed from the rat EF-1α  promoter as a polycistronic mRNA and    translated via the FMDV IRES.

• EF1 pAn is a strong polyadenylation signal.

• Lucia luciferase gene: Lucia is a secreted  luciferase with strong bioluminescent activity.