CpG-free SEAP

Synthetic SEAP reporter gene in expression plasmid

InvivoGen provides an optimized SEAP reporter gene that is entirely free of CpG motifs.

Secreted embryonic alkaline phosphatase (SEAP) is widely used to study promoter activity or gene expression. SEAP is secreted into cell culture supernatant and therefore offers many advantages over intracellular reporters.

It allows to determine reporter activity without disturbing the cells, does not require the preparation of cell lysates and can be used for kinetic studies.

SEAP activity that can be readily assessed qualitatively and quantitatively using HEK-Blue™ Detection or QUANTI-Blue™ Solution. 

The CpG-free SEAP gene is provided in the expression plasmid pSelect-zeo which is selectable with Zeocin® in both mammalian and E.coli cells.

Specifications

pSELECT-zeo-hSEAP-CpG-free

Gene: CpG-free SEAP

Description: Synthetic secreted embryonic alkaline phosphatase (SEAP) gene

Species: Human

Backbone: pSELECT-zeo

Selection: Zeocin®

109 CpG in the native gene

pSELECT-zeo-mSEAP-CpG-free

Gene: CpG-free SEAP

Description: Synthetic secreted embryonic alkaline phosphatase (SEAP) gene

Species: Mouse

Backbone: pSELECT-zeo

Selection: Zeocin®

65 CpG in the native gene

Contents

• 20 µg of lyophilized DNA
• 1 ml of Zeocin® (100 mg/ml)

Product is shipped at room temperature

Description

PLASMID FEATURES
First expression cassette
• hEF1-htLV prom is a composite promoter comprising the Elongation Factor-1alpha (EF-1α) core promoter [1 ]and the R segment and part of the U5 sequence (R-U5’) of the Human T-Cell Leukemia Virus (HTLV) Type 1 Long Terminal Repeat [2]. The EF-1α promoter exhibits a strong activity and yields long lasting expression of a transgene in vivo. The R-U5’ has been coupled to the EF-1α core promoter to enhance stability of RNA.
• CpG-free SEAP: Synthetic secreted alkaline phosphatase gene. InvivoGen has synthesied CpG-free human and mouse SEAP genes. The native hSEAP gene contains 109 CpG motifs, the native mSEAP has 65.
• SV40 pAn: the Simian Virus 40 late polyadenylation signal enables efficient cleavage and polyadenylation reactions resulting in high levels of steady-state mRNA [3].
• ori: a minimal E. coli origin of replication to limit vector size, but with the same activity as the longer Ori.
Second expression cassette
• CMV enh/prom: The human cytomegalovirus immediate-early gene 1 promoter/enhancer was originally isolated from the Towne strain and was found to be stronger than any other viral promoters.
• EM7 is a bacterial promoter that enables the constitutive expression of the antibiotic resistance gene in E. coli.
• Zeo: Resistance to Zeocin® is conferred by the Sh ble gene from Streptoalloteichus hindustanus The Sh ble gene is driven by the CMV enhancer/promoter in tandem with the bacterial EM7 promoter allowing selection in both mammalian cells and E. coli.
• ßGlo pAn: The human beta-globin 3’UTR and polyadenylation sequence allows efficient arrest of the transgene transcription [4].

1. Kim, D.W. et al. (1990). Gene 2: 217-223.
2. Takebe, Y. et al. (1988). Mol. Cell Biol. 1: 466-472.
3. Carswell, S. & Alwine, J.C. (1989). Mol. Cell Biol. 10: 4248-4258.
4. Yu J & Russell JE. (2001). Mol Cell Biol, 21(17):5879-88.