CpG-free LacZ

Synthetic LacZ reporter gene in expression plasmid

The synthetic LacZ gene is a humanized and CpG-free allele of the LacZ gene.

The lacZ gene codes for the enzyme β-galactosidase which catalyzes the hydrolysis of the substrate X-Gal to produce a blue color that is easily visualized under a microscope.

This CpG-free gene is ten times more active than the wild-type gene in mammalian cells.

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

References:

1. Kim, D.W. et al. (1990).Use of the human elongation factor 1 alpha promoter as a versatile and efficient expression system.Gene 2: 217-223.
2. Takebe, Y. et al. (1988).SR alpha promoter: an efficient and versatile mammalian cDNA expression system composed of the simian virus 40 early promoter and the R-U5 segment of human T-cell leukemia virus type 1 long terminal repeat.Mol. Cell Biol. 1: 466-472.
3. Carswell, S., & Alwine, J.C. (1989).Efficiency of utilization of the simian virus 40 late polyadenylation site: effects of upstream sequences.Mol. Cell Biol. 10: 4248-4258.
4. Yu J & Russell JE. (2001).Structural and functional analysis of an mRNP complex that mediates the high stability of human beta-globin mRNA.Mol Cell Biol, 21(17):5879-88.

Specifications

Gene: CpG-free LacZ

Description: Synthetic gene encoding β-Galactosidase

Backbone: pSELECT-zeo

Selection: Zeocin™

289 CpG in native gene

Contents

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

Product is shipped at room temperature

Description

First expression cassette

• hEF1-HTLV prom is a composite promoter comprising the Elongation Factor-1α (EF-1α) core promoter 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. 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.
• lacZΔCpG gene: a humanized and CpG-free allele of the LacZ gene. This CpG-free gene is ten times more active than the wild-type gene in mammalian cells. It can be used for in vitro or in vivo applications.
• SV40 pan: the Simian Virus 40 late polyadenylation signal enables efficient cleavage and polyadenylation reactions resulting in high levels of steady-state mRNA.
• 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.