pSELECT-zeo-LucSh plasmid哺乳动物长效Luc荧光表达质粒载体 BioVector NTCC质粒载体菌种细胞基因保藏中心

pSELECT-zeo-LucSh plasmid哺乳动物长效Luc荧光表达质粒载体

pSelect-zeo plasmids contain genes that have been chemically
synthesized. The DNA sequence of these genes was modified by
optimizing the codon usage, reducing or eliminating the CpG motifs and
avoiding secondary DNA structures without changing the amino acid
sequence of the wild type proteins.
pSelect-zeo plasmids may be used:
to subclone the synthetic gene into another vector. To facilitate
subcloning, the LucSh gene is flanked by two unique restriction sites:
Nco I at the 5’ end that encompasses the Start codon, and Nhe I at the
3’end.
As a gene reporter plasmid. pSelect-zeo is a mammalian expression
plasmid selectable in E. coli and mammalian cells with Zeocin™, as the
Sh ble gene in the second expression casssette is driven by the eukaryote
CMV enhancer/promoter in tandem with the bacterial EM7 promoter.

PLASMid FEAturES
First expression cassette
• hEF1-htLV prom is a composite promoter comprising the Elongation
Factor-1alpha (EF-1a) core promoter1 and the R segment and part of the
U5 sequence (R-U5’) of the Human T-Cell Leukemia Virus (HTLV) Type
1 Long Terminal Repeat2
. The EF-1a promoter exhibits a strong activity
and yields long lasting expression of a transgene in vivo. The R-U5’ has
been coupled to the EF-1a core promoter to enhance stability of RNA.
• LucSh: Synthetic LucSh fusion gene (LucSh-ΔCpG): BioVector NTCC Inc. has
engineered a fusion between the firefly luciferase gene and the Sh ble gene
conferring Zeocin™ resistance. Both genes have been modified and contain
no CpG, whereas their wildtype counterparts contain 95 and 50 CpG motifs
respectively. This fusion exhibits a higher luciferase activity and enables a
better and faster selection of Zeocin™ resistant clones.
• SV40 pAn: the Simian Virus 40 late polyadenylation signal enables
efficient cleavage and polyadenylation reactions resulting in high levels
of steady-state mRNA3
.
• 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 transcription4
.
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.

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