pVITRO2-neo-mcs
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Cat.code:
pvitro2-nmcs
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ABOUT
Dual expression plasmids - Ferritin promoters - two multiple cloning sites (MCS)
Expression plasmids pVITRO2-MCS are developed mainly for in vitro studies.
pVITRO2-MCS allows the ubiquitous and constitutive co-expression of two genes of interest of your choice. They can be stably transfected in E. coli or mammalian cells, where the genes of interest are expressed at high levels.
These plasmids contain two multiple cloning sites (MCS) for the convenient cloning of two cDNAs. pVITRO2-MCS plasmids carry two human ferritin composite promoters, FerH (heavy chain) and FerL (light chain). To eliminate the iron regulation, their 5’UTRs have been replaced by the 5’UTR of the mouse and chimpanzee EF-1α genes.
pVITRO2-MCS plasmids are available with different selectable markers that are active both in E. coli and mammalian cells.
InvivoGen also provides Dual reporter pVITRO plasmids containing two reporter genes, including GFP, Lucia®, SEAP, or LacZ as a control.
All products are for research use only, and not for human or veterinary use.
SPECIFICATIONS
Specifications
Robust in vitro expression of one or two genes
In vitro transfection
Plasmid construct is confirmed by restriction analysis and full-length open reading frame (ORF) sequencing.
CONTENTS
Contents
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Product:pVITRO2-neo-mcs
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Cat code:pvitro2-nmcs
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Quantity:20 µg
Shipping & Storage
- Shipping method: Room temperature
- -20°C
- Avoid repeated freeze-thaw cycles
Storage:
Caution:
Details
• hFerH and hFerL composite promoters: Ferritin is a 24 subunit protein composed of two subunit types, termed H (heavy) and L (light). Ferritin is ubiquitously expressed. Its synthesis is highly regulated by the iron status of the cell. To eliminate the iron regulation of the ferritin promoters, the 5’UTR of FerH and FerL have been replaced by the 5’UTR of the mouse and chimpanzee elongation factor 1 (EF1) genes, respectively.
• 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.
• 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 severalfold 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: 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.
• MCS1 and MCS2: Each multiple cloning site contains several restriction sites that are compatible with many other enzymes, thus facilitating cloning.
MCS1 includes the following restriction sites:
Age I, Eco RV, Bam HI, Sal I and Avr II
- Age I is compatible with Bsp EI and Sgr AI.
- Eco RV (blunt-end restriction enzyme)
- Bam HI is compatible with Bgl II, Bst YI and Bcl I.
- Sal I is compatible with Ava I and Xho I.
- Avr II is compatible with Xba I, Spe I and Nhe I.
MCS2 includes the following restriction sites:
Sgr AI, Bgl II, Xho I and Nhe I
- Sgr AI is compatible with Bsp EI and Age I.
- Bgl II is compatible with Bam HI, Bst YI and Bcl I.
- Xho I is compatible with Ava I and Sal I.
- Nhe I is compatible with Xba I, Spe I and Avr II.
DOCUMENTS
Documents
Technical Data Sheet
Plasmid Sequence
Safety Data Sheet
Certificate of analysis
Need a CoA ?