Products | Vector Systems

CMV-based constitutive Expression Vectors
phCMV1, phCMV2, phCMV3

CMV-based constitutive Expression Vectors
		Introduction
		Order Information
		Literature
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The phCMV series of vectors are designed to achieve significantly higher expression levels than traditional human cytomegalovirus (CMV) promoter-based constitutive expression vectors. This makes the phCMV vectors ideal tools for routine protein expression studies as well as animal injection experiments.
The phCMV vectors consistently deliver superior expression levels when compared with other commercially available CMV promoter-based expression vectors. It is not unusual to obtain protein expression levels from phCMV that are severalfold higher than those obtained from other popularly used vectors. In addition, because phCMV vectors use the Kan/Neo gene for selection in both bacteria and cultured cell lines, they offer smaller vector sizes and improved transfection efficiency.

In summary, the three phCMV vectors offer the following benefits:

Maximized high-level expression with optimized CMV promoter
G418 resistance gene for the selection of stable cell lines
Optional N- or C-terminal HA fusion tags for simplified protein detection and purification with
anti-HA antibodies and affinity resins
Extensive multiple cloning region for convenient and easy cloning
Small vector sizes for efficient transfection

phCMV Vector Features

Feature	Function
Modified Human cytomegalovirus (CMV) immediate-early promoter/enhancer/intron	High-level expression promoter in vitro and in vivo.
T7 promoter priming site in phCMV, phCMV3	For sequencing of the inserted DNA fragment
SV40 polyadenylation signal	Efficient transcription termination and polyadenylation of mRNA.
SV40 promoter	High-level expression of the neomycin resistance gene for selection of stably transfected cells.
Kanamycin resistance gene	Efficient selection of vector in E. coli cells
pUC origin	High copy number replication of vector in E. coli cells.
Multiple cloning site and HA tags	For cloning a gene into vector with either wild type (phCMV), N-terminal HA tag (phCMV2), and C-terminal HA

Available Kits and Contents

The following phCMV vectors are currently available from MoBiTec:

phCMV1 CMV-based constitutive expressionvector For encoding recombinant proteins in their native form, without any tags
Kit contents	• 25 µg supercoiled phCMV1 Vector, 1 µg/µl solution. • 10 µg phCMV1/CAT positive Control vector, 1 µg/µl solution
phCMV2 CMV-based constitutive expressionvector For encoding recombinant proteins with an N-terminal HAfusion tag
Kit contents	• 25 µg supercoiled phCMV2 Vector, 1 µg/µl solution. • 10 µg phCMV2/CAT positive Control vector, 1 µg/µl solution
phCMV3 CMV-based constitutive expressionvector For encoding recombinant proteins with a C-terminal HAfusion tag
Kit contents	• 25 µg supercoiled phCMV3 Vector, 1 µg/µl solution. • 10 µg phCMV3/CAT positive Control vector, 1 µg/µl solution

Order Information

Please click on the respective order # for further details.

ORDER INFORMATION, SHIPPING & STORAGE:
order#	description	amount	Vector Map	Sequence
VS-EXC00100-01	phCMV1 CMV-based constitutive expression vector	kit	map	txt
VS-EXC00200-01	phCMV2 CMV-based constitutive expression vector	kit	map	txt
VS-EXC00300-01	phCMV3 CMV-based constitutive expression vector	kit	map	txt
Shipped frozen; store at -20 °C. Stable for 6 months when stored properly.

Accessory Products

MoBiTec offers the following products for use in conjunction with the phCMV vectors:

3-minute transformation and efficient propagation of phCMV vectors
order#	description	amount
CB-CEC30100-01	QuickCells Competent E. coli	20 x 50 µl
CB-CEC30150-01	QuickCells F' Competent E. coli	20 x 50 µl

For sensitive and highly-specific detection of HA-tagged proteins
order#	description	amount
AB-PTN01010-01	Anti-HA Polyclonal Antibody	500 µl
AB-PTN01011-01	Anti-HA Polyclonal Antibody, HRP Conjugate	250 µl

For efficient transfection and high-level expression
order#	description	amount
0201B	GeneTrans II Transfection Reagent	75 reactions (0.75 ml)
0202B	GeneTrans II Transfection Reagent	150 reactions (1.5 ml)
0203B	GeneTrans II Transfection Reagent	75 reactions (5 x 1.5 ml)
CB-TDT10666-01	GenePusher Reagent Kit	3 boosters (1.5 ml each)

Literature

Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A., and Struhl, K. (1994). Current Protocols in Molecular Biology (New York: Greene Publishing Associates and Wiley-Interscience).
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual, Second Edition (Plainview, New York: Cold Spring Harbor Laboratory Press).