pFLAG-CMV-6a Expression Vector
Catalog Number E1900
Store at –20 °C
Product Description
The pFLAG-CMV-6a expression vector is a 4.7 kb
1
derivative of pCMV5 used to establish transient
intracellular expression of N-terminal Met-FLAG fusion
proteins in mammalian cells. pFLAG-CMV-6a,
pFLAG-CMV-6b, Catalog No. E2025, and
pFLAG-CMV-6c, Catalog No. E2150, are designed for
expression in each of the three translational reading
frames.
The promoter-regulatory region of the human
cytomegalovirus2 drives transcription of FLAG-fusion
constructs.
pFLAG-CMV-6a expression vector is a shuttle vector
for E. coli and mammalian cells. Efficiency of
replication is optimal when using an SV40 T antigenexpressing host, such as COS cells.
pFLAG-CMV-6a expression vector is supplied in
1 mM Tris, 0.1 mM EDTA, pH 8.
Precautions and Disclaimer
This product is for R&D use only, not for drug,
household, or other uses. Please consult the Material
Safety Data Sheet for information regarding hazards
and safe handling practices.
References
1. Andersson, S., et al., J. Biol Chem., 264,
8222-8229 (1989)
2. Thomsen, D.R., et al., Proc. Natl. Acad. Sci. USA,
81, 659-663 (1984)
pFLAG-CMV-6a Features
Hind III
Met
Not I
Bgl II EcoRV
Bam HI Cla I
Pst I Sal I
Kpn I
EcoRI
Multiple Cloning Region
FLAG
f1 ori
CMV Promoter
CMV 30
Amp
pFLAG-CMV-6a
CMV 24
4693 bp
hGH polyA
pBR322 ori
SV40 ori
Sma I
Feature
Map Position
CMV promoter
419 - 910
CMV 30 sequencing primer
818 - 847
Translational initiation
926 - 928
FLAG sequence
929 - 952
Multiple cloning region
956 - 1033
hGH polyA
1030 - 1655
CMV 24 sequencing primer
1091 - 1114
SV40 ori
1674 - 1996
pBR322 ori/ampicillin resistance
2046 - 4236
f 1 ori
4237 - 4687
2
Nucleotide Sequence of the Multiple Cloning Region
of the pFLAG-CMV-6a Expression Vector
Sequence Range: 923 to 1036 bp
Translational initiation
_(1)_
Hind III
↓
ACC ATG GAC TAC AAA GAC GAT GAC GAC AAG CTC A AG CT T
TGG TAC CTG ATG TTT CTG CTA CTG CTG T TC GAG T T C GA↑A
Asp Tyr Lys Asp Asp Asp Asp Lys
← Multiple
← FLAG Coding Sequence →
Cloning Region →
Not I
Eco RI
Bgl II
Pst I
Eco RV
Sal I
_________
___________
__________
↓
↓
↓
↓
↓
↓
GC G GCC G CG AAT T CA GAT C T C TGC A GG AT A TCG TCG ACG
CG C CGG ↑CGC TTA A↑GT CTA G↑AG↑ ACG T CC TA ↑T AGC AGC T↑GC
← Multiple Cloning Region →
Bam HI
Kpn I
Cla I
2
STOP
2
STOP
STOP
2
Sma I
___________
____ ____ ___
↓
↓
↓
GAT C CG GTA C CA T CG ATA GAT AAT TGA CCC GGG TGG
CTA G↑GC↑ CAT G GT A GC↑ TAT CTA TTA ACT GGG↑ CCC ACC
↓
← Multiple Cloning Region →
Note:
(1) pFLAG-CMV-6a-specific spacer sequence.
(2) The Sma I site may be used in combination with other sites for directed cloning. However, using
Sma I alone for cloning purposes will result in failure to express FLAG-fusion proteins.
FLAG is a registered trademark, and pFLAG-CMV is a trademark, of Sigma-Aldrich Biotechnology LP and Sigma-Aldrich Co..
AH,RS,PHC 08/10-1
Sigma brand products are sold through Sigma-Aldrich, Inc.
Sigma-Aldrich, Inc. warrants that its products conform to the information contained in this and other Sigma-Aldrich publications.
Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and conditions may apply.
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