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pFusionRed-actinin vector

cat.# FP426

The vector sequence has been compiled using the information from sequence databases, published literature, and other sources, together with partial sequences obtained by Evrogen. This vector has not been completely sequenced.

vector information:
pFusionRed-actininFP42620 μg€ 480 / 240*
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Vector typemammalian expression vector
Reporter codon usagemammalian
Promoter for FusionRedPCMV IE
Host cellsmammalian
Selectionprokaryotic – kanamycin
eukaryotic – neomycin (G418)
Replicationprokaryotic – pUC ori
eukaryotic – SV40 ori
Use red fluorescent labeling of α-actinin

Vector description

pFusionRed-actinin is a mammalian expression vector encoding FusionRed-actinin fusion protein (see reporter description). The vector can be used for fluorescent labeling of α-actinin in living cells.

African green monkey kidney fibroblast cell expressing fusion of human α-actinin with FusionRed.

In addition to high concentrations of α-actinin being visualized in ruffles at the periphery of the cell, the fusion is also observed in focal adhesions and decorating stress fibers. Movie from Shemiakina et al., 2012.

FusionRed codon usage is optimized for high expression in mammalian cells (humanized) [Haas et al., 1996]. Human α-actinin is fused to the FusionRed N-terminus.

pFusionRed-actinin vector can be used as a source of FusionRed-actinin hybrid sequence. The vector backbone contains unique restriction sites that permit its excision and further insertion into expression vector of choice.

Note: The plasmid DNA was isolated from dam+-methylated E. coli. Therefore some restriction sites are blocked by methylation. If you wish to digest the vector using such sites you will need to transform the vector into a dam- host and make fresh DNA.

The vector backbone contains immediate early promoter of cytomegalovirus (PCMV IE) for protein expression, SV40 origin for replication in mammalian cells expressing SV40 T-antigen, pUC origin of replication for propagation in E. coli, and f1 origin for single-stranded DNA production. SV40 polyadenylation signals (SV40 poly A) direct proper processing of the 3'-end of the reporter mRNA.

SV40 early promoter (PSV40) provides neomycin resistance gene (Neor) expression to select stably transfected eukaryotic cells using G418. Bacterial promoter (P) provides kanamycin resistance gene expression (Kanr) in E. coli. Kanr/Neor gene is linked with herpes simplex virus (HSV) thymidine kinase (TK) polyadenylation signals.

Expression in mammalian cells

pFusionRed-actinin vector can be transfected into mammalian cells by any known transfection method. CMV promoter provides strong, constitutive expression of the FusionRed-actinin fusion in eukaryotic cells. If required, stable transformants can be selected using G418 [Gorman, 1985].

Propagation in E. coli

Suitable host strains for propagation in E. coli include DH5alpha, HB101, XL1-Blue, and other general purpose strains. Plasmid incompatibility group is pMB1/ColE1. The vector confers resistance to kanamycin (30 μg/ml) to E. coli hosts. Copy number in E. coli is about 500.

Location of features

PCMV IE: 1-589
Enhancer region: 59-465
TATA box: 554-560
Transcription start point: 583
alpha-actinin-FusionRed fusion: 637-4068
alpha-actinin: 637-3312
Start codon (ATG): 637-639
Last amino acid in alpha-actinin: 3310-3312
FusionRed: 3370-4068
Stop codon: 4066-4068
SV40 early mRNA polyadenylation signal
Polyadenylation signals: 4221-4226 & 4250-4255
mRNA 3' ends: 4259 & 4271
f1 single-strand DNA origin: 4318-4773
Bacterial promoter for expression of Kanr gene
-35 region: 4835-4840
-10 region: 4858-4863
Transcription start point: 4870
SV40 origin of replication: 5114-5249
SV40 early promoter
Enhancer (72-bp tandem repeats): 4947-5018 & 5019-5090
21-bp repeats: 5094-5114, 5115-5135 & 5137-5157
Early promoter element: 5170-5176
Major transcription start points: 5166, 5204, 5210 & 5215
Kanamycin/neomycin resistance gene
Neomycin phosphotransferase coding sequences:
Start codon (ATG): 5298-5300
Stop codon: 6090-6092
G->A mutation to remove Pst I site: 5480
C->A (Arg to Ser) mutation to remove BssH II site: 5826
Herpes simplex virus (HSV) thymidine kinase (TK) polyadenylation signal
Polyadenylation signals: 6328-6333 & 6341-6346
pUC plasmid replication origin: 6677-7320


  • Gorman C. High efficiency gene transfer into mammalian cells. In DNA cloning: A Practical Approach, Vol. II. Ed. D. M. Glover. (IRL Press, Oxford, U.K.). 1985; 143-90.
  • Haas J, Park EC, Seed B. Codon usage limitation in the expression of HIV-1 envelope glycoprotein. Curr Biol. 1996; 6 (3):315-24. / pmid: 8805248
  • Shemiakina II, Ermakova GV, Cranfill PJ, Baird MA, Evans RA, Souslova EA, Staroverov DB, Gorokhovatsky AY, Putintseva EV, Gorodnicheva TV, Chepurnykh TV, Strukova L, Lukyanov S, Zaraisky AG, Davidson MW, Chudakov DM, Shcherbo D. A monomeric red fluorescent protein with low cytotoxicity. Nat Commun. 2012; 3 :1204. doi: 10.1038/ncomms2208 / pmid: 23149748

Notice to Purchaser:

FusionRed-related materials (also referred to as "Products") are intended for research use only. The Products are covered by European Pat. 1994149 and other Evrogen Patents and/or Patent applications pending. By use of these Products, you accept the terms and conditions of the applicable Limited Use Label License.

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