pmKate2-profilin is a mammalian expression vector encoding mKate2-profilin fusion protein (see reporter description). The vector can be used for fluorescent labeling of profilin in living cells.
Expression of mKate2 fusion with profilin in HeLa cells.
Scale bar represents 10 μm.
mKate2 codon usage is optimized for high expression in mammalian cells (humanized) [Haas et al., 1996]. Mouse profilin is fused to the mKate2 C-terminus. To increase mRNA translation efficiency, Kozak consensus translation initiation site is generated upstream of the mKate2-profilin coding sequence [Kozak, 1987].
pmKate2-profilin vector can be used as a source of mKate2-profilin hybrid sequence. The vector backbone contains unique restriction sites that permit its excision and further insertion into expression vector of choice.
Expression in mammalian cells
pmKate2-profilin vector can be transfected into mammalian cells by any known transfection method. CMV promoter provides strong, constitutive expression of the mKate2-profilin fusion in eukaryotic cells. If required, stable transformants can be selected using G418 [Gorman, 1985].
Location of features
PCMV IE: 1-589
Enhancer region: 59-465
TATA box: 554-560
Transcription start point: 583
mKate2-profilin fusion: 613-1764
Start codon (ATG): 613-615
Last amino acid in mKate2: 1312-1314
Stop codon: 1765-1767
SV40 early mRNA polyadenylation signal
Polyadenylation signals: 1928-1933 & 1957-1962
mRNA 3' ends: 1966 & 1978
f1 single-strand DNA origin: 2025-2480
Bacterial promoter for expression of Kanr gene
-35 region: 2542-2547
-10 region: 2565-2570
Transcription start point: 2577
SV40 origin of replication: 2821-2956
SV40 early promoter
Enhancer (72-bp tandem repeats): 2654-2725 & 2726-2797
21-bp repeats: 2801-2821, 2822-2842 & 2844-2864
Early promoter element: 2877-2883
Major transcription start points: 2873, 2911, 2917 & 2922
Kanamycin/neomycin resistance gene
Neomycin phosphotransferase coding sequences:
Start codon (ATG): 3005-3007
Stop codon: 3797-3799
G->A mutation to remove Pst I site: 3187
C->A (Arg to Ser) mutation to remove BssH II site: 3533
Herpes simplex virus (HSV) thymidine kinase (TK) polyadenylation signal
Polyadenylation signals: 4035-4040 & 4048-4053
pUC plasmid replication origin: 4384-5027
mKate2-related materials (also referred to as "Products") are intended for research use only. The Products are covered by U.S. Pat. 7,638,615; 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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