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.
|Bgl II||Sac I||Hind III||EcoR I||Sal I||Kpn I||Apa I||BamH I||Age I||TurboRFP|
|Afe I||Xho I||Pst I||Sac II|
pTurboRFP-PRL is a promoterless vector encoding red (orange) fluorescent protein TurboRFP, which can be used as in vivo reporter of promoter activity (see reporter description). TurboRFP codon usage is optimized for high expression in mammalian cells (humanized) [Haas et al., 1996]. To increase mRNA translation efficiency, Kozak consensus translation initiation site is generated upstream of the TurboRFP coding sequence [Kozak, 1987].
Multiple cloning site (MCS) is located upstream of the Kozak consensus translation initiation site and can be used to clone a promoter or a promoter/enchancer combination of interest. Without the addition of a functional promoter, this vector will not express TurboRFP.
The vector backbone contains SV40 origin for replication in mammalian cells expressing SV40 T-antigen, pUC origin of replication for propagation in
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
Note: The plasmid DNA was isolated from dam+-methylated
Suitable host strains for propagation in
Location of features
Kozak consensus translation initiation site: 90-100
Start codon (ATG): 97-99
Stop codon: 790-792
SV40 early mRNA polyadenylation signal
Polyadenylation signals: 946-951 & 975-980
mRNA 3' ends: 984 & 996
f1 single-strand DNA origin: 1043-1498
Bacterial promoter for expression of Kanr gene
-35 region: 1560-1565
-10 region: 1583-1588
Transcription start point: 1595
SV40 origin of replication: 1839-1974
SV40 early promoter
Enhancer (72-bp tandem repeats): 1672-1743 & 1744-1815
21-bp repeats: 1819-1839, 1840-1860 & 1862-1882
Early promoter element: 1895-1901
Major transcription start points: 1891, 1929, 1935 & 1940
Kanamycin/neomycin resistance gene
Neomycin phosphotransferase coding sequences:
Start codon (ATG): 2023-2025
Stop codon: 2815-2817
G->A mutation to remove Pst I site: 2205
C->A (Arg to Ser) mutation to remove BssH II site: 2551
Herpes simplex virus (HSV) thymidine kinase (TK) polyadenylation signal
Polyadenylation signals: 3053-3058 & 3066-3071
pUC plasmid replication origin: 3402-4045
TurboRFP-related materials (also referred to as "Products") are intended for research use only. The Products are covered by U.S. Pat. 8,138,320; 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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