pTurboFP602-PRL

pTurboFP602-PRL vector

cat.# FP715

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.


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vector information:
ProductCat.#SizePrice
pTurboFP602-PRLFP71520 μg€ 320 / 160*
*50% discount on the second and subsequent vectors encoding same fluorescent protein or sensor ordered by the same customer.
The price does not include delivery. The price varies in different countries. Please contact your local distributor for exact prices and delivery information.

Vector typepromoterless expression vector
ReporterTurboFP602
Reporter codon usagemammalian
Promoter for TurboFP602NO
Host cellsmammalian, prokaryotic
Selectionprokaryotic – kanamycin
eukaryotic – neomycin (G418)
Replicationprokaryotic – pUC ori
eukaryotic – SV40 ori
Use Monitoring of activity of different promoters and promoter/enhancer combinations
Multiple cloning site (MCS)
Bgl II Sac I Hind III EcoR I Sal I Kpn I Apa I BamH I Age I TurboFP602
Afe I Xho I Pst I Sac II Sma I/Xma I Nco I*
ACT A.GC G.CT A.CCG.GAC.TC A.GAT. CT C. GAG. CTC. AAG.CTT. C GA.ATT. C TG.CA G. TCG.AC G.GTA. CC G.C GG. G CC.C G G.G AT.CC A.CCG.GT C.GCC.A CC. ATG.G TG.GGT

* – not unique site.

Vector description

pTurboFP602-PRL is a promoterless vector encoding true-red fluorescent protein TurboFP602, which can be used as in vivo reporter of promoter activity (see reporter description). TurboFP602 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 TurboFP602 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 TurboFP602.

The vector backbone contains 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.

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.


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

MCS: 12-89
TurboFP602
Kozak consensus translation initiation site: 90-100
Start codon (ATG): 97-99
Stop codon: 802-804
SV40 early mRNA polyadenylation signal
Polyadenylation signals: 958-963 & 987-992
mRNA 3' ends: 996 & 1008
f1 single-strand DNA origin: 1055-1510
Bacterial promoter for expression of Kanr gene
-35 region: 1572-1577
-10 region: 1595-1600
Transcription start point: 1607
SV40 origin of replication: 1851-1986
SV40 early promoter
Enhancer (72-bp tandem repeats): 1684-1755 & 1756-1827
21-bp repeats: 1831-1851, 1852-1872 & 1874-1894
Early promoter element: 1907-1913
Major transcription start points: 1903, 1941, 1947 & 1952
Kanamycin/neomycin resistance gene
Neomycin phosphotransferase coding sequences:
Start codon (ATG): 2035-2037
Stop codon: 2827-2829
G->A mutation to remove Pst I site: 2217
C->A (Arg to Ser) mutation to remove BssH II site: 2563
Herpes simplex virus (HSV) thymidine kinase (TK) polyadenylation signal
Polyadenylation signals: 3065-3070 & 3078-3083
pUC plasmid replication origin: 3414-4057


References:

  • 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
  • Kozak M. An analysis of 5'-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res. 1987; 15 (20):8125-48. / pmid: 3313277

Notice to Purchaser:

TurboFP602-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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