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LMBP plasmid details

Last data update: 25 September 2020 04:24 CEST

Plasmid name: pLenti6c-HA-hFTH1 (LMBP 9530)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p9530.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human ferritin, heavy polypeptide 1 cDNA (FTH1, FTHL6, FHC, FTH, PIG15, PLIF)
Influenza HA epitope encoding the haemagglutinin tagging peptide; C-terminal
Promoter: Rous sarcoma virus/human immunodeficiency virus hybrid long terminal repeat (RSV/HIV-1 5' LTR)
Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer
Escherichia coli lac operon promoter
Simian virus 40 early promoter (SV40 early)
Phage T3 promoter
Phage T7 gene 10 promoter (T7g10)
Synthetic prokaryotic EM7 promoter
Ribosome
binding site:
-
Terminator: Rous sarcoma virus/human immunodeficiency virus hybrid long terminal repeat (RSV/HIV-1 3' LTR); modified HIV-1 3' LTR with deleted U3 region (ΔU3/3' LTR)
Simian virus 40 polyadenylation signal (SV40 polyA)
Selection marker: Ampicillin (amp)
Blasticidin (bsd)
Replicon: Escherichia coli plasmid pMB1 origin
Phage f1 origin
Simian virus 40 bidirectional origin (SV40)
Host range: Escherichia coli
Mammalian cells; SV40 permissive cells
Parental clone: pENTR223-hFTH1; pLenti6(Blast)HA-Dest
Further information: This lentiviral expression vector was constructed by cloning the human FTH1 coding sequence from pENTR223-hFTH1 into the pLenti6(Blast)HA-Dest vector via Gateway recombination.
This is a lentiviral Gateway expression vector designed for ViraPower II lentiviral-based expression of the human FTH1 gene in dividing and non-dividing mammalian cells.
The ViraPowerLentiviral Expression System (Invitrogen) possesses features which enhance its biosafety while allowing high-level gene expression in a wider range of cell types than traditional retroviral systems.
The vector carries the human CMV-IE promoter for driving constitutive expression of the human FTH1 gene. Although highly active in most mammalian cell lines, activity of the viral promoter can be down-regulated in some cell lines due to methylation, histone deacetylation, or both.
The U3 region of the HIV-1 3' LTR is deleted (ΔU3 or dU3) and facilitates self-inactivation of the HIV-1 5' LTR after transduction to enhance the biosafety of the vector. The element also contains a polyadenylation signal for transcription termination and polyadenylation of mRNA in transduced cells.
The vector contains the psi packaging signal, allowing viral packaging, and the rev response element (RRE) from HIV-1, permitting Rev-dependent nuclear export of unspliced viral mRNA.
Because the risk for recombination after each subcultivation is high, this plasmid is only available under the format of isolated plasmid DNA.
The nucleotide sequence of the human FTH1 coding sequence corresponds with the Genbank accession number NM_002032.2.
Other name of the plasmid is pLenti6c-HA-FTH1.
EMBL Accession number: NM_002032.2, view at GenBank
Latest sequence update: 05/02/2016
Authenticity test: The plasmid still needs to be subjected to the authenticity test.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr R. Beyaert(1)(2). It was constructed by Dr C. Müller(1)(2).
(1) Inflammation Research Center, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: -
Restricted distribution: - BCCM MTA
Distributed as: plasmid DNA
Host for distribution: Escherichia coli K12 DH5α
Host reference: Focus 8 (1986), 9
Related host reference: Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1 in E. coli; L2 in mammalian cells
Other culture collection numbers: -

Note: Up-to-date, validated data are enclosed with the biological material. Nevertheless, these data are a snapshot at a given moment; further updates are always possible.

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