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GeneCorner plasmid details

Last data update: 24 January 2024 16:39 CET

Plasmid name: pDG2i-hSFRS2-S101A-Flag puro (LMBP 9355)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p9355.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human serine and arginine rich splicing factor 2 cDNA (SRSF2, SFRS2, PR264, SC-35, SC35, SFRS2A, GeneID 6427); mutated sequence
Reverse tetracycline-responsive transcriptional activator (rtTA); variant 3 (rtTA3)
FLAG epitope tag; C-terminal
Promoter: Human cytomegalovirus immediate early promoter (CMV-IE); minimal promoter with tet responsive element (CMVTRE)
Rous sarcoma virus/human immunodeficiency virus hybrid long terminal repeat (RSV/HIV-1 5' LTR)
Simian virus 40 early promoter (SV40 early)
Escherichia coli lac operon promoter
Phage T7 gene 10 promoter (T7g10)
Phage T3 promoter
Ribosome
binding site:
Internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV) polyprotein gene
Terminator: Simian virus 40 polyadenylation signal (SV40 polyA)
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)
Selection marker: Ampicillin (amp)
Puromycin (puro)
Replicon: Simian virus 40 bidirectional origin (SV40)
Escherichia coli plasmid pMB1 origin
Phage f1 origin
Host range: Escherichia coli
Mammalian cells; SV40 permissive cells
Parental clone: pDG2i-Flag puro; pENTR3C-hSFRS2-S101A
Further information: The plasmid was constructed by cloning the mutant hSFRS2 coding sequence from pENTR3C-hSFRS2-S101A into the pDG2i-Flag puro vector via Gateway LR recombination.
The human SFRS2 coding sequence contains an S101A mutation. It does not contain a termination codon, and is fused with a C-terminal FLAG epitope tag.
This is a lentiviral Gateway expression vector, with the tet-inducible minimal human CMV-IE promotor and rtTA3 on the same plasmid.
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. Plasmid integrity should be checked via PvuII restriction digest after each subcultivation.
The nucleotide sequence of the pDG2i-Flag puro vector corresponds with the EMBL Nucleotide Sequence Database accession number LT009444.1.
Other name of the plasmid is pDG2-Flag-rtTA3puro - hSFRS2-S101A - stop.
EMBL Accession number: LT009444.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 28/04/2017
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 P. Vandenabeele(1) (2).
(1) Center for Inflammation Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: -
Related plasmid reference: De Groote et al., BioTechniques 60 (2016), 252-259 [PMID: 27177818] [DOI: 10.2144/000114417]
Restricted distribution: - BCCM MTA
Distributed as: plasmid DNA
Host for distribution: Escherichia coli K12 DH5α
Host reference: Focus 8 (1986), 9
Related host reference: Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791] [DOI: 10.1016/s0022-2836(83)80284-8]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 28°C
Biosafety level: L1 in E. coli; L2 in mammalian cells
Other culture collection numbers: -

Refer in your Materials and Methods:

pDG2i-hSFRS2-S101A-Flag puro (LMBP 9355) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr P. Vandenabeele .

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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