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GeneCorner plasmid details

Last data update: 23 September 2021 14:51 CEST

Plasmid name: pSV23SMTNFm84V (LMBP 2172)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p2172.gb (View with Genome Compiler)
p2172.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Mouse tumor necrosis factor cDNA (Tnf, TNF); A84V mutant
Promoter: Simian virus 40 early promoter (SV40 early)
Ribosome
binding site:
-
Terminator: Simian virus 40 polyadenylation signal (SV40 polyA)
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Simian virus 40 bidirectional origin (SV40)
Host range: Escherichia coli
Mammalian cells; SV40 permissive cells
Parental clone: pSV23SMTNF; pMcMTNFm84V
Further information: The plasmid was constructed as follows: the BstXI fragment of mouse TNF was replaced by the BstXI-mouse TNF fragment from pMcMTNFm84V.
This plasmid contains mutant, mouse TNF; the sequence of amino acids 84 (GCT) coding for Ala was mutated to GTG, coding for Val.
This mutation is described as A84V, according to the numbering of the codons in the human TNF coding sequence, although the mutated mouse codon is located on position 83.
The plasmid contains the SV40 small t-antigen splicing signal.
Restriction enzyme digestions revealed that approx. 60 nucleotides are missing from the 3' untranslated region of the mTNF gene, including the 3'-terminal (supposedly restored) EcoRI site.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727629.1.
The nucleotide sequence of the wild type mouse TNF corresponds with the EMBL Nucleotide Sequence Database accession number M11731.1.
EMBL Accession number: M11731.1, view at EMBL, GenBank, DDBJ
LT727629.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 17/01/1992
Sequence detail:
mouse TNF wild type:

    80  81  82  83  84  85  86  87
5' GTC AGC CGA TTT GCT ATC TCA TAC 3'
   Val Ser Arg Phe Ala Ile Ser Tyr

mouse TNF mutant:

    80  81  82  83  84  85  86  87
5' GTC AGC CGA TTT GTG ATC TCA TAC 3'
   Val Ser Arg Phe Val Ile Ser Tyr
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BamHI/XbaI, BglII/NcoI, BstXI/SalI, EcoRI/PstI, HaeII, HindIII/SpeI, TaqI and XmnI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr W. Fiers(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
It was constructed by X. Van Ostade, B. Vanhaesebroeck and E. Decoster.
Plasmid reference: -
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 MC1061
Host reference: Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
Related host reference: Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Other culture collection numbers: -

Refer in your Materials and Methods:

pSV23SMTNFm84V (LMBP 2172) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr W. Fiers .

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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