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GeneCorner plasmid details

Last data update: 05 December 2020 04:15 CET

Plasmid name: pCAGGSMtPHGPx (LMBP 4432)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p4432.gb (View with Genome Compiler)
p4432.txt
p4432.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human manganese superoxide dismutase cDNA (MnSOD); mitochondrial targeting sequence (MTS)
Pig phospholipid hydroperoxide glutathion peroxidase cDNA (PHGPx, GPx4)
Promoter: Chicken β-actin/rabbit β-globin hybrid promoter (AG)
Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only
Ribosome
binding site:
-
Terminator: Rabbit β-globin polyadenylation signal (β-globin polyA)
Simian virus 40 polyadenylation signal (SV40 polyA)
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Simian virus 40 bidirectional origin (SV40)
Polyoma virus origin
Host range: Escherichia coli
Mammalian cells; SV40 permissive cells
Parental clone: pCAGGSPy; pCAGGSpPHGPx; pCAGGShMnSOD
Further information: The plasmid was constructed as follows: (i) the MTS of the human MnSOD cDNA was amplified by PCR on pCAGGShMnSOD; (ii) the pig PHGPx cDNA was obtained from pCAGGSpPHGPx; (iii) the human MnSOD MTS and the pig PHGPx cDNA were fused by PCR and the amplicon was inserted into the pCAGGSPy vector.
pCAGGSMtPHGPx is useful for highly efficient expression of pig mitochondrial PHGPx under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells.
The nucleotide sequence of the human MnSOD cDNA corresponds with the EMBL Nucleotide Sequence Database accession number Y00472.1, with minor corrections in the 5' UTR and with the Ala->Val polymorphism in the MTS. The differences were confirmed by sequence analysis at the Department for Molecular Biomedical Research (VIB, Belgium).
The nucleotide sequence of the pig PHGPx cDNA corresponds with the EMBL Nucleotide Sequence Database accession number X76009.1.
Name mentioned in Rubio et al. (2012) is pCAGGS_ph-mt-GPX4.
Other name of the plasmid is pCAGGS/MtPHGPx.
EMBL Accession number: X76009.1, view at EMBL, GenBank, DDBJ
Y00472.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 11/01/2012
Sequence detail:
Mitochondrial targeting sequence (MTS) of the human MnSOD cDNA fused to the pig PHGPx cDNA:
    
5'...ATG.TTG.AGC.CGG.GCA.GTG.TGC.GGC.ACC.AGC.AGG.CAG.CTG.GCT.CCG  
     MET Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro 

                                 MTS -->|--> pPHGPx
     GTT.TTG.GGG.TAT.CTG.GGC.TCC.AGG.CAG.ATG.TGC.GCG.TCC.CGC.GAC...3' 
     Val Leu Gly Tyr Leu Gly Ser Arg Gln MET Cys Ala Ser Arg Asp
      *

*: polymorphism (wild type = GCT (Ala)).
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: ApaI, BamHI, HindIII, NcoI, PvuII and XhoI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr J. Grooten(1) (2). It was constructed by V. Goossens(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Rubio et al., Autophagy 8 (2012), 1312-1324 [PMID: 22889744]
Related plasmid reference: Brigelius-Flohé et al., J. Biol. Chem. 269 (1994), 7342-7348 [PMID: 8125951]
Heirman et al., Free Radic. Biol. Med. 40 (2006), 285-294 [PMID: 16413410]
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
Cultivation remark: -
Other culture collection numbers: -

Refer in your Materials and Methods:

pCAGGSMtPHGPx (LMBP 4432) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr J. Grooten and was published in Rubio et al., 2012.

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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