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GeneCorner plasmid details

Last data update: 28 November 2020 04:24 CET

Plasmid name: pCAGGS-E-hbeclin-1-C (LMBP 7286)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p7286.gb (View with Genome Compiler)
p7286.txt
p7286.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human beclin 1 cDNA (BECN1, ATG6, VPS30); fragment
E-tag; N-terminal
Promoter: Chicken β-actin/rabbit β-globin hybrid promoter (AG)
Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only
Escherichia coli lac operon promoter
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)
Host range: Escherichia coli
Mammalian cells; SV40 permissive cells
Parental clone: pCAGGS-E
Further information: The plasmid contains a C-terminal fragment of the human BECN1 cDNA, encoding amino acids 150 up to and including 450.
pCAGGS-E-hbeclin-1-C is useful for highly efficient expression of partial human BECN1 under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells.
When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT726851.1.
The nucleotide sequence of the human BECN1 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number AF077301.1, except for the A nucleotide at position 1164 of the full coding sequence, which was analysed by the depositor as a T, resulting in a silent point mutation.
Other names of the plasmid are pCAGGS-E-Beclin-C#1 and pCAGGS-E-Beclin-C1.
EMBL Accession number: AF077301.1, view at EMBL, GenBank, DDBJ
LT726851.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 07/07/2011
Sequence detail:
Primers used to PCR amplify the hBECN1-C fragment:

Forward: 5' ATAAGAATGCGGCCGCA.ATG.ACT.CAG.CTC.A 3'
                    NotI      ***
  
Reverse: 5' TGCGGTCGAC.TCA.TTT.GTT.ATA.AAA.TTG.TGA.GGA.CAC.C 3'
                SalI   +++

The forward primer as mentioned in Wirawan et al. (2010) contains an extra A nucleotide, disrupting the reading frame of hBECN1-C.

***: start codon.
+++: termination codon.
Punctuation indicates reading frame.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BglII, HincII, KpnI/NotI and NcoI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by E. Wirawan(1) (2) and Prof. Dr P. Vandenabeele(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Wirawan et al., Cell Death Dis 1 (2010), e18 [PMID: 21364619] [DOI: 10.1038/cddis.2009.16]
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:

pCAGGS-E-hbeclin-1-C (LMBP 7286) is available at BCCM/GeneCorner. This plasmid was deposited by E. Wirawan and Prof. Dr P. Vandenabeele and was published in Wirawan et al., 2010.

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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