GeneCorner plasmid details

Last data update: 22 May 2023 13:52 CEST

Plasmid name: pET-28a-MBP-TEV-Derp26 (LMBP 12823)

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Price category: Cat. 2 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: (View with Genome Compiler)
analysis results

Sanger: ef32243621.fasta

Sanger: ef32243606.fasta

Cloned DNA: Dermatophagoides pteronyssinus allergen Der p 26 cDNA (Der p 26, Der p26)
Escherichia coli maltose ABC transporter periplasmic binding protein cDNA (malE, GeneID 948538); mature sequence, N-terminal
Thrombin recognition site; N-terminal
Tobacco etch virus (TEV) protease recognition site; N-terminal
Histidine tag (His-tag); N-terminal
Escherichia coli DNA-binding transcriptional repressor LacI cDNA (lacI, GeneID 945007)
Histidine tag (His-tag); C-terminal
Promoter: Phage T7 gene 10 promoter (T7g10) with lac operator (T7lac)
Escherichia coli lac repressor promoter (lacI)
binding site:
Ribosome binding site (RBS) of the phage T7 gene 10 (T7g10)
Terminator: Phage T7 gene 10 terminator (T7g10)
Selection marker: Neomycin (neo; kanamycin (kan))
Replicon: Escherichia coli plasmid pMB1 origin
Phage f1 origin
Host range: Escherichia coli
Parental clone: pET28a-MBP-TEV
Further information: The plasmid was constructed by cloning the house dust mite Der p 26 coding sequence, optimised for E. coli, into the BamHI/XhoI opened pET28a-MBP-TEV vector.
The plasmid was designed to express the malE-Der p 26 fusion protein in E. coli BL21(DE3). The plasmid encodes for the mature form of the E. coli maltose binding protein, which does not contain a signal sequence, hence the protein will remain in the cytosol.
The N-terminal His-tag can be removed via a thrombin recognition site, while the maltose binding protein can be separated from the house dust mite Der p 26 allergen via the TEV protease recognition site. The His-tag at the C-terminal end of the allergen is not functional in this plasmid.
The amino acid sequence of the house dust mite Der p 26 allergen corresponds with GenBank accession number QAT18638.1.
Other name of the plasmid is pET-28a(+)-MBP-TEV-Derp26.
EMBL Accession number: QAT18638.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 14/06/2021
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BamHI/XhoI, BglI/NcoI, BglII/EcoRV, PvuII and XbaI.

The house dust mite Der p 26 coding sequence has also been sequenced at BCCM/GeneCorner.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr. G. Zvirblis (1).
(1) Institute of Biotechnology, Life Science Center, Vilnius University, Vilnius, Lithuania
Plasmid reference: -
Restricted distribution: - The depositor will be informed of the customer's identity upon release of a sample outside the depositor's department or outside the departments in which BCCM/GeneCorner is embedded, namely UGent-DBMB and VIB-IRC.
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 DH5αT1R
Host reference: -
Related host reference: Killmann et al., J. Bacteriol. 178 (1996), 6913-6920 [PMID: 8955314] [DOI: 10.1128/jb.178.23.6913-6920.1996]
Cultivation medium: LB-Lennox + kanamycin (50 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Other culture collection numbers: -

Refer in your Materials and Methods:

pET-28a-MBP-TEV-Derp26 (LMBP 12823) is available at BCCM/GeneCorner. This plasmid was deposited by Dr. G. Zvirblis .

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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