GeneCorner plasmid details

Last data update: 22 May 2023 13:52 CEST

Plasmid name: pET-28a-MBP-TEV-Cypc2 (LMBP 12818)

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

Sanger: ef32089258.fasta

Sanger: ef32243599.fasta

Cloned DNA: Cyprinus carpio beta-enolase cDNA (Cyp c 2, Cyp c2, GeneID 122144012)
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 carp Cyp c2 coding sequence as a BglII/SalI fragment into the BamHI/XhoI opened pET28a-MBP-TEV vector.
The plasmid was designed to express the malE-Cyp c2 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 carp Cyp c2 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 nucleotide sequence of the carp Cyp c2 gene corresponds with the EMBL Nucleotide Sequence Database accession number MH255788.1.
Other name of the plasmid is pET-28a(+)-MBP-TEV-Cypc2.
EMBL Accession number: MH255788.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 29/09/2021
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BglI/NcoI, BglII/EcoRV, PstI, PvuII and SacI.

The carp Cyp c2 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: Sliziene et al., Food Agric. Immunol. 33 (2022), 129-149 [DOI: 10.1080/09540105.2022.2028741]
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-Cypc2 (LMBP 12818) is available at BCCM/GeneCorner. This plasmid was deposited by Dr. G. Zvirblis and was published in Sliziene et al., 2022.

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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