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GeneCorner plasmid details

Last data update: 04 July 2022 09:25 CEST

Plasmid name: pET-28a-MBP-TEV-Scos1 (LMBP 12832)

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Price category: Cat. 2 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p12832.gb
Sequence
analysis results
Genecorner:

Sanger: ef32243627.fasta

Sanger: ef32243613.fasta

Cloned DNA: Scomber scombrus parvalbumin beta cDNA (pvalb1, Sco s 1, Sco s1)
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)
Ribosome
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 S. scombrus pvalb1 coding sequence, optimised for E. coli, into the BamHI/XhoI opened pET28a-MBP-TEV vector.
The plasmid was designed to express the malE-pvalb1 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 S. scombrus pvalb1 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 S. scombrus pvalb1 allergen corresponds with GenBank accession number CAX32965.1.
Other name of the plasmid is pET-28a(+)-MBP-TEV-Scos1.
EMBL Accession number: CAX32965.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 15/06/2021
Authenticity test: The plasmid still needs to be subjected to the authenticity test.
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.
- BCCM MTA
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-Scos1 (LMBP 12832) 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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