Last data update: 22 May 2023 13:52 CEST
Plasmid name: pET-28a-MBP-TEV-Shark (LMBP 12829)
New search | Print data sheet |
Price category: | Cat. 2 (cf. price list) |
Status: | GeneCorner non-core plasmid |
Depositor's sequence: | p12829.gb |
Sequence analysis results Genecorner: |
Sanger: ef32243624.fasta Sanger: ef32243610.fasta |
Cloned DNA: |
Triakis semifasciata parvalbumin alpha cDNA (pvalb1) 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 shark pvalb1 coding sequence 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 shark 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. Other name of the plasmid is pET-28a(+)-MBP-TEV-Shark. |
EMBL Accession number: | - |
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-Shark (LMBP 12829) 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.