Last data update: 22 May 2023 13:52 CEST
Plasmid name: pET-28a-MBP-TEV-Cluh1 (LMBP 12820)
| New search | Print data sheet |
| Price category: | Cat. 2 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p12820.gb
(View with Genome Compiler) p12820.txt p12820.pdf |
| Sequence analysis results Genecorner: |
Sanger: ef32243618.fasta Sanger: ef32243603.fasta |
| Cloned DNA: |
Clupea harengus parvalbumin beta 2 cDNA (pvalb2, pvalb1, Clu h 1, Clu h1, GeneID 105910226) 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 C. harengus pvalb2 coding sequence, optimised for E. coli, into the BamHI/XhoI opened pET28a-MBP-TEV vector. The plasmid was designed to express the malE-pvalb2 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 C. harengus pvalb2 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 C. harengus pvalb2 allergen corresponds with GenBank accession number NP_001296764.1. Other name of the plasmid is pET-28a(+)-MBP-TEV-Cluh1. |
| EMBL Accession number: | NP_001296764.1, 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, PstI and PvuII. The C. harengus pvalb2 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. - 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-Cluh1 (LMBP 12820) 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.