Last data update: 22 May 2023 13:52 CEST
Plasmid name: pET-28a-MBP-TEV-Derp26 (LMBP 12823)
New search | Print data sheet |
Price category: | Cat. 2 (cf. price list) |
Status: | GeneCorner core plasmid |
GeneCorner sequence: |
p12823.gb
(View with Genome Compiler) p12823.txt p12823.pdf |
Sequence analysis results Genecorner: |
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) |
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 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. - 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-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.