Last data update: 24 January 2024 16:39 CET
Plasmid name: pET-28a-MBP-TEV-Cypc3 (LMBP 12819)
| New search | Print data sheet |
| Price category: | Cat. 2 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p12819.gb
(View with Genome Compiler) p12819.txt p12819.pdf |
| Sequence analysis results Genecorner: |
Sanger: ef32243617.fasta Sanger: ef32243601.fasta |
| Cloned DNA: |
Cyprinus carpio fructose-bisphosphate aldolase A cDNA (Cyp c 3, Cyp c3, GeneID 109108793) 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 carp Cyp c3 coding sequence into the BamHI/XhoI opened pET28a-MBP-TEV vector. The plasmid was designed to express the malE-Cyp c3 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 c3 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 c3 gene corresponds with the EMBL Nucleotide Sequence Database accession number MH248133.1. Other name of the plasmid is pET-28a(+)-MBP-Cypc3 |
| EMBL Accession number: | MH248133.1, 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 carp Cyp c3 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-Cypc3 (LMBP 12819) 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.