Last data update: 24 January 2024 16:39 CET
Plasmid name: pMSCV-His-AU-Ub-K63 (LMBP 6594)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p6594.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human ubiquitin B cDNA (UBB, Ub, HEL-S-50, GeneID 7314); mutated sequence Histidine tag (His-tag); N-terminal AU1-tag; N-terminal |
| Promoter: | Mouse stem cell PCMV virus long terminal repeat (5' MSCV-PCMV LTR) Mouse phosphoglycerate kinase 1 promoter (PGK1) Escherichia coli lac operon promoter |
| Ribosome binding site: |
- |
| Terminator: | Mouse stem cell PCMV virus long terminal repeat (3' MSCV-PCMV LTR) |
| Selection marker: | Ampicillin (amp) Puromycin (puro) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Mammalian cells |
| Parental clone: | pMSCVpuro; pET-His-AU-Ub-K63 |
| Further information: | The plasmid was constructed by PCR amplifying the mutated human UBB cds from pET-His-AU-Ub-K63 and ligating it into the HindIII/ApaI opened pMSCVpuro plasmid. This plasmid is generated for high-affinity isolation of ubiquitinated proteins of a specific linkage. For even greater specificity, this plasmid can be combined with the corresponding bio-tagged ubiquitin for 2D (first His-purification, then biotin precipitation) isolation of ubiquitinated proteins under denaturing conditions, for example for proteomic studies. Denaturing (8M Urea, 2% SDS) conditions also ensures that no deubiquitinase activity is present during purification. |
| EMBL Accession number: | - |
| Latest sequence update: | 04/02/2010 |
| Sequence detail: | Primers used to amplify the His-tagged human UBB:
Forward: 5' CATAAGCTTAAA.ATG.GTA.CAT.CAC.CAT.CAC.CAT.CAC
HindIII ***
Reverse: 5' TAAGGGCCC.CTA.CCC.ACC.TCT.GAG.AC
ApaI +++
***: start codon.
+++: termination codon.
Punctuation indicates reading frame. |
| 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 Prof. Dr R. Beyaert(1) (2). (1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 MC1061 |
| Host reference: | Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493] |
| Related host reference: | Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pMSCV-His-AU-Ub-K63 (LMBP 6594) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert . |
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.