Last data update: 24 January 2024 16:39 CET
Plasmid name: pcDNA3-HA-hUb-3KR (LMBP 5895)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p5895.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human ubiquitin B cDNA (UBB, Ub, HEL-S-50, GeneID 7314); 3KR mutated sequence Influenza HA epitope encoding the haemagglutinin tagging peptide; N-terminal |
| Promoter: | Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer Phage SP6 promoter Phage T7 gene 10 promoter (T7g10) Simian virus 40 early promoter (SV40 early) |
| Ribosome binding site: |
- |
| Terminator: | Bovine growth hormone polyadenylation signal (BGH polyA) Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Ampicillin (amp) Neomycin (neo; G418) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin Simian virus 40 bidirectional origin (SV40) |
| Host range: | Escherichia coli Mammalian cells; SV40 permissive cells |
| Parental clone: | pcDNA3-HA-hUb |
| Further information: | The plasmid is meant for eukaryotic expression of the human Ub gene. The human ubiquitin 3KR cDNA contains mutations converting the lysine numbers 29, 48 and 63 to arginine. There is uncertainty about the presence of a second enhancer in the SV40 early promoter. The absence of a second enhancer in this promoter may cause bacterial leakage expression of the Tn5 neomycin resistance gene. This would cause transformed E. coli cells to be resistant to kanamycin, although growth should be reduced compared to growth on medium containing ampicillin. The hUB cDNA was sequenced by BCCM/GeneCorner, it differs from the Genbank accession number NM_002954.4 in two nucleotides. |
| EMBL Accession number: | NM_002954.4, view at GenBank BT020104.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 10/02/2010 |
| Sequence detail: | Schematic representation of the human ubiquitin 3KR mutant:
6 11 27 29 33 48 63
wild type: 5' --K--K--K--K--K--K--K-- 3'
3KR mutant: 5' --K--K--K--R--K--R--R-- 3' |
| 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 I. Dikic(1). It was constructed by Dr. I. Szymkiewicz(1). (1) Institute of Biochemistry II, Goethe University School of Medicine, Frankfurt, Germany |
| Plasmid reference: | - |
| Related plasmid reference: | Haglund et al., Nat. Cell Biol. 5 (2003), 461-166 [PMID: 12717448] |
| 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: |
pcDNA3-HA-hUb-3KR (LMBP 5895) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr I. Dikic. |
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.