Last data update: 24 January 2024 16:39 CET
Plasmid name: pcDNA3-HA-hUb-K48R (LMBP 5898)
New search | Print data sheet |
Price category: | Cat. 1 (cf. price list) |
Status: | GeneCorner core plasmid |
GeneCorner sequence: |
p5898.gb
(View with Genome Compiler) p5898.txt p5898.pdf |
Sequence analysis results Genecorner: |
- |
Cloned DNA: |
Human ubiquitin-40S ribosomal protein S27a cDNA (RPS27A, UBA80, UBCEP80); mutated mature ubiquitin 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) Escherichia coli lac operon promoter |
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 ubiquitin gene. The human ubiquitin K48R cDNA contains a mutation converting the lysine number 48 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 nucleotide sequence of the human ubiquitin cDNA corresponds with the Genbank accession number NM_002954.5, except for three additional nucleotides. |
EMBL Accession number: | NM_002954.5, view at GenBank |
Latest sequence update: | 31/05/2018 |
Sequence detail: | Schematic representation of the human ubiquitin K48R mutant: 6 11 27 29 33 48 63 wild type: 5' --K--K--K--K--K--K--K-- 3' K48R mutant:5' --K--K--K--K--K--R--K-- 3' |
Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: EcoRI, HaeII, HindIII/XhoI and SspI. |
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: | Tanghe et al., Cell. Mol. Life Sci. 75 (2018), 2827-2841 [PMID: 29435596] [DOI: 10.1007/s00018-018-2763-6] 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-K48R (LMBP 5898) 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.