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GeneCorner plasmid details

Last data update: 26 November 2020 04:30 CET

Plasmid name: pET3E-His-AU1-hUb-K63 (LMBP 5913)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p5913.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human ubiquitin B cDNA (UBB, Ub, HEL-S-50, GeneID 7314); K63 mutated mature sequence
Histidine tag (His-tag); N-terminal
AU1-tag; N-terminal
Promoter: Phage T7 gene 10 promoter (T7g10)
Ribosome
binding site:
Ribosome binding site (RBS) of the phage T7 gene 10 (T7g10)
Terminator: Phage T7 gene 10 terminator (T7g10)
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Parental clone: pET3E
Further information: The human ubiquitin K63 cDNA only encodes the wild type lysine number 63; the other six lysine codons are mutated to arginine codons.
This plasmid contains the T7 gene 10 promoter whose transcription is initiated by T7 RNA polymerase, making the vector useful for in vitro preparation of single-stranded RNAs from cloned human UB gene.
EMBL Accession number: BT020104, view at EMBL, GenBank, DDBJ
Latest sequence update: 06/03/2009
Sequence detail:
Schematic representation of the human ubiquitin K63 mutant:
 
                 6  11 27 29 33 48 63
wild type:  5' --K--K--K--K--K--K--K-- 3'

K63 mutant: 5' --R--R--R--R--R--R--K-- 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 K.-L. Guan(1). It was received from Dr. E. Tang(1).
(1) Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, Michigan, USA
Plasmid reference: -
Related plasmid reference: Tang et al., J. Biol. Chem. 278 (2003), 37297-37305 [PMID: 12867425]
Restricted distribution: - BCCM/MTA adapted by Michigan University
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:

pET3E-His-AU1-hUb-K63 (LMBP 5913) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr K.-L. Guan.

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.

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