GeneCorner plasmid details

Last data update: 23 July 2021 09:43 CEST

Plasmid name: AU2 (LMBP 7766)

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Price category: Cat. 3 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: (View with Genome Compiler)
analysis results

NGS: 7766-gc-feb2020.fasta

Cloned DNA: Phage MS2 assembly protein gene (A)
Phage MS2 coat protein gene (COAT)
Histidine tag (His-tag); N-terminal
Thrombin recognition site; N-terminal
S-tag; N-terminal
Histidine tag (His-tag); C-terminal
Escherichia coli lac repressor gene (lacI)
Promoter: Escherichia coli lac repressor promoter (lacI)
Phage T7 gene 10 promoter (T7g10) with lac operator (T7lac)
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: pET-30b(+); pMS27
Further information: The plasmid was created by PCR amplifying a pMS27 DNA fragment, consisting of the A protein gene and the COAT protein gene and encoding the phage MS2 maturase, and ligating the KpnI-HindIII amplicon into the KpnI-HindIII opened pET30b(+) vector.
The His-tags, the thrombin recognition site and the S-tag are not in frame with the phage MS2 A and COAT genes.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727565.1.
The nucleotide sequence of the pET-30b(+) vector part corresponds with the pET-30b(+) sequence file from Novagen (version 25/5/2002).
The nucleotide sequence of the pMS27 insert corresponds with the BCCM/GeneCorner sequence analysis results obtained from plasmid BH20 (LMBP 7765) and with the sequence analysis results from the depositor. However, the 'C' nucleotide at position 291 of the COAT protein cds was originally a 'T' in the sequence file of pMS27 (LMBP 5349). This nucleotide change concerns a silent mutation.
EMBL Accession number: LT727565.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 01/12/2020
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BglI, HincII, HindIII/KpnI, PvuII and XbaI.

The plasmid has also been fully sequenced.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr V. Jestin(1).
(1) VIPAC unit, National reference laboratory on avian influenza & Newcastle disease, Ploufragan, France
Plasmid reference: -
Related plasmid reference: Mikel et al., Front Microbiol 7 (2016), 1911 [PMID: 28133456] [DOI: 10.3389/fmicb.2016.01911]
Yu et al., J. Clin. Microbiol. 46 (2008), 837-841 [PMID: 18160451]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli B BL21(DE3)
Host reference: Studier and Moffatt, J. Mol. Biol. 189 (1986), 113-130 [PMID: 3537305] [DOI: 10.1016/0022-2836(86)90385-2]
Related host reference: Daegelen et al., J. Mol. Biol. 394 (2009), 634-643 [PMID: 19765591] [DOI: 10.1016/j.jmb.2009.09.022]
Jeong et al., J. Mol. Biol. 394 (2009), 644-652 [PMID: 19786035] [DOI: 10.1016/j.jmb.2009.09.052]
Studier et al., J. Mol. Biol. 394 (2009), 653-680 [PMID: 19765592] [DOI: 10.1016/j.jmb.2009.09.021]
Helper plasmid: -
Cultivation medium: LB-Lennox + kanamycin (50 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: -
Other culture collection numbers: -

Refer in your Materials and Methods:

AU2 (LMBP 7766) is available at BCCM/GeneCorner. This plasmid was deposited by Dr V. Jestin.

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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