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GeneCorner plasmid details

Last data update: 25 April 2019 04:11 CEST  


Name: pRMCE-DV2   Add to cart
Accession number: LMBP 8190
Sequence file: Depositor's sequence: p8190.gb     View with Genome Compiler
Status: GeneCorner non-core plasmid
Cloned DNA: B gene of the control of cell death locus of the Escherichia coli F plasmid (ccdB, lethal gene)
Promoter: Mouse phosphoglycerate kinase 1 promoter (PGK1)
Escherichia coli lac operon promoter
Escherichia coli lac operon promoter; mutant (lacUV5)
Ribosome
binding site:
-
Terminator: -
Selection marker: Ampicillin (amp)
Chloramphenicol (cam)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli; use a ccdB-resistant strain for propagation
Mammalian cells
Parental clone: pROSA26-DV2; pROSALUC
Further information: The plasmid was constructed by replacing the 5' UTR and 3' UTR regions of mouse ROSA26 and the neomycin resistance gene in pROSA26-DV2 with the heterospecific FRT sites present in pROSALUC. A PGK-ATG sequence was included to ensure a proper trap-coupled RMCE reaction when targeted into mouse G4 ROSALUC ES cells.
pRMCE-DV2 is a highly efficient Recombinase-Mediated Cassette Exchange (RMCE)-compatible, conditional Gateway destination vector.
This multisite Gateway destination vector contains the ccdB gene inserted between the attenuator sites attR3 and attR4.
The plasmid contains:
- two heterospecific Saccharomyces cerevisiae 2μ FLP Recombinase Target sites (one wt and one mutant FRT site)
- two adjacent copies of the chicken β-globin 5' DNase I hypersensitive site 4 (5'HS4) insulator core sequence. Insulators are DNA sequences which possess the ability to protect expressing genes from inappropriate signals emanating from their surrounding environment by acting as barriers that prevent the advance of nearby condensed chromatin that may otherwise silence expression.
The resulting multisite Gateway expression vector, by co-transfection of a FlpE plasmid, can subsequently be targeted to the ROSA26 locus of G4 ROSALUC ES cells using a trap-coupled RMCE approach, hereby restoring neomycin resistance.
The G4 ROSALUC ES cells are available at BCCM/GeneCorner as well.
Other name of the plasmid is pRMCE DV2.
EMBL Accession number: -
Latest sequence update: 01/12/2016
Authenticity test: The restriction enzyme pattern has still to be analysed.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr J. Haigh(1).
(1) Australian Centre for Blood Diseases, Central Clinical School, Monash University, Melbourne, Australia
Plasmid reference: PhD thesis Lieven Haenebalcke
Related plasmid reference: Nyabi et al., Nucleic Acids Res. 37 (2009), e55 [PMID: 19279185] [DOI: 10.1093/nar/gkp112]
Restricted distribution: - VIB/BCCM MTA
- The depositor will be informed of the customer's identity upon release of a sample outside the depositor's department or outside the departments in which BCCM/GeneCorner is embedded, namely UGent-DBMB and VIB-IRC.
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 DH5α
Host reference: Focus 8 (1986), 9
Related host reference: Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Other culture collection numbers: -

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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