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GeneCorner plasmid details

Last data update: 24 January 2024 16:39 CET

Plasmid name: pEntry-attL4-rtTA-IRES-Puro-pA-attR1 (LMBP 8213)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p8213.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Reverse tetracycline-responsive transcriptional activator (rtTA)
Promoter: -
Ribosome
binding site:
Internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV) polyprotein gene
Terminator: Bovine growth hormone polyadenylation signal (BGH polyA)
Escherichia coli rrnB operon T1 terminator
Escherichia coli rrnB operon T2 terminator
Selection marker: Neomycin (neo; G418; kanamycin (kan))
Puromycin (puro)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Mammalian cells
Parental clone: pENTRY-L4-R1
Further information: The plasmid was constructed by cloning the rtTA-IRES-Puro-pA cassette into the pENTRY-L4-R1 vector.
The plasmid is a multisite Gateway entry vector, designed as a 5' entry clone (with attL4-attR1 sites) for the multisite approach using the pCOIN DV vector.
The plasmid contains:
- a reverse tetracyclin transactivator (rtTA)
- an IRES-puromycin-pA drug-selectable cassette to select for rtTA expressing cells after Cre excision of the pCOIN DV vector.
The G4 ROSALUC ES cells (LMBP 10507CB) are available at BCCM/GeneCorner as well.
The nucleotide sequence of the rtTA corresponds with the EMBL Nucleotide Sequence Database accession number U89930.1.
Other name of the plasmid is pEntry L4 rtTA-IRES-Puro-pA R1.
EMBL Accession number: U89930.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 01/12/2016
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 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: Haenebalcke et al., Stem Cell Rev. Rep. 9 (2013), 774-785 [PMID: 23877658] [DOI: 10.1007/s12015-013-9458-z]
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: Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791] [DOI: 10.1016/s0022-2836(83)80284-8]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Cultivation medium: LB-Lennox + kanamycin (50 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Other culture collection numbers: -

Refer in your Materials and Methods:

pEntry-attL4-rtTA-IRES-Puro-pA-attR1 (LMBP 8213) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr J. Haigh and was published in Lieven Haenebalcke, .

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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