Last data update: 24 January 2024 16:39 CET
Plasmid name: pEntry-attL4-rtTA-IRES-Puro-pA-TRE-attR1 (LMBP 8214)
| New search | Print data sheet |
| Price category: | Cat. 2 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p8214.gb
(View with Genome Compiler) p8214.txt p8214.pdf |
| Sequence analysis results Genecorner: |
Sanger: ef32769313.fasta Sanger: ef32769312.fasta Sanger: ef32769311.fasta |
| Cloned DNA: |
Reverse tetracycline-responsive transcriptional activator (rtTA) |
| Promoter: | Human cytomegalovirus immediate early promoter (CMV-IE); minimal promoter with tet operator (CMVT) |
| Ribosome binding site: |
Internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV) polyprotein gene |
| Terminator: | Escherichia coli rrnB operon T1 terminator Escherichia coli rrnB operon T2 terminator Bovine growth hormone polyadenylation signal (BGH polyA) |
| Selection marker: | Neomycin (neo; kanamycin (kan)) Puromycin (puro) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli Mammalian cells |
| Parental clone: | pENTR-L4-R1 |
| Further information: | The plasmid was constructed by cloning the rtTA-IRES-Puro-pA-TRE cassette into the pENTR-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. - a tetracycline responsive element (TRE), consisting of the minimal human CMV promoter, combined with the tet operator. 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-TRE R1. |
| EMBL Accession number: | U89930.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 28/07/2021 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BamHI/DraI, PvuI, SacI and XbaI. Additionally, most of the IRES-puro-pA-hCMVT-attR1-neo-pMB1 region was sequenced at BCCM/GeneCorner |
| 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-TRE-attR1 (LMBP 8214) 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.