Last data update: 24 January 2024 16:39 CET
Plasmid name: pEntry-attL4-rtTA-IRES-Puro-pA-Ins/Ins-attR1 (LMBP 8211)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p8211.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-Ins/Ins 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. - 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 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-Ins/Ins 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-Ins/Ins-attR1 (LMBP 8211) 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.