Last data update:
15 July 2019 04:12 CEST
|Name:||pROSA26-RMCE-COIN-DV Add to cart|
|Accession number:||LMBP 8188|
|Sequence file:||Depositor's sequence: p8188.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)
Mouse gene trap ROSA 26 gene (Gtrosa26, Gt(ROSA)26Sor, beta geo, Gtrgeo26, R26, ROSA26, Thumpd3as1, GeneID 14910); 5' UTR and 3' UTR
|Promoter:||Mouse phosphoglycerate kinase 1 promoter (PGK1)
Escherichia coli lac operon promoter
Phage T7 gene 10 promoter (T7g10)
|Terminator:||Simian virus 40 polyadenylation signal (SV40 polyA)
Bovine growth hormone polyadenylation signal (BGH polyA)
Mouse phosphoglycerate kinase 1 polyadenylation signal (PGK1 polyA)
Herpes simplex virus (HSV) thymidine kinase polyadenylation signal (TK polyA)
|Selection marker:||Ampicillin (amp)
Neomycin (neo; G418)
|Replicon:||Escherichia coli plasmid pMB1 origin
Phage f1 origin
|Host range:||Escherichia coli; use a ccdB-resistant strain for propagation
|Further information:||This multisite Gateway Destination vector was constructed by cloning an FRT-loxP-pPGK1-neo-pA-loxP cassette between the 5' UTR of ROSA26 and the attR4 site, and a pA-FRT-neo cassette between the attR3 site and the 3' UTR of ROSA26 of pROSA26-DV3.
The plasmid is a multisite Gateway destination vector, containing the ccdB gene inserted between the attenuator sites R4 and R3. Next to this plasmid, three entry clones are needed in the multisite approach to target the gene of interest to the endogeneous ROSA26 locus in mouse ES cells: 1) a 5' entry clone with attL4-R1 sites and containing the rtTA transactivator, an IRES-Puro-pA selection cassette and a CMVTRE promoter (e.g. pEntry-attL4-rtTA-IRES-Puro-pA-TRE-attR1 (LMBP 8214)) ; 2) a middle entry clone with attL1-L2 sites and containing the gene of interest and 3) a 3' entry clone with attR2-L3 sites and containing the IRES-EGFP reporter (e.g. pEntry-attR2-IRES-eGFP-luc+-pA-Ins/Ins-attL3 (LMBP 8199)). In this approach, an exogeneous promoter is used to drive transgene expression, resulting in higher expression levels as compared to the monosite approach using the endogeneous ROSA26 promoter (see pROSA26-DV1 (LMBP 6451)).
The G4 ROSALUC ES cells are available at BCCM/GeneCorner as well.
Other name of the plasmid is pROSA26 RMCE COIN DV.
|EMBL Accession number:||-|
|Latest sequence update:||01/12/2016|
|Authenticity test:||The restriction enzyme pattern has still to be analysed.|
|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
|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 K12xB DB3.1|
|Related host reference:||Bernard et al., J. Mol. Biol. 226 (1992), 735-745 [PMID: 1324324]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml)|
|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.