GREAT AT SMALL THINGS

LMBP plasmid details

Last data update: 20 October 2018 04:23 CEST  


Name: pLenti6-eGFP-V5 puro   Add to cart
Accession number: LMBP 8402
Sequence file: LMBP sequence: p8402.gb, p8402.txt     View with Genome Compiler
Circular map: p8402.pdf
Status: LMBP core plasmid
Cloned DNA: B gene of the control of cell death locus of the Escherichia coli F plasmid (ccdB, lethal gene)
Aequorea victoria green fluorescent protein DNA (GFP); enhanced red-shifted variant (EGFP)
V5 epitope; C-terminal
Promoter: Rous sarcoma virus/human immunodeficiency virus hybrid long terminal repeat (RSV/HIV-1 5' LTR)
Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer
Escherichia coli lac operon promoter
Simian virus 40 early promoter (SV40 early)
Phage T3 promoter
Phage T7 gene 10 promoter (T7g10)
Escherichia coli lac operon promoter; mutant (lacUV5)
Ribosome
binding site:
-
Terminator: Rous sarcoma virus/human immunodeficiency virus hybrid long terminal repeat (RSV/HIV-1 3' LTR); modified HIV-1 3' LTR with deleted U3 region (ΔU3/3' LTR)
Simian virus 40 polyadenylation signal (SV40 polyA)
Selection marker: Ampicillin (amp)
Chloramphenicol (cam)
Puromycin (puro)
Replicon: Escherichia coli plasmid pMB1 origin
Phage f1 origin
Simian virus 40 bidirectional origin (SV40)
Host range: Escherichia coli; use a ccdB-resistant strain for propagation
Mammalian cells; SV40 permissive cells
Parental clone: pLenti6-V5 puro; pEGFP-C1
Further information: The construction of this plasmid is described in De Groote et al. (2016).
This is a lentiviral Gateway destination vector.
The plasmid is suitable for microRNA expression.
The U3 region of the HIV-1 3' LTR is deleted (ΔU3 or dU3) and facilitates self-inactivation of the HIV-1 5' LTR after transduction to enhance the biosafety of the vector. The element also contains a polyadenylation signal for transcription termination and polyadenylation of mRNA in transduced cells.
The vector contains the psi packaging signal, allowing viral packaging, and the rev response element (RRE) from HIV-1, permitting Rev-dependent nuclear export of unspliced viral mRNA.
Because of the high risk for recombination, plasmid integrity should be checked via PvuII restriction digest after each subcultivation.
This Gateway destination vector is compatible with the human orfeome library which is also available at BCCM/GeneCorner.
This vector can not be used to express short hairpin RNA (shRNA), which requires an RNA-polymerase III promotor. However, this vector can be used for knock-down experiments if the hairpin is expressed in a micro-RNA context (eg. Block-iT PolII miR RNAi kit).
The nucleotide sequence of the pLenti6-eGFP-V5 puro vector corresponds with the EMBL Nucleotide Sequence Database accession number LT009450.1.
Other names of the plasmid are pLenti6(puro)-EGFP-V5-DEST cl.1, pLenti6Fluo-puro and pLenti6-eGFP-V5-puro.
EMBL Accession number: LT009450.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 18/08/2015
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: ApaI/SpeI, HindIII, PvuII and XhoI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: The plasmid was deposited by Dr P. De Groote(1)(2) and Prof. Dr W. Declercq(1)(2).
(1) Inflammation Research Center, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: De Groote et al., BioTechniques 60 (2016), 252-259 [PMID: 27177818] [DOI: 10.2144/000114417]
Related plasmid reference: PhD thesis Philippe De Groote (2012)
Restricted distribution: - BCCM MTA
Distributed as: plasmid DNA
Host for distribution: Escherichia coli K12xB DB3.1
Host reference: -
Related host reference: Bernard et al., J. Mol. Biol. 226 (1992), 735-745 [PMID: 1324324]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml) + chloramphenicol (34 μg/ml)*
Cultivation temperature: 28°C
Biosafety level: L1 in E. coli; L2 in mammalian cells
Cultivation remark: *: selection of transformants on chloramphenicol; subsequent cultivation of a single colony in liquid medium with ampicillin
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.

 

New search