GREAT AT SMALL THINGS

0

GeneCorner plasmid details

Last data update: 21 October 2021 10:13 CEST

Plasmid name: pdcDNA-Myc (LMBP 7212)

Add to cart

New search Print data sheet
Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p7212.gb (View with Genome Compiler)
p7212.txt
p7212.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: B gene of the control of cell death locus of the Escherichia coli F plasmid (ccdB, lethal gene)
Myc epitope; N-terminal
Promoter: Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer
Simian virus 40 early promoter (SV40 early)
Escherichia coli lac operon promoter; mutant (lacUV5)
Escherichia coli lac operon promoter
Phage T7 gene 10 promoter (T7g10)
Phage SP6 promoter
Ribosome
binding site:
-
Terminator: Simian virus 40 polyadenylation signal (SV40 polyA)
Bovine growth hormone polyadenylation signal (BGH polyA)
Selection marker: Ampicillin (amp)
Chloramphenicol (cam)
Neomycin (neo; G418)
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: pdcDNA-FLAG; pDEST-Myc
Further information: The plasmid was constructed by cutting out the FLAG-tagged Gateway reading frame cassette A (rfA, from Invitrogen) of pdcDNA-FLAG, as a BstXI-HindIII fragment, and replacing it by the BstXI-HindIII restricted, Myc-tagged rfA cassette of pDEST-Myc.
pdcDNA-Myc is a Gateway destination vector, containing the the ccdB gene flanked by the bacteriophage λ attR recombination sites.
pdcDNA-Myc is designed for fusing a gene of interest to the 6 N-terminal Myc epitopes according to the Gateway Cloning Technology (Invitrogen) and for high-level, constitutive, native expression of this gene in mammalian cells under control of the human CMV-IE promoter and enhancer.
A Gateway entry clone, containing the gene of interest flanked by the λ attL recombination sites, recombines with the attR sites of the destination vector, replacing the ccdB gene. Standard E. coli strains with the unreacted destination vector or with the by-product plasmid retaining the ccdB gene will fail to grow. Thus, this negative selection provides high-efficiency recovery of only the desired clones. The chloramphenicol resistance gene located between the two attR sites permits counterselection.
This vector has been shown to provide resistance to kanamycin, although growth was reduced compared to growth on medium containing ampicillin.
This Gateway destination vector is compatible with the human orfeome library which is also available at BCCM/GeneCorner.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT726849.1.
The absence of a second enhancer in the SV40 early promoter was shown by sequence analysis at BCCM/GeneCorner.
Other name of the plasmid is pdcDNAmyc.
EMBL Accession number: LT726849.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 05/09/2011
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BamHI, HincII, NcoI/XmnI and PvuI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by K. Staes(1) (2) and Prof. Dr F. Van Roy(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: -
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or 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 (25 μg/ml)*
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: *: selection of transformants on chloramphenicol; subsequent cultivation of a single colony in liquid medium with ampicillin. This culture grows very slow: take 24 hours into account for saturated growth!
Other culture collection numbers: -

Refer in your Materials and Methods:

pdcDNA-Myc (LMBP 7212) is available at BCCM/GeneCorner. This plasmid was deposited by K. Staes and Prof. Dr F. Van Roy .

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