GeneCorner plasmid details

Last data update: 09 March 2021 04:23 CET

Plasmid name: pcDNA3.1-Flag (LMBP 5351)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: (View with Genome Compiler)
analysis results

Sanger: 144928.fasta

Cloned DNA: FLAG epitope tag; N-terminal
Promoter: Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer
Phage T7 gene 10 promoter (T7g10)
Simian virus 40 early promoter (SV40 early)
Escherichia coli lac operon promoter
binding site:
Terminator: Bovine growth hormone polyadenylation signal (BGH polyA)
Simian virus 40 polyadenylation signal (SV40 polyA)
Selection marker: Ampicillin (amp)
Neomycin (neo; G418)
Replicon: Escherichia coli plasmid pMB1 origin
Phage f1 origin
Simian virus 40 bidirectional origin (SV40)
Host range: Escherichia coli
Mammalian cells; SV40 permissive cells
Parental clone: pcDNA3.1/HisB
Further information: The plasmid was constructed by inserting the 39 bp HindIII/KpnI fragment, containing the FLAG epitope tag, between the HindIII and KpnI sites of pcDNA3.1/HisB.
The plasmid is designed for high-level expression of a recombinant protein, fused to the N-terminal FLAG epitope tag, under control of the strong hCMV-IE promoter and enhancer in mammalian cells.
There is uncertainty about the presence of a second enhancer in the SV40 early promoter. The absence of a second enhancer in this promoter may cause bacterial leakage expression of the Tn5 neomycin resistance gene. This would cause transformed E. coli cells to be resistant to kanamycin, although growth should be reduced compared to growth on medium containing ampicillin.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727011.1.
Other name of the plasmid is pcDNA3.1-flag b.
EMBL Accession number: LT727011.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 28/11/2007
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: Alw44I/BamHI, AvaI/XmnI, HincII, HindIII/PvuII, KpnI, NaeI/SacI and NdeI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr M. Baens(1).
(1) Department of Human Genetics, KULeuven, Belgium
Plasmid reference: -
Related plasmid reference: Noels et al., J. Biol. Chem. 282 (2007), 10180-10189 [PMID: 17287209]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 MC1061
Host reference: Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
Related host reference: Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: -
Other culture collection numbers: -

Refer in your Materials and Methods:

pcDNA3.1-Flag (LMBP 5351) is available at BCCM/GeneCorner. This plasmid was deposited by Dr M. Baens.

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.

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