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GeneCorner plasmid details

Last data update: 19 October 2020 04:30 CEST

Plasmid name: pCAGGS-E-SopE-G168V (LMBP 6116)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p6116.gb (View with Genome Compiler)
p6116.txt
p6116.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Salmonella enterica serovar Typhimurium SL1344 sopE cDNA; G168V mutant
E-tag; N-terminal
Promoter: Chicken β-actin/rabbit β-globin hybrid promoter (AG)
Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only
Escherichia coli lac operon promoter
Ribosome
binding site:
-
Terminator: Rabbit β-globin polyadenylation signal (β-globin polyA)
Simian virus 40 polyadenylation signal (SV40 polyA)
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Simian virus 40 bidirectional origin (SV40)
Host range: Escherichia coli
Mammalian cells; SV40 permissive cells
Parental clone: pCAGGS-E-sopE
Further information: The plasmid contains the S. typhimurium SL1344 sopE mutated sequence fused in phase to the N-terminal E-tag.
The mutated sequence of sopE was obtained by PCR using pCAGGS-E-SopE as template. As compared to the wt sopE cDNA, the glycine (G) codon at position 168 was replaced by a valine (V) codon, creating an additional BalI site.
pCAGGS-E-sopE-G168V is useful for highly efficient expression of mutated sopE under the control of the AG promoter and the hCMV-IE enhancer in various mammalian cells.
When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT726823.1.
The nucleotide sequence of the wt sopE cDNA corresponds with the EMBL Nucleotide Sequence Database accession number AF043239.1.
Other names of the plasmid are pCAGGSE-SopE G168V and pCAGGS-SopE-G/V.
EMBL Accession number: AF043239.1, view at EMBL, GenBank, DDBJ
LT726823.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 07/09/2007
Sequence detail:
Primers used to introduce the G168V mutation into the sopE coding sequence:

                               168
forward: 5' CG.CCA.AGC.GGC.GCA.GTG.GCC.AAT.CCT.TTT.ATA.ACT.C 3'
                                BalI
                                ^
                                      168
reverse: 5' G.AGT.TAT.AAA.AGG.ATT.GGC.CAC.TGC.GCC.GCT.TGG.CG 3'
                                BalI   ^

^  : mutated nucleotide.
Punctuation indicates reading frame.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BalI, BalI/NarI, EcoRI/HincII and HindIII/SacII.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr R. Beyaert(1) (2). It was constructed by A. Van Den Broeke(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: -
Related plasmid reference: Schlumberger et al., J. Biol. Chem. 278 (2003), 27149-27159 [PMID: 12719429]
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: LMBP 04876

Refer in your Materials and Methods:

pCAGGS-E-SopE-G168V (LMBP 6116) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert .

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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