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GeneCorner plasmid details

Last data update: 27 October 2020 04:27 CET

Plasmid name: pCAGGS-EYopE-R144A (LMBP 4665)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p4665.gb (View with Genome Compiler)
p4665.txt
p4665.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Yersinia enterocolitica outer membrane virulence protein E cDNA (yopE); R144A 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-EYopE
Further information: YopE-R144A is an inactive mutant of the yopE cDNA, the latter being involved in the type III secretion system (T3SS) and encoding a GTPase activating protein.
pCAGGS-EYopE-R144A is useful for highly efficient expression of YopE-R144A under the control of the AG promoter and the human CMV-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 LT727157.1.
The nucleotide sequence of the wild type yopE cDNA corresponds with the EMBL Nucleotide Sequence Database accession number M92066.1.
Other name of the plasmid is pCAGGSE-YopE M.
EMBL Accession number: M92066.1, view at EMBL, GenBank, DDBJ
LT727157.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 12/12/2007
Sequence detail:
E-tag
                    ---------------------------------------------------
5'...GAATTCCACC ATG.GGT.GCG.CCG.GTG.CCG.TAT.CCG.GAC.CCG.CTG.GAA.CCG.CGT.GCG.GCC.
                Met Gly Ala Pro Val Pro Tyr Pro Asp Pro Leu Glu Pro Arg Ala Ala 
     EcoRI                                BspMII                        NotI

   |--> YopE
   | 2           5                           144    
GCT.AAA.ATA.TCA.TCA.TTT..........GGC.CCT.CTT.GCC.GGC.AGT.ATC...
    Lys Ile Ser Ser Phe          Gly Pro Leu Ala Gly Ser Ile      
                                             ***
                                             NaeI

   215             219 
...TTA.CTG.TCA.TTG.ATG.TGA.CCCGGGACATTAAACGCGTGGGTACCGGATCC...3'
   Leu Leu Ser Leu Met     SmaI                KpnI  BamHI

***: mutated nucleotides (CGT = wt).
Punctuation indicates reading frame.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BamHI, NaeI, NaeI/SacII and NotI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr R. Beyaert(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Schotte et al., J. Biol. Chem. 279 (2004), 25134-25142 [PMID: 15060067]
Related plasmid reference: Michiels et al., Infect. Immun. 58 (1990), 2840-2849 [PMID: 2129533]
Sory et al., Proc. Natl. Acad. Sci. U.S.A. 92 (1995), 11998-12002 [PMID: 8618831]
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:

pCAGGS-EYopE-R144A (LMBP 4665) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert and was published in Schotte et al., 2004.

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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