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GeneCorner plasmid details

Last data update: 26 November 2020 04:30 CET

Plasmid name: pCAGGS-EYopT (LMBP 4666)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p4666.gb (View with Genome Compiler)
p4666.txt
p4666.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Yersinia enterocolitica outer membrane virulence protein T cDNA (yopT)
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-EmCASP-1; pYV40
Further information: The plasmid was constructed as follows: wild type yopT from the pYV40 plasmid of strain E40 was amplified (PCR), and cloned in frame with the N-terminal E-tag of pCAGGS-EmCASP-1 as a NotI/XmaI fragment.
YopT is involved in the type III secretion system (T3SS) and encodes a cysteine protease.
pCAGGS-EYopT is useful for highly efficient expression of YopT 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 LT727158.1.
The nucleotide sequence of the yopT cDNA corresponds with the EMBL Nucleotide Sequence Database accession number AF102990.1.
Other name of the plasmid is pCAGGSE-YopTwt.
EMBL Accession number: AF102990.1, view at EMBL, GenBank, DDBJ
LT727158.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 11/09/2006
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

   |--> YopT
   | 2           5                           320 321  
GCT.GAC.AGT.ATT.CAC.GGA..........GAC.TCC.AAG.GAG.GTT.TGA.CCCGGGACATTAAACGCGTGGGTACCGGATCC...3'
    Asp Ser Ile His Gly          Asp Ser Lys Glu Val     XmaI                KpnI  BamHI
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BamHI, EcoRI, NcoI and NotI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by P. Schotte(1) (2) and 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-EYopT (LMBP 4666) is available at BCCM/GeneCorner. This plasmid was deposited by P. Schotte and 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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