Last data update: 24 January 2024 16:39 CET
Plasmid name: pCAGGS-EYopE (LMBP 4664)
New search | Print data sheet |
Price category: | Cat. 1 (cf. price list) |
Status: | GeneCorner core plasmid |
GeneCorner sequence: |
p4664.gb
(View with Genome Compiler) p4664.txt p4664.pdf |
Sequence analysis results Genecorner: |
- |
Cloned DNA: |
Yersinia enterocolitica outer membrane virulence protein E cDNA (yopE) 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 yopE 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. YopE is involved in the type III secretion system (T3SS) and encodes a GTPase activating protein. pCAGGS-EYopE is useful for highly efficient expression of YopE 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 LT727156.1. The nucleotide sequence of the yopE cDNA corresponds with the EMBL Nucleotide Sequence Database accession number M92066.1. Other name of the plasmid is pCAGGSE-YopEwt. |
EMBL Accession number: | M92066.1, view at EMBL, GenBank, DDBJ LT727156.1, 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 |--> YopE | 2 5 215 219 GCT.AAA.ATA.TCA.TCA.TTT..........TTA.CTG.TCA.TTG.ATG.TGA.CCCGGGACATTAAACGCGTGGGTACCGGATCC...3' Lys Ile Ser Ser Phe Leu Leu Ser Leu Met XmaI KpnI BamHI |
Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BamHI, NcoI, NotI and PstI. |
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: | Trosky et al., Cell. Microbiol. 10 (2008), 557-565 [PMID: 18081726] [DOI: 10.1111/j.1462-5822.2007.01109.x] Sory et al., Proc. Natl. Acad. Sci. U.S.A. 92 (1995), 11998-12002 [PMID: 8618831] Michiels et al., Infect. Immun. 58 (1990), 2840-2849 [PMID: 2129533] |
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 (LMBP 4664) 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.