Last data update: 24 January 2024 16:39 CET
Plasmid name: pCAGGS-EYopH (LMBP 4663)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p4663.gb
(View with Genome Compiler) p4663.txt p4663.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Yersinia enterocolitica outer membrane virulence protein H cDNA (yopH) 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 yopH 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. YopH is involved in the type III secretion system (T3SS). pCAGGS-EYopH is useful for highly efficient expression of YopH 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 LT727155.1. The nucleotide sequence of the yopH cDNA corresponds with the EMBL Nucleotide Sequence Database accession number M30457.1. Other name of the plasmid is pCAGGSE-YopH. |
| EMBL Accession number: | M30457.1, view at EMBL, GenBank, DDBJ LT727155.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 11/08/2006 |
| 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
|--> YopH
| 2 5 465 468
GCT.AAC.TTA.TCA.TTA.AGC..........CCA.TTA.TTA.AAT.AGC.TAA.CCCGGGACATTAAACGCGTGGGTACCGGATCC...3'
Asn Leu Ser Leu Ser Pro Leu Leu Asn Ser XmaI KpnI BamHI
Primers used to amplify YopH:
Forward primer: 5'...CGCAAGGAAGCGGCCGCTAACTTATCATTAAGCGATCTTCATCGTCAGG...3'
NotI
Reverse primer: 5'...GTCGCCCCGGGTTAGCTATTTAATAATGGTCGCCC...3'
XmaI |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: KpnI, NcoI, NotI, PstI and XbaI. |
| 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-EYopH (LMBP 4663) 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.