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GeneCorner plasmid details

Last data update: 28 October 2020 04:17 CET

Plasmid name: pCAGGS-E-crmA (LMBP 4888)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p4888.gb (View with Genome Compiler)
p4888.txt
p4888.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Cowpox virus cytokine response modifier A cDNA (crmA)
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: pCAGGScrmA; pCAGGS-EmCASP-1
Further information: The plasmid was constructed as follows: (i) the crmA coding sequence was PCR amplified using pCAGGScrmA as template, (ii) the NotI/XmaI amplicon was then inserted in frame to the N-terminal E-tag between the NotI and XmaI sites of pCAGGS-EmCASP-1.
pCAGGS-E-crmA is useful for highly efficient expression of crmA under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells. CrmA inhibits caspases, especially caspase-1 and caspase-8.
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 LT727057.1.
The nucleotide sequence of the crmA cDNA corresponds with the EMBL Nucleotide Sequence Database accession number M14217.1.
Name mentioned in Maelfait et al. (2008) is pCAGGS-E-CrmA.
EMBL Accession number: M14217.1, view at EMBL, GenBank, DDBJ
LT727057.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 01/02/2005
Sequence detail:
Nucleotide sequence at the start of crmA:

                   ---------------------- E-tag --------------------->
5' ... TTCCACC.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
               ***                                                     NotI
            NcoI

          |--> crmA
          | 2           5
       GCT.GAT.ATC.TTC.AGG.GAA.ATC ... 3'
       Ala Asp Ile Phe Arg Glu Ile
      

Nucleotide sequence at the end of crmA:

5' ... TCT.CCA.ACA.ACT.AAT.TAA.CCCGGGACATTAAACGCGTGGGTACCGGATCC … 3'
       Ser Pro Thr Thr Asn     SmaI                KpnI  BamHI      
      

***: start codon.
Punctuation indicates reading frame.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: NcoI, NotI and SnaBI.
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: -
Related plasmid reference: Maelfait et al., J. Exp. Med. 205 (2008), 1967-1973 [PMID: 18725521] [DOI: 10.1084/jem.20071632]
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-E-crmA (LMBP 4888) is available at BCCM/GeneCorner. This plasmid was deposited by P. Schotte and 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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