Last data update: 30 July 2021 10:22 CEST
Plasmid name: pCAGGS-E-sifA (LMBP 6774)
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|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
Salmonella enterica serovar Typhimurium LT2 sifA cDNA|
|Promoter:||Chicken β-actin/rabbit β-globin hybrid promoter (AG)
Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only
Escherichia coli lac operon promoter
|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
|Further information:||The plasmid was constructed by replacing the hA20 cds from the NotI/XhoI opened pCAGGSEhA20 by the PCR amplified sifA cds. As a result, the sifA coding sequence is fused in phase to the N-terminal E-tag.
pCAGGS-E-sifA is useful for highly efficient expression of sifA under the control of the AG promoter and the hCMV-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.
Protein expression of this vector in HEK293T cells was tested at GeneCorner.
The nucleotide sequence of the sifA cds corresponds with the EMBL Nucleotide Sequence Database accession number AE006468.1.
|EMBL Accession number:||AE006468.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||07/09/2010|
Primers used to isolate the SifA cDNA: Forward primer: 5' AAGGAAGCGGCCGCT.CCG.ATT.ACT.ATA.GGG.AAT.GG NotI AA2 Reverse primer: 5' GTCGCCTCGAG.TTA.TAA.AAA.ACA.ACA.TAA.ACA.GC XhoI +++ +++: termination codon Punctuation indicates reading frame.
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: HincII, NcoI, NotI/XhoI and PvuI.|
|History of deposit:||This plasmid was deposited by W. De Schamphelaire(1) and Prof. Dr R. Beyaert(1).
(1) BCCM/GeneCorner, Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|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)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pCAGGS-E-sifA (LMBP 6774) is available at BCCM/GeneCorner. This plasmid was deposited by W. De Schamphelaire 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.