GREAT AT SMALL THINGS

0

GeneCorner plasmid details

Last data update: 03 December 2020 04:26 CET

Plasmid name: pCAGGS-E-sipA (LMBP 6772)

Add to cart

New search

Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p6772.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Salmonella enterica serovar Typhimurium LT2 sipA cDNA
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: pCAGGSEhA20
Further information: The plasmid was constructed by replacing the hA20 cds from the NotI/XhoI opened pCAGGSEhA20 by the PCR amplified sipA cds. As a result, the sipA coding sequence is fused in phase to the N-terminal E-tag.
pCAGGS-E-sipA is useful for highly efficient expression of sipA 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 sipA 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: 07/09/2007
Sequence detail:
Primers used to isolate the SipA cDNA:

Forward primer: 5' AAGGAAGCGGCCGCT.GTT.ACA.AGT.GTA.AGG.ACT.CAG
                         NotI      AA2  
Reverse primer: 5' GTCGCCTCGAG.TTA.ACG.CTG.CAT.GTG.CA
                        XhoI   +++ 

+++: termination codon
Punctuation indicates reading frame.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: HincII, NcoI, NotI/XhoI and PvuI.
Class: Recombinant plasmid
Type: Plasmid
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
Plasmid reference: -
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-sipA (LMBP 6772) 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.

New search