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GeneCorner plasmid details

Last data update: 05 December 2020 04:15 CET

Plasmid name: pCAGGS-E-sptP (LMBP 4878)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p4878.gb (View with Genome Compiler)
p4878.txt
p4878.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Salmonella enterica serovar Typhimurium LT2 sptP 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: pCAGGS-E-tag
Further information: The plasmid was constructed by inserting a NotI-XmaI fragment, obtained by PCR using genomic DNA of the S. typhimurium LT2 strain (gift from M. McClelland, Sidney Kimmel Cancer Center, San Diego, USA) as template and containing the S. typhimurium LT2 sptP coding sequence, into the NotI-XmaI opened pCAGGS-E-tag vector. As a result, the sptP coding sequence is fused in phase to the N-terminal E-tag.
pCAGGS-E-sptP is useful for highly efficient expression of sptP 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.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727054.1.
The nucleotide sequence of the sptP cDNA corresponds with the Genbank accession number NC_003197.1.
Other name of the plasmid is pCAGGSE-Sptp.
EMBL Accession number: NC_003197.1, view at EMBL, GenBank, DDBJ
LT727054.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 07/09/2007
Sequence detail:
Primers used to amplify the sptP coding sequence:

forward: 5' CGCAAGGAAGCGGCCGCT.ATG.CTA.AAG.TAT.GAG.GAG.AG 3'
                     NotI      ***
                               
reverse: 5' GTCGCCCCGGG.TCA.GCT.TGC.CGT.CGT.CAT.AAG.C 3'
                 XmaI   +++

***: start codon.
+++: termination codon.
Punctuation indicates reading frame.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: AccIII, Alw44I/BamHI, ApaI, ApaI/EcoRV, BstXI/NotI, EcoRI/HincII, NcoI, NcoI/XmaI and XmaI.

As compared to the compiled nucleotide sequence, the restriction site analysis pattern revealed that ApaI only cuts once instead of twice. The ApaI site at position 5299 could not be experimentally confirmed.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr R. Beyaert(1) (2). It was constructed by Dr P. Schotte(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) 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-sptP (LMBP 4878) is available at BCCM/GeneCorner. This plasmid was deposited by 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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