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GeneCorner plasmid details

Last data update: 07 December 2022 12:22 CET

Plasmid name: pCAGGS (LMBP 2453)

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Price category: Cat. 3 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p2453.gb (View with Genome Compiler)
p2453.txt
p2453.pdf
Sequence
analysis results
Genecorner:

NGS: a04-gc-dec2018-partial.fasta

NGS: 2453-gc-jun2020.fasta

NGS: 2453-ps-gc-mar2022.fasta

Cloned DNA: -
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)
Copy number: High copy number
Host range: Escherichia coli
Mammalian cells; SV40 permissive cells
Parental clone: pUC13; pAGS-3
Further information: This plasmid is useful for highly efficient expression of genes under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells.
The AG promoter sequence consists of the chicken β-actin promoter, the first exon and part of the first intron (that seems to have a strong enhancer-like activity) linked to a rabbit β-globin fragment, consisting of a 3' part of the second intron (inclusive a branch point which is required for normal splicing reactions) and a 5' part of the third exon.
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 LT727518.1 except for the adjustments due to the implementation of the next-generation sequencing results.
EMBL Accession number: LT727518.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 05/11/2019
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BglII, EcoRI, NcoI, SacII and XhoI.

This plasmid has also been fully sequenced. For some nucleotides, different NGS results contradict each other, in which case the sequence info from the depositor was kept.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr J. Miyazaki(1).
(1) University of Tokyo, Japan
Plasmid reference: Niwa et al., Gene 108 (1991), 193-200 [PMID: 1660837]
Related plasmid reference: Lee et al., Mol. Cancer 10 (2011), 113 [PMID: 21917150] [DOI: 10.1186/1476-4598-10-113]
Thorrez et al., Mol. Ther. 14 (2006), 442-451 [PMID: 16750937] [DOI: 10.1016/j.ymthe.2006.03.019]
Verhelst et al., EMBO J. 31 (2012), 3845-3855 [PMID: 23032186] [DOI: 10.1038/emboj.2012.240]
Staal et al., bioRxiv (2017), 1-25 [DOI: 10.1101/228767]
Pattabriaman et al., J. Cell Biol. 208 (2015), 53-69 [PMID: 25547156] [DOI: 10.1083/jcb.201404109]
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 (LMBP 2453) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr J. Miyazaki and was published in Niwa et al., 1991.

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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