Last data update: 22 October 2020 04:25 CEST
Plasmid name: pCAGGS-GFP (LMBP 4800)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Aequorea victoria green fluorescent protein DNA (GFP); mutated sequence
|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
|Parental clone:||pCAGGS; pGFP-C1mut|
|Further information:||The plasmid was constructed by inserting the blunted BspHI-BsmI fragment of pGFP-C1mut, containing the mutated A. victoria GFP coding sequence, into the blunted XhoI site of pCAGGS (this information was obtained from Dr K. Heyninck and differs from the description in Heyninck et al. (1999); it was confirmed by our restriction enzyme analysis).
The GFP insert contains a Ser/Thr mutation at codon position 65.
pCAGGS-GFP is useful for C-terminal fusion of a gene of interest to the mutated GFP gene and for highly efficient expression of the fusion protein under the control of the AG promoter and the human CMV-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 LT727037.1.
Since the nucleotide sequence of the 5' untranslated region of the GFP gene could not be traced back, there is uncertainty as to the cutting frequency of the restriction enzymes, except for the sites that were analysed in the authenticity test.
|EMBL Accession number:||LT727037.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||02/01/2004|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: BalI, BamHI, EcoRI, HindII, HindIII, NcoI, NheI and XhoI.|
|History of deposit:||This plasmid was deposited by Dr K. Heyninck(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:||Heyninck et al., FEBS Lett. 442 (1999), 147-150 [PMID: 9928991]
|Related plasmid reference:||Heyninck et al., J. Cell Biol. 145 (1999), 1471-1482 [PMID: 10385526]
Vinograd_Byk et al., J. Neurosci. 35 (2015), 936-942 [PMID: 25609612] [DOI: 10.1523/JNEUROSCI.1998-14.2015]
|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-GFP (LMBP 4800) is available at BCCM/GeneCorner. This plasmid was deposited by Dr K. Heyninck and Prof. Dr R. Beyaert and was published in Heyninck et al., 1999.|
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.