Last data update: 20 October 2020 04:14 CEST
Plasmid name: pCAGGS-NLS-Cre-PGK-Puro (LMBP 7779)
|Price category:||Cat. 2 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||SV40 large T-antigen nuclear localization signal (NLS); N-terminal
Phage P1 cre gene encoding recombinase; with Kozak consensus sequence
|Promoter:||Chicken β-actin/rabbit β-globin hybrid promoter (AG)
Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only
Mouse phosphoglycerate kinase 1 promoter (PGK1)
Escherichia coli lac operon promoter
|Terminator:||Rabbit β-globin polyadenylation signal (β-globin polyA)
Simian virus 40 polyadenylation signal (SV40 polyA)
Mouse phosphoglycerate kinase 1 polyadenylation signal (PGK1 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 is meant to be used in puromycin-sensitive cells for the excision of loxP-site flanked (floxed) DNA sequences.
The nucleotide sequence of the cre gene corresponds with the EMBL Nucleotide Sequence Database accession number X03453.1.
The region between the AG promoter and the rabbit β-globin polyA was sequenced by LMBP.
Since the nucleotide sequence between the 3' flanking region of rabbit β-globin and the lac promoter could not be completely 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.
Name mentioned in Nyabi et al. (2009) is pCAGG-NLS-Cre, PGK-puro.
Other names of the plasmid are pCAGGS-NLS-Cre-PGKpuro/cg and pCAGGS-nls-Cre; PGK-puro.
|EMBL Accession number:||X03453.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||21/08/2012|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: AgeI, EcoRI/PvuI, EcoRV/SacII, HincII, NotI/XbaI.
As compared to the compiled nucleotide sequence, two additional HincII sites are present, likely around positions 1700 and 3300. Additionally, the EcoRI site at position 7631 could not be experimentally confirmed, while this site was confirmed via sequence analysis at GeneCorner.
|History of deposit:||This plasmid was deposited by Dr A. Nagy(1).
(1) Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Canada
|Related plasmid reference:||Nyabi et al., Nucleic Acids Res. 37 (2009), e55 [PMID: 19279185] [DOI: 10.1093/nar/gkp112]
|Restricted distribution:||- BCCM/MTA adapted by Mount Sinai Hospital|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 DH5α|
|Host reference:||Focus 8 (1986), 9
|Related host reference:||Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pCAGGS-NLS-Cre-PGK-Puro (LMBP 7779) is available at BCCM/GeneCorner. This plasmid was deposited by Dr A. Nagy.|
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.