Last data update: 24 January 2024 16:39 CET
Plasmid name: pCAGGST7 (LMBP 5216)
New search | Print data sheet |
Price category: | Cat. 1 (cf. price list) |
Status: | GeneCorner core plasmid |
GeneCorner sequence: |
p5216.gb
(View with Genome Compiler) p5216.txt p5216.pdf |
Sequence analysis results Genecorner: |
- |
Cloned DNA: |
- |
Promoter: | Chicken β-actin/rabbit β-globin hybrid promoter (AG) Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only Phage T7 gene 10 promoter (T7g10) 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 |
Further information: | This plasmid was constructed as follows: 1) a PCR was performed using pCAGGS as template; 2) the PCR fragment was cloned as a SnaBI(Eco105I)/XhoI fragment in the SnaBI/XhoI opened pCAGGS. As a result, the T7 promoter was inserted downstream of the AG promoter. 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 LT726974.1. |
EMBL Accession number: | LT726974.1, view at EMBL, GenBank, DDBJ |
Latest sequence update: | 17/01/2007 |
Sequence detail: | Primers used to introduce the T7 promoter: Forward : 5' GTCATCTACGTATTAGTCATCGCTATTACC 3' SnaBI Reverse : 5' CCGCTCGAGCCCTATAGTGAGTCGTATTATTCTTTGCCAAAATGATGAGAAG 3' XhoI >>>T7 promoter>>> |
Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BsrDI, Eco105I/XhoI and XhoI. |
Class: | Recombinant plasmid |
Type: | Plasmid |
History of deposit: | This plasmid was deposited by Dr B. Schepens(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: | - |
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: |
pCAGGST7 (LMBP 5216) is available at BCCM/GeneCorner. This plasmid was deposited by Dr B. Schepens 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.