Last data update: 24 January 2024 16:39 CET
Plasmid name: pEGFPC2-haTctn(179-2860) (LMBP 4550)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p4550.gb
(View with Genome Compiler) p4550.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Aequorea victoria green fluorescent protein DNA (GFP); enhanced red-shifted variant (EGFP) Human αT-catenin cDNA (aTctn, αTctn, CTNNA3) |
| Promoter: | Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer Escherichia coli β-lactamase promoter (amp) Simian virus 40 early promoter (SV40 early) |
| Ribosome binding site: |
Ribosome binding site (RBS) of the Escherichia coli ampicillin resistance gene (amp) |
| Terminator: | Herpes simplex virus (HSV) thymidine kinase polyadenylation signal (TK polyA) Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Neomycin (neo; G418; kanamycin (kan)) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin Simian virus 40 bidirectional origin (SV40) |
| Host range: | Escherichia coli Mammalian cells; SV40 permissive cells |
| Parental clone: | pEGFP-C2 |
| Further information: | The plasmid was constructed as follows: cDNA of the human αT-catenin (haTctn) was isolated from a human testis Marathon cDNA preparation by the use of primers. The coding sequence (from nucleotide position 179 to 2860 of the cDNA) was inserted between the EcoRI and SalI sites of pEGFP-C2. As a result human aTctn is N-terminally fused to the enhanced red-shifted variant of the Aequorea victoria green fluorescent protein gene (EGFP). The fusion gene is expressed under control of the human cytomegalovirus immediate early promoter (hCMV-IE) and enhancer. EGFP is a red-shifted variant of wild-type GFP that has been optimized for brighter fluorescence and higher expression in mammalian cells (excitation maximum = 488 nm; emission maximum = 507 nm). EGFP encodes the GFPmut1 variant, which contains the amino acid substitutions Phe-64 to Leu and Ser-65 to Thr. The neomycin resistance cassette, consisting of the SV40 early promoter, the neomycin/kanamycin resistance gene of Tn5, and polyadenylation signals of the HSV-TK gene, allows stably transfected eukaryotic cells to be selected using G418. The E. coli β-lactamase promoter upstream of this cassette expresses kanamycin resistance in E. coli. The PstI site and the BssHII site in the neomycin phosphotransferase coding sequence have been removed. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727261.1. The nucleotide sequence of the human aTctn cDNA corresponds with the EMBL Nucleotide Sequence Database accession number AF091606.1. Other name of the plasmid is pEGFP2-hαTctn(179-2860). |
| EMBL Accession number: | AF091606.1, view at EMBL, GenBank, DDBJ LT727261.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 03/07/2002 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BglII/SalI, EcoRI and NcoI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr B. Janssens(1) (2) and Prof. Dr F. Van Roy(1) (2). (1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | Janssens et al., J. Cell Sci. 114 (2001), 3177-3188 [PMID: 11590244] |
| Related plasmid reference: | Lippincott-Schwartz et al., Science 300 (2003), 87-91 [PMID: 12677058] |
| Restricted distribution: | - BCCM MTA |
| 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: | Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660] Rodriguez-Quinones et al., Focus 15 (1993), 110-112 Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791] [DOI: 10.1016/s0022-2836(83)80284-8] Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187] Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051] |
| Cultivation medium: | LB-Lennox + kanamycin (30 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pEGFPC2-haTctn(179-2860) (LMBP 4550) is available at BCCM/GeneCorner. This plasmid was deposited by Dr B. Janssens and Prof. Dr F. Van Roy and was published in Janssens et al., 2001. |
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.