Last data update: 24 January 2024 16:39 CET
Plasmid name: pEF1mtBIDHis (LMBP 4323)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p4323.gb
(View with Genome Compiler) p4323.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Mouse BH3 interacting domain death agonist (BID); truncated sequence (tBID) V5 epitope; C-terminal Histidine tag (His-tag); C-terminal |
| Promoter: | Human elongation factor 1α promoter (EF1α) Phage T7 gene 10 promoter (T7g10) Simian virus 40 early promoter (SV40 early) |
| Ribosome binding site: |
- |
| Terminator: | Bovine growth hormone polyadenylation signal (BGH polyA) Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Ampicillin (amp) Neomycin (neo; G418) |
| 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: | pEF1/V5-HisC; pGAD424mBID |
| Further information: | The plasmid was constructed as follows: the truncated mouse BH3 interacting domain death agonist (mtBID), containing the coding sequence for amino acid 60 up to and including 195, was isolated from pGAD424mBID by the use of primers including an initiation codon and inserted as an EcoRI/AspI(Tth111I, filled in) fragment between the EcoRI and NotI (filled in) sites of pEF1/V5-HisC. As a result, mtBID was fused in phase to the C-terminal V5 epitope and histidine tag (His-tag). There is uncertainty about the presence of a second enhancer in the SV40 early promoter. The absence of a second enhancer in this promoter may cause bacterial leakage expression of the Tn5 neomycin resistance gene. This would cause transformed E. coli cells to be resistant to kanamycin, although growth should be reduced compared to growth on medium containing ampicillin. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727336.1. Other name of the plasmid is pEF1/V5-HisC mBIDtruncated with Histag. |
| EMBL Accession number: | LT727336.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 14/06/2004 |
| Sequence detail: | Forward primer: 5' G.GAA.TTC.ATG.GGC.AGC.CAG.GCC.AGC.CGC.TCC.TTC. 3'
EcoRI ***
Reverse primer: 5' T.GCG.GTC.GAC.TCA.GTC.CAT.CTC.GTT.TCT.AAC.CAA.GTT.C 3'
Tth111I
Nucleotide sequence at the start of the mBID cDNA:
--------------- mBID -------------->
60
5' ... TCGGATCCACTAGTCCAGTGTGGTGGAATTC.ATG.GGC.AGC.CAG.GCC.AGC.CGC.TCC.TTC.AAC ... 3'
BamHI SpeI BstXI EcoRI *** Gly Ser Gln Ala Ser Arg Ser Phe Asn
Nucleotide sequence at the end of the mBID cDNA:
------- mBID ------>
195 ---------------
5' ... AGA.AAC.GAG.ATG.GAC.TGG.GCC.GCT.CGA.GGT.CAC.CCA.TTC.GAA.GGT.AAG.CCT.ATC.
Arg Asn Glu Met Asp Trp Ala Ala Arg Gly His Pro Phe Glu Gly Lys Pro Ile
XhoI BstEII AsuII
V5 epitope ---------------------------> ----------- His-tag
CCT.AAC.CCT.CTC.CTC.GGT.CTC.GAT.TCT.ACG.CGT.ACC.GGT.CAT.CAT.CAC.CAT.CAC.
Pro Asn Pro Leu Leu Gly Leu Asp Ser Thr Arg Thr Gly His His His His His
AgeI
-->
CAT.TGA.GTTTAAACCCGCTGATCAGCCTCGA ... 3'
His *** PmeI
***: Start codon.
+++: Termination codon.
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BssHII, BssHII/EcoRI, EcoRI, XhoI and XmnI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr P. Vandenabeele(1)(2) and onstructed by M. Van Gurp. (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: |
pEF1mtBIDHis (LMBP 4323) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr P. Vandenabeele and onstructed by M. Van Gurp. Department for Molecular Biomedical Research, VIB, Ghent, Belgium Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium. |
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.