Last data update: 24 January 2024 16:39 CET
Plasmid name: pSCGALyEGFPmBAX (LMBP 4203)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p4203.gb
(View with Genome Compiler) p4203.txt p4203.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Aequorea victoria green fluorescent protein DNA (GFP); yeast enhanced sequence (yEGFP) Mouse Bcl2-associated X cDNA (Bax, BAX); α variant |
| Promoter: | Saccharomyces cerevisiae galactokinase promoter (GAL1) |
| Ribosome binding site: |
- |
| Terminator: | Saccharomyces cerevisiae 2 micron plasmid (2μ) FLP terminator Saccharomyces cerevisiae iso-1-cytochrome C terminator (CYC1) |
| Selection marker: | Ampicillin (amp) LEU2; auxotrophic |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli Saccharomyces cerevisiae; leu2(-), integrative |
| Parental clone: | pSCGAL1-L; pUC18-GFP; pCR-BluntII-TOPO-Bax |
| Further information: | The plasmid was constructed as follows: the BglII-XbaI fragment from pUC18-GFP, containing the yEGFP coding sequence, and the BglII-HindIII fragment from pCR-BluntII-TOPO-Bax, containing the mBAX α variant coding sequence, were inserted into the XbaI-HindIII opened pSCGAL1-L plasmid. pSCGALyEGFPmBAX is an integrative reporter plasmid for the yEGFP/mBax-α fusion protein in S. cerevisiae. After BstEII linearisation in its LEU2 element, single-copy genomical integration leads to stable expression. The nucleotide sequence of the mBAX cDNA corresponds with the EMBL Nucleotide Sequence Database accession number L22472.1. The nucleotide sequence of the yEGFP DNA corresponds with the EMBL Nucleotide Sequence Database accession number U73901.1. Other name of the plasmid is YIpUTL-GFP/Bax. |
| EMBL Accession number: | L22472.1, view at EMBL, GenBank, DDBJ U73901.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 12/03/2003 |
| Sequence detail: | Primers used for amplification of the yEGFP fragment from pCAPSelect1 as template:
(this yEGFP fragment was inserted in the PstI-XbaI opened pUC18)
forward primer: 5' GCTCTAGA.ATG.TCT.AAA.GGT.GAA.GAA.TTA.TTC 3'
XbaI ***
reverse primer: 5' AACTGCAGGGA.AGA.TCT.TCC.TTT.GTA.CAA.TTC.ATC.CAT.ACC 3'
PstI BglII
Primers used for amplification of the mBAX fragment from pSCGAL1mBAX-L as template:
(this mBAX fragment was inserted in pCR-BluntII-TOPO (Invitrogen))
forward primer: 5' GGA.AGA.TCT.TCC.ATG.GAC.GGG.TCC.GGG.GAG.C 3'
BglII ***
reverse primer: 5' TTGGATCATAAGCTTGCATGCCTGCAGGTC.TCA.GCC.C 3'
HindIII +++
***: start codon.
+++: termination codon.
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BamHI, BglII/HindIII, BglII/XbaI, BstEII/XmnI, HindII and MluI. The XmnI site at position 4966 could not be experimentally confirmed. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr R. Contreras(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: |
pSCGALyEGFPmBAX (LMBP 4203) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Contreras . |
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.