Last data update: 24 January 2024 16:39 CET
Plasmid name: pSCTPIMF1 (LMBP 2894)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p2894.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Saccharomyces cerevisiae α-mating factor 1 gene (MFα1); prepro secretion signal sequence (ppMF) |
| Promoter: | Saccharomyces cerevisiae triosephosphate isomerase promoter (TPI) Escherichia coli lac operon promoter |
| Ribosome binding site: |
Ribosome binding site (RBS) of the Escherichia coli lac Z gene (lacZ) |
| Terminator: | - |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli |
| Parental clone: | pSCMF3 |
| Further information: | The Saccharomyces cerevisiae triosephosphate isomerase (TPI) promoter was isolated as a BamHI-XhoI fragment (TPI-II), and with a XhoI-PstI linker cloned into the PstI and BglII sites of pSCMF3. As a result the TPI promoter was correctly inserted in front of the preMFα1 gene. The segment from residue 149 to 401 corresponds in anticlockwise orientation to the 3' end of the lacZα fragment. ATTENTION: the vector part of the plasmid is probably not correct: restriction sites were not experimentally verified. 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. Other name of the plasmid is pTPIMA1. |
| EMBL Accession number: | - |
| Latest sequence update: | 16/11/1989 |
| Sequence detail: | Nucleotide sequence at the end of the prepro sequence:
-- ppMF --> |
5' GGG.GTA.TCT.TTG.GAT.AAA.AGG|CCT.CTT.GAA.GCT.TGG 3'
Gly Val Ser Leu Asp Lys Arg|Pro Ala Glu Ala Trp
*|*** **
StuI HindIII
*: mutated nucleotide
|: Cutting site of the alpha mating factor Kex2 endoprotease. |
| Authenticity test: | The plasmid still needs to be subjected to the authenticity test. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by J. Zhu(1) and Prof. Dr R. Contreras(1). (1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | PhD thesis Zhu Jingdong |
| 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 |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pSCTPIMF1 (LMBP 2894) is available at BCCM/GeneCorner. This plasmid was deposited by J. Zhu and Prof. Dr R. Contreras and was published in Zhu Jingdong, . |
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.