PSF-OXB20-DAGFP - BACTERIAL DAGFP PLASMID OGS245-5UG
The expression of the green fluorescent reporter gene daGFP in bacterial cells. The daGFP gene is within the primary multiple cloning site.
Promoter Expression Level: This plasmid contains a constitutive bacterial promoter that does not require induction. It is the strongest bacterial promoter we sell and this can cause solubility and expression problems with some proteins. We also offer a range of other bacterial promoters that are compatible with this plasmid and are available on request.
Cloning in a gene: This plasmid contains a gene within the main multiple cloning site (NotI-ClaI). Any plasmid that we sell where the gene is this configuration will be located in the exact same position in relation to the start and stop codon of the gene. The only exceptions to this rule are fusions proteins where the fusion gene may be positioned at the front or end of the MCS to allow gene fusion.
By positioning all of our genes in the same location it allows them to be transferred between plasmids using the same cloning method and restriction sites regardless of the plasmid being used from our product range. Inserting a new gene into this plasmid should be easily possible using a range of standard restriction enzyme sites that flank the gene currently in the vector.
Multiple cloning site notes: In the multiple cloning site there are two important restriction sites called BsgI and BseRI sites. These sites both cut the DNA at the same position and cleave the stop codon of the gene in the multiple cloning site in this plasmid thereby producing a TA overhang. This overhang is compatible with any of our peptide or reporter fusion tag plasmids also cut with either of these enzymes. This allows seamless C-terminal fusions to be made with the gene in this multiple cloning site using a single cloning step from our C-terminal peptide and reporter tag product range. Normally the easiest method is to clone the C-terminal tag from our other plasmid products into this plasmid using BsgI or BseRI and the downstream ClaI restriction site.
BseRI and BsgI sites are non-palindromic and cleave a defined number of bases away from their binding sites. This allows them to cut the upstream stop codon in the gene in this plasmid regardless of the gene sequence.
Sequence
Please select the file type you require. For reference most cloning programs will import a .gb (Genbank) file and will show all of the plasmids features automatically when downloaded and imported.
Genebank Vector Sequence File,
FASTA Vector Sequence File,
Full Plasmid Map,
Quick-reference Plasmid Map,
Analysis Note
Looking for more vector options to move your experiments forward faster? Consider a custom cloning vector designed and built by Oxford Genetics™. Find out more at Oxford Genetics, - Sigma's partner for cloning and expression vectors for molecular biology and synthetic biology applications.
Oxford Genetics is a trademark of Oxford Genetics Ltd
form | buffered aqueous solution |
mol wt | size 4533 bp |
Origin of replication | pUC (500 copies) |
Peptide cleavage | no cleavage |
Promoter | Promoter name: OXB20 Promoter activity: constitutive Promoter type: bacterial |
bacteria selection | kanamycin |
reporter gene | GFP |
shipped in | ambient |
storage temp. | 20°C |
RIDADR | NONH for all modes of transport |