OGS38
PSF-SP6 - SP6 PROMOTER PLASMID
plasmid vector for molecular cloning
Synonyme(s) :
cloning vector, expression vector, molecular cloning vector, plasmid, plasmid vector, snapfast vector, vector
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About This Item
Code UNSPSC :
12352200
Nomenclature NACRES :
NA.85
Produits recommandés
Forme
buffered aqueous solution
Poids mol.
size 3862 bp
Sélection de bactéries
kanamycin
Origine de la réplication
pUC (500 copies)
Clivage des peptides
no cleavage
Gène reporter
none
Conditions d'expédition
ambient
Température de stockage
−20°C
Description générale
This vector allows transcription of un-polyadenylated and un-capped RNA using the SP6 bacteriophage polymerase. Some in vitro transcription kits will enable the production poly-adenylated and capped RNA using this vector; however this will depend on the kit used. SnapFast vectors do not contain an SP6 terminator and so to prevent the polymerase from circling the plasmid it must be cleaved at a restriction site that is 3 prime to the end of your gene to allow run-off of the polymerase. If you are expressing your gene in a vaccinia virus-free mammalian SP6 system it will require an IRES upstream of your gene because of the absence of a 5 prime cap on the RNA produced.
Promoter Expression Level:
Promoter Expression Level:
Application
Cloning in a gene: This plasmid has been designed to be compatible with a range of cloning techniques. The multiple cloning site contains a range of standard commonly used restriction sites for cloning. Using these sites genes can be inserted using standard cloning methods with DNA ligase. Other methods such as ligase independent cloning (LIC) Gibson Assembly InFusionHD or Seamless GeneArt can also be used and because all of our plasmids are based on the same backbone the same method can be used for cloning into all of our catalogue vectors.
Multiple cloning site notes: There are a few important sites within the MCS. These include the NcoI site the XbaI site and the BsgI and BseRI sites. The NcoI site contains a start codon that is immediately downstream of both a Kozak and Shine-Dalgarno ribosomal binding site. These allow for optimal positioning of genes when the start codon is placed in this location. If this is not required and you wish to use a downstream site for gene cloning you can remove the NcoI site by cleaving the plasmid with KpnI.
The XbaI site contains a stop codon. This stop codon is positioned in a specific position in relation to the BsgI and BseRI sites that are immediately downstream. When either BseRI or BsgI cleave the plasmid they produce a TA overhang from the stop codon in the XbaI site that is compatible with all of our peptide tag plasmids cut with the same sites. BseRI and BsgI sites are non-palindromic and cleave a defined number of bases away from their binding site.
Whenever we clone a gene into our multiple cloning site we always position the start and stop codon in the same positions in the MCS. If the start and ends of the genes are not compatible with NcoI and XbaI we extend the sequence to the nearest external sites but keep the start and stop codons locations consistent.
Multiple cloning site notes: There are a few important sites within the MCS. These include the NcoI site the XbaI site and the BsgI and BseRI sites. The NcoI site contains a start codon that is immediately downstream of both a Kozak and Shine-Dalgarno ribosomal binding site. These allow for optimal positioning of genes when the start codon is placed in this location. If this is not required and you wish to use a downstream site for gene cloning you can remove the NcoI site by cleaving the plasmid with KpnI.
The XbaI site contains a stop codon. This stop codon is positioned in a specific position in relation to the BsgI and BseRI sites that are immediately downstream. When either BseRI or BsgI cleave the plasmid they produce a TA overhang from the stop codon in the XbaI site that is compatible with all of our peptide tag plasmids cut with the same sites. BseRI and BsgI sites are non-palindromic and cleave a defined number of bases away from their binding site.
Whenever we clone a gene into our multiple cloning site we always position the start and stop codon in the same positions in the MCS. If the start and ends of the genes are not compatible with NcoI and XbaI we extend the sequence to the nearest external sites but keep the start and stop codons locations consistent.
Séquence
To view sequence information for this product, please visit the product page
Remarque sur l'analyse
To view the Certificate of Analysis for this product, please visit www.oxgene.com
Produit(s) apparenté(s)
Réf. du produit
Description
Tarif
Code de la classe de stockage
12 - Non Combustible Liquids
Point d'éclair (°F)
Not applicable
Point d'éclair (°C)
Not applicable
Certificats d'analyse (COA)
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