Merck Mammalian 10His Tag Vector Set

Mammalian 10His Tag Vector Set

plasmid vectors for molecular cloning

expression vector, molecular cloning vector, vector, snapfast vector, plasmid vector, cloning vector, plasmid

태그

10-His tagged

form

buffered aqueous solution

박테리아 선정

kanamycin

포유류 세포 선정

puromycin

복제개시점

pUC (500 copies)

펩타이드 절단

TEV
no cleavage

펩타이드 태그 위치

C-terminal
N-terminal

프로모터

Promoter name: CMV
Promoter activity: constitutive
Promoter type: mammalian

배송 상태

ambient

저장 온도

−20°C

Molecular cloning often benefits from optimizing the vector used for expression.

This pack enables you to compare placing decahistidine (10His) affinity tags at either the N or C terminus of your gene of interest (inserted into the MCS, under transcriptional control of the CMV promoter) with, and also without a TEV (Tobacco Etch Virus) protease cleavage site. The TEV site enables removal of the 10His tag from the protein following purification. Comparing these four configurations should enable you to evaluate how best to express and purify your gene of interest from mammalian cells. We also provide many other functional tags and cleavage sites, if required.This plasmid set 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.

Transcription Termination: These plasmids contains three alternative transcription terminators for mammalian bacterial and bacteriophage (T7) expression. This means that only the promoter needs to be changed to alter the expression system you are using. We sell multiple promoters that can be used in each of these systems. The presence of each terminator does not reduce expression in the alternative systems.