タグ
FLAG® tagged
形状
buffered aqueous solution
微生物選択
kanamycin
複製起点
pUC (500 copies)
ペプチド切断
TEV
no cleavage
ペプチドタグ位置
C-terminal
N-terminal
プロモーター
Promoter name: OXB20
Promoter activity: constitutive
Promoter type: bacterial
輸送温度
ambient
保管温度
−20°C
詳細
Molecular cloning often benefits from optimizing the vector used for expression.
This pack enables you to compare placing FLAG epitope tags at either the N or C terminus of your gene of interest (inserted into the MCS, under transcriptional control of the OXB20 strong bacterial promoter) with, and also without a TEV (Tobacco Etch Virus) protease cleavage site. The TEV site enables removal of the FLAG tag from the protein following production. Comparing these four configurations should enable you to evaluate how best to express and detect your gene of interest from bacterial 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.
This pack enables you to compare placing FLAG epitope tags at either the N or C terminus of your gene of interest (inserted into the MCS, under transcriptional control of the OXB20 strong bacterial promoter) with, and also without a TEV (Tobacco Etch Virus) protease cleavage site. The TEV site enables removal of the FLAG tag from the protein following production. Comparing these four configurations should enable you to evaluate how best to express and detect your gene of interest from bacterial 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.
シーケンス
For Genebank sequence, FASTA sequence, Quick-reference Plasmid Map, and Full Plasmid Map, see the individual vector product page for links.
アナリシスノート
この製品の分析証明書を確認するには、www.oxfordgenetics.comをご覧ください。
その他情報
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.
法的情報
These plasmids are sold free of reach-through rights and can be used to make commercial products. However the plasmids themselves (or derivatives) cannot be sold.
FLAG is a registered trademark of Merck KGaA, Darmstadt, Germany
Oxford Genetics is a trademark of Oxford Genetics Ltd
キットの構成要素のみ
製品番号
詳細
- PSF-OXB20-NH2-FLAG®-TEV - N-TERMINAL FLAG® TAG BACTERIAL PLASMID, plasmid vector for molecular cloning
- PSF-OXB20-COOH-TEV-FLAG® - C-TERMINAL FLAG® TAG BACTERIAL PLASMID, plasmid vector for molecular cloning
試験成績書（COA）
製品のロット番号・バッチ番号を入力して、試験成績書（COA） を検索できます。ロット番号・バッチ番号は、製品ラベルに「Lot」または「Batch」に続いて記載されています。
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