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平臺編號：bio-107412
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質(zhì)粒信息： pTWIN1
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規(guī)格：2ml甘油管 20ul質(zhì)粒
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用途：/
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服務(wù)費(fèi)用：
加載中…… - 訂購
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注意事項：僅用于科學(xué)研究或者工業(yè)應(yīng)用等非醫(yī)療目的不可用于人類或動物的臨床診斷或治療，非藥用，非食用（產(chǎn)品信息以出庫為準(zhǔn)）
pTWIN1
| 別稱 | pTWINI |
| 啟動子 | T7 |
| 復(fù)制子 | M13 |
| 質(zhì)粒分類 | 大腸桿菌載體；蛋白表達(dá)質(zhì)粒 |
| 質(zhì)粒大小 | 7375bp |
| 原核抗性 | Amp |
| 克隆菌株 | DH5a |
| 培養(yǎng)條件 | 37℃，有氧，LB |
| 表達(dá)宿主 | 大腸桿菌 |
| 誘導(dǎo)方式 | IPTG或乳糖及其類似物 |
| 備注 | 表達(dá)質(zhì)粒 |
| 載體宿主 | 大腸桿菌 |
| 載體用途 | 蛋白表達(dá) |
| 基因種屬 | 空載體 |
| 基因類型 | ORF |
| 原核抗性 | Amp |
| 質(zhì)粒簡介 | pTWIN1 is an E. coli plasmid cloning vector designed for recombinant protein expression, labeling, and cyclization using the IMPACT-TWIN Kit (1). It contains the pMB1 origin of replication from pBR322 and is maintained at a similar copy number to pBR322; in addition, pTWIN1 also contains an M13 origin of replication. The multiple cloning site (MCS) is positioned to allow translational fusion of an intein tag to the N-terminus, C-terminus, or both, of the cloned target protein. The mini-inteins encoded by pTWIN1, the Ssp DnaB intein and the Mxe GyrA intein, cleave the peptide bond at their C- and N-termini, respectively (1-4). The chitin binding domain (CBD) from B. circulans, fused to each intein, facilitates purification of the intein-target protein precursor. Transcription of the intein tags is controlled by the inducible T7 promoter, requiring E. coli strains containing integrated copies of the T7 RNA polymerase gene [e.g. BL21(DE3)] for expression. Basal expression from the T7 promoter is minimized by the binding of the Lac repressor, encoded by the lacI gene, to the lac operator immediately downstream of the T7 promoter (5). Translation of the intein tags utilizes the translation initiation signal (Shine Dalgarno sequence) from the strongly expressed T7 gene 10 protein (φ10). |
