■ 基本信息
別名: | PB513B-1；pB-CMV-MCS-ef1α-greenpuro |
啟動(dòng)子: | CMV |
復(fù)制子: | pUC |
終止子: | SV40 poly(A) signal |
質(zhì)粒分類: | 哺乳系列質(zhì)粒；哺乳編輯質(zhì)粒；哺乳轉(zhuǎn)座質(zhì)粒 |
質(zhì)粒大小: | 7258bp |
原核抗性: | Amp |
真核抗性: | Puro |
克隆菌株: | Stbl3 |
培養(yǎng)條件: | 37℃，5%CO2 |
表達(dá)宿主: | 293T等哺乳細(xì)胞 |
5'測序引物: | CMV-F（CGCAAATGGGCGGTAGGCGTG） |
3'測序引物: | Sv40-polyA-R（GAAATTTGTGATGCTATTGC） |
備注: | 哺乳細(xì)胞轉(zhuǎn)座表達(dá)載體 |
■ 質(zhì)粒屬性
質(zhì)粒宿主: | 哺乳細(xì)胞 |
質(zhì)粒用途: | 蛋白表達(dá),轉(zhuǎn)座 |
片段類型: | ORF
|
片段物種: | 空載體
|
原核抗性: | Amp |
真核抗性: | Puro
|
熒光標(biāo)記: | 綠色
|
■ 質(zhì)粒簡介
PiggyBac Dual promoter (PB513B-1)是用于哺乳細(xì)胞轉(zhuǎn)座實(shí)驗(yàn)的質(zhì)粒，位于CMV啟動(dòng)子下游的多克隆位點(diǎn)（MCS），使目的基因或microRNA更方便克隆。下游的EF-1α core promoter啟動(dòng)GFP的表達(dá)。
The PiggyBac (PB) transposon is a mobile genetic element that efficiently transposes between vectors and chromosomes via a "cut and paste" mechanism. During transposition, the PB transposase recognizes transposon-specific inverted terminal repeat sequences (ITRs) located on both ends of the transposon vector and efficiently moves the contents from the original sites and efficiently integrates them into TTAA chromosomal sites. The powerful activity of the piggyBac transposon system enables genes of interest between the two ITRs in the PB vector to be easily mobilized into target genomes.
The unique features of piggyBac transposons are that there is NO Cargo Limit and it is also Reversible. Genomes containing an inserted piggyBac vector can be transiently re-transfected with the PB transposase expression vector. The PB transposase will remove the transposons from the genome, footprint-free.The Super PiggyBac transposase transient expression vector and PB513B-1 were co-transfected into HeLa cells and puromycin selection applied for 10 days (10ug/ml). Cells efficiently transposed were Puro resistant and GFP positive.
■ 質(zhì)粒圖譜
■ 質(zhì)粒序列
質(zhì)粒序列請下載：
ZK564PiggyBac Dual promoter (PB513B-1)哺乳轉(zhuǎn)座質(zhì)粒.txt
