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一键询价-
HRMVPCS 永生化人视网膜周细胞 QCH791
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钦诚生物 | 一周
4000￥
获取电话pESC-URA酿酒酵母质粒
基本信息
启动子: GAL1,GAL10 promoter
复制子: 2μ ori,ori,FI ori
终止子: CYC1
质粒大小: 6631bp
质粒标签: C-Flag, C-Myc
原核抗性: 氨苄青霉素Amp
筛选标记: URA3
克隆菌株: DH5α
培养条件: 37℃，有氧，LB
表达宿主: 酵母细胞
质粒简介
The pESC vectors are a series of epitope-tagging vectors for expression of eukaryotic genes in the yeast S. cerevisiae. Each vector contains GAL1 and GAL10 yeast promoters in opposing orientations. With these vectors you can introduce one or two cloned genes into a yeast host strain under the control of an inducible promoter. These vectors also feature an extensive polylinker sequence and the ability to generate end-specific RNA transcripts from T3 and T7 promoters. Each of the pESC vectors contains one of four different yeast-selectable markers (HIS3, TRP1, LEU2, or URA3) in the same vector backbone.
The pESC vectors contain DNA sequences coding for epitope peptides that can be specifically recognized by monoclonal antibodies. A sequence for the FLAG® epitope (DYKDDDDK) is located in the multiple cloning site (MCS) downstream of the GAL10 promoter; a sequence for the c-Myc epitope (EQKLISEEDL) is located in the MCS downstream of the GAL1 promoter. You can insert your gene of interest in front of the epitope sequence to generate C-terminal tagging or after the epitope sequence for N-terminal tagging. These tags allow the protein of interest to be studied without generating a specific antibody to that protein. The epitope-tagged fusion proteins can be studied in transformed cells using well-characterized antibodies.
• Express two different genes simultaneously in S. cerevisiae
• Proteins tagged with unique epitope tags
• Fast and easy immunoprecipitations
质粒图谱
pESC-URA酿酒酵母质粒使用说明：
1、收到质粒干粉后请先5000rpm离心1min，再加入20μl无菌水溶解质粒，室温放置1min；
2、从-80℃冰箱中取出相应的感受态，置于冰盒上解冻，并做好标记；
3、取2μl质粒加至100μl感受态中，冰浴30min；
4、42℃热激90s，再冰浴2min；
5、加入900μl无抗的LB液体培养基，180rpm震荡培养45min；
6、6000rpm离心5min，仅留100ul上清混匀菌体沉淀；
7、混匀后的菌液加至对应抗性的LB平板上，倒入适量玻璃珠，涂匀液体；
8、将平板正向培养1h，再倒置培养12h~16h；
9、挑取单克隆菌落至对应抗性的LB液体培养基中，震荡培养12h~16h，根据实验需要提取质粒。
pESC-URA酿酒酵母质粒注意事项：
1、如果您收到的是甘油菌种，请先四区划线，挑取单克隆培养。
2、如果第二天转化平板长的过多，请将质粒按比例稀释后再转化。
