pCMV3-N-FLAG (适合细胞内蛋白的表达)

pCMV3-N-FLAG 质粒图谱
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pCMV3-N-FLAG Physical Map

pCMV3-N-FLAG:

  • • CMV 启动子: 碱基 250-837
  • • 增强子: 碱基 838-1445
  • • SV40 早期启动子: 碱基 2339-2708
  • • 潮霉素 ORF: 碱基 2726-3751
  • • pUC 原点: 碱基 4394-5067
  • • 卡那霉素 ORF: 碱基 5141-5956

ORF 插入位点

Cloning Sites for ORF Inserting

pCMV3-N-FLAG 载体基本信息

载体名称 pCMV3-N-Flag
载体大小 6098bp
载体类型 哺乳动物表达载体
表达方式 组成型, 稳定 / 瞬时
启动子 CMV
抗生素抗性 卡那霉素
在哺乳动物细胞中的筛选 潮霉素
标签 FLAG
测序引物 Forward: T7(TAATACGACTCACTATAGGG); Reverse: BGH(TAGAAGGCACAGTCGAGG)

Flag Tag Information

FLAG-tag, or FLAG octapeptide, is a polypeptide protein tag that can be added to a protein using recombinant DNA technology. It can be used for affinity chromatography, then used to separate recombinant, overexpressed protein from wild-type protein expressed by the host organism. It can also be used in the isolation of protein complexes with multiple subunits.

A FLAG-tag can be used in many different assays that require recognition by an antibody. If there is no antibody against the studied protein, adding a FLAG-tag to this protein allows one to follow the protein with an antibody against the FLAG sequence. Examples are cellular localization studies by immunofluorescence or detection by SDS PAGE protein electrophoresis.

The peptide sequence of the FLAG-tag from the N-terminus to the C-terminus is: DYKDDDDK (1012 Da). It can be used in conjunction with other affinity tags, for example a polyhistidine tag (His-tag), HA-tag or Myc-tag. It can be fused to the C-terminus or the N-terminus of a protein. Some commercially available antibodies (e.g., M1/4E11) recognize the epitope only when it is present at the N-terminus. However, other available antibodies (e.g., M2) are position-insensitive.

哺乳动物表达载体

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