BioVector® pENTR11Dual 入门克隆载体 / BioVector® pENTR11Dual Entry Vector

通用定义 / General Definition:

BioVector® pENTR11Dual 是一种高度灵活的 Gateway® 入门载体，旨在通过 $attL1$ 和 $attL2$ 重组位点高效构建供体克隆。其“双重（Dual）”特性通常体现在其多克隆位点（MCS）的设计上，允许使用传统的限制性内切酶克隆方法插入目的基因，同时其两侧的 Gateway 位点支持后续通过 LR 反应将基因快速转移至各类表达载体（Destination Vectors）。它常用于高通量蛋白表达、多基因共表达以及跨物种的基因功能研究。

BioVector® pENTR11Dual is a highly versatile Gateway® Entry Vector designed for the efficient construction of donor clones via $attL1$ and $attL2$ recombination sites. Its "Dual" characteristic typically refers to its multiple cloning site (MCS) design, which allows for the insertion of target genes using traditional restriction enzyme-based cloning, while the flanking Gateway sites facilitate the rapid transfer of the gene into various Destination Vectors through LR reactions. It is widely used in high-throughput protein expression, multi-gene co-expression, and cross-species functional genomics.

BioVector® pENTR11Dual 载体技术说明书 (Plasmid Datasheet)

中文版说明书

1. 产品基本信息

• 产品名称： BioVector® pENTR11Dual 入门载体

• 克隆方法： 限制性内切酶克隆 或 Gateway® 重组技术

• 载体类型： 入门载体 (Entry Vector)

• 抗性基因： 卡那霉素 (Kanamycin)

• 筛选标记：$ccdB$ 致死基因（用于阴性筛选）

2. 培养与扩增条件

• 宿主菌： BioVector® cccB 敏感型菌株（如 DH5$\alpha$）用于含有目的基因的克隆；BioVector® $ccdB$ 抗性菌株（如 $ccdB$ Survival™）用于空载体扩增。

• 培养基： BioVector® LB 培养基

• 抗生素添加： BioVector® 卡那霉素 (50 $\mu$g/mL)

• 培养环境： 37°C，恒温振荡培养

3. 载体应用

• 基因穿梭： 将目的基因从 BioVector® pENTR11Dual 转移至各种含有 $attR$ 位点的表达载体中。

• 双启动子系统： 支持在不同的宿主系统中通过重组实现高效表达。

• 高通量克隆： 适用于大规模功能基因组筛选。

4. 注意事项

• 负向筛选： 载体含有 $ccdB$ 基因，常规 $E. coli$ 转化前请确保目的基因已成功替换该致死基因。

• 序列验证： 建议使用 BioVector® M13 Forward 和 M13 Reverse 引物对插入片段进行测序验证。

English Datasheet

1. General Product Information

• Product Name: BioVector® pENTR11Dual Entry Vector

• Cloning Method: Restriction Enzyme Cloning or Gateway® Recombination Technology

• Vector Type: Entry Vector

• Antibiotic Resistance: Kanamycin

• Selection Marker:$ccdB$ lethal gene (for negative selection)

2. Culture and Amplification Conditions

• Host Strains: BioVector® cccB-sensitive strains (e.g., DH5$\alpha$) for recombinants; BioVector® $ccdB$ Survival™ strains for empty vector propagation.

• Culture Medium: BioVector® LB Medium

• Antibiotic Supplement: BioVector® Kanamycin (50 $\mu$g/mL)

• Incubation: 37°C, constant shaking

3. Applications

• Gene Shuttling: Transferring target genes from BioVector® pENTR11Dual into various Destination Vectors containing $attR$ sites.

• Dual Promoter Systems: Facilitating high-efficiency expression across different host systems via recombination.

• High-Throughput Cloning: Ideal for large-scale functional genomics screening.

4. Key Usage Notes

• Negative Selection: The vector contains the $ccdB$ gene; ensure the target gene has successfully replaced this lethal cassette before transformation into standard $E. coli$ competent cells.

• Sequence Verification: It is recommended to verify the insert using BioVector® M13 Forward and M13 Reverse sequencing primers.

Note: For optimal cloning efficiency and plasmid yield, use BioVector® LB Medium and BioVector® competent cells. Always perform restriction enzyme digestion or sequencing to confirm the orientation and integrity of your insert within the BioVector® pENTR11Dual backbone.

Supplier生产厂家: BioVector NTCC典型培养物保藏中心

E-mail: BioVector@163.com

http://www.biovector.net