pCAS-RK2T BioVector® Plasmid假单胞菌基因组编辑和转录调控质粒载体-BioVector NTCC典型培养物保藏中心

价　　格：¥49950
货　　号：BioVector® pCAS-RK2T
产　　地：北京

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Product Name / 产品名称:
BioVector® pCAS-RK2T Plasmid
BioVector® pCAS-RK2T质粒

Description / 描述:
The BioVector® pCAS-RK2T plasmid is an E. coli-Pseudomonas shuttle vector designed for CRISPR-based genome editing and transcriptional regulation in Pseudomonas species, particularly Pseudomonas putida. It serves as the backbone for a series of CRISPR tools and enables stable maintenance and expression of CRISPR components in pseudomonads -1.
BioVector® pCAS-RK2T 质粒是一种大肠杆菌-假单胞菌穿梭载体，专为在假单胞菌属（特别是恶臭假单胞菌）中进行基于CRISPR的基因组编辑和转录调控而设计。它是系列CRISPR工具的骨架载体，能够在假单胞菌中稳定维持和表达CRISPR组件 -1。

Depositing Lab / 保藏实验室:
Sheng Yang Lab, Center for Excellence in Molecular Plant Sciences (CEMPS), CAS / 杨晟实验室，中国科学院分子植物科学卓越创新中心 -2

Plasmid Type / 质粒类型:

E. coli-Pseudomonas shuttle vector / 大肠杆菌-假单胞菌穿梭载体
CRISPR/Cas9 vector backbone / CRISPR/Cas9载体骨架
Broad-host-range vector / 广宿主范围载体

Species / 物种:

Escherichia coli (for propagation) / 大肠杆菌（用于扩增）
Pseudomonas putida and related pseudomonads (for application) / 恶臭假单胞菌及相关假单胞菌（用于应用）

Key Features / 关键特征:

Broad-host-range RK2 replication origin / 广宿主范围RK2复制起点: Enables stable maintenance in diverse Gram-negative bacteria including Pseudomonas species.
Tetracycline resistance marker / 四环素抗性标记: For selection in both E. coli and Pseudomonas hosts -2.
Shuttle vector design / 穿梭载体设计: Allows for convenient cloning in E. coli followed by transfer to Pseudomonas for functional studies.
Low copy number in E. coli / 大肠杆菌中低拷贝数: Facilitates stable maintenance of potentially toxic CRISPR components -2.

Vector Backbone / 载体骨架:
pCAS-RK2T (derived from RK2 broad-host-range plasmid) / pCAS-RK2T（来源于RK2广宿主范围质粒）

Total Vector Size / 总载体大小:
~13.9 kb (based on derivative pCAS-ZE3 size) / 约13.9 kb（基于衍生质粒pCAS-ZE3的大小） -2

Key Components / 关键组件:

RK2 replicon / RK2复制子: For broad-host-range replication
TetR / 四环素抗性基因: Selection marker (tetracycline resistance) / 四环素抗性标记 -2
Multiple Cloning Site (MCS) / 多克隆位点: For insertion of CRISPR components (Cas9, sgRNA, etc.) / 用于插入CRISPR组件
Origin of transfer (oriT) / 转移起始点: For conjugation-based plasmid transfer

Growth Conditions in Bacteria / 细菌生长条件:

Parameter / 参数	Specification / 规格
Bacterial Resistance / 细菌抗性	BioVector® Tetracycline (10-25 μg/mL for E. coli; 50-100 μg/mL for Pseudomonas)
Copy Number / 拷贝数	Low in E. coli / 大肠杆菌中低拷贝数 -2
Growth Strain / 生长菌株	Standard cloning strains (e.g., DH5α, TOP10) / 标准克隆菌株
Growth Temperature / 培养温度	37°C

Applications / 应用:

CRISPR Vector Backbone: Serves as the foundation for constructing CRISPR-Cas9 tools in Pseudomonas species -1
Genome Editing: Used in conjunction with Cas9 and sgRNA expression cassettes for targeted mutagenesis
Transcriptional Regulation: Backbone for dCas9-based CRISPR interference (CRISPRi) systems -1
Metabolic Engineering: Enables genetic modification of Pseudomonas for bioproduction applications
Synthetic Biology: Platform for constructing genetic circuits in pseudomonads

Related Plasmids / 相关质粒:

Product Name / 产品名称	Description / 描述
BioVector® pCAS-ZE3 Plasmid	Derivative containing dCas9 and sgRNA targeting J5 constitutive promoter for transcriptional repression in P. putida-2 含有dCas9和靶向J5组成型启动子的sgRNA的衍生质粒，用于恶臭假单胞菌中的转录抑制
BioVector® pCAS-Cas9 Plasmid	Derivative containing wild-type Cas9 for genome editing applications 含有野生型Cas9的衍生质粒，用于基因组编辑应用

Quality Control / 质量控制:
Plasmid is verified by restriction digestion and/or sequencing. Full sequence information is available from the depositor.
质粒经过限制性酶切和/或测序验证。完整序列信息可向保藏者获取。

Storage / 存储:

Plasmid is sent as a bacterial stab in agar or as purified DNA. Upon receipt, streak onto a plate with BioVector® Tetracycline, grow a single colony, and prepare a glycerol stock for long-term storage at -80°C.
质粒以细菌穿刺菌或纯化DNA形式提供。收到后，应在含 BioVector® 四环素的平板上划线，挑取单克隆，并制备甘油菌于-80°C长期保存。

Citation / 引用:
When using this plasmid, please cite the following article:
使用此质粒时，请引用以下文献：

Sun J, Wang Q, Jiang Y, Wen Z, Yang L, Wu J, Yang S. Genome editing and transcriptional repression in Pseudomonas putida KT2440 via the type II CRISPR system. Microb Cell Fact. 2018 Mar 17;17(1):41. -1

Important Note / 重要提示:

The pCAS-RK2T plasmid is a broad-host-range vector specifically optimized for Pseudomonas species but can be maintained in other Gram-negative bacteria.
pCAS-RK2T质粒是一种广宿主范围载体，特别针对假单胞菌属优化，但也能在其他革兰氏阴性菌中维持。
Due to its low copy number in E. coli, larger culture volumes may be needed for plasmid preparation -2.
由于其在大肠杆菌中为低拷贝数，质粒制备时可能需要较大培养体积。
The tetracycline resistance marker is functional in both E. coli and Pseudomonas, but optimal concentrations may vary between species -2.
四环素抗性标记在大肠杆菌和假单胞菌中均具有功能，但最适浓度可能因物种而异。