Product Name / 產(chǎn)品名稱:
BioVector? pCAS-RK2T Plasmid
BioVector? pCAS-RK2T質(zhì)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 質(zhì)粒是一種大腸桿菌-假單胞菌穿梭載體，專為在假單胞菌屬（特別是惡臭假單胞菌）中進(jìn)行基于CRISPR的基因組編輯和轉(zhuǎn)錄調(diào)控而設(shè)計(jì)。它是系列CRISPR工具的骨架載體，能夠在假單胞菌中穩(wěn)定維持和表達(dá)CRISPR組件 -1。

Depositing Lab / 保藏實(shí)驗(yàn)室:
Sheng Yang Lab, Center for Excellence in Molecular Plant Sciences (CEMPS), CAS / 楊晟實(shí)驗(yàn)室，中國(guó)科學(xué)院分子植物科學(xué)卓越創(chuàng)新中心 -2

Plasmid Type / 質(zhì)粒類型:

• E. coli-Pseudomonas shuttle vector / 大腸桿菌-假單胞菌穿梭載體

• CRISPR/Cas9 vector backbone / CRISPR/Cas9載體骨架

• Broad-host-range vector / 廣宿主范圍載體

Species / 物種:

• Escherichia coli (for propagation) / 大腸桿菌（用于擴(kuò)增）

• Pseudomonas putida and related pseudomonads (for application) / 惡臭假單胞菌及相關(guān)假單胞菌（用于應(yīng)用）

Key Features / 關(guān)鍵特征:

• Broad-host-range RK2 replication origin / 廣宿主范圍RK2復(fù)制起點(diǎn): Enables stable maintenance in diverse Gram-negative bacteria including Pseudomonas species.

• Tetracycline resistance marker / 四環(huán)素抗性標(biāo)記: For selection in both E. coli and Pseudomonas hosts -2.

• Shuttle vector design / 穿梭載體設(shè)計(jì): Allows for convenient cloning in E. coli followed by transfer to Pseudomonas for functional studies.

• Low copy number in E. coli / 大腸桿菌中低拷貝數(shù): Facilitates stable maintenance of potentially toxic CRISPR components -2.

Vector Backbone / 載體骨架:
pCAS-RK2T (derived from RK2 broad-host-range plasmid) / pCAS-RK2T（來(lái)源于RK2廣宿主范圍質(zhì)粒）

Total Vector Size / 總載體大小:
~13.9 kb (based on derivative pCAS-ZE3 size) / 約13.9 kb（基于衍生質(zhì)粒pCAS-ZE3的大小） -2

Key Components / 關(guān)鍵組件:

• RK2 replicon / RK2復(fù)制子: For broad-host-range replication

• TetR / 四環(huán)素抗性基因: Selection marker (tetracycline resistance) / 四環(huán)素抗性標(biāo)記 -2

• Multiple Cloning Site (MCS) / 多克隆位點(diǎn): For insertion of CRISPR components (Cas9, sgRNA, etc.) / 用于插入CRISPR組件

• Origin of transfer (oriT) / 轉(zhuǎn)移起始點(diǎn): For conjugation-based plasmid transfer

Growth Conditions in Bacteria / 細(xì)菌生長(zhǎng)條件:

Applications / 應(yīng)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 / 相關(guān)質(zhì)粒:

Quality Control / 質(zhì)量控制:
Plasmid is verified by restriction digestion and/or sequencing. Full sequence information is available from the depositor.
質(zhì)粒經(jīng)過(guò)限制性酶切和/或測(cè)序驗(yàn)證。完整序列信息可向保藏者獲取。

Storage / 存儲(chǔ):

• 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.

• 質(zhì)粒以細(xì)菌穿刺菌或純化DNA形式提供。收到后，應(yīng)在含 BioVector? 四環(huán)素 的平板上劃線，挑取單克隆，并制備甘油菌于-80°C長(zhǎng)期保存。

Citation / 引用:
When using this plasmid, please cite the following article:
使用此質(zhì)粒時(shí)，請(qǐng)引用以下文獻(xiàn)：

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質(zhì)粒是一種廣宿主范圍載體，特別針對(duì)假單胞菌屬優(yōu)化，但也能在其他革蘭氏陰性菌中維持。

• Due to its low copy number in E. coli, larger culture volumes may be needed for plasmid preparation -2.

• 由于其在大腸桿菌中為低拷貝數(shù)，質(zhì)粒制備時(shí)可能需要較大培養(yǎng)體積。

• The tetracycline resistance marker is functional in both E. coli and Pseudomonas, but optimal concentrations may vary between species -2.

• 四環(huán)素抗性標(biāo)記在大腸桿菌和假單胞菌中均具有功能，但最適濃度可能因物種而異。

BioVector? Tetracycline
BioVector? 四環(huán)素

For selection of pCAS-RK2T plasmid and its derivatives in both E. coli and Pseudomonas hosts.
用于在大腸桿菌和假單胞菌宿主中選擇pCAS-RK2T質(zhì)粒及其衍生質(zhì)粒。

BioVector NTCC質(zhì)粒載體菌株細(xì)胞蛋白抗體基因保藏中心

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