| 出品公司: | ZYbscience |
|---|---|
| 载体名称: | pGuide-it-tdTomato |
| 质粒类型: | 慢病毒载体；CRISPR/Cas9载体；荧光报告载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 克隆方法: | 限制性内切酶，多克隆位点 |
| 启动子: | Minimal CMV |
| 载体大小: | 9038 bp |
| 5' 测序引物及序列: | -- |
| 3' 测序引物及序列: | -- |
| 载体标签: | -- |
| 载体抗性: | 氨苄青霉素 |
| 筛选标记: | tdTomato红色荧光蛋白 |
| 克隆菌株: | Stbl3、Stellar |
| 宿主细胞（系）: | 常规细胞系（293、CV-1、CHO等） |
| 备注: | 慢病毒载体pGuide-it-tdTomato是CRISPR/Cas9载体，Clontech提供的是线性化的质粒，Biofeng提供的是环状质粒。 |
| 产品目录号: | ZY632604 |
| 稳定性: | 稳表达 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 慢病毒 |
- ¥8000
- ZYbscience
- 中国/美国
- ZY632604
- 2025年07月15日
11 年钻石会员
企业认证
万千商家帮你免费找货
0 人在求购买到急需产品
- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20℃低温保存
- 保质期：
三年
- 英文名：
pGuide-IT-tdTomato
- 库存：
20
- 供应商：
泽叶生物
载体基本信息
载体质粒图谱和多克隆位点信息
载体简介
CRISPR/Cas9 技术
The Guide-it CRISPR/Cas9 System (Green, Cat. No. 632601; Red, Cat. No. 632602) is a complete system for the cloning and expression of a target single guide RNA (sgRNA) for mammalian gene modification studies using CRISPR/Cas9 technology. The included vector is used to simultaneously express Cas9, a target-specific sgRNA, and an exceptionally bright fluorescent protein (either ZsGreen1 or tdTomato) for monitoring transfection efficiency and/or for further enriching/isolating transfected cells by flow cytometry. The vector is pre-linearized for simple insertion of a target sequence for sgRNA expression from the human U6 promoter; Cas9 nuclease and either ZsGreen1 or tdTomato are co-expressed from a bidirectional CMV promoter. Sufficient reagents are provided in this kit for construction of 10 different target (sgRNA) expression plasmids. To construct the vector, a pair of user-provided oligos corresponding to the target genomic sequence of interest are annealed to form a duplex. Then, the duplexed DNA is cloned into the pre-linearized vector using the included high-efficiency ligation mix. This kit also contains necessary controls and Stellar? Competent Cells.
Cloning and Expression System for sgRNAs
The Guide-it CRISPR/Cas9 Systems are kits for the cloning and expression of target single guide RNAs (sgRNAs) for mammalian genome editing using CRISPR/Cas9 technology. The vector in this system simultaneously expresses Cas9 nuclease, a target-specific sgRNA, and an exceptionally bright fluorescent protein for monitoring transfection efficiency and/or for further enriching/isolating transfected cells by flow cytometry (ZsGreen1 and tdTomato versions are available). Generating a plasmid that expresses a sequence-specific sgRNA with this system is simple; a pair of user-provided oligos corresponding to the target genomic sequence of interest are annealed to form a duplex, and the duplexed oligos are inserted into the pre-linearized vector using the included high-efficiency ligation mix. The kit also includes Stellar competent cells to ensure high efficiency transformation.
Features
A single vector expresses your target-specific sgRNA from the human U6 promoter, and Cas9 nuclease and a bright fluorescent marker (either ZsGreen1 or tdTomato) are expressed from a bidirectional CMV promoter tdTomato and ZsGreen1 are exceptionally bright fluorescent proteins, 6x and 2.5x brighter than EGFP, respectively; these markers can be used to monitor transfection efficiency and/or to enrich/isolate transfected cells by flow cytometry Simple plasmid construction—design oligos specific to your target sequence, and clone into the pre-linearized plasmid using the included ligation reagents and high- efficiency competent cells Includes sufficient reagents for construction of 10 different target (sgRNA) expression plasmids
Applications
Cloning and expression of a target single guide RNA (sgRNA) for mammalian genome editing using CRISPR/Cas9 technology
载体序列
LOCUS pGuide-it-tdTomato Vector (Linear) 9042 bp ds-DNA linear SYN 05-DEC-2014
COMMENT Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT Cat No. 632602, 632604
SOURCE synthetic DNA construct
FEATURES Location/Qualifiers
misc_feature 1..93
/label="sgRNA"
/note="single-guide RNA scaffold segment; user clones guide sequence"
rep_origin complement(147..746)
/label="pUC origin"
/note="ori;high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(908..1768)
/label="AmpR"
/note="beta-lactamase, confers resistance to ampicillin, carbenicillin, and
related antibiotics"
polyA_signal complement(1904..2091)
/label="SV40 polyA signal"
CDS complement(2129..3556)
/label="tdTomato"
promoter complement(3574..3642)
/label="Minimal CMV Promoter 2"
misc_feature 3706..4115
/label="Enhancer"
promoter 4116..4241
/label="Minimal CMV Promoter 1"
CDS 4286..8389
/label="Cas9"
CDS 8405..8425
/label="NLS"
/note="nuclear localization signal of SV40 large T antigen"
CDS 8426..8449
/label="DYKDDDDK tag"
misc_feature 8488..8675
/label="SV40 polyA signal"
promoter 8792..9032
/label="U6 Promoter"
/note="U6 promoter;RNA polymerase III promoter for human U6
snRNA"
ORIGIN
1 gtttaagagc tatgctggaa acagcatagc aagtttaaat aaggctagtc cgttatcaac
61 ttgaaaaagt ggcaccgagt cggtgctttt tttgagcaaa aggccagcaa aaggccagga
121 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
181 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
241 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
301 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
361 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
421 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
481 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
541 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
601 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
661 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
721 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
781 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
841 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
901 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
961 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
1021 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
1081 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
1141 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
1201 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
1261 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
1321 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
1381 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
1441 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
1501 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
1561 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
1621 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
1681 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
1741 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
1801 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
1861 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcggcagt gaaaaaaatg
1921 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
1981 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
2041 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcctctaga
2101 ctgcagcctc aggagatctg ggcccctact tgtacagctc gtccatgccg tacaggaaca
2161 ggtggtggcg gccctcggag cgctcgtact gttccacgat ggtgtagtcc tcgttgtggg
2221 aggtgatgtc cagcttggtg tccacgtagt agtagccggg cagttgcacg ggcttcttgg
2281 ccatgtagat ggtcttgaac tccaccaggt agtggccgcc gtccttcagc ttcagggcct
2341 ggtggatctc gcccttcagc acgccgtcgc gggggtacag gcgctcggtg gaggcctccc
2401 agcccatggt cttcttctgc attacggggc cgtcgggggg gaagttggtg ccgcgcatct
2461 tcaccttgta gatcagcgtg ccgtcctgca gggaggagtc ctgggtcacg gtcaccagac
2521 cgccgtcctc gaagttcatc acgcgctccc acttgaagcc ctcggggaag gacagcttct
2581 tgtaatcggg gatgtcggcg gggtgcttca cgtacgcctt ggagccgtac atgaactggg
2641 gggacaggat gtcccaggcg aagggcaggg ggccgccctt ggtcaccttc agcttggcgg
风险提示：丁香通仅作为第三方平台，为商家信息发布提供平台空间。用户咨询产品时请注意保护个人信息及财产安全，合理判断，谨慎选购商品，商家和用户对交易行为负责。对于医疗器械类产品，请先查证核实企业经营资质和医疗器械产品注册证情况。
文献和实验相关实验
1，tdTomato，GFP，mcherry，GFP，FP635等。 «贵公司的慢病毒载体可以插入多大的目的片段? 我们采用独特的载体设计，最大可高效包装4kb的目的片段。 «贵公司的慢病毒的安全性如何? 我们的慢病毒载体经过了一系列的改造，病毒在感染细胞中并不能自我复制，从而大大降低了其危险性。尤其是我们使用了现在最新型第四代载体系统，是目前重组系数最低的一个慢病毒包装系统，至今为止未见重组报道。慢病毒对热、去垢剂、医用消毒水、福尔马林等均很敏感，但UV照射无法
3.你们的慢病毒载体上有什么报告基因? 答：目前我们主要提供的报告基因是绿色荧光蛋白EGFP和红色荧光蛋白tdTomato的载体系统。我们所选的这两个报告基因的荧光亮度都很高，同时是以单体的形式存在。此外，我们还可以根据客户的需要提供包含其它报告基因(如半乳糖苷酶lacZ或荧光素酶luciferase等)的载体系统。 4.我以前用过慢病毒感染造血细胞，但是表达不理想，你们有没有好的解决办法? 答：影响慢病毒载体介导的目的蛋白表达的因素主要是病毒的感染能力和特定的启动
特异性或广泛表达启动子与hACE2基因构建的表达载体，通过原核注射的方法，获得随机插入的转基因小鼠模型，比如，细胞角蛋白（上皮细胞特异性表达-K18）启动子；CAG启动子（广泛表达）或小鼠ACE2基因启动子。应用这些ACE2人源化小鼠模型的研究结果表明，hACE2的表达水平高低与其疾病的严重程度直接相关。虽然，这些小鼠模型都表现有呼吸道上皮细胞感染，但同时在小鼠脑部也可见高表达的hACE2基因，并在SARS-CoV感染模型中发现，病毒量在小鼠脑部的增加与广泛扩散，最终导致小鼠发生脑炎而死亡
技术资料暂无技术资料 索取技术资料
pGuide-IT-tdTomato载体
¥8000
