我们所展示的质粒图谱主要是从文献和开放数据库中收集而来,主要是为了方便研究工作,其中一小部分质粒进行了质量控制,可供科学家使用。
所有的产品都严格仅供科学研究使用,不能应用于临床试验,包括人体摄入、注射或外用的药理学用途。
确保质粒的关键元件正确,但是我们并不能保证实验结果。页面展示的图谱序列为理论序列,可能与测序结果不一致,请自行比对后确定是否满足要求。(如果测过序,本页面一般会提供下载)
开放数据库中的大多数载体序列都没有被完全测序。如果实际序列与参考序列的相似度超过99%,则将其视为正确。
由于科学研究是在探索未知,具有很大的不确定性,在任何情况下,我们都不承担超出质粒本身的额外经济损失或责任。
- 载体名称:
- pGL3-3'UTR
- 载体抗性:
- Ampicillin
- 载体长度:
- 6248 bp
- 载体类型:
- Reporter vector
- 复制子:
- ori
- 载体来源:
- Bi Y, Qiao X, Hua Z, Zhang L, Liu X, Li L, Hua W, Xiao H, Zhou J, Wei Q, Zheng X.
pGL3-3'UTR 载体图谱
质粒操作方法
1. 发货形式:质粒干粉(常温运输,存于-20度,请务必先转化提质粒后使用)
2. 收到质粒干粉后请先5000rpm离心1min,再加入20μl ddH2O溶解质粒;(质粒复测的浓度有时候与标称值差距较大,这可能是因为冻干质粒在管中的位置、复溶效率、测量偏差以及管壁的吸附导致,因此建议先转化提质粒后再使用)
3. 取1支100μl 感受态于冰上解冻10min,加入2μl质粒,再冰浴30min后,42℃热激60s,不要搅动,再冰浴2min;
4. 加入900μl无抗的LB液体培养基,180rpm震荡37℃培养45min (30℃培养1-1.5小时);
5. 6000rpm离心5min,仅留100μl上清液重悬细菌沉淀,并涂布至目标质粒抗性的LB平板上;
6. 将平板倒置37℃培养14h,如果要求是30℃则培养20h; (菌落过多则将质粒稀释后再转化;没有菌落则加入10μl质粒转化;建议不要直接转表达感受态, 要先转克隆感受态,重提质粒后再导入表达感受态);
7. 挑取单菌落至LB液体培养基中,加入对应抗生素,220rpm震荡培养14h,根据实验需要和质粒提取试剂盒说明书提取质粒。
pGL3-3'UTR 载体序列
LOCUS 40924_21948 6248 bp DNA circular SYN 18-DEC-2018 DEFINITION Reporter vector pGL3-3'UTR, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6248) AUTHORS Bi Y, Qiao X, Hua Z, Zhang L, Liu X, Li L, Hua W, Xiao H, Zhou J, Wei Q, Zheng X. TITLE An asymmetric PCR-based, reliable and rapid single-tube native DNA engineering strategy JOURNAL BMC Biotechnol. 12, 39 (2012) PUBMED 22768962 REFERENCE 2 (bases 1 to 6248) AUTHORS Bi Y. TITLE Direct Submission JOURNAL Submitted (03-AUG-2011) Biotechnology Group, Institute of Animal Science and Veterinary, Hubei Academy of Agricultural Science, No. 1, Nanhu Yaoyuan, Hongshan District, Wuhan, Hubei 430064, P. R. China REFERENCE 3 (bases 1 to 6248) TITLE Direct Submission REFERENCE 4 (bases 1 to 6248) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "BMC Biotechnol. 12, 39 (2012)" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (03-AUG-2011) Biotechnology Group, Institute of Animal Science and Veterinary, Hubei Academy of Agricultural Science, No. 1, Nanhu Yaoyuan, Hongshan District, Wuhan, Hubei 430064, P. R. China" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..6248 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 48..244 /label=SV40 promoter /note="SV40 early promoter" CDS 280..1929 /label=luciferase /note="firefly luciferase" regulatory 1988..3433 /regulatory_class="terminator" regulatory 2170..2175 /regulatory_class="polyA_signal_sequence" regulatory 2584..2589 /regulatory_class="polyA_signal_sequence" regulatory 2908..2913 /regulatory_class="polyA_signal_sequence" regulatory 3269..3274 /regulatory_class="polyA_signal_sequence" rep_origin complement(3751..4339) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4513..5370) /label=AmpR /note="beta-lactamase" promoter complement(5371..5475) /label=AmpR promoter rep_origin 5502..5957 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" polyA_signal 6088..6136 /label=poly(A) signal /note="synthetic polyadenylation signal" misc_feature 6150..6241 /label=pause site /note="RNA polymerase II transcriptional pause signal from the human alpha-2 globin gene"