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。
pCP20 is an ampicillin and CmR plasmid that shows temperature-sensitive replication and thermal induction of FLP synthesis. CmR and KmR mutants were transformed with pCP20, and ampicillin-resistant transformants were selected at 30°C, after which a few were colony-purified once nonselectively at 43°C and then tested for loss of all antibiotic resistances. The majority lost the FRT-flanked resistance gene and the FLP helper plasmid simultaneously. This vector was first published in 1995, by Dr. W. Wackernagel, Genetik, Fachbereich Biologic, University Oldenburg.
- 载体名称:
- pCP20
- 载体抗性:
- Ampicillin
- 载体长度:
- 9332 bp
- 载体类型:
- Knockout Vectors
- 复制子:
- pSC101 ori
- 感受态:
- DH10B
- 培养温度:
- 37℃
pCP20 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。
pCP20 载体序列
LOCUS V001036 9332 bp DNA circular SYN 13-JAN-2022 DEFINITION Exported. ACCESSION V001036 VERSION V001036 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 9332) TITLE Direct Submission REFERENCE 2 (bases 1 to 9332) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..9332 /mol_type="other DNA" /organism="synthetic DNA construct" CDS complement(564..1220) /label="CmR" /note="chloramphenicol acetyltransferase" promoter complement(1221..1323) /label="cat promoter" /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" rep_origin 1875..2097 /label="pSC101 ori" /note="low-copy replication origin that requires the Rep101 protein" CDS 2145..3092 /label="Rep101(Ts)" /note="temperature-sensitive version of the RepA protein needed for replication with the pSC101 origin (Armstrong et al., 1984)" CDS complement(4016..4360) /gene="mbeC" /label="Mobilization protein MbeC" /note="Mobilization protein MbeC from Escherichia coli. Accession#: P13657" CDS complement(4560..5417) /label="AmpR" /note="beta-lactamase" promoter complement(5418..5522) /label="AmpR promoter" CDS complement(6559..7827) /label="FLP" /note="site-specific recombinase" CDS 7928..8638 /label="lambda repressor (ts)" /note="temperature-sensitive variant of the phage lambda repressor"