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

pCP209332 bp400800120016002000240028003200360040004400480052005600600064006800720076008000840088009200CmRcat promoterpSC101 oriRep101(Ts)Mobilization protein MbeCAmpRAmpR promoterFLPlambda repressor (ts)

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"