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:
- pWUR768
- 载体抗性:
- Kanamycin
- 载体长度:
- 10193 bp
- 载体类型:
- Regulator-reporter vector
- 复制子:
- ori
- 载体来源:
- van Rossum T, Muras A, Baur MJ, Creutzburg SC, van der Oost J, Kengen SW.
pWUR768 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。
pWUR768 载体序列
LOCUS 40924_46673 10193 bp DNA circular SYN 18-DEC-2018 DEFINITION Regulator-reporter vector pWUR768, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10193) AUTHORS van Rossum T, Muras A, Baur MJ, Creutzburg SC, van der Oost J, Kengen SW. TITLE A growth- and bioluminescence-based bioreporter for the in vivo detection of novel biocatalysts JOURNAL Microb Biotechnol (2017) In press PUBMED 28393499 REFERENCE 2 (bases 1 to 10193) AUTHORS van Rossum T, Muras A, Baur MJ, Creutzburg SC, van der Oost J, Kengen SW. TITLE Direct Submission JOURNAL Submitted (04-AUG-2016) Laboratory of Microbiology, Wageningen University and Research, Stippeneng 4, Wageningen 6708 WE, The Netherlands REFERENCE 3 (bases 1 to 10193) TITLE Direct Submission REFERENCE 4 (bases 1 to 10193) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Microb Biotechnol (2017) In press" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (04-AUG-2016) Laboratory of Microbiology, Wageningen University and Research, Stippeneng 4, Wageningen 6708 WE, The Netherlands" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..10193 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 10..87 /label=lacIq promoter /note="In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold." regulatory 95..100 /label=RBS /note="RBS" /regulatory_class="ribosome_binding_site" CDS 107..982 /label=araC /note="L-arabinose regulatory protein" CDS complement(995..1807) /label=KanR /note="aminoglycoside phosphotransferase" regulatory complement(1814..1819) /label=RBS /note="RBS" /regulatory_class="ribosome_binding_site" regulatory complement(1828..2138) /label=PBAD-adapt /note="PBAD-adapt" /regulatory_class="promoter" rep_origin complement(2289..2877) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" regulatory 2959..3269 /label=PBAD-adapt /note="PBAD-adapt" /regulatory_class="promoter" regulatory 3276..3281 /label=RBS /note="RBS" /regulatory_class="ribosome_binding_site" CDS 3288..4727 /label=LuxC /note="LuxC fatty acid reductase" CDS 4743..5663 /label=LuxD /note="LuxD acyltransferase" CDS 5715..6794 /label=LuxA /note="LuxA luciferase subunit" CDS 6812..7792 /label=LuxB /note="LuxB luciferase subunit" CDS 7974..9083 /label=LuxE /note="LuxE" terminator 9106..9192 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator complement(9218..9312) /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(9336..9992) /label=CmR /note="chloramphenicol acetyltransferase" promoter complement(9993..10095) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase"