pWUR768 载体 (V002116)

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

pWUR76810193 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000lacIq promoterRBSaraCKanRRBSPBAD-adaptoriPBAD-adaptRBSLuxCLuxDLuxALuxBLuxErrnB T1 terminatorlambda t0 terminatorCmRcat promoter

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"