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:
- pBG51
- 载体抗性:
- Kanamycin
- 载体长度:
- 3959 bp
- 载体类型:
- Cloning vector
- 复制子:
- R6K γ ori
- 载体来源:
- Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank LM.
- 启动子:
- Pc
pBG51 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。
pBG51 载体序列
LOCUS 40924_6302 3959 bp DNA circular SYN 17-DEC-2018 DEFINITION Cloning vector pBG51, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3959) AUTHORS Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank LM. TITLE A Tn7-based device for calibrated heterologous gene expression in Pseudomonas putida JOURNAL ACS Synth Biol (2015) In press PUBMED 26133359 REFERENCE 2 (bases 1 to 3959) AUTHORS Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank LM. TITLE Direct Submission JOURNAL Submitted (09-JUN-2015) System Biology, Centro Nacional de Biotecnologia, C/Darwin 3, Madrid 28049, Spain REFERENCE 3 (bases 1 to 3959) AUTHORS Zobel S, Benedetti I, Eisenbach L, de Lorenzo V, Wierckx N, Blank LM. TITLE Direct Submission JOURNAL Submitted (13-NOV-2015) System Biology, Centro Nacional de Biotecnologia, C/Darwin 3, Madrid 28049, Spain REFERENCE 4 (bases 1 to 3959) TITLE Direct Submission REFERENCE 5 (bases 1 to 3959) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth Biol (2015) In press" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (09-JUN-2015) System Biology, Centro Nacional de Biotecnologia, C/Darwin 3, Madrid 28049, Spain" COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted (13-NOV-2015) System Biology, Centro Nacional de Biotecnologia, C/Darwin 3, Madrid 28049, Spain" COMMENT SGRef: number: 4; type: "Journal Article" COMMENT ##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## On Nov 13, 2015 this sequence version replaced KT192133.1. FEATURES Location/Qualifiers source 1..3959 /mol_type="other DNA" /organism="synthetic DNA construct" regulatory 9..49 /label=promoter 14d /note="promoter 14d" /regulatory_class="promoter" misc_feature 56..132 /label=BCD2 bicistronic linker /note="BCD2 bicistronic linker" RBS 124..132 /label=Shine-Dalgarno sequence /note="full consensus sequence for ribosome-binding sites upstream of start codons in E. coli; complementary to a region in the 3' end of the 16S rRNA (Chen et al., 1994)" CDS 151..864 /label=superfolder GFP /note="GFP variant that folds robustly even when fused to poorly folded proteins (Pedelacq et al., 2006)" misc_feature 868..924 /label=MCS /note="pUC18/19 multiple cloning site" terminator 967..1061 /label=lambda t0 terminator /note="transcription terminator from phage lambda" misc_feature 1066..1264 /label=Tn7R transposase recognition site /note="Tn7R transposase recognition site" CDS 1374..2186 /label=KanR /note="aminoglycoside phosphotransferase" oriT 2345..2453 /label=oriT /note="incP origin of transfer" rep_origin 2472..2860 /label=R6K gamma ori /note="gamma replication origin from E. coli plasmid R6K; requires the R6K initiator protein pi for replication" mobile_element 2872..3037 /label=Tn7L /note="mini-Tn7 element (left end of the Tn7 transposon)" terminator complement(3051..3137) /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" CDS complement(3206..3736) /label=GmR /note="gentamycin acetyltransferase" promoter complement(3925..3953) /label=Pc promoter /note="class 1 integron promoter"