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。
For RNA production using in vivo transcription by Escherichia coli.
- 载体名称:
- pBSM
- 载体抗性:
- Ampicillin
- 载体长度:
- 3089 bp
- 载体类型:
- Bacterial Expression
- 载体来源:
- Luc Ponchon
- 拷贝数:
- High copy number
- 培养温度:
- 37℃
pBSM 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。
pBSM 载体序列
LOCUS Exported 3089 bp DNA circular SYN 23-AUG-2025 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBSM SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3089) AUTHORS Ponchon L, Catala M, Seijo B, El Khouri M, Dardel F, Nonin-Lecomte S, Tisne C TITLE Co-expression of RNA-protein complexes in Escherichia coli and applications to RNA biology. JOURNAL Nucleic Acids Res. 2013 Aug;41(15):e150. doi: 10.1093/nar/gkt576. Epub 2013 Jun 26. PUBMED 23804766 REFERENCE 2 (bases 1 to 3089) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic Acids Res."; date: "2013-08"; volume: "41(15):e150. doi"; pages: " 10.1093/nar/gkt576. Epub 2013 Jun 26" FEATURES Location/Qualifiers source 1..3089 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 1..20 /label=T7 /note="T7 promoter, forward primer" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(279..297) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(318..334) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(318..334) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(331..353) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 342..358 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(366..396) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 411..432 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(549..566) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(720..1308) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(800..819) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1479..2339) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 2102..2121 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2340..2444) /gene="bla" /label=AmpR promoter rep_origin complement(2470..2925) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2607..2628) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2819..2838 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3052..3074 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 3066..3083 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 3067..3083 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants"