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
- 感受态:
- Top10
- 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"