质粒编号
质粒名称
规格
价格
V018110
pBSM
5 μg (冻干粉)

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

pBSM3089 bp6001200180024003000T7T3 promoterIn lacZ genelac promoterCAP binding siteL4440oriAmpRAmpR promoterf1 oriIn lacZ gene

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"