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:
- pC1300intB-35SnosBK
- 载体抗性:
- Kanamycin
- 载体长度:
- 9946 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 宿主:
- Plants
- 载体来源:
- Ouwerkerk PBF., Meijer AH.
- 启动子:
- CaMV 35S (enhanced)
pC1300intB-35SnosBK 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。
pC1300intB-35SnosBK 载体序列
LOCUS 40924_7896 9946 bp DNA circular SYN 17-DEC-2018 DEFINITION Cloning vector pC1300intB-35SnosBK, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9946) AUTHORS Ouwerkerk PBF., Meijer AH. TITLE Binary expression vector for use in plants JOURNAL Unpublished REFERENCE 2 (bases 1 to 9946) AUTHORS Ouwerkerk PBF., Meijer AH. TITLE Direct Submission JOURNAL Submitted (26-FEB-2004) Molecular Cell Biology, Institute of Biology, Leiden University, Wassenaarseweg 64, Leiden 2333 AL, The Netherlands REFERENCE 3 (bases 1 to 9946) TITLE Direct Submission REFERENCE 4 (bases 1 to 9946) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (26-FEB-2004) Molecular Cell Biology, Institute of Biology, Leiden University, Wassenaarseweg 64, Leiden 2333 AL, The Netherlands" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..9946 /mol_type="other DNA" /organism="synthetic DNA construct" terminator complement(48..300) /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" misc_feature complement(318..356) /note="multiple cloning site; SalI, EcoRI, XhoI, BglII, KpnI, SpeI are unique" misc_feature complement(358..413) /label=TMV Omega /note="translational enhancer from the tobacco mosaic virus 5'-leader sequence (Gallie et al., 1988)" promoter complement(433..777) /label=CaMV 35S promoter /note="strong constitutive promoter from cauliflower mosaic virus" primer_bind complement(837..853) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" misc_feature 1056..1080 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 2380..3006 /label=pVS1 StaA /note="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS 3443..4507 /label=pVS1 RepA /note="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" rep_origin 4576..4770 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" misc_feature 5114..5254 /label=bom /note="basis of mobility region from pBR322" rep_origin complement(5440..6028) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6118..6909) /label=KanR /note="aminoglycoside phosphotransferase" misc_feature 7334..7358 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" polyA_signal complement(7436..7610) /label=CaMV poly(A) signal /note="cauliflower mosaic virus polyadenylation signal" exon complement(7661..8347) /gene="hptII" /number=1 intron complement(8348..8537) /label=cat1 intron /note="castor bean catalase intron, modified" exon complement(8538..8876) /gene="hptII" /number=2 promoter complement(8955..9632) /label=CaMV 35S promoter (enhanced) /note="cauliflower mosaic virus 35S promoter with a duplicated enhancer region" protein_bind 9823..9844 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 9859..9889 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9897..9913 /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)." primer_bind 9921..9937 /label=M13 rev /note="common sequencing primer, one of multiple similar variants"