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:
- pVMG-TnpR
- 载体抗性:
- Ampicillin
- 载体长度:
- 7264 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Oke V, Long SR.
pVMG-TnpR 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。
pVMG-TnpR 载体序列
LOCUS V002230 7264 bp DNA circular SYN 18-DEC-2018 DEFINITION Exported. ACCESSION V002230 VERSION V002230 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 7264) AUTHORS Oke V, Long SR. TITLE Bacterial genes induced within the nodule during the Rhizobium-legume symbiosis JOURNAL Mol. Microbiol. 32 (4), 837-849 (1999) PUBMED 10361286 REFERENCE 2 (bases 1 to 7264) AUTHORS Gao M, Teplitski M. TITLE RIVET-a tool for in vivo analysis of symbiotically relevant gene expression in Sinorhizobium meliloti JOURNAL Mol. Plant Microbe Interact. 21 (2), 162-170 (2008) PUBMED 18184060 REFERENCE 3 (bases 1 to 7264) AUTHORS Gao M, Oke V, Teplitski M. TITLE Direct Submission JOURNAL Submitted (17-OCT-2007) Soil and Water Science, University of REFERENCE 4 (bases 1 to 7264) TITLE Direct Submission REFERENCE 5 (bases 1 to 7264) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Microbiol."; date: "1999"; volume: "32"; issue: "4"; pages: "837-849" SGRef: number: 2; type: "Journal Article"; journalName: "Mol. Plant Microbe Interact."; date: "2008"; volume: "21"; issue: "2"; pages: "162-170" SGRef: number: 3; type: "Journal Article"; journalName: "Submitted (17-OCT-2007) Soil and Water Science, University of Florida-IFAS, 1376 Mowry Rd., Cancer " SGRef: number: 4; type: "Journal Article" FEATURES Location/Qualifiers source 1..7264 /mol_type="other DNA" /organism="synthetic DNA construct" regulatory 28..58 /label="Escherichia coli trpA" /note="Escherichia coli trpA" /regulatory_class="terminator" misc_feature 103..113 /label="contains stop codons in all three reading frames" /note="contains stop codons in all three reading frames" regulatory 189..195 /gene="tnpR" /regulatory_class="ribosome_binding_site" CDS 205..753 /gene="tnpR" /label="Transposon gamma-delta resolvase" /note="Transposon gamma-delta resolvase from Escherichia coli (strain K12). Accession#: P03012" regulatory 756..761 /gene="uidA" /regulatory_class="ribosome_binding_site" CDS 788..2596 /label="GUS" /note="beta-glucuronidase" CDS 3019..3810 /label="NeoR/KanR" /note="aminoglycoside phosphotransferase" primer_bind complement(4183..4199) /label="M13 fwd" /note="common sequencing primer, one of multiple similar variants" rep_origin 4412..4867 /label="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" oriT complement(5206..5315) /direction=LEFT /label="oriT" /note="incP origin of transfer" promoter 5485..5589 /label="AmpR promoter" CDS 5590..6447 /label="AmpR" /note="beta-lactamase" rep_origin 6621..7209 /direction=RIGHT /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"