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:
- pBMT-4
- 载体抗性:
- Tetracycline
- 载体长度:
- 4181 bp
- 载体类型:
- Broad host range vector
- 复制子:
- pBBR1 oriV
- 载体来源:
- Lynch MD, Gill RT.
pBMT-4 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。
pBMT-4 载体序列
LOCUS 40924_6856 4181 bp DNA circular SYN 17-DEC-2018 DEFINITION Broad host range vector pBMT-4, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4181) AUTHORS Lynch MD, Gill RT. TITLE Broad host range vectors for stable genomic library construction JOURNAL Biotechnol. Bioeng. 94 (1), 151-158 (2006) PUBMED 16496398 REFERENCE 2 (bases 1 to 4181) AUTHORS Lynch MD, Gill RT. TITLE Direct Submission JOURNAL REFERENCE 3 (bases 1 to 4181) TITLE Direct Submission REFERENCE 4 (bases 1 to 4181) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol. Bioeng."; date: "2006"; volume: "94"; issue: "1"; pages: "151-158" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (12-MAY-2005) Chemical " COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..4181 /mol_type="other DNA" /organism="synthetic DNA construct" terminator complement(60..96) /label=soxR terminator /note="bidirectional E. coli soxR transcription terminator" CDS complement(104..1123) /codon_start=1 /gene="mob" /product="Mob" /label=mob /note="required for mobilization" /protein_id="AAY99681.1" /translation="MAAYAIMRCKKLAKMGNVAASLKHAYRERETPNADASRTPENEHW AASSTDEAMGRLRELLPEKRRKDAVLAVEYVMTASPEWWKSASQEQQAAFFEKAHKWLA DKYGADRIVTASIHRDETSPHMTAFVVPLTQDGRLSAKEFIGNKAQMTRDQTTFAAAVA DLGLQRGIEGSKARHTRIQAFYEALERPPVGHVTISPQAVEPRAYAPQGLAEKLGISKR VETPEAVADRLTKAVRQGYEPALQAAAGAREMRKKADQAQETARDLRERLKPVLDALGP LNRDMQAKAAAIIKAVGEKLLTEQREVQRQKQAQRQQERGRAHFPEKCHLTSKKPLLS" gene complement(104..1123) /gene="mob" /label=mob /note="mob gene from pBBR1MCS, GenBank Accession Number U02374" rep_origin 1347..2116 /label=pBBR1 oriV /note="replication origin of the broad-host-range plasmid pBBR1 from Bordetella bronchiseptica; requires the pBBR1 Rep protein for replication" CDS 2117..2776 /codon_start=1 /label=pBBR1 Rep /note="replication protein for the broad-host-range plasmid pBBR1 from Bordetella bronchiseptica" /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR" CDS 2876..4063 /codon_start=1 /label=TcR /note="tetracycline efflux protein" /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST" terminator complement(4086..4117) /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator"