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:
- pSC101-PhaC-pct
- 载体抗性:
- Ampicillin
- 载体长度:
- 7358 bp
- 载体类型:
- Cloning vector
- 复制子:
- pSC101 ori
- 载体来源:
- Guo L.
- 启动子:
- tetR/tetAs
pSC101-PhaC-pct 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。
pSC101-PhaC-pct 载体序列
LOCUS 62056_19380 7358 bp DNA circular SYN 16-MAY-2020 DEFINITION Cloning vector pSC101-PhaC-pct, complete sequence. ACCESSION MN623110 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7358) AUTHORS Guo L. TITLE Direct Submission JOURNAL Submitted (28-OCT-2019) Jiangnan University, State Key Laboratory of Food Science and Technology, 1800 Lihu Avenue, Wuxi, Jiangsu 214122, China REFERENCE 2 (bases 1 to 7358) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Submitted (28-OCT-2019) Jiangnan University, State Key Laboratory of Food Science and Technology, 1800 Lihu Avenue, Wuxi, Jiangsu 214122, China" FEATURES Location/Qualifiers source 1..7358 /mol_type="other DNA" /organism="synthetic DNA construct" terminator complement(100..129) /label=T3Te terminator /note="phage T3 early transcription terminator" CDS complement(185..808) /label=TetR /note="tetracycline repressor TetR" promoter 824..879 /label=tetR/tetA promoters /note="overlapping promoters for bacterial tetR and tetA" gene 909..2588 /gene="phaC" /label=phaC CDS 909..2588 /codon_start=1 /transl_table=11 /gene="phaC" /product="polyhydroxyalkanoate synthase" /label=phaC /protein_id="QJR97783.1" /translation="MSNKNSDDLNRQASENTLGLNPVIGLRGKDLLTSARMVLTQAIKQ PIHSVKHVAHFGIELKNVMFGKSKLQPESDDRRFNDPAWSQNPLYKRYLQTYLAWRKEL HDWIGNSKLSEQDINRAHFVITLMTEAMAPTNSAANPAAVKRFFETGGKSLLDGLTHLA KDLVNNGGMPSQVDMGAFEVGKSLGTTEGAVVFRNDVLELIQYRPTTEQVHERPLLVVP PQINKFYVFDLSPDKSLARFCLSNNQQTFIVSWRNPTKAQREWGLSTYIDALKEAVDVV SAITGSKDINMLGACSGGITCTALLGHYAALGEKKVNALTLLVTVLDTTLDSQVALFVD EKTLEAAKRHSYQAGVLEGRDMAKVFAWMRPNDLIWNYWVNNYLLGNEPPVFDILFWNN DTTRLPAAFHGDLIEMFKNNPLVRANALEVSGTPIDLKQVTADIYSLAGTNDHITPWKS CYKSAQLFGGKVEFVLSSSGHIKSILNPPGNPKSRYMTSTDMPATANEWQENSTKHTDS WWLHWQAWQAERSGKLKKSPTSLGNKAYPSGEAAPGTYVHER" gene 2616..4169 /gene="pct" /label=pct CDS 2616..4169 /codon_start=1 /transl_table=11 /gene="pct" /product="propionyl-CoA transferase" /label=pct /protein_id="QJR97784.1" /translation="MRKVEIITAEQAAQLVKDNDTITSIGFVSSAHPEALTKALEKRFL DTNTPQNLTYIYAGSQDKRDGRAAEHLAHTGLLKRAIIGHWQTVPAIGKLAVENKIEAY NFSQGTLVHWFRALAGHKLGVFTDIGLETFLDPRQLGGKLNDVTKEDLVKLIEVDGHEQ LFYPTFPVNVAFLRGTYADESGNITMDEEIGPFESTSVAQAVHNCGGKVVVQVKDVVAH GSLDPRMVKIPGIYVDYVVVAAPEDHQQTYDCEYDPSLSGEHRAPEGATDAALPMSAKK IIGRRGALELTENAVVNLGVGAPEYVASVAGEEGIADTITLTVEGGAIGGVPQGGARFG SSRNADAIIDHTYQFDFYDGGGLDIAYLGLAQCDGSGNINVSKFGTNVAGCGGFPNISQ QTPNVYFCGTFTAGGLKIAVEDGKVKILQEGKAKKFIKAVDQITFNGSYAARNGKHVLY ITERCVFELTKEGLKLIEVAPGIDIEKDILAHMDFKPIIDNPKLMDARLFQDGPMGLKK " terminator 4217..4248 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" terminator 4373..4467 /label=lambda t0 terminator /note="transcription terminator from phage lambda" rep_origin 4959..5181 /label=pSC101 ori /note="low-copy replication origin that requires the Rep101 protein" CDS 5229..6176 /label=Rep101 /note="RepA protein needed for replication with the pSC101 origin" CDS 6468..7325 /label=AmpR /note="beta-lactamase" terminator 7339..7358 /label=his operon terminator /note="This putative transcriptin terminator from the E. coli his operon has a 2-bp deletion introduced during synthesis. Its efficiency has not been determined."