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:
- prHom-1
- 载体抗性:
- Ampicillin
- 载体长度:
- 5698 bp
- 载体类型:
- Mammalian Expression Vectors
- 复制子:
- ori
- 载体来源:
- Clontech
- 拷贝数:
- High copy number
- 启动子:
- SV40
prHom-1 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。
prHom-1 载体序列
LOCUS prHom-1. 5698 bp DNA circular SYN 01-JAN-1980 DEFINITION Mammalian vector encoding two FKBP homodimerizer domains, for creating cytoplasmic oligomers that can be dissociated with a drug. Also known as pC4EN-FM2E. ACCESSION . VERSION . KEYWORDS prHom-1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5698) AUTHORS Clontech TITLE Direct Submission REFERENCE 2 (bases 1 to 5698) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" COMMENT Clone into the XbaI site to fuse two copies of DmrD to the C-terminus of the partner protein. FEATURES Location/Qualifiers source 1..5698 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 19..322 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 323..526 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" 5'UTR 616..655 /label=HSV TK 5'-UTR /note="5' untranslated region from the herpes simplex virus thymidine kinase gene" CDS 676..678 /codon_start=1 /product="start codon" /label=start codon /note="ATG" /translation="M" CDS 688..1008 /label=DmrD /note="dimeric F36M mutant of FK506-binding protein FKBP12" CDS 1015..1335 /codon_start=1 /product="dimeric F36M mutant of FK506-binding protein FKBP12" /label=dimeric F36M mutant of FK506-binding protein FK /note="DmrD" /note="dimerization can be reversed by ligand binding" /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKMDSSRDRNKP FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL KLE" CDS 1342..1368 /label=HA /note="HA (human influenza hemagglutinin) epitope tag" intron 1400..1972 /label=beta-globin intron /note="intron from rabbit beta-globin gene" polyA_signal 2169..2224 /label=beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" rep_origin complement(2577..2712) /direction=LEFT /label=SV40 ori /note="SV40 origin of replication" rep_origin complement(2989..3577) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3748..4620) /codon_start=1 /product="beta-lactamase" /label=beta-lactamase /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFQVEVAMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDL VEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTR LDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAE IGASLIKHW" promoter complement(4621..4725) /label=AmpR promoter rep_origin complement(5057..5512) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 5657..5673 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 5680..5698 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase"