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:
- pXNS2pat-YFP
- 载体抗性:
- Ampicillin
- 载体长度:
- 6000 bp
- 载体类型:
- Plant binary vector
- 复制子:
- oriV
- 载体来源:
- Dahncke K, Witte CP.
pXNS2pat-YFP 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。
pXNS2pat-YFP 载体序列
LOCUS 40924_47243 6000 bp DNA circular SYN 18-DEC-2018 DEFINITION Plant binary vector pXNS2pat-YFP, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6000) AUTHORS Dahncke K, Witte CP. TITLE Plant purine nucleoside catabolism employs a guanosine deaminase required for the generation of xanthosine in Arabidopsis JOURNAL Plant Cell 25 (10), 4101-4109 (2013) PUBMED 24130159 REFERENCE 2 (bases 1 to 6000) AUTHORS Dahncke K, Witte C-P. TITLE Direct Submission JOURNAL Submitted (05-AUG-2013) Department of Plant Biochemistry, Freie Universitaet Berlin, Dahlem Centre of Plant Sciences, Koenigin-Luise-Str. 12-16, Berlin 14195, Germany REFERENCE 3 (bases 1 to 6000) TITLE Direct Submission REFERENCE 4 (bases 1 to 6000) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant Cell"; date: "2013"; volume: "25"; issue: "10"; pages: "4101-4109" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (05-AUG-2013) Department of Plant Biochemistry, Freie Universitaet Berlin, Dahlem Centre of Plant Sciences, Koenigin-Luise-Str. 12-16, Berlin 14195, Germany" COMMENT SGRef: number: 3; type: "Journal Article" COMMENT ##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..6000 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 30..746 /codon_start=1 /label=EYFP /note="enhanced YFP" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(778..794) /label=SK primer /note="common sequencing primer, one of multiple similar variants" misc_feature 1077..1101 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" rep_origin 1213..1922 /label=oriV /note="incP origin of replication" promoter 1996..2087 /label=AmpR promoter CDS 2088..2945 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 3119..3707 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature 4019..4043 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" terminator complement(4137..4389) /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" CDS complement(4412..4960) /codon_start=1 /label=BlpR /note="phosphinothricin acetyltransferase" /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF WQLDFSLPVPPRPVLPVTEI" promoter complement(5003..5182) /label=NOS promoter /note="nopaline synthase promoter" promoter 5326..5999 /label=CaMV 35S promoter (enhanced) /note="cauliflower mosaic virus 35S promoter with a duplicated enhancer region"