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:
- pCXUN-HA
- 载体抗性:
- Kanamycin
- 载体长度:
- 12038 bp
- 载体类型:
- Binary vector
- 复制子:
- ori
- 载体来源:
- Chen S, Songkumarn P, Liu J, Wang GL.
- 启动子:
- Ubi
pCXUN-HA 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。
pCXUN-HA 载体序列
LOCUS 40924_14045 12038 bp DNA circular SYN 17-DEC-2018 DEFINITION Binary vector pCXUN-HA, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12038) AUTHORS Chen S, Songkumarn P, Liu J, Wang GL. TITLE A versatile zero background T-vector system for gene cloning and functional genomics JOURNAL Plant Physiol. 150 (3), 1111-1121 (2009) PUBMED 19403729 REFERENCE 2 (bases 1 to 12038) AUTHORS Chen S, Songkumarn P, Liu J, Wang G-L. TITLE Direct Submission JOURNAL Submitted (09-APR-2009) Department of Plant Pathology, The Ohio State University, 201 Kottman Hall, 2021 Coffey Rd, Columbus, OH 43210, USA REFERENCE 3 (bases 1 to 12038) TITLE Direct Submission REFERENCE 4 (bases 1 to 12038) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant Physiol."; date: "2009"; volume: "150"; issue: "3"; pages: "1111-1121" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (09-APR-2009) Department of Plant Pathology, The Ohio State University, 201 Kottman Hall, 2021 Coffey Rd, Columbus, OH 43210, USA" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..12038 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 90..106 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" terminator complement(115..367) /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" CDS 728..1030 /label=ccdB /note="CcdB, a bacterial toxin that poisons DNA gyrase" CDS complement(1089..1115) /label=HA /note="HA (human influenza hemagglutinin) epitope tag" regulatory complement(1131..3121) /label=maize ubiquitin-1 promoter /note="maize ubiquitin-1 promoter" /regulatory_class="promoter" promoter complement(1131..3121) /label=Ubi promoter /note="maize polyubiquitin gene promoter" primer_bind complement(3143..3159) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" misc_feature 3362..3386 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 4685..5311 /label=pVS1 StaA /note="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS 5748..6812 /label=pVS1 RepA /note="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" rep_origin 6881..7075 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" misc_feature 7419..7559 /label=bom /note="basis of mobility region from pBR322" rep_origin complement(7745..8333) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(8423..9214) /label=KanR /note="aminoglycoside phosphotransferase" misc_feature 9639..9663 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" polyA_signal complement(9741..9915) /label=CaMV poly(A) signal /note="cauliflower mosaic virus polyadenylation signal" CDS complement(9958..10980) /label=HygR /note="aminoglycoside phosphotransferase from E. coli" promoter complement(11047..11724) /label=CaMV 35S promoter (enhanced) /note="cauliflower mosaic virus 35S promoter with a duplicated enhancer region" protein_bind 11915..11936 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 11951..11981 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 11989..12005 /label=lac operator /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 12013..12029 /label=M13 rev /note="common sequencing primer, one of multiple similar variants"