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:
- pRED-PT4exp
- 载体抗性:
- Kanamycin
- 载体长度:
- 12675 bp
- 载体类型:
- Binary vector
- 复制子:
- ori
- 宿主:
- Plants
- 载体来源:
- Krajinski F.
- 启动子:
- CaMV 35S
pRED-PT4exp 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。
pRED-PT4exp 载体序列
LOCUS 40924_36453 12675 bp DNA circular SYN 18-DEC-2018 DEFINITION Binary vector pRED-PT4exp, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12675) AUTHORS Krajinski F. TITLE A vector to express artificial microRNAs in Medicago truncatula JOURNAL Unpublished REFERENCE 2 (bases 1 to 12675) AUTHORS Krajinski F. TITLE Direct Submission JOURNAL Submitted (05-JUL-2012) Max Planck Institute of Molecular Plant Physiology, Am Muehlenberg 1, Potsdam (OT) Golm 14476, Germany REFERENCE 3 (bases 1 to 12675) TITLE Direct Submission REFERENCE 4 (bases 1 to 12675) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (05-JUL-2012) Max Planck Institute of Molecular Plant Physiology, Am Muehlenberg 1, Potsdam (OT) Golm 14476, Germany" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..12675 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 80..424 /label=CaMV 35S promoter /note="strong constitutive promoter from cauliflower mosaic virus" CDS 439..1239 /label=NeoR/KanR /note="aminoglycoside phosphotransferase from Tn5" regulatory 1255..1456 /regulatory_class="terminator" terminator complement(1469..1721) /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" CDS complement(1737..2414) /label=DsRed1 /note="wild-type DsRed" regulatory complement(2428..3907) /label=ubiquitin 10 promoter from Arabidopsis thaliana /note="ubiquitin 10 promoter from Arabidopsis thaliana" /regulatory_class="promoter" regulatory 3945..3954 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" regulatory complement(3956..4822) /label=MtPt4 promoter from Medicago truncatula /note="MtPt4 promoter from Medicago truncatula" /regulatory_class="promoter" regulatory complement(4870..5065) /regulatory_class="terminator" misc_feature 5149..5173 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" rep_origin 5818..6406 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature complement(6591..6731) /label=bom /note="basis of mobility region from pBR322" CDS 8066..8692 /label=pVS1 StaA /note="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS 9124..10194 /label=pVS1 RepA /note="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" rep_origin 10263..10457 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS 11062..11850 /label=SmR /note="aminoglycoside adenylyltransferase (Murphy, 1985)" misc_feature 12373..12397 /label=LB T-DNA repeat /note="left border repeat from octopine Ach5 T-DNA"