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JOURNAL Unpublished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"Unpublished"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"