pRED-PT4exp 载体 (V003814)

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

pRED-PT4exp12675 bp6001200180024003000360042004800540060006600720078008400900096001020010800114001200012600CaMV 35S promoterNeoR/KanRregulatoryNOS terminatorDsRed1ubiquitin 10 promoter from Arabidopsis thalianavertebrate consensus sequence for strong initiation of translation (Kozak, 1987)MtPt4 promoter from Medicago truncatularegulatoryRB T-DNA repeatoribompVS1 StaApVS1 RepApVS1 oriVSmRLB T-DNA repeat

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  JOURNAL   Unpublished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            "Unpublished"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"