pGL3-3'UTR 载体 (V005972)

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:
pGL3-3'UTR
载体抗性:
Ampicillin
载体长度:
6248 bp
载体类型:
Reporter vector
复制子:
ori
载体来源:
Bi Y, Qiao X, Hua Z, Zhang L, Liu X, Li L, Hua W, Xiao H, Zhou J, Wei Q, Zheng X.

pGL3-3'UTR 载体图谱

pGL3-3'UTR6248 bp30060090012001500180021002400270030003300360039004200450048005100540057006000SV40 promoterluciferaseregulatoryoriAmpRAmpR promoterf1 oripoly(A) signalpause site

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。

pGL3-3'UTR 载体序列

LOCUS       40924_21948        6248 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Reporter vector pGL3-3'UTR, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6248)
  AUTHORS   Bi Y, Qiao X, Hua Z, Zhang L, Liu X, Li L, Hua W, Xiao H, Zhou J, 
            Wei Q, Zheng X.
  TITLE     An asymmetric PCR-based, reliable and rapid single-tube native DNA 
            engineering strategy
  JOURNAL   BMC Biotechnol. 12, 39 (2012)
  PUBMED    22768962
REFERENCE   2  (bases 1 to 6248)
  AUTHORS   Bi Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-AUG-2011) Biotechnology Group, Institute of Animal 
            Science and Veterinary, Hubei Academy of Agricultural Science, No. 
            1, Nanhu Yaoyuan, Hongshan District, Wuhan, Hubei 430064, P. R. 
            China
REFERENCE   3  (bases 1 to 6248)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6248)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC 
            Biotechnol. 12, 39 (2012)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (03-AUG-2011) Biotechnology Group, Institute of Animal Science and 
            Veterinary, Hubei Academy of Agricultural Science, No. 1, Nanhu 
            Yaoyuan, Hongshan District, Wuhan, Hubei 430064, P. R. China"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..244
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     CDS             280..1929
                     /label=luciferase
                     /note="firefly luciferase"
     regulatory      1988..3433
                     /regulatory_class="terminator"
     regulatory      2170..2175
                     /regulatory_class="polyA_signal_sequence"
     regulatory      2584..2589
                     /regulatory_class="polyA_signal_sequence"
     regulatory      2908..2913
                     /regulatory_class="polyA_signal_sequence"
     regulatory      3269..3274
                     /regulatory_class="polyA_signal_sequence"
     rep_origin      complement(3751..4339)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4513..5370)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5371..5475)
                     /label=AmpR promoter
     rep_origin      5502..5957
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     polyA_signal    6088..6136
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     misc_feature    6150..6241
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"