Ganesh-R1 载体 (V009965)

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:
Ganesh-R1
载体抗性:
Ampicillin
载体长度:
11450 bp
载体类型:
Reporter vector
复制子:
ori
载体来源:
Swanson CI, Hinrichs T, Johnson LA, Zhao Y, Barolo S.
启动子:
lac UV5

Ganesh-R1 载体图谱

Ganesh-R111450 bp500100015002000250030003500400045005000550060006500700075008000850090009500100001050011000P element 3' endwhiteattR1lac UV5 promoterCmRccdBattR2spacer sequence includes partial KanR geneDrosophila melanogaster Hsp70 minimal promoterDsRed-ExpressDrosophila melanogaster tra gene NLSSV40 poly(A) signalP element 5' endoriAmpRAmpR promoter

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。

Ganesh-R1 载体序列

LOCUS       40924_1014       11450 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Reporter vector Ganesh-R1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11450)
  AUTHORS   Swanson CI, Hinrichs T, Johnson LA, Zhao Y, Barolo S.
  TITLE     A directional recombination cloning system for restriction- and 
            ligation-free construction of GFP, DsRed, and lacZ transgenic 
            Drosophila reporters
  JOURNAL   Gene 408 (1-2), 180-186 (2008)
  PUBMED    18077106
REFERENCE   2  (bases 1 to 11450)
  AUTHORS   Rogers CI, Hinrichs T, Johnson LA, Zhao Y, Barolo S.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 11450)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 11450)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene 408 
            (1-2), 180-186 (2008)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-FEB-2007) Cell "
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(1..233)
                     /label=P element 3' end
                     /note="P element 3' end"
     gene            242..4247
                     /gene="white"
                     /allele="+mC"
                     /label=white
     protein_bind    4255..4379
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        4404..4434
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             4488..5144
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             5489..5791
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(5835..5959)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    5965..6712
                     /label=spacer sequence includes partial KanR gene
                     /note="spacer sequence includes partial KanR gene"
     regulatory      6713..6846
                     /label=Drosophila melanogaster Hsp70 minimal promoter
                     /note="Drosophila melanogaster Hsp70 minimal promoter"
                     /regulatory_class="promoter"
     CDS             6920..7594
                     /label=DsRed-Express
                     /note="rapidly maturing tetrameric variant of DsRed
                     fluorescent protein (Bevis and Glick, 2002)"
     misc_feature    7598..7714
                     /gene="DsRed.T4-NLS-SV40t"
                     /label=Drosophila melanogaster tra gene NLS
                     /note="Drosophila melanogaster tra gene NLS"
     polyA_signal    7852..7986
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    complement(8105..8690)
                     /label=P element 5' end
     rep_origin      complement(8925..9513)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9687..10544)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(10545..10649)
                     /label=AmpR promoter