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:
- pBS 35S hRluc-attR
- 载体抗性:
- Ampicillin
- 载体长度:
- 6781 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 宿主:
- Plants
- 载体来源:
- Subramanian C, Woo J, Cai X, Xu X, Servick S, Johnson CH, Nebenfuhr A, von Arnim AG.
- 启动子:
- CaMV 35S (enhanced)
pBS 35S hRluc-attR 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。
pBS 35S hRluc-attR 载体序列
LOCUS 40924_7091 6781 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pBS 35S hRluc-attR, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6781)
AUTHORS Subramanian C, Woo J, Cai X, Xu X, Servick S, Johnson CH, Nebenfuhr
A, von Arnim AG.
TITLE A suite of tools and application notes for in vivo protein
interaction assays using bioluminescence resonance energy transfer
(BRET)
JOURNAL Plant J. 48 (1), 138-152 (2006)
PUBMED 16925598
REFERENCE 2 (bases 1 to 6781)
AUTHORS von Arnim AG, Subramanian C.
TITLE Direct Submission
JOURNAL Submitted (30-MAR-2005) BCMB, University of Tennessee Knoxville,
1406 Circle Drive, Hesler Biology Bldg 618, Knoxville, TN 37996, USA
REFERENCE 3 (bases 1 to 6781)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6781)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant J.";
date: "2006"; volume: "48"; issue: "1"; pages: "138-152"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(30-MAR-2005) BCMB, University of Tennessee Knoxville, 1406 Circle
Drive, Hesler Biology Bldg 618, Knoxville, TN 37996, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6781
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(6..461)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 603..619
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 626..644
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 669..685
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
promoter 789..1460
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
regulatory 1460..1602
/note="translational enhancer; 5' UTR from tobacco etch
virus"
/regulatory_class="enhancer"
CDS 1610..2542
/label=hRluc
/note="Renilla luciferase"
protein_bind 2549..2673
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 2698..2728
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 2782..3438
/label=CmR
/note="chloramphenicol acetyltransferase"
CDS 3783..4085
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(4129..4253)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
regulatory 4324..4534
/label=35S cauliflower mosaic virus terminator
/note="35S cauliflower mosaic virus terminator"
/regulatory_class="terminator"
promoter complement(4596..4614)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(4635..4651)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4659..4675)
/label=lac operator
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4683..4713)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4728..4749)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5037..5625)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5799..6656)
/label=AmpR
/note="beta-lactamase"
promoter complement(6657..6761)
/label=AmpR promoter