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:
- pUB-DEST
- 载体抗性:
- Streptomycin
- 载体长度:
- 10818 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 宿主:
- Plants
- 载体来源:
- Grefen C, Donald N, Hashimoto K, Kudla J, Schumacher K, Blatt MR.
- 启动子:
- NOS
pUB-DEST 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。
pUB-DEST 载体序列
LOCUS 40924_44799 10818 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pUB-DEST, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10818)
AUTHORS Grefen C, Donald N, Hashimoto K, Kudla J, Schumacher K, Blatt MR.
TITLE A ubiquitin-10 promoter-based vector set for fluorescent protein
tagging facilitates temporal stability and native protein
distribution in transient and stable expression studies
JOURNAL Plant J. 64 (2), 355-365 (2010)
PUBMED 20735773
REFERENCE 2 (bases 1 to 10818)
AUTHORS Grefen C, Donald N, Hashimoto K, Kudla J, Schumacher K, Blatt MR.
TITLE Direct Submission
JOURNAL Submitted (27-NOV-2012) Plant Sciences Group, University of Glasgow,
University Ave, Glasgow G12 8QQ, United Kingdom
REFERENCE 3 (bases 1 to 10818)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10818)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant J.";
date: "2010"; volume: "64"; issue: "2"; pages: "355-365"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(27-NOV-2012) Plant Sciences Group, University of Glasgow,
University Ave, Glasgow G12 8QQ, United Kingdom"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10818
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 1..639
/note="ubiquitin 10 promoter from Arabidopsis thaliana
Columbia-6 ecotype (At4g05050)"
/regulatory_class="promoter"
protein_bind 668..792
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 817..847
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 901..1557
/label=CmR
/note="chloramphenicol acetyltransferase"
CDS 1902..2204
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(2248..2372)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
regulatory 2449..2657
/label=T35S terminator
/note="T35S terminator"
/regulatory_class="terminator"
promoter 2794..2973
/label=NOS promoter
/note="nopaline synthase promoter"
CDS 3015..3563
/label=BlpR
/note="phosphinothricin acetyltransferase"
terminator 3586..3838
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
misc_feature complement(3931..3955)
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 4476..5264
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
rep_origin 5514..6102
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(6288..6428)
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(6772..6966)
/direction=LEFT
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(7035..8099)
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(8536..9162)
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
misc_feature complement(10492..10516)
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
protein_bind 10681..10702
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 10717..10747
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 10755..10771
/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)."
primer_bind 10779..10795
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"