质粒编号
质粒名称
规格
价格
V008081
pCXUN
5 μg (冻干粉)
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学术研究使用，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。
Note:
- 载体名称:
- pCXUN
- 载体抗性:
- Kanamycin
- 载体长度:
- 12008 bp
- 载体类型:
- Binary vector
- 复制子:
- ori
- 载体来源:
- Chen S, Songkumarn P, Liu J, Wang GL.
- 启动子:
- Ubi
pCXUN 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。
pCXUN 载体序列
LOCUS 40924_14055 12008 bp DNA circular SYN 17-DEC-2018
DEFINITION Binary vector pCXUN, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12008)
AUTHORS Chen S, Songkumarn P, Liu J, Wang GL.
TITLE A versatile zero background T-vector system for gene cloning and
functional genomics
JOURNAL Plant Physiol. 150 (3), 1111-1121 (2009)
PUBMED 19403729
REFERENCE 2 (bases 1 to 12008)
AUTHORS Chen S, Songkumarn P, Liu J, Wang G-L.
TITLE Direct Submission
JOURNAL Submitted (09-APR-2009) Department of Plant Pathology, The Ohio
State University, 201 Kottman Hall, 2021 Coffey Rd, Columbus, OH
43210, USA
REFERENCE 3 (bases 1 to 12008)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 12008)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant
Physiol."; date: "2009"; volume: "150"; issue: "3"; pages:
"1111-1121"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(09-APR-2009) Department of Plant Pathology, The Ohio State
University, 201 Kottman Hall, 2021 Coffey Rd, Columbus, OH 43210,
USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..12008
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 90..106
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
terminator complement(115..367)
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(442..744)
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
regulatory complement(1101..3091)
/label=maize ubiquitin-1 promoter
/note="maize ubiquitin-1 promoter"
/regulatory_class="promoter"
promoter complement(1101..3091)
/label=Ubi promoter
/note="maize polyubiquitin gene promoter"
primer_bind complement(3113..3129)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 3332..3356
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 4655..5281
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
CDS 5718..6782
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
rep_origin 6851..7045
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 7389..7529
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(7715..8303)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8393..9184)
/label=KanR
/note="aminoglycoside phosphotransferase"
misc_feature 9609..9633
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal complement(9711..9885)
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(9928..10950)
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
promoter complement(11017..11694)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
protein_bind 11885..11906
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 11921..11951
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 11959..11975
/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 11983..11999
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"