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:
- pXUN
- 载体抗性:
- Ampicillin
- 载体长度:
- 5632 bp
- 载体类型:
- Transient expression vector
- 复制子:
- ori
- 载体来源:
- Chen S, Songkumarn P, Liu J, Wang GL.
- 启动子:
- Ubi
pXUN 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。
pXUN 载体序列
LOCUS 40924_47368 5632 bp DNA circular SYN 18-DEC-2018
DEFINITION Transient expression vector pXUN, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5632)
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 5632)
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 5632)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5632)
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..5632
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator complement(9..261)
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS 622..924
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
promoter complement(995..2985)
/label=Ubi promoter
/note="maize polyubiquitin gene promoter"
primer_bind complement(3016..3032)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3040..3056)
/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(3064..3094)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3109..3130)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3418..4006)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4180..5037)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5038..5142)
/label=AmpR promoter
primer_bind 5616..5632
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"