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:
- pVMG-TnpR
- 载体抗性:
- Ampicillin
- 载体长度:
- 7264 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Oke V, Long SR.
pVMG-TnpR 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。
pVMG-TnpR 载体序列
LOCUS V002230 7264 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V002230
VERSION V002230
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 7264)
AUTHORS Oke V, Long SR.
TITLE Bacterial genes induced within the nodule during the
Rhizobium-legume symbiosis
JOURNAL Mol. Microbiol. 32 (4), 837-849 (1999)
PUBMED 10361286
REFERENCE 2 (bases 1 to 7264)
AUTHORS Gao M, Teplitski M.
TITLE RIVET-a tool for in vivo analysis of symbiotically relevant gene
expression in Sinorhizobium meliloti
JOURNAL Mol. Plant Microbe Interact. 21 (2), 162-170 (2008)
PUBMED 18184060
REFERENCE 3 (bases 1 to 7264)
AUTHORS Gao M, Oke V, Teplitski M.
TITLE Direct Submission
JOURNAL Submitted (17-OCT-2007) Soil and Water Science, University of
REFERENCE 4 (bases 1 to 7264)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 7264)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol.
Microbiol."; date: "1999"; volume: "32"; issue: "4"; pages:
"837-849"
SGRef: number: 2; type: "Journal Article"; journalName: "Mol. Plant
Microbe Interact."; date: "2008"; volume: "21"; issue: "2"; pages:
"162-170"
SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(17-OCT-2007) Soil and Water Science, University of Florida-IFAS,
1376 Mowry Rd., Cancer "
SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7264
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 28..58
/label="Escherichia coli trpA"
/note="Escherichia coli trpA"
/regulatory_class="terminator"
misc_feature 103..113
/label="contains stop codons in all three reading frames"
/note="contains stop codons in all three reading frames"
regulatory 189..195
/gene="tnpR"
/regulatory_class="ribosome_binding_site"
CDS 205..753
/gene="tnpR"
/label="Transposon gamma-delta resolvase"
/note="Transposon gamma-delta resolvase from Escherichia
coli (strain K12). Accession#: P03012"
regulatory 756..761
/gene="uidA"
/regulatory_class="ribosome_binding_site"
CDS 788..2596
/label="GUS"
/note="beta-glucuronidase"
CDS 3019..3810
/label="NeoR/KanR"
/note="aminoglycoside phosphotransferase"
primer_bind complement(4183..4199)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 4412..4867
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
oriT complement(5206..5315)
/direction=LEFT
/label="oriT"
/note="incP origin of transfer"
promoter 5485..5589
/label="AmpR promoter"
CDS 5590..6447
/label="AmpR"
/note="beta-lactamase"
rep_origin 6621..7209
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"