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:
- pRZ106
- 载体抗性:
- Kanamycin
- 载体长度:
- 4985 bp
- 载体类型:
- Transposon donor vector
- 复制子:
- R6K γ ori
- 载体来源:
- Zordan RE, Beliveau BJ, Trow JA, Craig NL, Cormack BP.
pRZ106 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。
pRZ106 载体序列
LOCUS V003532 4985 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V003532
VERSION V003532
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 4985)
AUTHORS Zordan RE, Beliveau BJ, Trow JA, Craig NL, Cormack BP.
TITLE Avoiding the ends: internal epitope tagging of proteins using
transposon Tn7
JOURNAL Genetics 200 (1), 47-58 (2015)
PUBMED 25745023
REFERENCE 2 (bases 1 to 4985)
AUTHORS Zordan RE, Cormack BP.
TITLE Direct Submission
JOURNAL Submitted (24-JAN-2015) Molecular Biology and Genetics, Johns
Hopkins School of Medicine, 725 N Wolfe St., Hunterian 609,
Baltimore, MD 21205, USA
REFERENCE 3 (bases 1 to 4985)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4985)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Genetics";
date: "2015"; volume: "200"; issue: "1"; pages: "47-58"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(24-JAN-2015) Molecular Biology and Genetics, Johns Hopkins School
of Medicine, 725 N Wolfe St., Hunterian 609, Baltimore, MD 21205,
USA"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4985
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 361..1152
/label="NeoR/KanR"
/note="aminoglycoside phosphotransferase"
protein_bind 1430..1451
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1466..1496
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 1504..1520
/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 1528..1544
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 1820..1849
/label="linker 1"
/note="linker 1"
CDS 1850..2563
/label="yeGFP"
/note="yeast-enhanced green fluorescent protein"
misc_feature 2564..2593
/label="linker 2"
/note="linker 2"
CDS 2772..3242
/gene="dhfrI"
/label="Dihydrofolate reductase type 1"
/note="Dihydrofolate reductase type 1 from Escherichia
coli. Accession#: P00382"
primer_bind complement(3473..3489)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(3924..4292)
/label="traJ"
/note="oriT-recognizing protein"
oriT complement(4325..4434)
/direction=LEFT
/label="oriT"
/note="incP origin of transfer"
rep_origin 4594..4982
/label="R6K gamma ori"
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"