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:
- pdLucFXR
- 载体抗性:
- Ampicillin
- 载体长度:
- 10961 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Houten SM, Volle DH, Cummins CL, Mangelsdorf DJ, Auwerx J.
pdLucFXR 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。
pdLucFXR 载体序列
LOCUS 40924_14755 10961 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pdLucFXR, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10961)
AUTHORS Houten SM, Volle DH, Cummins CL, Mangelsdorf DJ, Auwerx J.
TITLE In vivo imaging of farnesoid X receptor activity reveals the ileum
as the primary bile acid signaling tissue
JOURNAL Mol. Endocrinol. 21 (6), 1312-1323 (2007)
PUBMED 17426284
REFERENCE 2 (bases 1 to 10961)
AUTHORS Houten SM, Auwerx J.
TITLE Direct Submission
JOURNAL Submitted (21-APR-2004) Institut de Genetique et de Biologie
Moleculaire et Cellulaire, 1 rue Laurent Fries, Illkirch 67404,
France
REFERENCE 3 (bases 1 to 10961)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10961)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol.
Endocrinol."; date: "2007"; volume: "21"; issue: "6"; pages:
"1312-1323"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(21-APR-2004) Institut de Genetique et de Biologie Moleculaire et
Cellulaire, 1 rue Laurent Fries, Illkirch 67404, France"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10961
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(24..40)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 181..636
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 714..818
/label=AmpR promoter
CDS 819..1676
/label=AmpR
/note="beta-lactamase"
rep_origin 1850..2438
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2726..2747
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2762..2792
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2800..2816
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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 2824..2840
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 2858..2876
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
misc_feature 2902..4124
/label=beta-globin insulator
/note="beta-globin insulator"
misc_feature 4125..5334
/label=beta-globin insulator
/note="beta-globin insulator"
protein_bind 5385..5479
/label=5X UAS
/note="five tandem copies of the 'ScaI site' 17-mer
CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS)
that efficiently binds yeast Gal4 (Webster et al., 1988;
Pfeiffer et al., 2010)"
protein_bind 5510..5595
/label=FXR binding site
/bound_moiety="FXR"
regulatory 5614..6723
/label=Wnt-1 promoter
/note="Wnt-1 promoter"
/regulatory_class="promoter"
CDS 6765..8414
/label=luciferase
/note="firefly luciferase"
polyA_signal complement(8611..8732)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
misc_feature 8844..10934
/label=3' flanking sequence of Wnt-1
/note="3' flanking sequence of Wnt-1"