pFR-Luc 载体 (V018103)

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。

pFR-Luc is a mammalian reporter plasmid designed to study transcriptional regulation. It contains the firefly luciferase gene under the control of a minimal promoter with multiple repeats of specific response elements (e.g., Gal4 binding sites), allowing detection of transcription factor activity or promoter activation. The plasmid typically includes an antibiotic resistance marker (e.g., ampicillin) for selection and is widely used in signal transduction assays, promoter analysis, and high-throughput screening of regulatory elements.

载体名称:
pFR-Luc
载体抗性:
Ampicillin
载体长度:
5748 bp
载体类型:
Cloning vector
载体来源:
Zheng C.-F

pFR-Luc 载体图谱

pFR-Luc5748 bp60012001800240030003600420048005400ampicillin resistance proteinCol EI originfive 17nt GAL4 binding elementsregulatoryluciferase

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。

pFR-Luc 载体序列

LOCUS       AF058756                5748 bp    DNA     linear   SYN 26-JUL-2016
DEFINITION  Cloning vector pFR-Luc, complete sequence.
ACCESSION   AF058756
VERSION     AF058756.1
KEYWORDS    .
SOURCE      Cloning vector pFR-Luc
  ORGANISM  Cloning vector pFR-Luc
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 5748)
  AUTHORS   Zheng,C.-F.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-APR-1998) Technical Services, Stratagene, 11011 N.
            Torrey Pines Rd., La Jolla, CA 92037, USA
REFERENCE   2  (bases 1 to 5748)
  AUTHORS   Zheng,C.-F.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-OCT-1998) Technical Services, Stratagene, 11011 N.
            Torrey Pines Rd., La Jolla, CA 92037, USA
  REMARK    Sequence update by submitter
REFERENCE   3  (bases 1 to 5748)
  AUTHORS   Zheng,C.-F.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-JAN-1999) Technical Services, Stratagene, 11011 N.
            Torrey Pines Rd., La Jolla, CA 92037, USA
  REMARK    Sequence update by submitter
COMMENT     On Jan 24, 1999 this sequence version replaced gi:3769433.
FEATURES             Location/Qualifiers
     source          1..5748
                     /organism="Cloning vector pFR-Luc"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:75696"
                     /lab_host="Escherichia coli K12"
                     /plasmid="pFR-Luc"
     CDS             137..997
                     /codon_start=1
                     /transl_table=11
                     /product="ampicillin resistance protein"
                     /protein_id="AAD08914.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     rep_origin      998..1930
                     /note="Col EI origin"
     repeat_region   2692..2785
                     /note="five 17nt GAL4 binding elements"
                     /rpt_type=tandem
     regulatory      2804..2809
                     /regulatory_class="TATA_box"
     CDS             2868..4520
                     /note="from Photinus pyralis"
                     /codon_start=1
                     /transl_table=11
                     /product="luciferase"
                     /protein_id="AAD08913.1"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTD
                     AHIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAV
                     APANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQG
                     FQSMYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVR
                     FSHARDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRS
                     LQDYKIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLP
                     GIRQGYGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCV
                     RGPMIMSGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVA
                     PAELESILLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTT
                     AKKLRGGVVFVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"