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:
pDV-NYFP-CTAP
载体抗性:
Ampicillin
载体长度:
8021 bp
载体类型:
Cloning vector
复制子:
ori
载体来源:
Meima ME, Weening KE, Schaap P.

pDV-NYFP-CTAP 载体图谱

pDV-NYFP-CTAP8021 bp400800120016002000240028003200360040004400480052005600600064006800720076008000EYFPmultipe cloning siteCBPTEV siteProtAProtA2H3 terminatorNeoR/KanRAmpR promoterAmpRoriactin15 promoter

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。

pDV-NYFP-CTAP 载体序列

LOCUS       40924_16375        8021 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pDV-NYFP-CTAP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8021)
  AUTHORS   Meima ME, Weening KE, Schaap P.
  TITLE     Vectors for expression of proteins with single or combinatorial 
            fluorescent protein and tandem affinity purification tags in 
            Dictyostelium
  JOURNAL   Protein Expr. Purif. 53 (2), 283-288 (2007)
  PUBMED    17296313
REFERENCE   2  (bases 1 to 8021)
  AUTHORS   Meima ME, Weening KE, Schaap P.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-SEP-2006) School of Life Sciences, University of 
            Dundee, Dow Street, Dundee DD1 5EH, United Kingdom
REFERENCE   3  (bases 1 to 8021)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8021)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein 
            Expr. Purif."; date: "2007"; volume: "53"; issue: "2"; pages: 
            "283-288"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-SEP-2006) School of Life Sciences, University of Dundee, Dow 
            Street, Dundee DD1 5EH, United Kingdom"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             24..740
                     /label=EYFP
                     /note="enhanced YFP"
     misc_feature    741..764
                     /label=multipe cloning site
                     /note="multipe cloning site"
     CDS             768..845
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             873..893
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     CDS             933..1106
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             1110..1280
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNGAQAPK"
     regulatory      1331..2311
                     /label=2H3 terminator
                     /note="2H3 terminator"
                     /regulatory_class="terminator"
     CDS             3080..3871
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        4764..4868
                     /label=AmpR promoter
     CDS             4869..5726
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5900..6488
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      7713..8021
                     /label=actin15 promoter
                     /note="actin15 promoter"
                     /regulatory_class="promoter"