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:
pNL4-3
载体抗性:
Ampicillin
载体长度:
14825 bp
载体类型:
HIV-1 vector
复制子:
ori
载体来源:
Adachi A, Gendelman HE, Koenig S, Folks T, Willey R, Rabson A, Martin MA.
感受态:
DH5alpha
培养温度:
37℃

pNL4-3 载体图谱

pNL4-314825 bp7001400210028003500420049005600630070007700840091009800105001120011900126001330014000147005' long terminal repeatHIV-1 PsiHIV-1 polvpr proteinEnvelope glycoprotein gp160Protein Nef3' LTR3' cellular genomic sequenceoriAmpRAmpR promoter5' cellular genomic sequence

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。

pNL4-3 载体序列

LOCUS       V004299                14825 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004299
VERSION     V004299
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 14825)
  AUTHORS   Adachi A, Gendelman HE, Koenig S, Folks T, Willey R, Rabson A,
            Martin MA.
  TITLE     Production of acquired immunodeficiency syndrome-associated
            retrovirus in human and nonhuman cells transfected with an
            infectious molecular clone
  JOURNAL   J. Virol. 59 (2), 284-291 (1986)
   PUBMED   3016298
REFERENCE   2  (bases 1 to 14825)
  AUTHORS   Bosche WJ, Poon DTK., Ott DE, Hu W-S., Gorelick RJ.
  TITLE     Complete Plasmid Sequence of pNL4-3
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 14825)
  AUTHORS   Bosche WJ, Poon DTK., Ott DE, Hu W-S., Gorelick RJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-NOV-2000) AIDS Vaccine Program, SAIC Frederick and
            Drug Resistance Program, National Cancer Institute at Frederick, PO
            Box B, Frederick, Maryland 21702-1202, USA
REFERENCE   4  (bases 1 to 14825)
  AUTHORS   Strebel KJ, Martin MA.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-MAY-2010) NIAID, Viral Biochemistry, National
            Institutes of Health, Bethesda, MD, USA
REFERENCE   5  (bases 1 to 14825)
  TITLE     Direct Submission
REFERENCE   6  (bases 1 to 14825)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. Virol.";
            date: "1986"; volume: "59"; issue: "2"; pages: "284-291"
            SGRef: number: 2; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
            (28-NOV-2000) AIDS Vaccine Program, SAIC Frederick and Drug
            Resistance Program, National Cancer Institute at Frederick, PO Box
            B, Frederick, Maryland 21702-1202, USA"
            SGRef: number: 4; type: "Journal Article"; journalName: "Submitted
            (24-MAY-2010) NIAID, Viral Biochemistry, National Institutes of
            Health, Bethesda, MD, USA"
            SGRef: number: 5; type: "Journal Article"
            On May 24, 2010 this sequence version replaced AF324493.1.
FEATURES             Location/Qualifiers
     source          1..14825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..634
                     /label="5' long terminal repeat"
                     /note="5' long terminal repeat"
     LTR             377..634
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     repeat_region   454..550
                     /note="R; 5' copy"
     misc_feature    681..806
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     CDS             790..2289
                     /label="HIV-1 gag"
                     /note="gag protein from human immunodeficiency virus 1"
     CDS             2085..5093
                     /label="HIV-1 pol"
                     /note="pol protein from human immunodeficiency virus 1"
     CDS             5041..5616
                     /gene="vif"
                     /label="Virion infectivity factor"
                     /note="Virion infectivity factor from Human
                     immunodeficiency virus type 1 group M subtype B (isolate
                     NY5). Accession#: P12504"
     CDS             5559..5849
                     /codon_start=1
                     /product="vpr protein"
                     /label="vpr protein"
                     /protein_id="AAK08485.1"
                     /translation="MEQAPEDQGPQREPYNEWTLELLEELKSEAVRHFPRIWLHNLGQH
                     IYETYGDTWAGVEAIIRILQQLLFIHFRIGCRHSRIGVTRQRRARNGASRS"
     misc_feature    5743..5748
                     /label="EcoRI site of recombination"
                     /note="EcoRI site of recombination"
     CDS             5830..6003
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate BH5). Accession#: P04612"
                     /label="Protein Tat"
     CDS             6221..8779
                     /gene="env"
                     /label="Envelope glycoprotein gp160"
                     /note="Envelope glycoprotein gp160 from Human
                     immunodeficiency virus type 1 group M subtype B (isolate
                     MFA). Accession#: P19551"
     CDS             8787..8813
                     /gene="nef"
                     /label="Protein Nef"
                     /note="Protein Nef from Human immunodeficiency virus type 1
                     group M subtype D (isolate Z84). Accession#: P12481"
     LTR             9076..9709
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    9710..11283
                     /label="3' cellular genomic sequence"
                     /note="3' cellular genomic sequence"
     rep_origin      complement(11520..12108)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(12282..13139)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(13140..13244)
                     /label="AmpR promoter"
     misc_feature    13648..14825
                     /label="5' cellular genomic sequence"
                     /note="5' cellular genomic sequence"