质粒编号
质粒名称
规格
价格
V004299
pNL4-3
5 μg (冻干粉)
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学术研究使用，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。
Note:
- 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:
- stbl3
- 培养温度:
- 37℃
pNL4-3 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。
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JOURNAL Unpublished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"