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:
- pHR_pGK_LaG17_synNotch_Gal4VP64
- 载体抗性:
- Ampicillin
- 载体长度:
- 11116 bp
- 载体类型:
- Lentiviral
- 复制子:
- ori
- 拷贝数:
- High Copy
- 启动子:
- mPGK
pHR_pGK_LaG17_synNotch_Gal4VP64 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。
pHR_pGK_LaG17_synNotch_Gal4VP64 载体序列
LOCUS V007389 11116 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V007389
VERSION V007389
KEYWORDS pHR_pGK_LaG17_synNotch_Gal4VP64
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11116)
AUTHORS Morsut L, Roybal KT, Xiong X, Gordley RM, Coyle SM, Thomson M, Lim
WA
TITLE Engineering Customized Cell Sensing and Response Behaviors Using
Synthetic Notch Receptors.
JOURNAL Cell. 2016 Feb 11;164(4):780-91. doi: 10.1016/j.cell.2016.01.012.
Epub 2016 Jan 28.
PUBMED 26830878
REFERENCE 2 (bases 1 to 11116)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11116)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.cell.2016.01.012"; journalName: "Cell"; date:
"2016-02-11- 11"; volume: "164"; issue: "4"; pages: "780-91"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11116
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 1..634
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1721..1765
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
CDS 1914..1955
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2050..2167
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2227..2726
/label="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
regulatory 2774..2783
/label="Kozak sequence"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 2843..2872
/label="Myc"
/note="Myc (human c-Myc proto-oncogene) epitope tag"
CDS 4235..4675
/label="GAL4 DNA binding domain"
/note="DNA binding domain of the GAL4 transcriptional
activator"
CDS 4712..4861
/label="VP64"
/note="tetrameric repeat of the minimal activation domain
of herpes simplex virus VP16 (Beerli et al., 1998)"
misc_feature 4980..5568
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5571..5587)
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5572..5588)
/label="pBluescriptKS"
/note="For pBluescript vector"
LTR 5678..5911
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
promoter complement(5936..5954)
/label="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(6244..6263)
/label="pBRrevBam"
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(6477..6496)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6596..6618
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
primer_bind complement(6656..6674)
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6742..6846
/label="AmpR promoter"
CDS 6847..7704
/label="AmpR"
/note="beta-lactamase"
rep_origin 7878..8466
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8620..8637
/label="L4440"
/note="L4440 vector, forward primer"
promoter 8712..9041
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
intron 10175..10240
/label="small t intron"
/note="SV40 (simian virus 40) small t antigen intron"
CDS 10370..10390
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 10815..10949
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"