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pLVX-EF1α-IRES-mCherry 质粒 (编号: V012682)

Note: pLVX-EF1α-IRES-mCherry is a lentiviral bicistronic expression vector for constitutively co-expressing a protein of interest. It includes a mCherry from the EF-1α promoter.

pLVX-EF1α-IRES-mCherry8904 bp400800120016002000240028003200360040004400480052005600600064006800720076008000840088003' LTRHIV-1 PsiRREgp41 peptideProtein TatcPPT/CTSEF-1-alpha promotermCherryWPRE5' LTRM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoterSV40 poly(A) signal
复制序列 NovoBuilder中查看图谱

基本信息

载体名称:
pLVX-EF1α-IRES-mCherry
载体抗性:
Ampicillin
载体长度:
8904 bp
载体类型:
Viral Expression & Packaging Vectors
复制子:
ori
载体来源:
Clontech
筛选标记:
mCherry
拷贝数:
High copy number
启动子:
EF-1α
克隆方法:
Enzyme Cut
5'引物:
EF1a Forward: TCAAGCCTCAGACAGTGGTTC
3'引物:
IRES-R: CCTCACATTGCCAAAAGACG
感受态:
stbl3
培养温度:
37℃
表达方法:
Constiutive, Stable

产品信息

质粒编号:V012682
质粒名称:pLVX-EF1α-IRES-mCherry
规格:5 μg (冻干粉)

下载资源

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，供学术研究使用。

确保质粒的关键元件正确，但是我们并不能保证实验效果。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。

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。

参考文献

  • Shi Y, Yuan J, Rraklli V, Maxymovitz E, Cipullo M, Liu M, Li S, Westerlund I, Bedoya-Reina OC, Bullova P, Rorbach J, Juhlin CC, Stenman A, Larsson C, Kogner P, O'Sullivan MJ, Schlisio S, Holmberg J. Aberrant splicing in neuroblastoma generates RNA-fusion transcripts and provides vulnerability to spliceosome inhibitors. Nucleic Acids Res. 2021 Mar 18;49(5):2509-2521.

pLVX-EF1α-IRES-mCherry 质粒 (编号: V012682)序列

LOCUS       pLVX-EF1α-IRES-m        8904 bp    DNA     circular SYN 26-DEC-2025
DEFINITION  Exported.
ACCESSION   V012682
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8904)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8904)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8904)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8904
                     /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
                     /label=Protein Tat
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    2027..2144
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2338..3519
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     misc_feature    3575..4152
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             4149..4856
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     misc_feature    4873..5461
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5668..6301
                     /label=5' LTR
                     /note="5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6429..6445)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6453..6469)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(6477..6507)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6522..6543)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6831..7419)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7593..8450)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8451..8555)
                     /label=AmpR promoter
     polyA_signal    8603..8737
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"