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pCDH-CMV-MCS-EF1-Puro 质粒 (编号: V006738)

Note: The pCDH-CMV-MCS-EF1-Puro is a lentiviral expression plasmid. It includes a CMV promoter for high-level expression of a gene cloned into its multiple cloning site (MCS) and an EF1 promoter for a puromycin resistance gene, enabling stable cell line generation

pCDH-CMV-MCS-EF1-Puro7403 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300660069007200RSV promoter5' LTR (truncated)HIV-1 PsiRREgp41 peptideProtein TatcPPT/CTSCMV promoterEF-1-alpha core promoter5' LTR (truncated)PuroRWPRE3' LTR (Delta-U3)SV40 poly(A) signalSV40 oriM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoterM13 fwd
复制序列 NovoBuilder中查看图谱

基本信息

载体名称:
pCDH-CMV-MCS-EF1-Puro
载体抗性:
Ampicillin
载体长度:
7403 bp
载体类型:
Lentiviral vectors
复制子:
ori
筛选标记:
Puromycin
启动子:
RSV
克隆方法:
Enzyme digestion and ligation
5'引物:
CMV-F: CGC AAA TGG GCG GTA GGC GTG
感受态:
stbl3

产品信息

质粒编号:V006738
质粒名称:pCDH-CMV-MCS-EF1-Puro
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

  • Shuwen H, Yinhang W, Jing M, Gong C, Xiaohui H, Xi Y, Wei W. Open Reading Frame-3a gene of the 2019 novel coronavirus inhibits the occurrence and development of colorectal cancer. Discov Oncol. 2022 Mar 20;13(1):14. doi: 10.1007/s12672-022-00473-6. PMID: 35306605; PMCID: PMC8934246.

  • Fan T-J, Xie C, Li L, Jin X, Cui J, Wang J-H. HIV-1 Nef activates proviral DNA transcription by recruiting Src kinase to phosphorylate host protein Nef-associated factor 1 to compromise its viral restrictive function. J Virol. 2025 May 20;99(5):e0028025. doi: 10.1128/jvi.00280-25. Epub 2025 Apr 24. PMID: 40272155; PMCID: PMC12090801.

pCDH-CMV-MCS-EF1-Puro 质粒 (编号: V006738)序列

LOCUS       pCDH-CMV-MCS-EF1        7403 bp    DNA     circular SYN 26-DEC-2025
DEFINITION  Exported.
ACCESSION   V006738
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7403)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7403)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7403)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          4611..4631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..232
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             233..413
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    460..585
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1078..1311
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1496..1540
                     /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             1689..1730
                     /label=Protein Tat
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    1807..1923
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2013..2216
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        2357..2568
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     LTR             2581..2849
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2876..3472
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    3482..4070
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             4144..4377
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4449..4583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      4589..4745
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4779..4794)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4802..4818)
                     /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(4826..4856)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4871..4892)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5180..5768)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5942..6799)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6800..6904)
                     /label=AmpR promoter
     primer_bind     7378..7394
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"