质粒类型：慢病毒载体

高拷贝/低拷贝：高拷贝

启动子：CMV

克隆方法：多克隆位点，限制性内切酶

载体大小：8269bp

5' 测序引物及序列：CMV-F: CGCAAATGGGCGGTAGGCGTG(Invitrogen)

3' 测序引物及序列：IRES-R: CCTCACATTGCCAAAAGACG

载体抗性：Ampicillin (氨苄青霉素)

筛选标记：新霉素

pLVX-IRES-Neo is an HIV-1-based, lentiviral expression vector that allows the simultaneous expression of your protein of interest and G418 resistance (Neor ) in virtually any mammalian cell type, including primary cells. The vector expresses your protein of interest and Neor from a bicistronic mRNA transcript, allowing G418 resistance to be used as an indicator of transduction efficiency and a marker for selection.

Expression of the bicistronic transcript is driven by the constitutively active human cytomegalovirus immediate early promoter (PCMV IE) located just upstream of the MCS. An encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES), positioned between the MCS and Neor , facilitates cap-independent translation of Neor from an internal start site at the IRES/Neor junction (1).

pLVX-IRES-Neo contains all of the viral processing elements necessary for the production of replication-incompetent lentivirus, as well as elements to improve viral titer, transgene expression, and overall vector function. The woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) promotes RNA processing events and enhances nuclear export of viral RNA (2), leading to increased viral titers from packaging cells. In addition, the vector includes a Rev-response element (RRE), which further increases viral titers by enhancing the transport of unspliced viral RNA out of the nucleus (3). Finally, pLVX-IRES-Neo also contains a central polypurine tract/central termination sequence element (cPPT/CTS). During target cell infection, this element creates a central DNA flap that increases nuclear import of the viral genome, resulting in improved vector integration and more efficient transduction (4). The vector also contains a pUC origin of replication and an E. coli ampicillin resistance gene (Ampr ) for propagation and selection in bacteria.

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pLVX-IRES-Neo质粒序列.txt