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网站首页 >> pCDNA3.1(-)哺乳表达质粒 货号：P2003
pCDNA3.1(-)哺乳表达质粒 货号：P2003

质粒信息

别名 pCDNA3.1-
启动子 CMV
复制子 pUC
终止子 BGH poly（A）signal
质粒分类 哺乳系列质粒；哺乳表达质粒；pCDNA系列质粒
质粒大小 5427bp
原核抗性 Amp
真核抗性 G418
克隆菌株 DH5α
培养条件 37℃
表达宿主 293T等哺乳细胞
培养条件 37℃，5%CO2
诱导方式 无需诱导
5'测序引物 pCDNA3.1-F（CTAGAGAACCCACTGCTTAC）
3'测序引物 pCDNA3.1-R（TAGAAGGCACAGTCGAGG）
备注 需要添加起始密码子ATG
质粒宿主 哺乳细胞
质粒用途 蛋白表达
片段类型 基因CDS
片段物种 空载体
原核抗性 氨苄青霉素

质粒简介

          pCDNA3.1(-)是来自pCDNA3的5.4kb载体，设计用于哺乳动物宿主中的高水平稳定和瞬时表达。大多数哺乳动物细胞可以进行高水平的稳定和非复制性瞬时表达。人类巨细胞病毒立即早期（CMV）启动子，用于在广泛哺乳动物细胞中的高水平表达。前向（+）和反向（-）方向上的多个克隆位点，以促进克隆用于选择稳定细胞系的新霉素抗性基因。在潜伏感染SV40或表达SV40大T抗原（例如COS-1，COS-7）的细胞系中的异常复制。
          pCDNA 3.1(-) is 5.4 kb vector derived from pcDN3 and designed for high-level stable and transient expression in mammalian hosts. High-level stable and non-replicative transient expression can be carried out in most mammalian cells. The vectors contain the following elements:  
          Human cytomegalovirus immediate-early (CMV) promoter for high-level expression in a wide range of mammalian cells  
          Multiple cloning sites in the forward (+) and reverse (C) orientations to facilitate cloning  
          Neomycin resistance gene for selection of stable cell lines  
          Episomal replication in cells lines that are latently infected with SV40 or that express the SV40 large T antigen (e.g. COS-1, COS-7)
          The control plasmid, pcDNA3.1/CAT, is included for use as a positive control for transfection and expression in the cell line of choice.
Use the following outline to clone and express your gene of interest in pcDNA 3.1. 
          1. Design  the multiple cloning  
          2.  Ligate your insert into the appropriate vector and transform into E. coli. Select transformants on LB plates containing 100 g/ml ampicillin
          3.  Analyze your transformants for the presence of insert by restriction digestion. 
          4.  Select a transformant with the correct restriction pattern and use sequencing to confirm that your gene is cloned in the proper orientation. 
          5.  Transfect your construct into the mammalian cell line of interest using your own method of choice. Generate a stable cell line, if desired. 
          6.  Test for expression of your recombinant gene by western blot analysis or functional assay.

最常用的哺乳动物表达载体之一，CMV promoter强启动子调控外源基因表达。

载体骨架与pcDNA3.1(+)基本相同，只是MCS区的酶切位点是互相颠倒的。

载体拷贝数很高，表达量也很高。

无荧光标记，也没有携带tag。

Amp+原核筛选抗性，Neo+真核筛选抗性，可以利用G418筛选稳定细胞株。


图谱




序列 
1 GACGGATCGGGAGATCTCCCGATCCCCTATGGTGCACTCTCAGTACAATC 
51 TGCTCTGATGCCGCATAGTTAAGCCAGTATCTGCTCCCTGCTTGTGTGTT 
101 GGAGGTCGCTGAGTAGTGCGCGAGCAAAATTTAAGCTACAACAAGGCAAG 
151 GCTTGACCGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCG 
201 CTGCTTCGCGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGAC 
251 TAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATA 
301 TGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCG 
351 CCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGT 
401 AACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGT 
451 AAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCC 
501 CCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTA 
551 CATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCA 
601 TCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGA 
651 TAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAA 
701 TGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTA 
751 ACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAG 
801 GTCTATATAAGCAGAGCTCTCTGGCTAACTAGAGAACCCACTGCTTACTG 
851 GCTTATCGAAATTAATACGACTCACTATAGGGAGACCCAAGCTGGCTAGC 
901 GTTTAAACGGGCCCTCTAGACTCGAGCGGCCGCCACTGTGCTGGATATCT 
951 GCAGAATTCCACCACACTGGACTAGTGGATCCGAGCTCGGTACCAAGCTT 
1001 AAGTTTAAACCGCTGATCAGCCTCGACTGTGCCTTCTAGTTGCCAGCCAT 
1051 CTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACT 
1101 CCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAG 
1151 TAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGG 
1201 AGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCGGTGGGCTCTATG 
1251 GCTTCTGAGGCGGAAAGAACCAGCTGGGGCTCTAGGGGGTATCCCCACGC 
1301 GCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCG 
1351 TGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTC 
1401 CCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCG 
1451 GGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCA 
1501 AAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAG 
1551 ACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACT 
1601 CTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTG 
1651 ATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTG 
1701 ATTTAACAAAAATTTAACGCGAATTAATTCTGTGGAATGTGTGTCAGTTA 
1751 GGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCAT 
1801 GCATCTCAATTAGTCAGCAACCAGGTGTGGAAAGTCCCCAGGCTCCCCAG 
1851 CAGGCAGAAGTATGCAAAGCATGCATCTCAATTAGTCAGCAACCATAGTC 
1901 CCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCA 
1951 TTCTCCGCCCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGG 
2001 CCGCCTCTGCCTCTGAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGA 
2051 GGCCTAGGCTTTTGCAAAAAGCTCCCGGGAGCTTGTATATCCATTTTCGG 
2101 ATCTGATCAAGAGACAGGATGAGGATCGTTTCGCATGATTGAACAAGATG 
2151 GATTGCACGCAGGTTCTCCGGCCGCTTGGGTGGAGAGGCTATTCGGCTAT 
2201 GACTGGGCACAACAGACAATCGGCTGCTCTGATGCCGCCGTGTTCCGGCT 
2251 GTCAGCGCAGGGGCGCCCGGTTCTTTTTGTCAAGACCGACCTGTCCGGTG 
2301 CCCTGAATGAACTGCAGGACGAGGCAGCGCGGCTATCGTGGCTGGCCACG 
2351 ACGGGCGTTCCTTGCGCAGCTGTGCTCGACGTTGTCACTGAAGCGGGAAG 
2401 GGACTGGCTGCTATTGGGCGAAGTGCCGGGGCAGGATCTCCTGTCATCTC 
2451 ACCTTGCTCCTGCCGAGAAAGTATCCATCATGGCTGATGCAATGCGGCGG 
2501 CTGCATACGCTTGATCCGGCTACCTGCCCATTCGACCACCAAGCGAAACA 
2551 TCGCATCGAGCGAGCACGTACTCGGATGGAAGCCGGTCTTGTCGATCAGG 
2601 ATGATCTGGACGAAGAGCATCAGGGGCTCGCGCCAGCCGAACTGTTCGCC 
2651 AGGCTCAAGGCGCGCATGCCCGACGGCGAGGATCTCGTCGTGACCCATGG 
2701 CGATGCCTGCTTGCCGAATATCATGGTGGAAAATGGCCGCTTTTCTGGAT 
2751 TCATCGACTGTGGCCGGCTGGGTGTGGCGGACCGCTATCAGGACATAGCG 
2801 TTGGCTACCCGTGATATTGCTGAAGAGCTTGGCGGCGAATGGGCTGACCG 
2851 CTTCCTCGTGCTTTACGGTATCGCCGCTCCCGATTCGCAGCGCATCGCCT 
2901 TCTATCGCCTTCTTGACGAGTTCTTCTGAGCGGGACTCTGGGGTTCGAAA 
2951 TGACCGACCAAGCGACGCCCAACCTGCCATCACGAGATTTCGATTCCACC 
3001 GCCGCCTTCTATGAAAGGTTGGGCTTCGGAATCGTTTTCCGGGACGCCGG 
3051 CTGGATGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTCGCCCACC 
3101 CCAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATC 
3151 ACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTT 
3201 GTCCAAACTCATCAATGTATCTTATCATGTCTGTATACCGTCGACCTCTA 
3251 GCTAGAGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAATTGT 
3301 TATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAA 
3351 AGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCT 
3401 CACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGA 
3451 ATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGC 
3501 TTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGG 
3551 TATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGAT 
3601 AACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCG 
3651 TAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACG 
3701 AGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGA 
3751 CTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCC 
3801 TGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGG 
3851 GAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTG 
3901 TAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCC 
3951 CGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAA 
4001 GACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGA 
4051 GCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTA 
4101 CGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAG 
4151 TTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACC 
4201 GCTGGTAGCGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAA 
4251 AGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGT 
4301 GGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGG 
4351 ATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTA 
4401 AAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTG 
4451 AGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGA 
4501 CTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCC 
4551 CAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTAT 
4601 CAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCA 
4651 ACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGT 
4701 AAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAG 
4751 GCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGT 
4801 TCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGC 
4851 GGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAG 
4901 TGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATG 
4951 CCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATT 
5001 CTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATAC 
5051 GGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGA 
5101 AAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATC 
5151 CAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTA 
5201 CTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCA 
5251 AAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCT 
5301 TTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGAT 
5351 ACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACA 
5401 TTTCCCCGAAAAGTGCCACCTGACGTC

garmin 230