p3XFLAG-CMV-7
p3XFLAG-CMV-7
编号 | 载体名称 |
北京华越洋生物VECT6179 | p3XFLAG-CMV-7 |
p3XFLAG-CMV-7载体基本信息
载体名称: | p3XFLAG-CMV-7 |
质粒类型: | 哺乳动物表达载体 |
高拷贝/低拷贝: | 低拷贝 |
克隆方法: | 多克隆位点；限制性内切酶 |
启动子: | CMV |
载体大小: | 4690 bp |
5' 测序引物及序列: | CMV Forward：CGCAAATGGGCGGTAGGCGTG |
3' 测序引物及序列: | -- |
载体标签: | Met-3X FLAG (N-末端） |
载体抗性: | 氨苄青霉素 |
筛选标记: | 无 |
克隆菌株: | -- |
宿主细胞（系）: | 常用细胞系，如293、Hela等 |
备注: | CMV启动子驱动目的蛋白高水平表达；FLAG标签含8个氨基酸残基（DYKDDDDK），3XFLAG标签含22个氨基酸残基，一般位于融合蛋白的外表面，具有高度免疫灵 敏性，便于检测和纯化目的蛋白；用EK蛋白酶可将3XFLAG标签切除。 |
稳定性: | 瞬表达 |
组成型/诱导型: | 组成型 |
病毒/非病毒: | 非病毒 |
p3XFLAG-CMV-7载体质粒图谱和多克隆位点信息
p3XFLAG-CMV-7载体简介
p3XFLAG-CMV-7 Expression Vector is a 4.7 kb derivative of pCMV5 used to establish expression of N-terminal Met-FLAG fusion proteins in mammalian cells. The vector encodes the ANTI-FLAG M2 antibody epitope (Asp-Tyr-Lys-Xaa-Xaa-Asp) three times, upstream of the multiple cloning region. This results in increased detection sensitivity using ANTI-FLAG M2. The third epitope includes the enterokinase recognition sequence allowing cleavage of the 3XFLAG peptide from the purified fusion protein. Efficiency of replication is optimal when using an SV40 T antigen-expressing host.
p3XFLAG-CMV-7-BAP Control Plasmid is a 6.2 kb derivative of pCMV5 used for transient intracellular expression of N-terminal 3XFLAG bacterial alkaline phosphatase fusion protein in mammalian cells. The vector encodes three adjacent FLAG epitopes (Asp- Tyr-Lys-Xaa-Xaa-Asp) upstream of the multiple cloning region. This results in increased detection sensitivity using ANTI-FLAG M2 antibody.3 The third FLAG epitope includes the enterokinase recognition sequence, allowing cleavage of the 3XFLAG peptide from the purified fusion protein.
Principle
The promoter-regulatory region of the human cytomegalovirus drives transcription of FLAG-fusion constructs. p3XFLAG-CMV-7 Expression Vector is a shuttle vector for E. coli and mammalian cells.
p3XFLAG-CMV-7载体序列
ORIGIN
1 CCATTCGCCA TTCAGGCTGC GCAACTGTTG GGAAGGGCGA TCGGTGCGGG CCTCTTCGCT
61 ATTACGCCAG CTGGCGAAAG GGGGATGTGC TGCAAGGCGA TTAAGTTGGG TAACGCCAGG
121 GTTTTCCCAG TCACGACGTT GTAAAACGAC GGCCAGTGCC AAGCTGATCT ATACATTGAA
181 TCAATATTGG CAATTAGCCA TATTAGTCAT TGGTTATATA GCATAAATCA ATATTGGCTA
241 TTGGCCATTG CATACGTTGT ATCTATATCA TAATATGTAC ATTTATATTG GCTCATGTCC
301 AATATGACCG CCATGTTGAC ATTGATTATT GACTAGTTAT TAATAGTAAT CAATTACGGG
361 GTCATTAGTT CATAGCCCAT ATATGGAGTT CCGCGTTACA TAACTTACGG TAAATGGCCC
421 GCCTGGCTGA CCGCCCAACG ACCCCCGCCC ATTGACGTCA ATAATGACGT ATGTTCCCAT
481 AGTAACGCCA ATAGGGACTT TCCATTGACG TCAATGGGTG GAGTATTTAC GGTAAACTGC
541 CCACTTGGCA GTACATCAAG TGTATCATAT GCCAAGTCCG CCCCCTATTG ACGTCAATGA
601 CGGTAAATGG CCCGCCTGGC ATTATGCCCA GTACATGACC TTACGGGACT TTCCTACTTG
661 GCAGTACATC TACGTATTAG TCATCGCTAT TACCATGGTG ATGCGGTTTT GGCAGTACAC
721 CAATGGGCGT GGATAGCGGT TTGACTCACG GGGATTTCCA AGTCTCCACC CCATTGACGT
781 CAATGGGAGT TTGTTTTGGC ACCAAAATCA ACGGGACTTT CCAAAATGTC GTAATAACCC
841 CGCCCCGTTG ACGCAAATGG GCGGTAGGCG TGTACGGTGG GAGGTCTATA TAAGCAGAGC
901 TCGTTTAGTG AACCGTCAGA ATTCACCATG GACTACAAAG ACCATGACGG TGATTATAAA
961 GATCATGATA TCGATTACAA GGATGACGAT GACAAGCTTG CATGCCTGCA GGTCGACTCT
1021 AGAGGATCCC GGGTGGCATC CCTGTGACCC CTCCCCAGTG CCTCTCCTGG CCTTGGAAGT
1081 TGCCACTCCA GTGCCCACCA GCCTTGTCCT AATAAAATTA AGTTGCATCA TTTTGTCTGA
1141 CTAGGTGTCC TTCTATAATA TTATGGGGTG GAGGGGGGTG GTATGGAGCA AGGGGCAAGT
1201 TGGGAAGACA ACCTGTAGGG CCTGCGGGGT CTATTGGGAA CCAAGCTGGA GTGCAGTGGC
1261 ACAATCTTGG CTCACTGCAA TCTCCGCCTC CTGGGTTCAA GCGATTCTCC TGCCTCAGCC
1321 TCCCGAGTTG TTGGGATTCC AGGCATGCAT GACCAGGCTC AGCTAATTTT TGTTTTTTTG
1381 GTAGAGACGG GGTTTCACCA TATTGGCCAG GCTGGTCTCC AACTCCTAAT CTCAGGTGAT
1441 CTACCCACCT TGGCCTCCCA AATTGCTGGG ATTACAGGCG TGAACCACTG CTCCCTTCCC
1501 TGTCCTTCTG ATTTTAAAAT AACTATACCA GCAGGAGGAC GTCCAGACAC AGCATAGGCT
1561 ACCTGGCCAT GCCCAACCGG TGGGACATTT GAGTTGCTTG CTTGGCACTG TCCTCTCATG
1621 CGTTGGGTCC ACTCAGTAGA TGCCTGTTGA ATTGGGTACG CGGCCAGCTT GGCTGTGGAA
1681 TGTGTGTCAG TTAGGGTGTG GAAAGTCCCC AGGCTCCCCA GCAGGCAGAA GTATGCAAAG
1741 CATGCATCTC AATTAGTCAG CAACCAGGTG TGGAAAGTCC CCAGGCTCCC CAGCAGGCAG
1801 AAGTATGCAA AGCATGCATC TCAATTAGTC AGCAACCATA GTCCCGCCCC TAACTCCGCC
1861 CATCCCGCCC CTAACTCCGC CCAGTTCCGC CCATTCTCCG CCCCATGGCT GACTAATTTT
1921 TTTTATTTAT GCAGAGGCCG AGGCCGCCTC GGCCTCTGAG CTATTCCAGA AGTAGTGAGG
1981 AGGCTTTTTT GGAGGCCTAG GCTTTTGCAA AAAGCTCCTC GAGGAACTGA AAAACCAGAA
2041 AGTTAATTCC CTATAGTGAG TCGTATTAAA TTCGTAATCA TGTCATAGCT GTTTCCTGTG
2101 TGAAATTGTT ATCCGCTCAC AATTCCACAC AACATACGAG CCGGAAGCAT AAAGTGTAAA
2161 GCCTGGGGTG CCTAATGAGT GAGCTAACTC ACATTAATTG CGTTGCGCTC ACTGCCCGCT
2221 TTCCAGTCGG GAAACCTGTC GTGCCAGCTG CATTAATGAA TCGGCCAACG CGCGGGGAGA
2281 GGCGGTTTGC GTATTGGGCG CTCTTCCGCT TCCTCGCTCA CTGACTCGCT GCGCTCGGTC
2341 GTTCGGCTGC GGCGAGCGGT ATCAGCTCAC TCAAAGGCGG TAATACGGTT ATCCACAGAA
2401 TCAGGGGATA ACGCAGGAAA GAACATGTGA GCAAAAGGCC AGCAAAAGGC CAGGAACCGT
2461 AAAAAGGCCG CGTTGCTGGC GTTTTTCCAT AGGCTCCGCC CCCCTGACGA GCATCACAAA
2521 AATCGACGCT CAAGTCAGAG GTGGCGAAAC CCGACAGGAC TATAAAGATA CCAGGCGTTT
2581 CCCCCTGGAA GCTCCCTCGT GCGCTCTCCT GTTCCGACCC TGCCGCTTAC CGGATACCTG
2641 TCCGCCTTTC TCCCTTCGGG AAGCGTGGCG CTTTCTCATA GCTCACGCTG TAGGTATCTC
2701 AGTTCGGTGT AGGTCGTTCG CTCCAAGCTG GGCTGTGTGC ACGAACCCCC CGTTCAGCCC
2761 GACCGCTGCG CCTTATCCGG TAACTATCGT CTTGAGTCCA ACCCGGTAAG ACACGACTTA
2821 TCGCCACTGG CAGCAGCCAC TGGTAACAGG ATTAGCAGAG CGAGGTATGT AGGCGGTGCT
2881 ACAGAGTTCT TGAAGTGGTG GCCTAACTAC GGCTACACTA GAAGAACAGT ATTTGGTATC
2941 TGCGCTCTGC TGAAGCCAGT TACCTTCGGA AAAAGAGTTG GTAGCTCTTG ATCCGGCAAA
3001 CAAACCACCG CTGGTAGCGG TGGTTTTTTT GTTTGCAAGC AGCAGATTAC GCGCAGAAAA
3061 AAAGGATCTC AAGAAGATCC TTTGATCTTT TCTACGGGGT CTGACGCTCA GTGGAACGAA
3121 AACTCACGTT AAGGGATTTT GGTCATGAGA TTATCAAAAA GGATCTTCAC CTAGATCCTT
3181 TTAAATTAAA AATGAAGTTT TAAATCAATC TAAAGTATAT ATGAGTAAAC TTGGTCTGAC
3241 AGTTACCAAT GCTTAATCAG TGAGGCACCT ATCTCAGCGA TCTGTCTATT TCGTTCATCC
3301 ATAGTTGCCT GACTCCCCGT CGTGTAGATA ACTACGATAC GGGAGGGCTT ACCATCTGGC
3361 CCCAGTGCTG CAATGATACC GCGAGACCCA CGCTCACCGG CTCCAGATTT ATCAGCAATA
3421 AACCAGCCAG CCGGAAGGGC CGAGCGCAGA AGTGGTCCTG CAACTTTATC CGCCTCCATC
3481 CAGTCTATTA ATTGTTGCCG GGAAGCTAGA GTAAGTAGTT CGCCAGTTAA TAGTTTGCGC
3541 AACGTTGTTG CCATTGCTAC AGGCATCGTG GTGTCACGCT CGTCGTTTGG TATGGCTTCA
3601 TTCAGCTCCG GTTCCCAACG ATCAAGGCGA GTTACATGAT CCCCCATGTT GTGCAAAAAA
3661 GCGGTTAGCT CCTTCGGTCC TCCGATCGTT GTCAGAAGTA AGTTGGCCGC AGTGTTATCA
3721 CTCATGGTTA TGGCAGCACT GCATAATTCT CTTACTGTCA TGCCATCCGT AAGATGCTTT
3781 TCTGTGACTG GTGAGTACTC AACCAAGTCA TTCTGAGAAT AGTGTATGCG GCGACCGAGT
3841 TGCTCTTGCC CGGCGTCAAT ACGGGATAAT ACCGCGCCAC ATAGCAGAAC TTTAAAAGTG
3901 CTCATCATTG GAAAACGTTC TTCGGGGCGA AAACTCTCAA GGATCTTACC GCTGTTGAGA
3961 TCCAGTTCGA TGTAACCCAC TCGTGCACCC AACTGATCTT CAGCATCTTT TACTTTCACC
4021 AGCGTTTCTG GGTGAGCAAA AACAGGAAGG CAAAATGCCG CAAAAAAGGG AATAAGGGCG
4081 ACACGGAAAT GTTGAATACT CATACTCTTC CTTTTTCAAT ATTATTGAAG CATTTATCAG
4141 GGTTATTGTC TCATGAGCGG ATACATATTT GAATGTATTT AGAAAAATAA ACAAATAGGG
4201 GTTCCGCGCA CATTTCCCCG AAAAGTGCCA CCTGACGCGC CCTGTAGCGG CGCATTAAGC
4261 GCGGCGGGTG TGGTGGTTAC GCGCAGCGTG ACCGCTACAC TTGCCAGCGC CCTAGCGCCC
4321 GCTCCTTTCG CTTTCTTCCC TTCCTTTCTC GCCACGTTCG CCGGCTTTCC CCGTCAAGCT
4381 CTAAATCGGG GGCTCCCTTT AGGGTTCCGA TTTAGTGCTT TACGGCACCT CGACCCCAAA
4441 AAACTTGATT AGGGTGATGG TTCACGTAGT GGGCCATCGC CCTGATAGAC GGTTTTTCGC
4501 CCTTTGACGT TGGAGTCCAC GTTCTTTAAT AGTGGACTCT TGTTCCAAAC TGGAACAACA
4561 CTCAACCCTA TCTCGGTCTA TTCTTTTGAT TTATAAGGGA TTTTGCCGAT TTCGGCCTAT
4621 TGGTTAAAAA ATGAGCTGAT TTAACAAAAA TTTAACGCGA ATTTTAACAA AATATTAACG
4681 CTTACAATTT
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其他哺乳动物表达载体：
