pFLAG-CMV-4
pFLAG-CMV-4
编号 | 载体名称 |
北京华越洋生物VECT6177 | pFLAG-CMV-4 |
pFLAG-CMV-4载体基本信息
载体名称: | pFLAG-CMV-4 |
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: | 6271 bp |
5' 测序引物及序列: | CMV Forward：CGCAAATGGGCGGTAGGCGTG |
3' 测序引物及序列: | CCAGCTTGGTTCCCAATAGA |
载体标签: | FLAG tag (N-末端） |
载体抗性: | 氨苄青霉素 |
筛选标记: | Neomycin |
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），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 |
pFLAG-CMV-4载体质粒图谱和多克隆位点信息
pFLAG-CMV-4载体简介
pFLAG-CMV-4 is a 6.2 kb expression vector used for transient or stable expression in mammalian cells. The vector is a derivative of pCMV5 used for transient or stable expression of a properly inserted open reading frame as an N-terminal Met-FLAG fusion protein.
The pFLAG-CMV-4 Expression Vector is a shuttle vector containing both bacterial and SV40 origins of replication for propagation in both E.coli and mammalian cells.
Efficiency of replication and genomic integration is optimal when using host cells that express the SV40 large T antigen.
The FLAG epitope is a small, hydrophilic 8 amino acid tag (DYKDDDDK) that provides for sensitive detection and high quality purification using ANTI-FLAG products available for Sigma Aldrich.
pFLAG-CMV-4-BAP Control Plasmid is a 7.7 kb derivative of pCMV5 used for transient intracellular expression of N-terminal Met-FLAG bacterial alkaline phosphatase fusion protein in mammalian cells.
Principle
The promoter-regulatory region of the human cytomegalovirus2 drives transcription of FLAG-fusion constructs. The aminoglycoside phosphotransferase II gene (neo-r) confers resistance to aminoglycosides such as G 418 allowing for selection of stable transfectants.
pFLAG-CMV-4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 ATTAATTCAG ATCGATCTAC CATGGACTAC AAAGACGATG
961 ACGACAAGCT TGCGGCCGCG AATTCATCGA TAGATCTGAT ATCGGTACCA GTCGACTCTA
1021 GAGGATCCCG GGTGGCATCC CTGTGACCCC TCCCCAGTGC CTCTCCTGGC CCTGGAAGTT
1081 GCCACTCCAG TGCCCACCAG CCTTGTCCTA ATAAAATTAA GTTGCATCAT TTTGTCTGAC
1141 TAGGTGTCCT TCTATAATAT TATGGGGTGG AGGGGGGTGG TATGGAGCAA GGGGCAAGTT
1201 GGGAAGACAA CCTGTAGGGC CTGCGGGGTC TATTGGGAAC CAAGCTGGAG TGCAGTGGCA
1261 CAATCTTGGC TCACTGCAAT CTCCGCCTCC TGGGTTCAAG CGATTCTCCT GCCTCAGCCT
1321 CCCGAGTTGT TGGGATTCCA GGCATGCATG ACCAGGCTCA GCTAATTTTT GTTTTTTTGG
1381 TAGAGACGGG GTTTCACCAT ATTGGCCAGG CTGGTCTCCA ACTCCTAATC TCAGGTGATC
1441 TACCCACCTT GGCCTCCCAA ATTGCTGGGA TTACAGGCGT GAACCACTGC TCCCTTCCCT
1501 GTCCTTCTGA TTTTAAAATA ACTATACCAG CAGGAGGACG TCCAGACACA GCATAGGCTA
1561 CCTGGCCATG CCCAACCGGT GGGACATTTG AGTTGCTTGC TTGGCACTGT CCTCTCATGC
1621 GTTGGGTCCA CTCAGTAGAT GCCTGTTGAA TTGGGTACGC GGCCAGCTTG GCTGTGGAAT
1681 GTGTGTCAGT TAGGGTGTGG AAAGTCCCCA GGCTCCCCAG CAGGCAGAAG TATGCAAAGC
1741 ATGCATCTCA ATTAGTCAGC AACCAGGTGT GGAAAGTCCC CAGGCTCCCC AGCAGGCAGA
1801 AGTATGCAAA GCATGCATCT CAATTAGTCA GCAACCATAG TCCCGCCCCT AACTCCGCCC
1861 ATCCCGCCCC TAACTCCGCC CAGTTCCGCC CATTCTCCGC CCCATGGCTG ACTAATTTTT
1921 TTTATTTATG CAGAGGCCGA GGCCGCCTCG GCCTCTGAGC TATTCCAGAA GTAGTGAGGA
1981 GGCTTTTTTG GAGGAATTGA TCAGCTTGGG ATCTGATCAA GAGACAGGAT GAGGATCGTT
2041 TCGCATGATT GAACAAGATG GATTGCACGC AGGTTCTCCG GCCGCTTGGG TGGAGAGGCT
2101 ATTCGGCTAT GACTGGGCAC AACAGACAAT CGGCTGCTCT GATGCCGCCG TGTTCCGGCT
2161 GTCAGCGCAG GGGCGCCCGG TTCTTTTTGT CAAGACCGAC CTGTCCGGTG CCCTGAATGA
2221 ACTGCAGGAC GAGGCAGCGC GGCTATCGTG GCTGGCCACG ACGGGCGTTC CTTGCGCAGC
2281 TGTGCTCGAC GTTGTCACTG AAGCGGGAAG GGACTGGCTG CTATTGGGCG AAGTGCCGGG
2341 GCAGGATCTC CTGTCATCTC ACCTTGCTCC TGCCGAGAAA GTATCCATCA TGGCTGATGC
2401 AATGCGGCGG CTGCATACGC TTGATCCGGC TACCTGCCCA TTCGACCACC AAGCGAAACA
2461 TCGCATCGAG CGAGCACGTA CTCGGATGGA AGCCGGTCTT GTCGATCAGG ATGATCTGGA
2521 CGAAGAGCAT CAGGGGCTCG CGCCAGCCGA ACTGTTCGCC AGGCTCAAGG CGCGCATGCC
2581 CGACGGCGAG GATCTCGTCG TGACCCATGG CGATGCCTGC TTGCCGAATA TCATGGTGGA
2641 AAATGGCCGC TTTTCTGGAT TCATCGACTG TGGCCGGCTG GGTGTGGCGG ACCGCTATCA
2701 GGACATAGCG TTGGCTACCC GTGATATTGC TGAAGAGCTT GGCGGCGAAT GGGCTGACCG
2761 CTTCCTCGTG CTTTACGGTA TCGCCGCTCC CGATTCGCAG CGCATCGCCT TCTATCGCCT
2821 TCTTGACGAG TTCTTCTGAG CGGGACTCTG GGGTTCGAAA TGACCGACCA AGCGACGCCC
2881 AACCTGCCAT CACGAGATTT CGATTCCACC GCCGCCTTCT ATGAAAGGTT GGGCTTCGGA
2941 ATCGTTTTCC GGGACGCCGG CTGGATGATC CTCCAGCGCG GGGATCTCAT GCTGGAGTTC
3001 TTCGCCCACC CCGGGCTCGA TCCCCTCGCG AGTTGGTTCA GCTGCTGCCT GAGGCTGGAC
3061 GACCTCGCGG AGTTCTACCG GCAGTGCAAA TCCGTCGGCA TCCAGGAAAC CAGCAGCGGC
3121 TATCCGCGCA TCCATGCCCC CGAACTGCAG GAGTGGGGAG GCACGATGGC CGCTTTGGTC
3181 GACCCGGACG GGACGCTCCT GCGCCTGATA CAGAACGAAT TGCTTGCAGG CATCTCATGA
3241 GTGTGTCTTC CCGTTTTCCG CCTGAGGTCA CTGCGTGGAT GGAGCGCTGG CGCCTGCTGC
3301 GCGACGGCGA GCTGCTCACC ACCCACTCGC CAAGCTGGAA CCGTAAAAAG GCCGCGTTGC
3361 TGGCGTTTTT CCATAGGCTC CGCCGATCAT AATCAGCCAT ACCACATTTG TAGAGGTTTT
3421 ACTTGCTTTA AAAAACCTCC CACACCTCCC CCTGAACCTG AAACATAAAA TGAATGCAAT
3481 TGTTGTTGTT AACTTGTTTA TTGCAGCTTA TAATGGTTAC AAATAAAGCA ATAGCATCAC
3541 AAATTTCACA AATAAAGCAT TTTTTTCACT GCATTCTAGT TGTGGTTTGT CCAAACTCAT
3601 CAATGTATCT TATCATGTCT GGATCAATTC CCTATAGTGA GTCGTATTAA ATTCGTAATC
3661 ATGTCATAGC TGTTTCCTGT GTGAAATTGT TATCCGCTCA CAATTCCACA CAACATACGA
3721 GCCGGAAGCA TAAAGTGTAA AGCCTGGGGT GCCTAATGAG TGAGCTAACT CACATTAATT
3781 GCGTTGCGCT CACTGCCCGC TTTCCAGTCG GGAAACCTGT CGTGCCAGCT GCATTAATGA
3841 ATCGGCCAAC GCGCGGGGAG AGGCGGTTTG CGTATTGGGC GCTCTTCCGC TTCCTCGCTC
3901 ACTGACTCGC TGCGCTCGGT CGTTCGGCTG CGGCGAGCGG TATCAGCTCA CTCAAAGGCG
3961 GTAATACGGT TATCCACAGA ATCAGGGGAT AACGCAGGAA AGAACATGTG AGCAAAAGGC
4021 CAGCAAAAGG CCAGGAACCG TAAAAAGGCC GCGTTGCTGG CGTTTTTCCA TAGGCTCCGC
4081 CCCCCTGACG AGCATCACAA AAATCGACGC TCAAGTCAGA GGTGGCGAAA CCCGACAGGA
4141 CTATAAAGAT ACCAGGCGTT TCCCCCTGGA AGCTCCCTCG TGCGCTCTCC TGTTCCGACC
4201 CTGCCGCTTA CCGGATACCT GTCCGCCTTT CTCCCTTCGG GAAGCGTGGC GCTTTCTCAT
4261 AGCTCACGCT GTAGGTATCT CAGTTCGGTG TAGGTCGTTC GCTCCAAGCT GGGCTGTGTG
4321 CACGAACCCC CCGTTCAGCC CGACCGCTGC GCCTTATCCG GTAACTATCG TCTTGAGTCC
4381 AACCCGGTAA GACACGACTT ATCGCCACTG GCAGCAGCCA CTGGTAACAG GATTAGCAGA
4441 GCGAGGTATG TAGGCGGTGC TACAGAGTTC TTGAAGTGGT GGCCTAACTA CGGCTACACT
4501 AGAAGAACAG TATTTGGTAT CTGCGCTCTG CTGAAGCCAG TTACCTTCGG AAAAAGAGTT
4561 GGTAGCTCTT GATCCGGCAA ACAAACCACC GCTGGTAGCG GTGGTTTTTT TGTTTGCAAG
4621 CAGCAGATTA CGCGCAGAAA AAAAGGATCT CAAGAAGATC CTTTGATCTT TTCTACGGGG
4681 TCTGACGCTC AGTGGAACGA AAACTCACGT TAAGGGATTT TGGTCATGAG ATTATCAAAA
4741 AGGATCTTCA CCTAGATCCT TTTAAATTAA AAATGAAGTT TTAAATCAAT CTAAAGTATA
4801 TATGAGTAAA CTTGGTCTGA CAGTTACCAA TGCTTAATCA GTGAGGCACC TATCTCAGCG
4861 ATCTGTCTAT TTCGTTCATC CATAGTTGCC TGACTCCCCG TCGTGTAGAT AACTACGATA
4921 CGGGAGGGCT TACCATCTGG CCCCAGTGCT GCAATGATAC CGCGAGACCC ACGCTCACCG
4981 GCTCCAGATT TATCAGCAAT AAACCAGCCA GCCGGAAGGG CCGAGCGCAG AAGTGGTCCT
5041 GCAACTTTAT CCGCCTCCAT CCAGTCTATT AATTGTTGCC GGGAAGCTAG AGTAAGTAGT
5101 TCGCCAGTTA ATAGTTTGCG CAACGTTGTT GCCATTGCTA CAGGCATCGT GGTGTCACGC
5161 TCGTCGTTTG GTATGGCTTC ATTCAGCTCC GGTTCCCAAC GATCAAGGCG AGTTACATGA
5221 TCCCCCATGT TGTGCAAAAA AGCGGTTAGC TCCTTCGGTC CTCCGATCGT TGTCAGAAGT
5281 AAGTTGGCCG CAGTGTTATC ACTCATGGTT ATGGCAGCAC TGCATAATTC TCTTACTGTC
5341 ATGCCATCCG TAAGATGCTT TTCTGTGACT GGTGAGTACT CAACCAAGTC ATTCTGAGAA
5401 TAGTGTATGC GGCGACCGAG TTGCTCTTGC CCGGCGTCAA TACGGGATAA TACCGCGCCA
5461 CATAGCAGAA CTTTAAAAGT GCTCATCATT GGAAAACGTT CTTCGGGGCG AAAACTCTCA
5521 AGGATCTTAC CGCTGTTGAG ATCCAGTTCG ATGTAACCCA CTCGTGCACC CAACTGATCT
5581 TCAGCATCTT TTACTTTCAC CAGCGTTTCT GGGTGAGCAA AAACAGGAAG GCAAAATGCC
5641 GCAAAAAAGG GAATAAGGGC GACACGGAAA TGTTGAATAC TCATACTCTT CCTTTTTCAA
5701 TATTATTGAA GCATTTATCA GGGTTATTGT CTCATGAGCG GATACATATT TGAATGTATT
5761 TAGAAAAATA AACAAATAGG GGTTCCGCGC ACATTTCCCC GAAAAGTGCC ACCTGACGCG
5821 CCCTGTAGCG GCGCATTAAG CGCGGCGGGT GTGGTGGTTA CGCGCAGCGT GACCGCTACA
5881 CTTGCCAGCG CCCTAGCGCC CGCTCCTTTC GCTTTCTTCC CTTCCTTTCT CGCCACGTTC
5941 GCCGGCTTTC CCCGTCAAGC TCTAAATCGG GGGCTCCCTT TAGGGTTCCG ATTTAGTGCT
6001 TTACGGCACC TCGACCCCAA AAAACTTGAT TAGGGTGATG GTTCACGTAG TGGGCCATCG
6061 CCCTGATAGA CGGTTTTTCG CCCTTTGACG TTGGAGTCCA CGTTCTTTAA TAGTGGACTC
6121 TTGTTCCAAA CTGGAACAAC ACTCAACCCT ATCTCGGTCT ATTCTTTTGA TTTATAAGGG
6181 ATTTTGCCGA TTTCGGCCTA TTGGTTAAAA AATGAGCTGA TTTAACAAAA ATTTAACGCG
6241 AATTTTAACA AAATATTAAC GCTTACAATT T
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其他哺乳动物表达载体：
