p3xFLAG-CMV-10

编号	载体名称
YB6030	p3xFLAG-CMV-10

p3xFLAG-CMV-10载体基本信息

载体名称:	p3XFLAG-CMV-10
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:	6299 bp
5' 测序引物及序列:	CMV Forward：CGCAAATGGGCGGTAGGCGTG
3' 测序引物及序列:	--
载体标签:	3XFLAG 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），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:	瞬表达或稳表达
组成型/诱导型:	组成型
病毒/非病毒:	非病毒

p3xFLAG-CMV-10载体质粒图谱和多克隆位点信息

 

 

 

p3xFLAG-CMV-10载体简介

FLAG 和 3xFLAG 标签：

 

A Proven System for Detection and Purification of Proteins The FLAG Expression System is an established way to express, purify and detect recombinant fusion proteins. FLAG and 3xFLAG have proven utility in numerous applications such as Western blotting, immunocytochemistry, immunoprecipitation, flow cytometry, protein purification, and in the study of protein-protein interactions, cell ultrastructure and protein localization. These small hydrophilic tags facilitate superior detection and purification of recombinant fusion proteins when using our highly specific and sensitive ANTI-FLAG antibodies. Ideal Epitope Tags: Small, Hydrophilic and Cleavable The FLAG expression system utilizes a short, hydrophilic 8-amino acid peptide that is fused to the recombinant protein of interest. Because of its hydrophilic nature, the FLAG peptide is likely to be located on the surface of the fusion protein. As a result, the FLAG peptide is easily accessible for cleavage by enterokinase (Ek) and for detection with antibodies. In addition, because of the small size of the FLAG peptide tag, it is not likely to obscure other epitopes, domains, or alter function, secretion, or transport of the fusion protein. The 3xFLAG system is an improvement upon the original system by fusing 3 tandem FLAG epitopes (22 amino acids). Detection of fusion proteins containing 3xFLAG is enhanced up to 200 times more than any other system. Like the original FLAG tag, 3xFLAG is hydrophilic, contains an Ek cleavage site, and is relatively small. Therefore, the risk of altering protein function, blocking other epitopes or decreasing solubility is minimized.

3xFLAG System Expression Vectors for Ultra-Sensitive

 

重组蛋白的检测

 

The 3xFLAG system is an improvement upon the original system by fusing 3 tandem FLAG epitopes for a total of 22 amino acids. Detection of fusion proteins containing 3xFLAG is enhanced up to 200 times more than any other system. Like the original FLAG tag, 3xFLAG is hydrophilic, contains an Enterokinase cleavage site, and is relatively small. Therefore, the risk of altering protein function, blocking other epitopes or decreasing solubility is minimized. In cases of low-level expression, 3xFLAG is ideal. Our selection of CMV vectors provides options for transient or stable expression of 3xFLAG fusion proteins.

 

Ultrasensitive System - Detect < 10 femtomoles using ANTI-FLAG Antibodies; Ideal for cases of low-level expression in mammalian cells.

Superior Detection - 20-200x more sensitive than any other system.

Versatile - Enhanced detection for immunoprecipitation, Western blots and immunocytochemistry.

Wide Selection of Detection and Purification Products - ANTI-FLAG antibodies, resins, and affinity capture plates.

 

 

CMV启动子：

Our mammalian expression vectors contain the strong CMV promoter for high-level constitutive expression in mammalian cells. The strong human cytomegalovirus (CMV) promoter regulatory region drives constitutive protein expression levels as high as 1 mg/L in COS cells. For less potent cell lines, protein levels are typically ~0.1 mg/L. The presence of the SV40 replication origin will result in high levels of DNA replication in SV40 replication permissive COS cells. Vectors containing the preprotrypsin leader (PPT) sequence direct secretion of FLAG fusion proteins into the culture medium for purification using ANTI-FLAG antibodies, resins, and plates.

 

p3xFLAG-CMV-10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 ATTAACCATG GACTACAAAG ACCATGACGG TGATTATAAA

  961 GATCATGACA TCGATTACAA GGATGACGAT GACAAGCTTG CGGCCGCGAA TTCATCGATA

 1021 GATCTGATAT CGGTACCAGT CGACTCTAGA GGATCCCGGG TGGCATCCCT GTGACCCCTC

 1081 CCCAGTGCCT CTCCTGGCCC TGGAAGTTGC CACTCCAGTG CCCACCAGCC TTGTCCTAAT

 1141 AAAATTAAGT TGCATCATTT TGTCTGACTA GGTGTCCTTC TATAATATTA TGGGGTGGAG

 1201 GGGGGTGGTA TGGAGCAAGG GGCAAGTTGG GAAGACAACC TGTAGGGCCT GCGGGGTCTA

 1261 TTGGGAACCA AGCTGGAGTG CAGTGGCACA ATCTTGGCTC ACTGCAATCT CCGCCTCCTG

 1321 GGTTCAAGCG ATTCTCCTGC CTCAGCCTCC CGAGTTGTTG GGATTCCAGG CATGCATGAC

 1381 CAGGCTCAGC TAATTTTTGT TTTTTTGGTA GAGACGGGGT TTCACCATAT TGGCCAGGCT

 1441 GGTCTCCAAC TCCTAATCTC AGGTGATCTA CCCACCTTGG CCTCCCAAAT TGCTGGGATT

 1501 ACAGGCGTGA ACCACTGCTC CCTTCCCTGT CCTTCTGATT TTAAAATAAC TATACCAGCA

 1561 GGAGGACGTC CAGACACAGC ATAGGCTACC TGGCCATGCC CAACCGGTGG GACATTTGAG

 1621 TTGCTTGCTT GGCACTGTCC TCTCATGCGT TGGGTCCACT CAGTAGATGC CTGTTGAATT

 1681 GGGTACGCGG CCAGCTTGGC TGTGGAATGT GTGTCAGTTA GGGTGTGGAA AGTCCCCAGG

 1741 CTCCCCAGCA GGCAGAAGTA TGCAAAGCAT GCATCTCAAT TAGTCAGCAA CCAGGTGTGG

 1801 AAAGTCCCCA GGCTCCCCAG CAGGCAGAAG TATGCAAAGC ATGCATCTCA ATTAGTCAGC

 1861 AACCATAGTC CCGCCCCTAA CTCCGCCCAT CCCGCCCCTA ACTCCGCCCA GTTCCGCCCA

 1921 TTCTCCGCCC CATGGCTGAC TAATTTTTTT TATTTATGCA GAGGCCGAGG CCGCCTCGGC

 1981 CTCTGAGCTA TTCCAGAAGT AGTGAGGAGG CTTTTTTGGA GGAATTGATC AGCTTGGGAT

 2041 CTGATCAAGA GACAGGATGA GGATCGTTTC GCATGATTGA ACAAGATGGA TTGCACGCAG

 2101 GTTCTCCGGC CGCTTGGGTG GAGAGGCTAT TCGGCTATGA CTGGGCACAA CAGACAATCG

 2161 GCTGCTCTGA TGCCGCCGTG TTCCGGCTGT CAGCGCAGGG GCGCCCGGTT CTTTTTGTCA

 2221 AGACCGACCT GTCCGGTGCC CTGAATGAAC TGCAGGACGA GGCAGCGCGG CTATCGTGGC

 2281 TGGCCACGAC GGGCGTTCCT TGCGCAGCTG TGCTCGACGT TGTCACTGAA GCGGGAAGGG

 2341 ACTGGCTGCT ATTGGGCGAA GTGCCGGGGC AGGATCTCCT GTCATCTCAC CTTGCTCCTG

 2401 CCGAGAAAGT ATCCATCATG GCTGATGCAA TGCGGCGGCT GCATACGCTT GATCCGGCTA

 2461 CCTGCCCATT CGACCACCAA GCGAAACATC GCATCGAGCG AGCACGTACT CGGATGGAAG

 2521 CCGGTCTTGT CGATCAGGAT GATCTGGACG AAGAGCATCA GGGGCTCGCG CCAGCCGAAC

 2581 TGTTCGCCAG GCTCAAGGCG CGCATGCCCG ACGGCGAGGA TCTCGTCGTG ACCCATGGCG

 2641 ATGCCTGCTT GCCGAATATC ATGGTGGAAA ATGGCCGCTT TTCTGGATTC ATCGACTGTG

 2701 GCCGGCTGGG TGTGGCGGAC CGCTATCAGG ACATAGCGTT GGCTACCCGT GATATTGCTG

 2761 AAGAGCTTGG CGGCGAATGG GCTGACCGCT TCCTCGTGCT TTACGGTATC GCCGCTCCCG

 2821 ATTCGCAGCG CATCGCCTTC TATCGCCTTC TTGACGAGTT CTTCTGAGCG GGACTCTGGG

 2881 GTTCGAAATG ACCGACCAAG CGACGCCCAA CCTGCCATCA CGAGATTTCG ATTCCACCGC

 2941 CGCCTTCTAT GAAAGGTTGG GCTTCGGAAT CGTTTTCCGG GACGCCGGCT GGATGATCCT

 3001 CCAGCGCGGG GATCTCATGC TGGAGTTCTT CGCCCACCCC GGGCTCGATC CCCTCGCGAG

 3061 TTGGTTCAGC TGCTGCCTGA GGCTGGACGA CCTCGCGGAG TTCTACCGGC AGTGCAAATC

 3121 CGTCGGCATC CAGGAAACCA GCAGCGGCTA TCCGCGCATC CATGCCCCCG AACTGCAGGA

 3181 GTGGGGAGGC ACGATGGCCG CTTTGGTCGA CCCGGACGGG ACGCTCCTGC GCCTGATACA

 3241 GAACGAATTG CTTGCAGGCA TCTCATGAGT GTGTCTTCCC GTTTTCCGCC TGAGGTCACT

 3301 GCGTGGATGG AGCGCTGGCG CCTGCTGCGC GACGGCGAGC TGCTCACCAC CCACTCGCCA

 3361 AGCTGGAACC GTAAAAAGGC CGCGTTGCTG GCGTTTTTCC ATAGGCTCCG CCGATCATAA

 3421 TCAGCCATAC CACATTTGTA GAGGTTTTAC TTGCTTTAAA AAACCTCCCA CACCTCCCCC

 3481 TGAACCTGAA ACATAAAATG AATGCAATTG TTGTTGTTAA CTTGTTTATT GCAGCTTATA

 3541 ATGGTTACAA ATAAAGCAAT AGCATCACAA ATTTCACAAA TAAAGCATTT TTTTCACTGC

 3601 ATTCTAGTTG TGGTTTGTCC AAACTCATCA ATGTATCTTA TCATGTCTGG ATCAATTCCC

 3661 TATAGTGAGT CGTATTAAAT TCGTAATCAT GTCATAGCTG TTTCCTGTGT GAAATTGTTA

 3721 TCCGCTCACA ATTCCACACA ACATACGAGC CGGAAGCATA AAGTGTAAAG CCTGGGGTGC

 3781 CTAATGAGTG AGCTAACTCA CATTAATTGC GTTGCGCTCA CTGCCCGCTT TCCAGTCGGG

 3841 AAACCTGTCG TGCCAGCTGC ATTAATGAAT CGGCCAACGC GCGGGGAGAG GCGGTTTGCG

 3901 TATTGGGCGC TCTTCCGCTT CCTCGCTCAC TGACTCGCTG CGCTCGGTCG TTCGGCTGCG

 3961 GCGAGCGGTA TCAGCTCACT CAAAGGCGGT AATACGGTTA TCCACAGAAT CAGGGGATAA

 4021 CGCAGGAAAG AACATGTGAG CAAAAGGCCA GCAAAAGGCC AGGAACCGTA AAAAGGCCGC

 4081 GTTGCTGGCG TTTTTCCATA GGCTCCGCCC CCCTGACGAG CATCACAAAA ATCGACGCTC

 4141 AAGTCAGAGG TGGCGAAACC CGACAGGACT ATAAAGATAC CAGGCGTTTC CCCCTGGAAG

 4201 CTCCCTCGTG CGCTCTCCTG TTCCGACCCT GCCGCTTACC GGATACCTGT CCGCCTTTCT

 4261 CCCTTCGGGA AGCGTGGCGC TTTCTCATAG CTCACGCTGT AGGTATCTCA GTTCGGTGTA

 4321 GGTCGTTCGC TCCAAGCTGG GCTGTGTGCA CGAACCCCCC GTTCAGCCCG ACCGCTGCGC

 4381 CTTATCCGGT AACTATCGTC TTGAGTCCAA CCCGGTAAGA CACGACTTAT CGCCACTGGC

 4441 AGCAGCCACT GGTAACAGGA TTAGCAGAGC GAGGTATGTA GGCGGTGCTA CAGAGTTCTT

 4501 GAAGTGGTGG CCTAACTACG GCTACACTAG AAGAACAGTA TTTGGTATCT GCGCTCTGCT

 4561 GAAGCCAGTT ACCTTCGGAA AAAGAGTTGG TAGCTCTTGA TCCGGCAAAC AAACCACCGC

 4621 TGGTAGCGGT GGTTTTTTTG TTTGCAAGCA GCAGATTACG CGCAGAAAAA AAGGATCTCA

 4681 AGAAGATCCT TTGATCTTTT CTACGGGGTC TGACGCTCAG TGGAACGAAA ACTCACGTTA

 4741 AGGGATTTTG GTCATGAGAT TATCAAAAAG GATCTTCACC TAGATCCTTT TAAATTAAAA

 4801 ATGAAGTTTT AAATCAATCT AAAGTATATA TGAGTAAACT TGGTCTGACA GTTACCAATG

 4861 CTTAATCAGT GAGGCACCTA TCTCAGCGAT CTGTCTATTT CGTTCATCCA TAGTTGCCTG

 4921 ACTCCCCGTC GTGTAGATAA CTACGATACG GGAGGGCTTA CCATCTGGCC CCAGTGCTGC

 4981 AATGATACCG CGAGACCCAC GCTCACCGGC TCCAGATTTA TCAGCAATAA ACCAGCCAGC

 5041 CGGAAGGGCC GAGCGCAGAA GTGGTCCTGC AACTTTATCC GCCTCCATCC AGTCTATTAA

 5101 TTGTTGCCGG GAAGCTAGAG TAAGTAGTTC GCCAGTTAAT AGTTTGCGCA ACGTTGTTGC

 5161 CATTGCTACA GGCATCGTGG TGTCACGCTC GTCGTTTGGT ATGGCTTCAT TCAGCTCCGG

 5221 TTCCCAACGA TCAAGGCGAG TTACATGATC CCCCATGTTG TGCAAAAAAG CGGTTAGCTC

 5281 CTTCGGTCCT CCGATCGTTG TCAGAAGTAA GTTGGCCGCA GTGTTATCAC TCATGGTTAT

 5341 GGCAGCACTG CATAATTCTC TTACTGTCAT GCCATCCGTA AGATGCTTTT CTGTGACTGG

 5401 TGAGTACTCA ACCAAGTCAT TCTGAGAATA GTGTATGCGG CGACCGAGTT GCTCTTGCCC

 5461 GGCGTCAATA CGGGATAATA CCGCGCCACA TAGCAGAACT TTAAAAGTGC TCATCATTGG

 5521 AAAACGTTCT TCGGGGCGAA AACTCTCAAG GATCTTACCG CTGTTGAGAT CCAGTTCGAT

 5581 GTAACCCACT CGTGCACCCA ACTGATCTTC AGCATCTTTT ACTTTCACCA GCGTTTCTGG

 5641 GTGAGCAAAA ACAGGAAGGC AAAATGCCGC AAAAAAGGGA ATAAGGGCGA CACGGAAATG

 5701 TTGAATACTC ATACTCTTCC TTTTTCAATA TTATTGAAGC ATTTATCAGG GTTATTGTCT

 5761 CATGAGCGGA TACATATTTG AATGTATTTA GAAAAATAAA CAAATAGGGG TTCCGCGCAC

 5821 ATTTCCCCGA AAAGTGCCAC CTGACGCGCC CTGTAGCGGC GCATTAAGCG CGGCGGGTGT

 5881 GGTGGTTACG CGCAGCGTGA CCGCTACACT TGCCAGCGCC CTAGCGCCCG CTCCTTTCGC

 5941 TTTCTTCCCT TCCTTTCTCG CCACGTTCGC CGGCTTTCCC CGTCAAGCTC TAAATCGGGG

 6001 GCTCCCTTTA GGGTTCCGAT TTAGTGCTTT ACGGCACCTC GACCCCAAAA AACTTGATTA

 6061 GGGTGATGGT TCACGTAGTG GGCCATCGCC CTGATAGACG GTTTTTCGCC CTTTGACGTT

 6121 GGAGTCCACG TTCTTTAATA GTGGACTCTT GTTCCAAACT GGAACAACAC TCAACCCTAT

 6181 CTCGGTCTAT TCTTTTGATT TATAAGGGAT TTTGCCGATT TCGGCCTATT GGTTAAAAAA

 6241 TGAGCTGATT TAACAAAAAT TTAACGCGAA TTTTAACAAA ATATTAACGC TTACAATTT