p3XFLAG-CMV-7

价格:1700元

联系方式:I47-825O-882O

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载体基本信息

出品公司: SIGMA
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:
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。
产品目录号: 211173
稳定性: 瞬表达
组成型/诱导型: 组成型
病毒/非病毒: 非病毒

载体质粒图谱和多克隆位点信息

p3XFLAG-CMV-7载体图谱



p3XFLAG-CMV-7 多克隆位点

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.

载体序列

LOCUS       p3xFlag-CMV7	4690 bp 	DNA	SYN
DEFINITION  p3xFlag-CMV7
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
FEATURES             Location/Qualifiers
     source          1..4690
                     /organism="p3xFlag-CMV7"
                     /mol_type="other DNA"
     misc_feature    complement(12..155)
                     /label="lacZ_a"
     misc_feature    126..148
                     /label="M13_pUC_fwd_primer"
     promoter        141..157
                     /label="M13_forward20_primer"
     promoter        319..895
                     /label="CMV_immearly_promoter"
     misc_feature    398..685
                     /label="CAG_enhancer"
     promoter        841..912
                     /label="CMV_promoter"
     misc_feature    852..872
                     /label="CMV_fwd_primer"
     misc_feature    896..915
                     /label="pCEP_fwd_primer"
     misc_feature    982..996
                     /label="EK"
     terminator      1041..1507
                     /label="hGH_PA_terminator"
     misc_feature    complement(1677..1697)
                     /label="pBABE_3_primer"
     misc_feature    complement(1683..1898)
                     /label="SV40_enhancer"
     promoter        1695..1963
                     /label="SV40_promoter"
     rep_origin      1862..1939
                     /label="SV40_origin"
     misc_feature    1924..1943
                     /label="SV40pro_F_primer"
     promoter        complement(2050..2068)
                     /label="T7_promoter"
     misc_feature    complement(2095..2117)
                     /label="M13_pUC_rev_primer"
     promoter        complement(2131..2160)
                     /label="lac_promoter"
     gene            complement(3243..4103)
                     /label="Ampicillin"
                     /gene="Ampicillin"
     CDS             complement(3243..4103)
                     /label="ORF frame 3"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     promoter        complement(4145..4173)
                     /label="AmpR_promoter"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     rep_origin      4252..4558
                     /label="f1_origin"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
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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