pSecTag2 A

价格:1000元

联系方式:I47-825O-882O

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pSecTag2A载体基本信息

出品公司:
Invitrogen
载体名称: pSecTag2 A
质粒类型: 哺乳细胞表达载体;分泌表达载体
高拷贝/低拷贝: 高拷贝
启动子: CMV
克隆方法: 多克隆位点,限制性内切酶
载体大小: 5159bp
5' 测序引物及序列: T7 -F:5'd[TAATACGACTCACTATAGGG]3'
3' 测序引物及序列: --
载体标签: 6X His, myc
载体抗性: Ampicillin
筛选标记: Zeocin
备注: 含有前导序列IgK,引导重组蛋白分泌表达。
产品目录号: V90020
稳定性: 瞬表达
组成型: 组成型
病毒/非病毒: 非病毒

pSecTag2A载体质粒图谱和多克隆位点信息

pSecTag2 A载体图谱



pSecTag2 A多克隆位点

pSecTag2A载体简介

pSecTag2 and pSecTag2/Hygro are mammalian expression vectors designed for the secretion, purification, and detection of fusion proteins. 
Each vector has a large multiple cloning site in three reading frames to simplify cloning in frame with the N-terminal secretion signal. 
The vectors (Figure 1) offer the following features:

 Secretion signal from the V-J2-C region of the mouse Ig kappa-chain for efficient secretion of recombinant proteins (Figure 2)
 Cytomegalovirus (CMV) promoter for high-level constitutive expression
 C-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with an Anti-His(C-term) Antibody
 C-terminal c-myc epitope for detection with an Anti-myc Antibody
 Bovine Growth Hormone (BGH) polyadenylation signal and transcription termination sequence to enhance mRNA stability
 SV40 origin for episomal replication and simple vector rescue in cell lines expressing the large T antigen (e.g., COS-1, COS-7)

The pSecTag2 vectors carry the Zeocin resistance gene for cost-effective selection in mammalian cells. Zeocin selection can also be used in E. coli.
The pSecTag2/Hygro vectors have the Hygromycin B resistance gene for selection of stable mammalian cell lines.


引用及参考文献

Molecular dissection of the alpha-dystroglycan- and integrin-binding sites within the globular domain of human laminin-10.
Ido H, Harada K, Futaki S, Hayashi Y, Nishiuchi R, Natsuka Y, Li S, Wada Y, Combs AC, Ervasti JM, Sekiguchi K, J Biol Chem (2004) 279:10946-10954
Product usage: The 5 cDNA was inserted into the NheI/PmeI sites of pcDNA3.1. For site-directed mutagenesis of the LG3 module, the cDNA fragment encoding LG3 was excised from pcDNA-5LG4–5 with AscI and PmeI and recloned into pSecTag2A at the AscI/PmeI sites. Recombinant protein was produced usi
Prohaptoglobin is proteolytically cleaved in the endoplasmic reticulum by the complement C1r-like protein.Wicher KB, Fries E, Proc Natl Acad Sci U S A (2004) 101:14390-14395
Product usage: HEK293 and COS-1 cells were transiently transfected with C1r-LP cDNA cloned into pcDNA6-myc-his or with proHp cDNA cloned into pSecTag. After 24 h, the cells were rinsed twIce with PBS and grown for 48 h in a serum-free medium. The cell media were then collected.

pSecTag2A载体序列

LOCUS       pSecTag2 A	5159 bp 	DNA	SYN
DEFINITION  pSecTag2 A
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
FEATURES             Location/Qualifiers
     source          1..5159
                     /organism="pSecTag2 A"
                     /mol_type="other DNA"
     promoter        236..852
                     /label="CMV_immearly_promoter"
     misc_feature    315..602
                     /label="CAG_enhancer"
     misc_feature    769..789
                     /label="CMV_fwd_primer"
     promoter        863..881
                     /label="T7_promoter"
     misc_feature    905..967
                     /label="IgK_secretion"
     misc_feature    complement(1167..1184)
                     /label="BGH_rev_primer"
     terminator      1170..1397
                     /label="bGH_PA_terminator"
     rep_origin      1460..1766
                     /label="f1_origin"
     misc_feature    complement(1880..1900)
                     /label="pBABE_3_primer"
     misc_feature    complement(1886..2101)
                     /label="SV40_enhancer"
     promoter        1898..2166
                     /label="SV40_promoter"
     rep_origin      2065..2142
                     /label="SV40_origin"
     misc_feature    2127..2146
                     /label="SV40pro_F_primer"
     promoter        2260..2327
                     /label="EM7_promoter"
     gene            2328..2699
                     /label="bleo"
                     /gene="bleo"
     misc_feature    2328..2702
                     /label="sh_ble"
     terminator      2835..2954
                     /label="SV40_PA_terminator"
     misc_feature    2923..2942
                     /label="EBV_rev_primer"
     promoter        complement(2998..3016)
                     /label="M13_reverse_primer"
     misc_feature    complement(3015..3037)
                     /label="M13_pUC_rev_primer"
     promoter        complement(3051..3080)
                     /label="lac_promoter"
     rep_origin      complement(3389..4008)
                     /label="pBR322_origin"
     gene            complement(4163..5023)
                     /label="Ampicillin"
                     /gene="Ampicillin"
     CDS             complement(4163..5023)
                     /label="ORF frame 2"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     promoter        complement(5065..5093)
                     /label="AmpR_promoter"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
ORIGIN
    1 GACGGATCGG GAGATCTCCC GATCCCCTAT GGTCGACTCT CAGTACAATC TGCTCTGATG
   61 CCGCATAGTT AAGCCAGTAT CTGCTCCCTG CTTGTGTGTT GGAGGTCGCT GAGTAGTGCG
  121 CGAGCAAAAT TTAAGCTACA ACAAGGCAAG GCTTGACCGA CAATTGCATG AAGAATCTGC
  181 TTAGGGTTAG GCGTTTTGCG CTGCTTCGCG ATGTACGGGC CAGATATACG CGTTGACATT
  241 GATTATTGAC TAGTTATTAA TAGTAATCAA TTACGGGGTC ATTAGTTCAT AGCCCATATA
  301 TGGAGTTCCG CGTTACATAA CTTACGGTAA ATGGCCCGCC TGGCTGACCG CCCAACGACC
  361 CCCGCCCATT GACGTCAATA ATGACGTATG TTCCCATAGT AACGCCAATA GGGACTTTCC
  421 ATTGACGTCA ATGGGTGGAC TATTTACGGT AAACTGCCCA CTTGGCAGTA CATCAAGTGT
  481 ATCATATGCC AAGTACGCCC CCTATTGACG TCAATGACGG TAAATGGCCC GCCTGGCATT
  541 ATGCCCAGTA CATGACCTTA TGGGACTTTC CTACTTGGCA GTACATCTAC GTATTAGTCA
  601 TCGCTATTAC CATGGTGATG CGGTTTTGGC AGTACATCAA TGGGCGTGGA TAGCGGTTTG
  661 ACTCACGGGG ATTTCCAAGT CTCCACCCCA TTGACGTCAA TGGGAGTTTG TTTTGGCACC
  721 AAAATCAACG GGACTTTCCA AAATGTCGTA ACAACTCCGC CCCATTGACG CAAATGGGCG
  781 GTAGGCGTGT ACGGTGGGAG GTCTATATAA GCAGAGCTCT CTGGCTAACT AGAGAACCCA
  841 CTGCTTACTG GCTTATCGAA ATTAATACGA CTCACTATAG GGAGACCCAA GCTGGCTAGC
  901 CACCATGGAG ACAGACACAC TCCTGCTATG GGTACTGCTG CTCTGGGTTC CAGGTTCCAC
  961 TGGTGACGCG GCCCAGCCGG CCAGGCGCGC CGTACGAAGC TTGGTACCGA GCTCGGATCC
 1021 ACTCCAGTGT GGTGGAATTC TGCAGATATC CAGCACAGTG GCGGCCGCTC GAGGAGGGCC
 1081 CGAACAAAAA CTCATCTCAG AAGAGGATCT GAATAGCGCC GTCGACCATC ATCATCATCA
 1141 TCATTGAGTT TAAACCCGCT GATCAGCCTC GACTGTGCCT TCTAGTTGCC AGCCATCTGT
 1201 TGTTTGCCCC TCCCCCGTGC CTTCCTTGAC CCTGGAAGGT GCCACTCCCA CTGTCCTTTC
 1261 CTAATAAAAT GAGGAAATTG CATCGCATTG TCTGAGTAGG TGTCATTCTA TTCTGGGGGG
 1321 TGGGGTGGGG CAGGACAGCA AGGGGGAGGA TTGGGAAGAC AATAGCAGGC ATGCTGGGGA
 1381 TGCGGTGGGC TCTATGGCTT CTGAGGCGGA AAGAACCAGC TGGGGCTCTA GGGGGTATCC
 1441 CCACGCGCCC TGTAGCGGCG CATTAAGCGC GGCGGGTGTG GTGGTTACGC GCAGCGTGAC
 1501 CGCTACACTT GCCAGCGCCC TAGCGCCCGC TCCTTTCGCT TTCTTCCCTT CCTTTCTCGC
 1561 CACGTTCGCC GGCTTTCCCC GTCAAGCTCT AAATCGGGGC ATCCCTTTAG GGTTCCGATT
 1621 TAGTGCTTTA CGGCACCTCG ACCCCAAAAA ACTTGATTAG GGTGATGGTT CACGTAGTGG
 1681 GCCATCGCCC TGATAGACGG TTTTTCGCCC TTTGACGTTG GAGTCCACGT TCTTTAATAG
 1741 TGGACTCTTG TTCCAAACTG GAACAACACT CAACCCTATC TCGGTCTATT CTTTTGATTT
 1801 ATAAGGGATT TTGGGGATTT CGGCCTATTG GTTAAAAAAT GAGCTGATTT AACAAAAATT
 1861 TAACGCGAAT TAATTCTGTG GAATGTGTGT CAGTTAGGGT GTGGAAAGTC CCCAGGCTCC
 1921 CCAGCAGGCA GAAGTATGCA AAGCATGCAT CTCAATTAGT CAGCAACCAG GTGTGGAAAG
 1981 TCCCCAGGCT CCCCAGCAGG CAGAAGTATG CAAAGCATGC ATCTCAATTA GTCAGCAACC
 2041 ATAGTCCCGC CCCTAACTCC GCCCATCCCG CCCCTAACTC CGCCCAGTTC CGCCCATTCT
 2101 CCGCCCCATG GCTGACTAAT TTTTTTTATT TATGCAGAGG CCGAGGCCGC CTCTGCCTCT
 2161 GAGCTATTCC AGAAGTAGTG AGGAGGCTTT TTTGGAGGCC TAGGCTTTTG CAAAAAGCTC
 2221 CCGGGAGCTT GTATATCCAT TTTCGGATCT GATCAGCACG TGTTGACAAT TAATCATCGG
 2281 CATAGTATAT CGGCATAGTA TAATACGACA AGGTGAGGAA CTAAACCATG GCCAAGTTGA
 2341 CCAGTGCCGT TCCGGTGCTC ACCGCGCGCG ACGTCGCCGG AGCGGTCGAG TTCTGGACCG
 2401 ACCGGCTCGG GTTCTCCCGG GACTTCGTGG AGGACGACTT CGCCGGTGTG GTCCGGGACG
 2461 ACGTGACCCT GTTCATCAGC GCGGTCCAGG ACCAGGTGGT GCCGGACAAC ACCCTGGCCT
 2521 GGGTGTGGGT GCGCGGCCTG GACGAGCTGT ACGCCGAGTG GTCGGAGGTC GTGTCCACGA
 2581 ACTTCCGGGA CGCCTCCGGG CCGGCCATGA CCGAGATCGG CGAGCAGCCG TGGGGGCGGG
 2641 AGTTCGCCCT GCGCGACCCG GCCGGCAACT GCGTGCACTT CGTGGCCGAG GAGCAGGACT
 2701 GACACGTGCT ACGAGATTTC GATTCCACCG CCGCCTTCTA TGAAAGGTTG GGCTTCGGAA
 2761 TCGTTTTCCG GGACGCCGGC TGGATGATCC TCCAGCGCGG GGATCTCATG CTGGAGTTCT
 2821 TCGCCCACCC CAACTTGTTT ATTGCAGCTT ATAATGGTTA CAAATAAAGC AATAGCATCA
 2881 CAAATTTCAC AAATAAAGCA TTTTTTTCAC TGCATTCTAG TTGTGGTTTG TCCAAACTCA
 2941 TCAATGTATC TTATCATGTC TGTATACCGT CGACCTCTAG CTAGAGCTTG GCGTAATCAT
 3001 GGTCATAGCT GTTTCCTGTG TGAAATTGTT ATCCGCTCAC AATTCCACAC AACATACGAG
 3061 CCGGAAGCAT AAAGTGTAAA GCCTGGGGTG CCTAATGAGT GAGCTAACTC ACATTAATTG
 3121 CGTTGCGCTC ACTGCCCGCT TTCCAGTCGG GAAACCTGTC GTGCCAGCTG CATTAATGAA
 3181 TCGGCCAACG CGCGGGGAGA GGCGGTTTGC GTATTGGGCG CTCTTCCGCT TCCTCGCTCA
 3241 CTGACTCGCT GCGCTCGGTC GTTCGGCTGC GGCGAGCGGT ATCAGCTCAC TCAAAGGCGG
 3301 TAATACGGTT ATCCACAGAA TCAGGGGATA ACGCAGGAAA GAACATGTGA GCAAAAGGCC
 3361 AGCAAAAGGC CAGGAACCGT AAAAAGGCCG CGTTGCTGGC GTTTTTCCAT AGGCTCCGCC
 3421 CCCCTGACGA GCATCACAAA AATCGACGCT CAAGTCAGAG GTGGCGAAAC CCGACAGGAC
 3481 TATAAAGATA CCAGGCGTTT CCCCCTGGAA GCTCCCTCGT GCGCTCTCCT GTTCCGACCC
 3541 TGCCGCTTAC CGGATACCTG TCCGCCTTTC TCCCTTCGGG AAGCGTGGCG CTTTCTCAAT
 3601 GCTCACGCTG TAGGTATCTC AGTTCGGTGT AGGTCGTTCG CTCCAAGCTG GGCTGTGTGC
 3661 ACGAACCCCC CGTTCAGCCC GACCGCTGCG CCTTATCCGG TAACTATCGT CTTGAGTCCA
 3721 ACCCGGTAAG ACACGACTTA TCGCCACTGG CAGCAGCCAC TGGTAACAGG ATTAGCAGAG
 3781 CGAGGTATGT AGGCGGTGCT ACAGAGTTCT TGAAGTGGTG GCCTAACTAC GGCTACACTA
 3841 GAAGGACAGT ATTTGGTATC TGCGCTCTGC TGAAGCCAGT TACCTTCGGA AAAAGAGTTG
 3901 GTAGCTCTTG ATCCGGCAAA CAAACCACCG CTGGTAGCGG TGGTTTTTTT GTTTGCAAGC
 3961 AGCAGATTAC GCGCAGAAAA AAAGGATCTC AAGAAGATCC TTTGATCTTT TCTACGGGGT
 4021 CTGACGCTCA GTGGAACGAA AACTCACGTT AAGGGATTTT GGTCATGAGA TTATCAAAAA
 4081 GGATCTTCAC CTAGATCCTT TTAAATTAAA AATGAAGTTT TAAATCAATC TAAAGTATAT
 4141 ATGAGTAAAC TTGGTCTGAC AGTTACCAAT GCTTAATCAG TGAGGCACCT ATCTCAGCGA
 4201 TCTGTCTATT TCGTTCATCC ATAGTTGCCT GACTCCCCGT CGTGTAGATA ACTACGATAC
 4261 GGGAGGGCTT ACCATCTGGC CCCAGTGCTG CAATGATACC GCGAGACCCA CGCTCACCGG
 4321 CTCCAGATTT ATCAGCAATA AACCAGCCAG CCGGAAGGGC CGAGCGCAGA AGTGGTCCTG
 4381 CAACTTTATC CGCCTCCATC CAGTCTATTA ATTGTTGCCG GGAAGCTAGA GTAAGTAGTT
 4441 CGCCAGTTAA TAGTTTGCGC AACGTTGTTG CCATTGCTAC AGGCATCGTG GTGTCACGCT
 4501 CGTCGTTTGG TATGGCTTCA TTCAGCTCCG GTTCCCAACG ATCAAGGCGA GTTACATGAT
 4561 CCCCCATGTT GTGCAAAAAA GCGGTTAGCT CCTTCGGTCC TCCGATCGTT GTCAGAAGTA
 4621 AGTTGGCCGC AGTGTTATCA CTCATGGTTA TGGCAGCACT GCATAATTCT CTTACTGTCA
 4681 TGCCATCCGT AAGATGCTTT TCTGTGACTG GTGAGTACTC AACCAAGTCA TTCTGAGAAT
 4741 AGTGTATGCG GCGACCGAGT TGCTCTTGCC CGGCGTCAAT ACGGGATAAT ACCGCGCCAC
 4801 ATAGCAGAAC TTTAAAAGTG CTCATCATTG GAAAACGTTC TTCGGGGCGA AAACTCTCAA
 4861 GGATCTTACC GCTGTTGAGA TCCAGTTCGA TGTAACCCAC TCGTGCACCC AACTGATCTT
 4921 CAGCATCTTT TACTTTCACC AGCGTTTCTG GGTGAGCAAA AACAGGAAGG CAAAATGCCG
 4981 CAAAAAAGGG AATAAGGGCG ACACGGAAAT GTTGAATACT CATACTCTTC CTTTTTCAAT
 5041 ATTATTGAAG CATTTATCAG GGTTATTGTC TCATGAGCGG ATACATATTT GAATGTATTT
 5101 AGAAAAATAA ACAAATAGGG GTTCCGCGCA CATTTCCCCG AAAAGTGCCA CCTGACGTC

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