| 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 |
买家导航
pSecTag2A载体基本信息
pSecTag2A载体图谱质粒图谱和多克隆位点信息
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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pSecTag2A载体质粒应用举例
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