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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20℃
- 保质期：
6个月
- 英文名：
pTYB2
- 库存：
大量
- 供应商：
钰博生物
- 规格：
1ug/5ug
pTYB2
载体基本信息
| 出品公司: | Ybscience |
|---|---|
| 载体名称: | pTYB2 |
| 质粒类型: | 大肠杆菌表达载体；蛋白纯化 |
| 高拷贝/低拷贝: | -- |
| 启动子: | -- |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 7474 bp |
| 5' 测序引物及序列: | T7: TAATACGACTCACTATAGGG |
| 3' 测序引物及序列: | -- |
| 载体标签: | Intein |
| 载体抗性: | Ampicillin |
| 筛选标记: | -- |
| 备注: | -- |
| 稳定性: | -- |
| 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
载体简介
pTYB2 is an E. coli cloning and expression vector (7474 bp) used in the IMPACT™ Kit (NEB #E6901) which allows the overexpression of a target protein as a fusion to a self-cleavable affinity tag (1,2). This C-terminal fusion vector is designed for the in-frame insertion of a target gene into the polylinker upstream of an intein tag (the Sce VMA intein/chitin binding domain, 55 kDa) (1,2). This results in the fusion of the C-terminus of the target protein to the N-terminus of the intein tag. Thiol-induced self-cleavage of the intein releases the target protein from the chitin-bound intein tag, resulting in a single column purification of the target protein. For fusion of the N-terminus of the target protein to the intein tag, use pTYB21 (NEB #N6709) or pTYB22 (NEB #N6710). This vector can be used in conjuction with pTYB22 (NEB #N6710) to test which fusion constructin (N-terminal or C-terminal) maximizes the expression and yield of a target protein (3). Features The NdeI site in the polylinker contains an ATG sequence for translation initiation. The SmaI site is used for cloning the 3´ end of the target gene and will yield a target protein with a single glycine residue added to its C-terminus after cleavage of the intein. A pBR322 derivative with a ColE1 replication origin. Expression of the fusion gene is under the control of an IPTG-inducible T7 promoter (4). Expression requires an E. coli host that carries the T7 RNA Polymerase gene [e.g., T7 Express Competent E. coli (High Efficiency) NEB #C2566), BL21(DE3) Competent E. coli (NEB #C2527) and derivatives]. Ampicillin resistance. M13 allows for the production of single-stranded DNA by helper phage superinfection of cells bearing the plasmid. M13K07 helper phage is available from (NEB #N0315). Other IMPACT vectors are available which allow for fusion of a target gene to N- or C- terminus of an intein, the cleavage reaction may be induced by thiol reagent or temperature/pH shift. The sites in the polylinker region are identical to or compatible with (i.e., NheI of pTYB2 and SpeI of pTYB12) those of pTYB12 (NEB #N6902). This allows the same amplified target gene to be cloned into either vector for optimizing protein expression. Vector derived residues may be present at the N- and/or C-termini of the target proteins.
载体序列
LOCUS pTYB2 7474 bp DNA SYN
DEFINITION pTYB2
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..7474
/organism="pTYB2"
/mol_type="other DNA"
promoter 70..98
/label="AmpR_promoter"
gene 140..1000
/label="Ampicillin"
/gene="Ampicillin"
CDS 140..1000
/label="ORF frame 2"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
rep_origin complement(1232..1538)
/label="f1_origin"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
rep_origin 1650..2269
/label="pBR322_origin"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
misc_feature 2607..2629
/label="pGEX_3_primer"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
misc_feature complement(2623..2814)
/label="ROP"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
CDS 2953..3588
/label="ORF frame 1"
/translation="MDAAGPEKNHSGSMPAERQQDVAQRVGRHAGDNGLLLAETFGGG
TSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPERCRHLSYELHDKEDSHKCGDD
SHAPRPPEGADWVEGSQGHRSRSRCLMSELTYINCVALTARFPVGKPVVPAALMNRPT
RGERRFAYWAPGWFFFSPVRRATADCPSPPGPERVAASGPRWFAPAGENPV*"
gene complement(3010..3309)
/label="tet (611 - 312)"
/gene="tet (611 - 312)"
/translation="MDAAGPEKNHSGSMPAERQQDVAQRVGRHAGDNGLLLAETFGGG
TSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPERCRHLSYELHDKEDSHKCGDD
SHAPRPPEGADWVEGSQGHRSRSRCLMSELTYINCVALTARFPVGKPVVPAALMNRPT
RGERRFAYWAPGWFFFSPVRRATADCPSPPGPERVAASGPRWFAPAGENPV*"
misc_feature complement(3371..4462)
/label="lacI"
/translation="MDAAGPEKNHSGSMPAERQQDVAQRVGRHAGDNGLLLAETFGGG
TSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPERCRHLSYELHDKEDSHKCGDD
SHAPRPPEGADWVEGSQGHRSRSRCLMSELTYINCVALTARFPVGKPVVPAALMNRPT
RGERRFAYWAPGWFFFSPVRRATADCPSPPGPERVAASGPRWFAPAGENPV*"
CDS complement(3371..4330)
/label="ORF frame 1"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
CDS complement(4654..5415)
/label="ORF frame 2"
/translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT
TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH
RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE
SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP
TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"
terminator 4687..4730
/label="rrnB_T1_terminator"
/translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT
TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH
RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE
SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP
TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"
terminator 4870..4913
/label="rrnB_T1_terminator"
/translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT
TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH
RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE
SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP
TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"
terminator 5053..5096
/label="rrnB_T1_terminator"
/translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT
TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH
RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE
SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP
TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"
terminator 5236..5279
/label="rrnB_T1_terminator"
/translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT
TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH
RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE
SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP
TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"
terminator 5419..5462
/label="rrnB_T1_terminator"
/translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT
TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH
RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE
SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP
TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"
promoter 5637..5655
/label="T7_promoter"
/translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT
TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH
RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE
SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP
TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"
misc_feature 5655..5682
/label="lacO"
/translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT
TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH
RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE
SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP
TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"
misc_feature 5700..5716
/label="T7_transl_en_RBS"
/translation="MPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVHRQT
TDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQESVH
RQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICPTQE
SVHRQTTDKTKGPVFRLSLSFYLMPGELIPGSLWRASCPPPSGPLLRNVQIRSRRICP
TQESVHRQTTDKTKGPVFRLSLSFYLMPGELGINS*"
CDS 5725..7326
/label="ORF frame 1"
/translation="MASSRVDGGREFLEPGCFAKGTNVLMADGSIECIENIEVGNKVM
GKDGRPREVIKLPRGRETMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRTP
RSVRRLSRTIKGVEYFEVITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELVE
SYRKASNKAYFEWTIEARDLSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLTI
EGPKVLAYLLGLWIGDGLSDRATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQV
AKTVNLYSKVVRGNGIRNNLNTENPLWDAIVGLGFLKDGVKNIPSFLSTDNIGTRETF
LAGLIDSDGYVTDEHGIKATIKTIHTSVRDGLVSLARSLGLVVSVNAEPAKVDMNVTK
HKISYAIYMSGGDVLLNVLSKCAGSKKFRPAPAAAFARECRGFYFELQELKEDDYYGI
TLSDDSDHQFLLGSQVVVHACGGLTGLNSGLTTNPGVSAWQVNTAYTAGQLVTYNGKT
YKCLQPHTSLAGWEPSNVPALWQLQ*"
terminator 7346..7474
/label="T7_terminator"
/translation="MASSRVDGGREFLEPGCFAKGTNVLMADGSIECIENIEVGNKVM
GKDGRPREVIKLPRGRETMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRTP
RSVRRLSRTIKGVEYFEVITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELVE
SYRKASNKAYFEWTIEARDLSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLTI
EGPKVLAYLLGLWIGDGLSDRATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQV
AKTVNLYSKVVRGNGIRNNLNTENPLWDAIVGLGFLKDGVKNIPSFLSTDNIGTRETF
LAGLIDSDGYVTDEHGIKATIKTIHTSVRDGLVSLARSLGLVVSVNAEPAKVDMNVTK
HKISYAIYMSGGDVLLNVLSKCAGSKKFRPAPAAAFARECRGFYFELQELKEDDYYGI
TLSDDSDHQFLLGSQVVVHACGGLTGLNSGLTTNPGVSAWQVNTAYTAGQLVTYNGKT
YKCLQPHTSLAGWEPSNVPALWQLQ*"
misc_feature complement(7388..7406)
/label="T7_Terminal_primer"
/translation="MASSRVDGGREFLEPGCFAKGTNVLMADGSIECIENIEVGNKVM
GKDGRPREVIKLPRGRETMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRTP
RSVRRLSRTIKGVEYFEVITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELVE
SYRKASNKAYFEWTIEARDLSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLTI
EGPKVLAYLLGLWIGDGLSDRATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQV
AKTVNLYSKVVRGNGIRNNLNTENPLWDAIVGLGFLKDGVKNIPSFLSTDNIGTRETF
LAGLIDSDGYVTDEHGIKATIKTIHTSVRDGLVSLARSLGLVVSVNAEPAKVDMNVTK
HKISYAIYMSGGDVLLNVLSKCAGSKKFRPAPAAAFARECRGFYFELQELKEDDYYGI
TLSDDSDHQFLLGSQVVVHACGGLTGLNSGLTTNPGVSAWQVNTAYTAGQLVTYNGKT
YKCLQPHTSLAGWEPSNVPALWQLQ*"
ORIGIN
1 AACTACGTCA GGTGGCACTT TTCGGGGAAA TGTGCGCGGA ACCCCTATTT GTTTATTTTT
61 CTAAATACAT TCAAATATGT ATCCGCTCAT GAGACAATAA CCCTGATAAA TGCTTCAATA
121 ATATTGAAAA AGGAAGAGTA TGAGTATTCA ACATTTCCGT GTCGCCCTTA TTCCCTTTTT
181 TGCGGCATTT TGCCTTCCTG TTTTTGCTCA CCCAGAAACG CTGGTGAAAG TAAAAGATGC
241 TGAAGATCAG TTGGGTGCAC GAGTGGGTTA CATCGAACTG GATCTCAACA GCGGTAAGAT
301 CCTTGAGAGT TTTCGCCCCG AAGAACGTTC TCCAATGATG AGCACTTTTA AAGTTCTGCT
361 ATGTGGCGCG GTATTATCCC GTGTTGACGC CGGGCAAGAG CAACTCGGTC GCCGCATACA
421 CTATTCTCAG AATGACTTGG TTGAGTACTC ACCAGTCACA GAAAAGCATC TTACGGATGG
481 CATGACAGTA AGAGAATTAT GCAGTGCTGC CATAACCATG AGTGATAACA CTGCGGCCAA
541 CTTACTTCTG ACAACGATCG GAGGACCGAA GGAGCTAACC GCTTTTTTGC ACAACATGGG
601 GGATCATGTA ACTCGCCTTG ATCGTTGGGA ACCGGAGCTG AATGAAGCCA TACCAAACGA
661 CGAGCGTGAC ACCACGATGC CTGTAGCAAT GGCAACAACG TTGCGCAAAC TATTAACTGG
721 CGAACTACTT ACTCTAGCTT CCCGGCAACA ATTAATAGAC TGGATGGAGG CGGATAAAGT
781 TGCAGGACCA CTTCTGCGCT CGGCCCTTCC GGCTGGCTGG TTTATTGCTG ATAAATCTGG
841 AGCCGGTGAG CGTGGGTCTC GCGGTATCAT TGCAGCACTG GGGCCAGATG GTAAGCCCTC
901 CCGTATCGTA GTTATCTACA CGACGGGGAG TCAGGCAACT ATGGATGAAC GAAATAGACA
961 GATCGCTGAG ATAGGTGCCT CACTGATTAA GCATTGGTAA CTGTCAGACC AAGTTTACTC
1021 ATATATACTT TAGATTGATT TACCCCGGTT GATAATCAGA AAAGCCCCAA AAACAGGAAG
1081 ATTGTATAAG CAAATATTTA AATTGTAAAC GTTAATATTT TGTTAAAATT CGCGTTAAAT
1141 TTTTGTTAAA TCAGCTCATT TTTTAACCAA TAGGCCGAAA TCGGCAAAAT CCCTTATAAA
1201 TCAAAAGAAT AGCCCGAGAT AGGGTTGAGT GTTGTTCCAG TTTGGAACAA GAGTCCACTA
1261 TTAAAGAACG TGGACTCCAA CGTCAAAGGG CGAAAAACCG TCTATCAGGG CGATGGCCCA
1321 CTACGTGAAC CATCACCCAA ATCAAGTTTT TTGGGGTCGA GGTGCCGTAA AGCACTAAAT
1381 CGGAACCCTA AAGGGAGCCC CCGATTTAGA GCTTGACGGG GAAAGCCGGC GAACGTGGCG
1441 AGAAAGGAAG GGAAGAAAGC GAAAGGAGCG GGCGCTAGGG CGCTGGCAAG TGTAGCGGTC
1501 ACGCTGCGCG TAACCACCAC ACCCGCCGCG CTTAATGCGC CGCTACAGGG CGCGTAAAAG
1561 GATCTAGGTG AAGATCCTTT TTGATAATCT CATGACCAAA ATCCCTTAAC GTGAGTTTTC
1621 GTTCCACTGA GCGTCAGACC CCGTAGAAAA GATCAAAGGA TCTTCTTGAG ATCCTTTTTT
1681 TCTGCGCGTA ATCTGCTGCT TGCAAACAAA AAAACCACCG CTACCAGCGG TGGTTTGTTT
1741 GCCGGATCAA GAGCTACCAA CTCTTTTTCC GAAGGTAACT GGCTTCAGCA GAGCGCAGAT
1801 ACCAAATACT GTCCTTCTAG TGTAGCCGTA GTTAGGCCAC CACTTCAAGA ACTCTGTAGC
1861 ACCGCCTACA TACCTCGCTC TGCTAATCCT GTTACCAGTG GCTGCTGCCA GTGGCGATAA
1921 GTCGTGTCTT ACCGGGTTGG ACTCAAGACG ATAGTTACCG GATAAGGCGC AGCGGTCGGG
1981 CTGAACGGGG GGTTCGTGCA CACAGCCCAG CTTGGAGCGA ACGACCTACA CCGAACTGAG
2041 ATACCTACAG CGTGAGCTAT GAGAAAGCGC CACGCTTCCC GAAGGGAGAA AGGCGGACAG
2101 GTATCCGGTA AGCGGCAGGG TCGGAACAGG AGAGCGCACG AGGGAGCTTC CAGGGGGAAA
2161 CGCCTGGTAT CTTTATAGTC CTGTCGGGTT TCGCCACCTC TGACTTGAGC GTCGATTTTT
2221 GTGATGCTCG TCAGGGGGGC GGAGCCTATG GAAAAACGCC AGCAACGCGG CCTTTTTACG
2281 GTTCCTGGCC TTTTGCTGGC CTTTTGCTCA CATGTTCTTT CCTGCGTTAT CCCCTGATTC
2341 TGTGGATAAC CGTATTACCG CCTTTGAGTG AGCTGATACC GCTCGCCGCA GCCGAACGAC
2401 CGAGCGCAGC GAGTCAGTGA GCGAGGAAGC TATGGTGCAC TCTCAGTACA ATCTGCTCTG
2461 ATGCCGCATA GTTAAGCCAG TATACACTCC GCTATCGCTA CGTGACTGGG TCATGGCTGC
2521 GCCCCGACAC CCGCCAACAC CCGCTGACGC GCCCTGACGG GCTTGTCTGC TCCCGGCATC
2581 CGCTTACAGA CAAGCTGTGA CCGTCTCCGG GAGCTGCATG TGTCAGAGGT TTTCACCGTC
2641 ATCACCGAAA CGCGCGAGGC AGCTGCGGTA AAGCTCATCA GCGTGGTCGT GCAGCGATTC
2701 ACAGATGTCT GCCTGTTCAT CCGCGTCCAG CTCGTTGAGT TTCTCCAGAA GCGTTAATGT
2761 CTGGCTTCTG ATAAAGCGGG CCATGTTAAG GGCGGTTTTT TCCTGTTTGG TCACTGATGC
2821 CTCCGTGTAA GGGGGATTTC TGTTCATGGG GGTAATGATA CCGATGAAAC GAGAGAGGAT
2881 GCTCACGATA CGGGTTACTG ATGATGAACA TGCCCGGTTA CTGGAACGTT GTGAGGGTAA
2941 ACAACTGGCG GTATGGATGC GGCGGGACCA GAGAAAAATC ACTCAGGGTC AATGCCAGCC
3001 GAACGCCAGC AAGACGTAGC CCAGCGCGTC GGCCGCCATG CCGGCGATAA TGGCCTGCTT
3061 CTCGCCGAAA CGTTTGGTGG CGGGACCAGT GACGAAGGCT TGAGCGAGGG CGTGCAAGAT
3121 TCCGAATACC GCAAGCGACA GGCCGATCAT CGTCGCGCTC CAGCGAAAGC GGTCCTCGCC
3181 GAAAATGACC CAGAGCGCTG CCGGCACCTG TCCTACGAGT TGCATGATAA AGAAGACAGT
3241 CATAAGTGCG GCGACGATAG TCATGCCCCG CGCCCACCGG AAGGAGCTGA CTGGGTTGAA
3301 GGCTCTCAAG GGCATCGGTC GAGATCCCGG TGCCTAATGA GTGAGCTAAC TTACATTAAT
3361 TGCGTTGCGC TCACTGCCCG CTTTCCAGTC GGGAAACCTG TCGTGCCAGC TGCATTAATG
3421 AATCGGCCAA CGCGCGGGGA GAGGCGGTTT GCGTATTGGG CGCCAGGGTG GTTTTTCTTT
3481 TCACCAGTGA GACGGGCAAC AGCTGATTGC CCTTCACCGC CTGGCCCTGA GAGAGTTGCA
3541 GCAAGCGGTC CACGCTGGTT TGCCCCAGCA GGCGAAAATC CTGTTTGATG GTGGTTAACG
3601 GCGGGATATA ACATGAGCTG TCTTCGGTAT CGTCGTATCC CACTACCGAG ATATCCGCAC
3661 CAACGCGCAG CCCGGACTCG GTAATGGCGC GCATTGCGCC CAGCGCCATC TGATCGTTGG
3721 CAACCAGCAT CGCAGTGGGA ACGATGCCCT CATTCAGCAT TTGCATGGTT TGTTGAAAAC
3781 CGGACATGGC ACTCCAGTCG CCTTCCCGTT CCGCTATCGG CTGAATTTGA TTGCGAGTGA
3841 GATATTTATG CCAGCCAGCC AGACGCAGAC GCGCCGAGAC AGAACTTAAT GGGCCCGCTA
3901 ACAGCGCGAT TTGCTGGTGA CCCAATGCGA CCAGATGCTC CACGCCCAGT CGCGTACCGT
3961 CTTCATGGGA GAAAATAATA CTGTTGATGG GTGTCTGGTC AGAGACATCA AGAAATAACG
4021 CCGGAACATT AGTGCAGGCA GCTTCCACAG CAATGGCATC CTGGTCATCC AGCGGATAGT
4081 TAATGATCAG CCCACTGACG CGTTGCGCGA GAAGATTGTG CACCGCCGCT TTACAGGCTT
4141 CGACGCCGCT TCGTTCTACC ATCGACACCA CCACGCTGGC ACCCAGTTGA TCGGCGCGAG
4201 ATTTAATCGC CGCGACAATT TGCGACGGCG CGTGCAGGGC CAGACTGGAG GTGGCAACGC
4261 CAATCAGCAA CGACTGTTTG CCCGCCAGTT GTTGTGCCAC GCGGTTGGGA ATGTAATTCA
4321 GCTCCGCCAT CGCCGCTTCC ACTTTTTCCC GCGTTTTCGC AGAAACGTGG CTGGCCTGGT
4381 TCACCACGCG GGAAACGGTC TGATAAGAGA CACCGGCATA CTCTGCGACA TCGTATAACG
4441 TTACTGGTTT CACATTCACC ACCCTGAATT GACTCTCTTC CGGGCGCTAT CATGCCATAC
4501 CGCGAAAGGT TTTGCGCCAT TCGATGGTGT CCGGGATCTC GACGCTCTCC CTTATGCGAC
4561 TCCTGCATTA GGAAGCAGCC CAGTAGTAGG TTGAGGCCGT TGAGCACCGC CGCCGCAAGG
4621 AATGGTGCAT GCCGGCATGC CGCCCTTTCG TCTTCAAGAA TTAATTCCCA ATTCCCCAGG
4681 CATCAAATAA AACGAAAGGC TCAGTCGAAA GACTGGGCCT TTCGTTTTAT CTGTTGTTTG
4741 TCGGTGAACG CTCTCCTGAG TAGGACAAAT CCGCCGGGAG CGGATTTGAA CGTTGCGAAG
4801 CAACGGCCCG GAGGGTGGCG GGCAGGACGC CCGCCATAAA CTGCCAGGAA TTAATTCCCC
4861 AGGCATCAAA TAAAACGAAA GGCTCAGTCG AAAGACTGGG CCTTTCGTTT TATCTGTTGT
4921 TTGTCGGTGA ACGCTCTCCT GAGTAGGACA AATCCGCCGG GAGCGGATTT GAACGTTGCG
4981 AAGCAACGGC CCGGAGGGTG GCGGGCAGGA CGCCCGCCAT AAACTGCCAG GAATTAATTC
5041 CCCAGGCATC AAATAAAACG AAAGGCTCAG TCGAAAGACT GGGCCTTTCG TTTTATCTGT
5101 TGTTTGTCGG TGAACGCTCT CCTGAGTAGG ACAAATCCGC CGGGAGCGGA TTTGAACGTT
5161 GCGAAGCAAC GGCCCGGAGG GTGGCGGGCA GGACGCCCGC CATAAACTGC CAGGAATTAA
5221 TTCCCCAGGC ATCAAATAAA ACGAAAGGCT CAGTCGAAAG ACTGGGCCTT TCGTTTTATC
5281 TGTTGTTTGT CGGTGAACGC TCTCCTGAGT AGGACAAATC CGCCGGGAGC GGATTTGAAC
5341 GTTGCGAAGC AACGGCCCGG AGGGTGGCGG GCAGGACGCC CGCCATAAAC TGCCAGGAAT
5401 TAATTCCCCA GGCATCAAAT AAAACGAAAG GCTCAGTCGA AAGACTGGGC CTTTCGTTTT
5461 ATCTGTTGTT TGTCGGTGAA CGCTCTCCTG AGTAGGACAA ATCCGCCGGG AGCGGATTTG
5521 AACGTTGCGA AGCAACGGCC CGGAGGGTGG CGGGCAGGAC GCCCGCCATA AACTGCCAGG
5581 AATTGGGGAT CGGAATTAAT TCCCGGTTTA AACCGGGGAT CTCGATCCCG CGAAATTAAT
5641 ACGACTCACT ATAGGGGAAT TGTGAGCGGA TAACAATTCC CCTCTAGAAA TAATTTTGTT
5701 TAACTTTAAG AAGGAGATAT ACATATGGCT AGCTCGCGAG TCGACGGCGG CCGCGAATTC
5761 CTCGAGCCCG GGTGCTTTGC CAAGGGTACC AATGTTTTAA TGGCGGATGG GTCTATTGAA
5821 TGTATTGAAA ACATTGAGGT TGGTAATAAG GTCATGGGTA AAGATGGCAG ACCTCGTGAG
5881 GTAATTAAAT TGCCCAGAGG AAGAGAAACT ATGTACAGCG TCGTGCAGAA AAGTCAGCAC
5941 AGAGCCCACA AAAGTGACTC AAGTCGTGAA GTGCCAGAAT TACTCAAGTT TACGTGTAAT
6001 GCGACCCATG AGTTGGTTGT TAGAACACCT CGTAGTGTCC GCCGTTTGTC TCGTACCATT
6061 AAGGGTGTCG AATATTTTGA AGTTATTACT TTTGAGATGG GCCAAAAGAA AGCCCCCGAC
6121 GGTAGAATTG TTGAGCTTGT CAAGGAAGTT TCAAAGAGCT ACCCAATATC TGAGGGGCCT
6181 GAGAGAGCCA ACGAATTAGT AGAATCCTAT AGAAAGGCTT CAAATAAAGC TTATTTTGAG
6241 TGGACTATTG AGGCCAGAGA TCTTTCTCTG TTGGGTTCCC ATGTTCGTAA AGCTACCTAC
6301 CAGACTTACG CTCCAATTCT TTATGAGAAT GACCACTTTT TCGACTACAT GCAAAAAAGT
6361 AAGTTTCATC TCACCATTGA AGGTCCAAAA GTACTTGCTT ATTTACTTGG TTTATGGATT
6421 GGTGATGGAT TGTCTGACAG GGCAACTTTT TCGGTTGATT CCAGAGATAC TTCTTTGATG
6481 GAACGTGTTA CTGAATATGC TGAAAAGTTG AATTTGTGCG CCGAGTATAA GGACAGAAAA
6541 GAACCACAAG TTGCCAAAAC TGTTAATTTG TACTCTAAAG TTGTCAGAGG TAATGGTATT
6601 CGCAATAATC TTAATACTGA GAATCCATTA TGGGACGCTA TTGTTGGCTT AGGATTCTTG
6661 AAGGACGGTG TCAAAAATAT TCCTTCTTTC TTGTCTACGG ACAATATCGG TACTCGTGAA
6721 ACATTTCTTG CTGGTCTAAT TGATTCTGAT GGCTATGTTA CTGATGAGCA TGGTATTAAA
6781 GCAACAATAA AGACAATTCA TACTTCTGTC AGAGATGGTT TGGTTTCCCT TGCTCGTTCT
6841 TTAGGCTTAG TAGTCTCGGT TAACGCAGAA CCTGCTAAGG TTGACATGAA TGTCACCAAA
6901 CATAAAATTA GTTATGCTAT TTATATGTCT GGTGGAGATG TTTTGCTTAA CGTTCTTTCG
6961 AAGTGTGCCG GCTCTAAAAA ATTCAGGCCT GCTCCCGCCG CTGCTTTTGC ACGTGAGTGC
7021 CGCGGATTTT ATTTCGAGTT ACAAGAATTG AAGGAAGACG ATTATTATGG GATTACTTTA
7081 TCTGATGATT CTGATCATCA GTTTTTGCTT GGATCCCAGG TTGTCGTCCA TGCATGCGGT
7141 GGCCTGACCG GTCTGAACTC AGGCCTCACG ACAAATCCTG GTGTATCCGC TTGGCAGGTC
7201 AACACAGCTT ATACTGCGGG ACAATTGGTC ACATATAACG GCAAGACGTA TAAATGTTTG
7261 CAGCCCCACA CCTCCTTGGC AGGATGGGAA CCATCCAACG TTCCTGCCTT GTGGCAGCTT
7321 CAATGACTGC AGGAAGGGGA TCCGGCTGCT AACAAAGCCC GAAAGGAAGC TGAGTTGGCT
7381 GCTGCCACCG CTGAGCAATA ACTAGCATAA CCCCTTGGGG CCTCTAAACG GGTCTTGAGG
7441 GGTTTTTTGC TGAAAGGAGG AACTATATCC GGAT
//
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文献和实验相关实验
-CN蛋白纯化 系统,可利用还原剂DTT使标签自动裂解,无需外加蛋白酶切割,可以在4度条件下进行,小分子的还原剂也很容易去除;IMPACT-CN蛋白纯化系统包含的表达载体(pTYB载体)有两种,内含肽分别与目的蛋白的C末端(pTYB1和 pTYB2)或N末端(pTYB11和pTYB12)融合.这种在融合蛋白构建方面的机动性极大优化了目的蛋白成功表达和纯化的能力.pTYB2和 pTYB12载体的多克隆位点区设计了相同或者可兼容的酶切位点,使得目的蛋白可选择任一种融合构建载体.而pTYB1及pTYB
是IMPACT-CN蛋白纯化系统,可利用还原剂DTT使标签自动裂解,无需外加蛋白酶切割,可以在4度条件下进行,小分子的还原剂也很容易去除;IMPACT-CN蛋白纯化系统包含的表达载体(pTYB载体)有两种,内含肽分别与目的蛋白的C末端(pTYB1和 pTYB2)或N末端(pTYB11和pTYB12)融合.这种在融合蛋白构建方面的机动性极大优化了目的蛋白成功表达和纯化的能力.pTYB2和 pTYB12载体的多克隆位点区设计了相同或者可兼容的酶切位点,使得目的蛋白可选择任一种融合构建载体.而pTYB1及pTYB
载体（pTYB载体）有两种，内含肽分别与目的蛋白的C末端（pTYB1和pTYB2）或N末端（pTYB11和pTYB12）融合。这种在融合蛋白构建方面的机动性极大优化了目的蛋白成功表达和纯化的能力。pTYB2和pTYB12载体的多克隆位点区设计了相同或者可兼容的酶切位点，使得目的蛋白可选择任一种融合构建载体。而pTYB1及pTYB11载体可让目的基因与内含肽的裂解位点直接相连，在4℃利用 DTT 诱导内含肽的肽键裂解活性，使得最后纯化得到的天然蛋白不带有源自于载体的额外的蛋白残基，同时低温也最大程度地保
技术资料暂无技术资料 索取技术资料
pTYB2
¥800 - 1200
