| 出品公司: | |
|---|---|
| 别名: | pQE81L, pQE 81L |
| 质粒类型: | 大肠杆菌蛋白表达载体 |
| 表达水平: | 低拷贝 |
| 启动子: | T5 |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 4753 bp |
| 5' 测序引物及序列: | pQE30-F: AGCGGATAACAATTTCACACAG |
| 3' 测序引物及序列: | pQE30-R: TTCTGAGGTCATTACTGGATC |
| 载体标签: | N-His |
| 载体抗性: | Ampicillin |
| 备注: |
QIAexpress pQE vectors and constructs can be maintained in any E. coli strain that is ampicillin-sensitive and carries the pREP4 repressor plasmid, or harbors the lacIq gene on the F-factor episome. Alternatively, thepQE-80L series of expression vectors which encodes a lacIq repression module, allows use of any E. coli host strain. 配套表达菌株为M15[pREP4].
|
| 产品目录号: | ZY32923 |
| 稳定性: | 瞬时表达 Transient |
| 组成型: | 组成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20℃低温保存
- 保质期：
三年
- 英文名：
pQE-81L
- 库存：
20
- 供应商：
泽叶生物
pQE81L载体基本信息
pQE81L载体质粒图谱和多克隆位点信息
pQE81L载体简介
pQE81L载体序列
LOCUS pQE-81L 4753 bp DNA Circular SYN
DEFINITION pQE-81L
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTOAUTHORAD1|Novagen Inc.|
FEATURES Location/Qualifiers
source 1..4753
/organism="pQE-81L"
/mol_type="other DNA"
gene 348..1007
/label="CAT/CamR"
/gene="CAT/CamR"
CDS 348..1007
/label="ORF frame 3"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAF
LKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETF
SSLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNV
ANMDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG
A*"
terminator 1075..1118
/label="rrnB_T1_terminator"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAF
LKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETF
SSLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNV
ANMDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG
A*"
misc_feature complement(1251..2342)
/label="lacI"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAF
LKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETF
SSLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNV
ANMDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG
A*"
CDS complement(1251..2210)
/label="ORF frame 3"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
misc_feature complement(2495..2517)
/label="pGEX_3_primer"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
gene complement(3688..4548)
/label="Ampicillin"
/gene="Ampicillin"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
CDS complement(3688..4548)
/label="ORF frame 2"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
promoter complement(4590..4618)
/label="AmpR_promoter"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
ORIGIN
1 CTCGAGAAAT CATAAAAAAT TTATTTGCTT TGTGAGCGGA TAACAATTAT AATAGATTCA
61 ATTGTGAGCG GATAACAATT TCACACAGAA TTCATTAAAG AGGAGAAATT AACTATGAGA
121 GGATCTCACC ATCACCATCA CCATACGGAT CCGCATGCGA GCTCGGTACC CCGGGTCGAC
181 CTGCAGCCAA GCTTAATTAG CTGAGCTTGG ACTCCTGTTG ATAGATCCAG TAATGACCTC
241 AGAACTCCAT CTGGATTTGT TCAGAACGCT CGGTTGCCGC CGGGCGTTTT TTATTGGTGA
301 GAATCCAAGC TAGCTTGGCG AGATTTTCAG GAGCTAAGGA AGCTAAAATG GAGAAAAAAA
361 TCACTGGATA TACCACCGTT GATATATCCC AATGGCATCG TAAAGAACAT TTTGAGGCAT
421 TTCAGTCAGT TGCTCAATGT ACCTATAACC AGACCGTTCA GCTGGATATT ACGGCCTTTT
481 TAAAGACCGT AAAGAAAAAT AAGCACAAGT TTTATCCGGC CTTTATTCAC ATTCTTGCCC
541 GCCTGATGAA TGCTCATCCG GAATTTCGTA TGGCAATGAA AGACGGTGAG CTGGTGATAT
601 GGGATAGTGT TCACCCTTGT TACACCGTTT TCCATGAGCA AACTGAAACG TTTTCATCGC
661 TCTGGAGTGA ATACCACGAC GATTTCCGGC AGTTTCTACA CATATATTCG CAAGATGTGG
721 CGTGTTACGG TGAAAACCTG GCCTATTTCC CTAAAGGGTT TATTGAGAAT ATGTTTTTCG
781 TCTCAGCCAA TCCCTGGGTG AGTTTCACCA GTTTTGATTT AAACGTGGCC AATATGGACA
841 ACTTCTTCGC CCCCGTTTTC ACCATGGGCA AATATTATAC GCAAGGCGAC AAGGTGCTGA
901 TGCCGCTGGC GATTCAGGTT CATCATGCCG TTTGTGATGG CTTCCATGTC GGCAGAATGC
961 TTAATGAATT ACAACAGTAC TGCGATGAGT GGCAGGGCGG GGCGTAATTT TTTTAAGGCA
1021 GTTATTGGTG CCCTTAAACG CCTGGGGTAA TGACTCTCTA GCTTGAGGCA TCAAATAAAA
1081 CGAAAGGCTC AGTCGAAAGA CTGGGCCTTT CGTTTTATCT GTTGTTTGTC GGTGAACGCT
1141 CTCCTGAGTA GGACAAATCC GCCCTCTAGA TTACGTGCAG TCGATGATAA GCTGTCAAAC
1201 ATGAGAATTG TGCCTAATGA GTGAGCTAAC TTACATTAAT TGCGTTGCGC TCACTGCCCG
1261 CTTTCCAGTC GGGAAACCTG TCGTGCCAGC TGCATTAATG AATCGGCCAA CGCGCGGGGA
1321 GAGGCGGTTT GCGTATTGGG CGCCAGGGTG GTTTTTCTTT TCACCAGTGA GACGGGCAAC
1381 AGCTGATTGC CCTTCACCGC CTGGCCCTGA GAGAGTTGCA GCAAGCGGTC CACGCTGGTT
1441 TGCCCCAGCA GGCGAAAATC CTGTTTGATG GTGGTTAACG GCGGGATATA ACATGAGCTG
1501 TCTTCGGTAT CGTCGTATCC CACTACCGAG ATATCCGCAC CAACGCGCAG CCCGGACTCG
1561 GTAATGGCGC GCATTGCGCC CAGCGCCATC TGATCGTTGG CAACCAGCAT CGCAGTGGGA
1621 ACGATGCCCT CATTCAGCAT TTGCATGGTT TGTTGAAAAC CGGACATGGC ACTCCAGTCG
1681 CCTTCCCGTT CCGCTATCGG CTGAATTTGA TTGCGAGTGA GATATTTATG CCAGCCAGCC
1741 AGACGCAGAC GCGCCGAGAC AGAACTTAAT GGGCCCGCTA ACAGCGCGAT TTGCTGGTGA
1801 CCCAATGCGA CCAGATGCTC CACGCCCAGT CGCGTACCGT CTTCATGGGA GAAAATAATA
1861 CTGTTGATGG GTGTCTGGTC AGAGACATCA AGAAATAACG CCGGAACATT AGTGCAGGCA
1921 GCTTCCACAG CAATGGCATC CTGGTCATCC AGCGGATAGT TAATGATCAG CCCACTGACG
1981 CGTTGCGCGA GAAGATTGTG CACCGCCGCT TTACAGGCTT CGACGCCGCT TCGTTCTACC
2041 ATCGACACCA CCACGCTGGC ACCCAGTTGA TCGGCGCGAG ATTTAATCGC CGCGACAATT
2101 TGCGACGGCG CGTGCAGGGC CAGACTGGAG GTGGCAACGC CAATCAGCAA CGACTGTTTG
2161 CCCGCCAGTT GTTGTGCCAC GCGGTTGGGA ATGTAATTCA GCTCCGCCAT CGCCGCTTCC
2221 ACTTTTTCCC GCGTTTTCGC AGAAACGTGG CTGGCCTGGT TCACCACGCG GGAAACGGTC
2281 TGATAAGAGA CACCGGCATA CTCTGCGACA TCGTATAACG TTACTGGTTT CACATTCACC
2341 ACCCTGAATT GACTCTCTTC CGGGCGCTAT CATGCCATAC CGCGAAAGGT TTTGCACCAT
2401 TCGATGGTGT CGGAATTTCG GGCAGCGTTG GGTCCTGGCC ACGGGTGCGC ATGATCTAGA
2461 GCTGCCTCGC GCGTTTCGGT GATGACGGTG AAAACCTCTG ACACATGCAG CTCCCGGAGA
2521 CGGTCACAGC TTGTCTGTAA GCGGATGCCG GGAGCAGACA AGCCCGTCAG GGCGCGTCAG
2581 CGGGTGTTGG CGGGTGTCGG GGCGCAGCCA TGACCCAGTC ACGTAGCGAT AGCGGAGTGT
2641 ATACTGGCTT AACTATGCGG CATCAGAGCA GATTGTACTG AGAGTGCACC ATATGCGGTG
2701 TGAAATACCG CACAGATGCG TAAGGAGAAA ATACCGCATC AGGCGCTCTT CCGCTTCCTC
2761 GCTCACTGAC TCGCTGCGCT CGGTCGTTCG GCTGCGGCGA GCGGTATCAG CTCACTCAAA
2821 GGCGGTAATA CGGTTATCCA CAGAATCAGG GGATAACGCA GGAAAGAACA TGTGAGCAAA
2881 AGGCCAGCAA AAGGCCAGGA ACCGTAAAAA GGCCGCGTTG CTGGCGTTTT TCCATAGGCT
2941 CCGCCCCCCT GACGAGCATC ACAAAAATCG ACGCTCAAGT CAGAGGTGGC GAAACCCGAC
3001 AGGACTATAA AGATACCAGG CGTTTCCCCC TGGAAGCTCC CTCGTGCGCT CTCCTGTTCC
3061 GACCCTGCCG CTTACCGGAT ACCTGTCCGC CTTTCTCCCT TCGGGAAGCG TGGCGCTTTC
3121 TCATAGCTCA CGCTGTAGGT ATCTCAGTTC GGTGTAGGTC GTTCGCTCCA AGCTGGGCTG
3181 TGTGCACGAA CCCCCCGTTC AGCCCGACCG CTGCGCCTTA TCCGGTAACT ATCGTCTTGA
3241 GTCCAACCCG GTAAGACACG ACTTATCGCC ACTGGCAGCA GCCACTGGTA ACAGGATTAG
3301 CAGAGCGAGG TATGTAGGCG GTGCTACAGA GTTCTTGAAG TGGTGGCCTA ACTACGGCTA
3361 CACTAGAAGG ACAGTATTTG GTATCTGCGC TCTGCTGAAG CCAGTTACCT TCGGAAAAAG
3421 AGTTGGTAGC TCTTGATCCG GCAAACAAAC CACCGCTGGT AGCGGTGGTT TTTTTGTTTG
3481 CAAGCAGCAG ATTACGCGCA GAAAAAAAGG ATCTCAAGAA GATCCTTTGA TCTTTTCTAC
3541 GGGGTCTGAC GCTCAGTGGA ACGAAAACTC ACGTTAAGGG ATTTTGGTCA TGAGATTATC
3601 AAAAAGGATC TTCACCTAGA TCCTTTTAAA TTAAAAATGA AGTTTTAAAT CAATCTAAAG
3661 TATATATGAG TAAACTTGGT CTGACAGTTA CCAATGCTTA ATCAGTGAGG CACCTATCTC
3721 AGCGATCTGT CTATTTCGTT CATCCATAGT TGCCTGACTC CCCGTCGTGT AGATAACTAC
3781 GATACGGGAG GGCTTACCAT CTGGCCCCAG TGCTGCAATG ATACCGCGAG ACCCACGCTC
3841 ACCGGCTCCA GATTTATCAG CAATAAACCA GCCAGCCGGA AGGGCCGAGC GCAGAAGTGG
3901 TCCTGCAACT TTATCCGCCT CCATCCAGTC TATTAATTGT TGCCGGGAAG CTAGAGTAAG
3961 TAGTTCGCCA GTTAATAGTT TGCGCAACGT TGTTGCCATT GCTACAGGCA TCGTGGTGTC
4021 ACGCTCGTCG TTTGGTATGG CTTCATTCAG CTCCGGTTCC CAACGATCAA GGCGAGTTAC
4081 ATGATCCCCC ATGTTGTGCA AAAAAGCGGT TAGCTCCTTC GGTCCTCCGA TCGTTGTCAG
4141 AAGTAAGTTG GCCGCAGTGT TATCACTCAT GGTTATGGCA GCACTGCATA ATTCTCTTAC
4201 TGTCATGCCA TCCGTAAGAT GCTTTTCTGT GACTGGTGAG TACTCAACCA AGTCATTCTG
4261 AGAATAGTGT ATGCGGCGAC CGAGTTGCTC TTGCCCGGCG TCAATACGGG ATAATACCGC
4321 GCCACATAGC AGAACTTTAA AAGTGCTCAT CATTGGAAAA CGTTCTTCGG GGCGAAAACT
4381 CTCAAGGATC TTACCGCTGT TGAGATCCAG TTCGATGTAA CCCACTCGTG CACCCAACTG
4441 ATCTTCAGCA TCTTTTACTT TCACCAGCGT TTCTGGGTGA GCAAAAACAG GAAGGCAAAA
4501 TGCCGCAAAA AAGGGAATAA GGGCGACACG GAAATGTTGA ATACTCATAC TCTTCCTTTT
4561 TCAATATTAT TGAAGCATTT ATCAGGGTTA TTGTCTCATG AGCGGATACA TATTTGAATG
4621 TATTTAGAAA AATAAACAAA TAGGGGTTCC GCGCACATTT CCCCGAAAAG TGCCACCTGA
4681 CGTCTAAGAA ACCATTATTA TCATGACATT AACCTATAAA AATAGGCGTA TCACGAGGCC
4741 CTTTCGTCTT CAC
//
DEFINITION pQE-81L
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTOAUTHORAD1|Novagen Inc.|
FEATURES Location/Qualifiers
source 1..4753
/organism="pQE-81L"
/mol_type="other DNA"
gene 348..1007
/label="CAT/CamR"
/gene="CAT/CamR"
CDS 348..1007
/label="ORF frame 3"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAF
LKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETF
SSLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNV
ANMDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG
A*"
terminator 1075..1118
/label="rrnB_T1_terminator"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAF
LKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETF
SSLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNV
ANMDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG
A*"
misc_feature complement(1251..2342)
/label="lacI"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAF
LKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETF
SSLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNV
ANMDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG
A*"
CDS complement(1251..2210)
/label="ORF frame 3"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
misc_feature complement(2495..2517)
/label="pGEX_3_primer"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
gene complement(3688..4548)
/label="Ampicillin"
/gene="Ampicillin"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
CDS complement(3688..4548)
/label="ORF frame 2"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
promoter complement(4590..4618)
/label="AmpR_promoter"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
ORIGIN
1 CTCGAGAAAT CATAAAAAAT TTATTTGCTT TGTGAGCGGA TAACAATTAT AATAGATTCA
61 ATTGTGAGCG GATAACAATT TCACACAGAA TTCATTAAAG AGGAGAAATT AACTATGAGA
121 GGATCTCACC ATCACCATCA CCATACGGAT CCGCATGCGA GCTCGGTACC CCGGGTCGAC
181 CTGCAGCCAA GCTTAATTAG CTGAGCTTGG ACTCCTGTTG ATAGATCCAG TAATGACCTC
241 AGAACTCCAT CTGGATTTGT TCAGAACGCT CGGTTGCCGC CGGGCGTTTT TTATTGGTGA
301 GAATCCAAGC TAGCTTGGCG AGATTTTCAG GAGCTAAGGA AGCTAAAATG GAGAAAAAAA
361 TCACTGGATA TACCACCGTT GATATATCCC AATGGCATCG TAAAGAACAT TTTGAGGCAT
421 TTCAGTCAGT TGCTCAATGT ACCTATAACC AGACCGTTCA GCTGGATATT ACGGCCTTTT
481 TAAAGACCGT AAAGAAAAAT AAGCACAAGT TTTATCCGGC CTTTATTCAC ATTCTTGCCC
541 GCCTGATGAA TGCTCATCCG GAATTTCGTA TGGCAATGAA AGACGGTGAG CTGGTGATAT
601 GGGATAGTGT TCACCCTTGT TACACCGTTT TCCATGAGCA AACTGAAACG TTTTCATCGC
661 TCTGGAGTGA ATACCACGAC GATTTCCGGC AGTTTCTACA CATATATTCG CAAGATGTGG
721 CGTGTTACGG TGAAAACCTG GCCTATTTCC CTAAAGGGTT TATTGAGAAT ATGTTTTTCG
781 TCTCAGCCAA TCCCTGGGTG AGTTTCACCA GTTTTGATTT AAACGTGGCC AATATGGACA
841 ACTTCTTCGC CCCCGTTTTC ACCATGGGCA AATATTATAC GCAAGGCGAC AAGGTGCTGA
901 TGCCGCTGGC GATTCAGGTT CATCATGCCG TTTGTGATGG CTTCCATGTC GGCAGAATGC
961 TTAATGAATT ACAACAGTAC TGCGATGAGT GGCAGGGCGG GGCGTAATTT TTTTAAGGCA
1021 GTTATTGGTG CCCTTAAACG CCTGGGGTAA TGACTCTCTA GCTTGAGGCA TCAAATAAAA
1081 CGAAAGGCTC AGTCGAAAGA CTGGGCCTTT CGTTTTATCT GTTGTTTGTC GGTGAACGCT
1141 CTCCTGAGTA GGACAAATCC GCCCTCTAGA TTACGTGCAG TCGATGATAA GCTGTCAAAC
1201 ATGAGAATTG TGCCTAATGA GTGAGCTAAC TTACATTAAT TGCGTTGCGC TCACTGCCCG
1261 CTTTCCAGTC GGGAAACCTG TCGTGCCAGC TGCATTAATG AATCGGCCAA CGCGCGGGGA
1321 GAGGCGGTTT GCGTATTGGG CGCCAGGGTG GTTTTTCTTT TCACCAGTGA GACGGGCAAC
1381 AGCTGATTGC CCTTCACCGC CTGGCCCTGA GAGAGTTGCA GCAAGCGGTC CACGCTGGTT
1441 TGCCCCAGCA GGCGAAAATC CTGTTTGATG GTGGTTAACG GCGGGATATA ACATGAGCTG
1501 TCTTCGGTAT CGTCGTATCC CACTACCGAG ATATCCGCAC CAACGCGCAG CCCGGACTCG
1561 GTAATGGCGC GCATTGCGCC CAGCGCCATC TGATCGTTGG CAACCAGCAT CGCAGTGGGA
1621 ACGATGCCCT CATTCAGCAT TTGCATGGTT TGTTGAAAAC CGGACATGGC ACTCCAGTCG
1681 CCTTCCCGTT CCGCTATCGG CTGAATTTGA TTGCGAGTGA GATATTTATG CCAGCCAGCC
1741 AGACGCAGAC GCGCCGAGAC AGAACTTAAT GGGCCCGCTA ACAGCGCGAT TTGCTGGTGA
1801 CCCAATGCGA CCAGATGCTC CACGCCCAGT CGCGTACCGT CTTCATGGGA GAAAATAATA
1861 CTGTTGATGG GTGTCTGGTC AGAGACATCA AGAAATAACG CCGGAACATT AGTGCAGGCA
1921 GCTTCCACAG CAATGGCATC CTGGTCATCC AGCGGATAGT TAATGATCAG CCCACTGACG
1981 CGTTGCGCGA GAAGATTGTG CACCGCCGCT TTACAGGCTT CGACGCCGCT TCGTTCTACC
2041 ATCGACACCA CCACGCTGGC ACCCAGTTGA TCGGCGCGAG ATTTAATCGC CGCGACAATT
2101 TGCGACGGCG CGTGCAGGGC CAGACTGGAG GTGGCAACGC CAATCAGCAA CGACTGTTTG
2161 CCCGCCAGTT GTTGTGCCAC GCGGTTGGGA ATGTAATTCA GCTCCGCCAT CGCCGCTTCC
2221 ACTTTTTCCC GCGTTTTCGC AGAAACGTGG CTGGCCTGGT TCACCACGCG GGAAACGGTC
2281 TGATAAGAGA CACCGGCATA CTCTGCGACA TCGTATAACG TTACTGGTTT CACATTCACC
2341 ACCCTGAATT GACTCTCTTC CGGGCGCTAT CATGCCATAC CGCGAAAGGT TTTGCACCAT
2401 TCGATGGTGT CGGAATTTCG GGCAGCGTTG GGTCCTGGCC ACGGGTGCGC ATGATCTAGA
2461 GCTGCCTCGC GCGTTTCGGT GATGACGGTG AAAACCTCTG ACACATGCAG CTCCCGGAGA
2521 CGGTCACAGC TTGTCTGTAA GCGGATGCCG GGAGCAGACA AGCCCGTCAG GGCGCGTCAG
2581 CGGGTGTTGG CGGGTGTCGG GGCGCAGCCA TGACCCAGTC ACGTAGCGAT AGCGGAGTGT
2641 ATACTGGCTT AACTATGCGG CATCAGAGCA GATTGTACTG AGAGTGCACC ATATGCGGTG
2701 TGAAATACCG CACAGATGCG TAAGGAGAAA ATACCGCATC AGGCGCTCTT CCGCTTCCTC
2761 GCTCACTGAC TCGCTGCGCT CGGTCGTTCG GCTGCGGCGA GCGGTATCAG CTCACTCAAA
2821 GGCGGTAATA CGGTTATCCA CAGAATCAGG GGATAACGCA GGAAAGAACA TGTGAGCAAA
2881 AGGCCAGCAA AAGGCCAGGA ACCGTAAAAA GGCCGCGTTG CTGGCGTTTT TCCATAGGCT
2941 CCGCCCCCCT GACGAGCATC ACAAAAATCG ACGCTCAAGT CAGAGGTGGC GAAACCCGAC
3001 AGGACTATAA AGATACCAGG CGTTTCCCCC TGGAAGCTCC CTCGTGCGCT CTCCTGTTCC
3061 GACCCTGCCG CTTACCGGAT ACCTGTCCGC CTTTCTCCCT TCGGGAAGCG TGGCGCTTTC
3121 TCATAGCTCA CGCTGTAGGT ATCTCAGTTC GGTGTAGGTC GTTCGCTCCA AGCTGGGCTG
3181 TGTGCACGAA CCCCCCGTTC AGCCCGACCG CTGCGCCTTA TCCGGTAACT ATCGTCTTGA
3241 GTCCAACCCG GTAAGACACG ACTTATCGCC ACTGGCAGCA GCCACTGGTA ACAGGATTAG
3301 CAGAGCGAGG TATGTAGGCG GTGCTACAGA GTTCTTGAAG TGGTGGCCTA ACTACGGCTA
3361 CACTAGAAGG ACAGTATTTG GTATCTGCGC TCTGCTGAAG CCAGTTACCT TCGGAAAAAG
3421 AGTTGGTAGC TCTTGATCCG GCAAACAAAC CACCGCTGGT AGCGGTGGTT TTTTTGTTTG
3481 CAAGCAGCAG ATTACGCGCA GAAAAAAAGG ATCTCAAGAA GATCCTTTGA TCTTTTCTAC
3541 GGGGTCTGAC GCTCAGTGGA ACGAAAACTC ACGTTAAGGG ATTTTGGTCA TGAGATTATC
3601 AAAAAGGATC TTCACCTAGA TCCTTTTAAA TTAAAAATGA AGTTTTAAAT CAATCTAAAG
3661 TATATATGAG TAAACTTGGT CTGACAGTTA CCAATGCTTA ATCAGTGAGG CACCTATCTC
3721 AGCGATCTGT CTATTTCGTT CATCCATAGT TGCCTGACTC CCCGTCGTGT AGATAACTAC
3781 GATACGGGAG GGCTTACCAT CTGGCCCCAG TGCTGCAATG ATACCGCGAG ACCCACGCTC
3841 ACCGGCTCCA GATTTATCAG CAATAAACCA GCCAGCCGGA AGGGCCGAGC GCAGAAGTGG
3901 TCCTGCAACT TTATCCGCCT CCATCCAGTC TATTAATTGT TGCCGGGAAG CTAGAGTAAG
3961 TAGTTCGCCA GTTAATAGTT TGCGCAACGT TGTTGCCATT GCTACAGGCA TCGTGGTGTC
4021 ACGCTCGTCG TTTGGTATGG CTTCATTCAG CTCCGGTTCC CAACGATCAA GGCGAGTTAC
4081 ATGATCCCCC ATGTTGTGCA AAAAAGCGGT TAGCTCCTTC GGTCCTCCGA TCGTTGTCAG
4141 AAGTAAGTTG GCCGCAGTGT TATCACTCAT GGTTATGGCA GCACTGCATA ATTCTCTTAC
4201 TGTCATGCCA TCCGTAAGAT GCTTTTCTGT GACTGGTGAG TACTCAACCA AGTCATTCTG
4261 AGAATAGTGT ATGCGGCGAC CGAGTTGCTC TTGCCCGGCG TCAATACGGG ATAATACCGC
4321 GCCACATAGC AGAACTTTAA AAGTGCTCAT CATTGGAAAA CGTTCTTCGG GGCGAAAACT
4381 CTCAAGGATC TTACCGCTGT TGAGATCCAG TTCGATGTAA CCCACTCGTG CACCCAACTG
4441 ATCTTCAGCA TCTTTTACTT TCACCAGCGT TTCTGGGTGA GCAAAAACAG GAAGGCAAAA
4501 TGCCGCAAAA AAGGGAATAA GGGCGACACG GAAATGTTGA ATACTCATAC TCTTCCTTTT
4561 TCAATATTAT TGAAGCATTT ATCAGGGTTA TTGTCTCATG AGCGGATACA TATTTGAATG
4621 TATTTAGAAA AATAAACAAA TAGGGGTTCC GCGCACATTT CCCCGAAAAG TGCCACCTGA
4681 CGTCTAAGAA ACCATTATTA TCATGACATT AACCTATAAA AATAGGCGTA TCACGAGGCC
4741 CTTTCGTCTT CAC
//
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文献和实验相关实验
蛋白为44KD，融合后应为73KD，但是没有见到表达. 综合分析会是什么原因？该蛋白国外已在pQE30载体上表达成功，应该不会是稀有密码的原因造成表达终止吧？第1号和3号克隆已测序，表达框没有移码，基因全长正确。测序结果如下，请参考： AAATCGGATCTGGTTCCGCGTGGATCCCCGGAATTCCCGGGTCGACATGATACAAGTACTGATGGCTGCTGCAAGCTTTG
蛋白是相当重要的；同时可以纯化出那些完整表达的蛋白，一些在翻译过程中容易造成“暂停”现象的蛋白就不用怕纯化产物里含有不完整的产物。融合了6×组氨酸标签的有pQE-60和pQE-70，融合了6×His-DHFR的有pQE-16。 顺式抑制载体 pQE系列的表达载体一般情况下需要在M15菌株中进行表达的，但是设计的pQE80L系列就没有这种约束，它可以在任意大肠杆菌株中进行表达。这与一般的pQE载体利用M15中质粒pREP4的反式lacI抑制子调控不同，它是利用顺式lacIq调控产生大量lac抑制
8.0） PCI （25:24:1） RNase A （1 mg/mL） 1% agarose gel 方法步骤： 1） 由前一天转形所得的转形株中，挑选12~24 个单一菌落，在LB/Amp plate上划数道短的横线或斜线，同时也须要接种带有载体pQE31 的菌株，于37℃培养过夜或直至菌体长出。 2） 以接种环将菌体刮下，悬浊在30 μL 1×NET 中，震荡混合均匀。 ◆ 不要忘了pQE31/JM109! 3） 加入30 μL PCI,剧烈震荡。 4） 以12,000 rpm 离心
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pQE-81L载体
¥1100
