| 出品公司: | |
|---|---|
| 别名: | pET45b, pET 45b |
| 质粒类型: | 大肠杆菌蛋白表达 |
| 表达水平: | 高 |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 5260 bp |
| 5' 测序引物: | T7 |
| 5' 测序引物序列: | T7: 5'-TAATACGACTCACTATAGGG-3' |
| 3' 测序引物: | T7t |
| 3' 测序引物序列: | T7t: 5'-TGCTAGTTATTGCTCAGCGG-3' |
| 载体标签: | N-His,N-EK, C-S |
| 载体抗性: | Ampicillin |
| 表达宿主菌: | BL21(DE3) 、 Rosetta2(DE3)、C43(DE3) |
| 备注: |
N-term HisTag with minimal extraneous sequences.
|
| 产品目录号: | ZY71327-3 |
| 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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- 技术资料
- 保存条件：
-20℃低温保存
- 保质期：
三年
- 英文名：
pET-45b(+)
- 库存：
20
- 供应商：
泽叶生物
pET45b载体基本信息
pET45b载体质粒图谱和多克隆位点信息
| Feature Name | Start | End | |
|---|---|---|---|
| Ampicillin | 1442 | 582 | |
| AmpR_promoter | 1513 | 1485 | |
| pBR322_origin | 1600 | 2219 | |
| pGEX_3_primer | 2618 | 2640 | |
| ROP | 2825 | 2634 | |
| lacI | 4488 | 3397 | |
| tet (887 - 636) | 4831 | 4580 | |
| pBRrevBam_primer | 4763 | 4782 | |
| T7_promoter | 4870 | 4888 | |
| lacO | 4888 | 4915 | |
| T7_transl_en_RBS | 4933 | 4949 | |
| EK | 4999 | 5013 | |
| T7_Terminal_primer | 5192 | 5174 | |
| T7_terminator | 5193 | 5228 |
| ORF | Start | End | |
|---|---|---|---|
| ORF frame 3 | 1442 | 582 | |
| ORF frame 2 | 4356 | 3397 | |
| ORF frame 1 | 4729 | 4229 |
| Enzyme Name | Cut |
|---|---|
| FspI | 877 |
| HpaI | 3622 |
| ApaI | 3921 |
| BclI | 4110 |
| ClaI | 4849 |
| XbaI | 4916 |
| NcoI | 4955 |
| PmlI | 4980 |
| KpnI | 4988 |
| AgeI | 4987 |
| BstBI | 4993 |
| BamHI | 5019 |
| SacI | 5036 |
| AscI | 5047 |
| PstI | 5057 |
| SalI | 5059 |
| HindIII | 5065 |
| NotI | 5072 |
| EagI | 5072 |
| XhoI | 5080 |
| PacI | 5155 |
pET45b载体简介
The pET-45b(+) vector carries an N-terminal His•Tag® coding sequence followed by an enterokinase cleavage site and an optional C-terminal S•TagTM sequence. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the vector map. The f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand. Therefore, single-stranded sequencing should be performed using the T7 Terminator Primer (Cat. No. 69337-3).
pET45b载体序列
LOCUS pET-45b(+) 5260 bp DNA Circular SYN
DEFINITION pET-45b(+)
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..5260
/organism="pET-45b(+)"
/mol_type="other DNA"
gene complement(582..1442)
/label="Ampicillin"
/gene="Ampicillin"
CDS complement(582..1442)
/label="ORF frame 3"
promoter complement(1485..1513)
/label="AmpR_promoter"
rep_origin 1600..2219
/label="pBR322_origin"
misc_feature 2618..2640
/label="pGEX_3_primer"
misc_feature complement(2634..2825)
/label="ROP"
misc_feature complement(3397..4488)
/label="lacI"
CDS complement(3397..4356)
/label="ORF frame 2"
CDS complement(4229..4729)
/label="ORF frame 1"
gene complement(4580..4831)
/label="tet (887 - 636)"
/gene="tet (887 - 636)"
misc_feature 4763..4782
/label="pBRrevBam_primer"
promoter 4870..4888
/label="T7_promoter"
misc_feature 4888..4915
/label="lacO"
misc_feature 4933..4949
/label="T7_transl_en_RBS"
misc_feature 4999..5013
/label="EK"
misc_feature complement(5174..5192)
/label="T7_Terminal_primer"
terminator 5193..5228
/label="T7_terminator"
ORIGIN
1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttctg gcggcacgat
481 ggcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
541 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
601 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
661 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
721 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
781 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
841 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
901 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
961 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
1021 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
1081 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
1141 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
1201 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
1261 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
1321 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
1381 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
1441 atactcttcc tttttcaatc atgattgaag catttatcag ggttattgtc tcatgagcgg
1501 atacatattt gaatgtattt agaaaaataa acaaataggt catgaccaaa atcccttaac
1561 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
1621 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
1681 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
1741 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
1801 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
1861 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
1921 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
1981 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
2041 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
2101 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
2161 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
2221 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
2281 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
2341 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
2401 attttctcct tacgcatctg tgcggtattt cacaccgcat atatggtgca ctctcagtac
2461 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
2521 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
2581 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
2641 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
2701 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
2761 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
2821 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
2881 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
2941 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
3001 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
3061 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
3121 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
3181 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
3241 cgccagccta gccgggtcct caacgacagg agcacgatca tgctagtcat gccccgcgcc
3301 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
3361 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
3421 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
3481 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
3541 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
3601 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
3661 gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
3721 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
3781 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
3841 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
3901 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
3961 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
4021 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
4081 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
4141 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
4201 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
4261 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
4321 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
4381 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
4441 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
4501 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
4561 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
4621 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
4681 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
4741 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
4801 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc
4861 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
4921 aaataatttt gtttaacttt aagaaggaga tataccatgg cacatcacca ccaccatcac
4981 gtgggtaccg gttcgaatga tgacgacgac aagagtccgg atcccaattg ggagctcgtg
5041 tacacggcgc gcctgcaggt cgacaagctt gcggccgcac tcgagtctgg taaagaaacc
5101 gctgctgcga aatttgaacg ccagcacatg gactcgtcta ctagcgcagc ttaattaacc
5161 taggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
5221 ttgaggggtt ttttgctgaa aggaggaact atatccggat
DEFINITION pET-45b(+)
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..5260
/organism="pET-45b(+)"
/mol_type="other DNA"
gene complement(582..1442)
/label="Ampicillin"
/gene="Ampicillin"
CDS complement(582..1442)
/label="ORF frame 3"
promoter complement(1485..1513)
/label="AmpR_promoter"
rep_origin 1600..2219
/label="pBR322_origin"
misc_feature 2618..2640
/label="pGEX_3_primer"
misc_feature complement(2634..2825)
/label="ROP"
misc_feature complement(3397..4488)
/label="lacI"
CDS complement(3397..4356)
/label="ORF frame 2"
CDS complement(4229..4729)
/label="ORF frame 1"
gene complement(4580..4831)
/label="tet (887 - 636)"
/gene="tet (887 - 636)"
misc_feature 4763..4782
/label="pBRrevBam_primer"
promoter 4870..4888
/label="T7_promoter"
misc_feature 4888..4915
/label="lacO"
misc_feature 4933..4949
/label="T7_transl_en_RBS"
misc_feature 4999..5013
/label="EK"
misc_feature complement(5174..5192)
/label="T7_Terminal_primer"
terminator 5193..5228
/label="T7_terminator"
ORIGIN
1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttctg gcggcacgat
481 ggcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
541 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
601 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
661 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
721 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
781 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
841 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
901 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
961 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
1021 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
1081 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
1141 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
1201 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
1261 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
1321 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
1381 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
1441 atactcttcc tttttcaatc atgattgaag catttatcag ggttattgtc tcatgagcgg
1501 atacatattt gaatgtattt agaaaaataa acaaataggt catgaccaaa atcccttaac
1561 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
1621 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
1681 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
1741 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
1801 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
1861 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
1921 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
1981 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
2041 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
2101 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
2161 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
2221 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
2281 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
2341 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
2401 attttctcct tacgcatctg tgcggtattt cacaccgcat atatggtgca ctctcagtac
2461 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
2521 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
2581 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
2641 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
2701 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
2761 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
2821 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
2881 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
2941 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
3001 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
3061 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
3121 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
3181 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
3241 cgccagccta gccgggtcct caacgacagg agcacgatca tgctagtcat gccccgcgcc
3301 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
3361 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
3421 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
3481 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
3541 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
3601 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
3661 gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
3721 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
3781 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
3841 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
3901 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
3961 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
4021 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
4081 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
4141 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
4201 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
4261 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
4321 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
4381 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
4441 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
4501 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
4561 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
4621 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
4681 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
4741 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
4801 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc
4861 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
4921 aaataatttt gtttaacttt aagaaggaga tataccatgg cacatcacca ccaccatcac
4981 gtgggtaccg gttcgaatga tgacgacgac aagagtccgg atcccaattg ggagctcgtg
5041 tacacggcgc gcctgcaggt cgacaagctt gcggccgcac tcgagtctgg taaagaaacc
5101 gctgctgcga aatttgaacg ccagcacatg gactcgtcta ctagcgcagc ttaattaacc
5161 taggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
5221 ttgaggggtt ttttgctgaa aggaggaact atatccggat
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文献和实验相关实验
表达载体的构建是生物技术和所需蛋白质产生的基本工具，本文总结了pET-32α（+）载体技术的构建，该技术通常用于研究实验室。该方法包括获得用于构建载体的外源DNA片段，将DNA片段亚克隆到pET-32α（+）表达载体中，在大肠杆菌 BL21（DE3）中进行蛋白质表达以及在大肠杆菌中在天然条件下进行蛋白质纯化。裂解液。介绍作为分子生物学实验室中非常有用的实践，重组DNA技术（或基因克隆）是指将DNA片段从一种生物体转移到自我复制的遗传元件如表达载体。然后插入的DNA可以在外源宿主细胞中繁殖
相关专题 1)噬菌体DE3溶源化的菌株如BL21(DE3)是最常用的表达菌株，噬菌体DE3是λ噬菌体的衍生株，构建好的表达载体 可以直接转入表达菌株中，诱导蛋白的表达方式与lac启动子一样都是IPTG诱导。 2)表达菌株BL21(DE3)上带有lacI基因，T7 RNA 聚合酶编码基因以及lacUV5启动子，lacUV5启动子可以启动T7 RNA 聚合酶的表达; 3)表达质粒 如pET-28质粒带有T7启动子和编码阻遏
Expression System、HisPatch ThioFusion System、pBAD Inducible Bacterial System、pL System和T7-based Systems。这四个系列的载体可谓把原核表达的发展史中出现过的几种启动子都囊括进去了，包括T7、pL、pBAD和trc启动子，同时几种经典的调控方式也都出现了，包括有乳糖操纵子、色氨酸操纵子和阿拉伯糖操纵子。 冠军系列 Champion™ pET Expression System之所以取
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pET-45b(+)载体
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