| 出品公司: | EMD Biosciences (Novagen) |
|---|---|
| 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 |
| 备注: |
N-term HisTag with minimal extraneous sequences.
|
| 产品目录号: | 71327-3 |
| 稳定性: | 瞬时表达 Transient |
| 组成型: | 组成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
| 表达宿主菌: | BL21(DE3) 、BLR(DE3)、Rosetta(DE3)、OrigamiB(DE3)、C43(DE3) |
| pET表达手册: |
pET表达手册英文版 pET表达手册中文版 |
| pET载体汇总: |
pET-3a pET-11a pET-12a pET-14b pET-15b pET-16b pET-17b pET-19b pET-20b pET-21a pET-22b pET-23a pET-24a pET-25b pET-26b pET-27b pET-28a pET-29a pET-30a pET-31b pET-32a pET-33b pET-34b pET-35b pET-37b pET-39b pET-40b pET-41a pET-42a pET-43.1a pET-44a pET-45b pET-47b pET-48b pET-49b pET-50b pET-51b pET-52b pETDuet-1 pACYCDuet-1 pCDFDuet-1 pRSFDuet-1 pColaDuet-1 |
买家导航

pET-45b(+)载体 pET45b质粒基本信息
pET-45b(+)质粒图谱 pET45b载体图谱和pET-45b(+)载体序列 pET45b质粒序列多克隆位点信息
| 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 |
pET-45b(+)质粒 pET45b载体简介
pET-45b载体是带有N-His蛋白标签,EK蛋白酶位点和C-S标签,可以在大肠杆菌中高表达目的蛋白。独特的酶切位点在载体图谱中被标注出来。需要注意的是,这些载体序列是依据载体pBR322的方向来进行编号的,所以T7标签表达区域在载体图谱上是反向的。
T7 RNA聚合酶启动的克隆和表达区域在质粒图谱中也被标注了出来。质粒的F1复制子是被定向的,所以在T7噬菌体聚合酶的作用下,包含有蛋白编码序列的病毒 粒子能够产生,并启动蛋白表达,同时蛋白表达将被T7终止子序列(Cat. No. 69337-3)的作用下终止蛋白翻译。 pET-45b载体上面含有一个EK蛋白酶切位点,当将目的基因使用载体上面的PshAI限制性内切酶位点插入进去时,可以通过EK蛋白酶将载体上的融合的所有氨基酸序列包括标签序列,全部切除掉。

pET-45b(+)质粒序列 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
//
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pET-45b(+)质粒 pET45b载体应用举例
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