| 出品公司: | EMD Biosciences (Novagen) |
|---|---|
| 別名: | pET49b, pET 49b |
| 質粒類型: | 大腸桿菌蛋白表達 |
| 表達水平: | 高 |
| 克隆方法: | 多克隆位點，限制性內切酶 |
| 載體大小: | 5926 bp |
| 5' 測序引物: | T7或者pGEX 5' |
| 5' 測序引物序列: | T7: 5'-TAATACGACTCACTATAGGG-3'; pGEX 5': 5'-GGGCTGGCAAGCCACGTTTGGTG-3' |
| 3' 測序引物: | T7t |
| 3' 測序引物序列: | T7t: 5'-TGCTAGTTATTGCTCAGCGG-3' |
| 載體標簽: | N-GST, N-His, N-HRV 3C, C-S, C-Thrombin |
| 載體抗性: | Kanamycin |
| 備注: |
Same as pET47 but also has Nterm GST Tag; contains HRV 3C Protease cleavage site for fusion tag removal at low temperatures; Cterm thrombin cleavage site.
|
| 產品目錄號: | 71463-3 |
| 穩定性: | 瞬時表達 Transient |
| 組成型: | 組成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
| 表達宿主菌: | BL21(DE3) 、BLR(DE3)、Rosetta(DE3)、OrigamiB(DE3)、C43(DE3) |
| pET表達手冊： |
pET表達手冊英文版 pET表達手冊中文版 |
| pET載體匯總: |
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 |
買家導航
pET-49b(+)載體 pET49b質粒基本信息
pET-49b(+)質粒圖譜 pET49b載體圖譜和pET-49b(+)載體序列 pET49b質粒序列多克隆位點信息
| Feature Name | Start | End | |
|---|---|---|---|
| T7_terminator | 120 | 1 | |
| T7_Terminal_primer | 69 | 87 | |
| S15 | 212 | 168 | |
| 6xHIS | 359 | 342 | |
| GST (variant) | 1088 | 438 | |
| T7_transl_en_RBS | 1113 | 1097 | |
| lacO | 1158 | 1131 | |
| T7_promoter | 1176 | 1158 | |
| tet (300 - 563) | 1212 | 1475 | |
| pBRrevBam_primer | 1283 | 1264 | |
| lacI | 1558 | 2649 | |
| ROP | 3221 | 3412 | |
| pGEX_3_primer | 3428 | 3406 | |
| pBR322_origin | 4446 | 3827 | |
| KanR2 | 4552 | 5367 | |
| f1_origin | 5898 | 5592 |
| ORF | Start | End | |
|---|---|---|---|
| ORF frame 3 | 1088 | 141 | |
| ORF frame 3 | 1089 | 1817 | |
| ORF frame 1 | 1690 | 2649 | |
| ORF frame 1 | 4552 | 5367 |
| Enzyme Name | Cut |
|---|---|
| PacI | 141 |
| XhoI | 147 |
| SacI | 238 |
| NotI | 242 |
| EagI | 242 |
| HindIII | 249 |
| SalI | 255 |
| AatII | 266 |
| AscI | 270 |
| StuI | 280 |
| EcoRI | 289 |
| BamHI | 295 |
| KpnI | 306 |
| XmaI | 306 |
| SmaI | 308 |
| SacII | 338 |
| SpeI | 417 |
| MscI | 880 |
| NdeI | 1087 |
| XbaI | 1125 |
| ApaI | 2128 |
| HpaI | 2423 |
| NruI | 4640 |
pET-49b(+)質粒 pET49b載體簡介
pET-49b載體是帶有N-His和N-GST蛋白標簽，HRV3C蛋白酶位點和C-S標簽，可以在大腸桿菌中高表達目的蛋白。獨特的酶切位點在載體圖譜中被標注出來。需要注意的是，這些載體序列是依據載體pBR322的方向來進行編號的，所以T7標簽表達區域在載體圖譜上是反向的。
T7 RNA聚合酶啟動的克隆和表達區域在質粒圖譜中也被標注了出來。質粒的F1復制子是被定向的，所以在T7噬菌體聚合酶的作用下，包含有蛋白編碼序列的病毒 粒子能夠產生，并啟動蛋白表達，同時蛋白表達將被T7終止子序列(Cat. No. 69337-3)的作用下終止蛋白翻譯。
pET-49b載體上面含有一個EK蛋白酶切位點，當將目的基因使用載體上面的PshAI限制性內切酶位點插入進去時，可以通過EK蛋白酶將載體上的融合的所有氨基酸序列包括GST標簽序列，全部切除掉。
pET-49b(+)質粒序列 pET49b載體序列
LOCUS pET-49b(+) 5926 bp DNA circular 11-SEP-2008
DEFINITION pET-49b(+).
SOURCE
ORGANISM
COMMENT This file is created by Vector NTI
//gt12365.cn/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|gt12365.cn|
COMMENT VNTOAUTHORAD1|Novagen Inc.|
COMMENT VNTREPLTYPE|Plasmid
COMMENT VNTEXTCHREPL|Bacteria
FEATURES Location/Qualifiers
rep_origin complement(5467..5914)
/vntifkey="33"
/label=f1\ori
misc_RNA complement(3843..4398)
/vntifkey="53"
/label=RNA\II
rep_origin complement(3843..3843)
/vntifkey="33"
/label=ColE1,\pBR\ori
misc_RNA 4283..4390
/vntifkey="53"
/label=RNA\I
CDS 3221..3412
/vntifkey="4"
/label=ROP\gene
CDS 1567..2649
/vntifkey="4"
/label=lacI
/note="lac repressor"
promoter complement(1160..1176)
/vntifkey="30"
/label=T7\promoter
protein_bind complement(1135..1155)
/vntifkey="31"
/label=lacO
CDS complement(429..1088)
/vntifkey="4"
/label=GST
protein_bind complement(342..359)
/vntifkey="31"
/label=His-Tag
terminator complement(26..73)
/vntifkey="43"
/label=T7\terminator
CDS 4552..5367
/vntifkey="4"
/label=Kan
misc_feature complement(213..230)
/vntifkey="21"
/label=Thrombin\site
protein_bind complement(168..212)
/vntifkey="31"
/label=S-Tag
protein_bind complement(310..332)
/vntifkey="31"
/label=3c
primer complement(300..336)
/vntifkey="27"
/label=S-SacII/Bam\+SanDI\-dcm
primer complement(374..421)
/vntifkey="27"
/label=S-Tag\Spacer-Spe\I
BASE COUNT 1485 a 1486 c 1520 g 1435 t
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgtttagcag cctaggttaa ttaagcctcg agagcagcag aagtagagct gtccatgtgc
181 tggcgttcga atttagcagc agcggtttct ttactaccgc gtggcaccag agcgagctct
241 gcggccgcaa gcttgtcgac ggacgtcggg cgcgccaagg cctgtacaga attcggatcc
301 tggtacccgg gtccctgaaa gaggacttca agagccgcgg agtgatggtg atggtgatga
361 ccagaaccac tagatggagt aggagtagga ggattgttat tagaaccgcc accaccacta
421 gttgaaccat ccgattttgg aggatggtcg ccaccaccaa acgtggcttg ccagccctgc
481 aaaggccatg ctatatactt gctggatttc aagtacttat caatttgtgg gatagcttca
541 atacgttttt taaaacaaac taattttggg aacgcatcca ggcacattgg gtccatgtat
601 aaaacaacat caagagcgtc atacaacatg aagtcaggat gggttacatg atcaccattt
661 aaatatgttt tatgacataa acgatcttcg aacattttca gcatttcagg tagcttgcta
721 agaaaatcaa ctttgagagt ttcaaagtct ttactatatg caattctcga aacaccgtat
781 ctaatatcca aaaccgctcc ttcaagcatt gaaatctctg cacgctcttt tggacaacca
841 cccaacatgt tgtgcttgtc agctatataa cgtatgatgg ccatagactg tgttaattta
901 acatcaccat caatataata aggaagattg ggaaactcca aacccaattc aaactttttg
961 tttcgccatt tatcaccttc atcgcgctca tacaaatgct cttcatattt ttcttcaaga
1021 tattccaaaa gaagtcgagt gggttgcaca aggcccttaa ttttccaata acctagtata
1081 ggggacatat gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt
1141 tatccgctca caattcccct atagtgagtc gtattaattt cgcgggatcg agatcgatct
1201 cgatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg
1261 gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga
1321 gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca
1381 tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg
1441 gctgcttcct aatgcaggag tcgcataagg gagagcgtcg agatcccgga caccatcgaa
1501 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
1561 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
1621 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
1681 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
1741 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
1801 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
1861 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc
1921 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
1981 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
2041 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
2101 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg
2161 gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc
2221 gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt
2281 cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc
2341 gagtccgggc tgcgcgttgg tgcggacatc tcggtagtgg gatacgacga taccgaagac
2401 agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa
2461 accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg
2521 ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct
2581 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
2641 gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cgggatctcg
2701 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
2761 tagcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg
2821 ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc
2881 tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac
2941 gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct
3001 accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt
3061 tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg
3121 ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat
3181 tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac
3241 cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa
3301 cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga
3361 gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
3421 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
3481 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
3541 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
3601 catatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
3661 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
3721 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
3781 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
3841 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
3901 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
3961 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
4021 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
4081 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
4141 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
4201 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
4261 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
4321 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
4381 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
4441 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
4501 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
4561 attcaacggg aaacgtcttg ctctaggccg cgattaaatt ccaacatgga tgctgattta
4621 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg
4681 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
4741 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
4801 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggc
4861 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
4921 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagt
4981 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
5041 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
5101 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
5161 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
5221 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
5281 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
5341 catttgatgc tcgatgagtt tttctaagaa ttaattcatg agcggataca tatttgaatg
5401 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
5461 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
5521 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
5581 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
5641 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
5701 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
5761 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
5821 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
5881 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgcca
//
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pET-49b(+)質粒 pET49b載體應用舉例
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