出品公司: | EMD Biosciences (Novagen) |
---|---|
別名: | pET44c, pET 44c |
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: | 7309 bp |
5' 測序引物: | T7 |
5' 測序引物序列: | T7: 5'-TAATACGACTCACTATAGGG-3' |
3' 測序引物: | ColiDOWN |
3' 測序引物序列: | ColiDOWN: 5'-TTCACTTCTGAGTTCGGCATG-3' |
載體標簽: | C-HSV, N-His, C-His, N-Thrombin, N-Nus, N-EK |
載體抗性: | Ampicillin |
備注: |
Both Nterm and Cterm His tags; Nterm thrombin cleavage site; Nterm enterokinase cleavage site; a,b,c vary by MCS.
|
產品目錄號: | 71124-3 |
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-44c(+)載體 pET44c質粒基本信息
pET-44c(+)質粒圖譜 pET44c載體圖譜和pET-44c(+)載體序列 pET44c質粒序列多克隆位點信息
Feature Name | Start | End | |
---|---|---|---|
T7_Terminal_primer | 69 | 87 | |
rrnB_T2_terminator | 168 | 141 | |
rrnB_T1_terminator | 343 | 300 | |
rrnB_terminator | 466 | 309 | |
EK | 693 | 679 | |
S15 | 789 | 745 | |
6xHIS | 2340 | 2323 | |
T7_transl_en_RBS | 2377 | 2361 | |
lacO | 2422 | 2395 | |
T7_promoter | 2440 | 2422 | |
tet (300 - 563) | 2476 | 2739 | |
pBRrevBam_primer | 2547 | 2528 | |
lacI | 2822 | 3913 | |
ROP | 4485 | 4676 | |
pGEX_3_primer | 4692 | 4670 | |
pBR322_origin | 5710 | 5091 | |
AmpR_promoter | 5797 | 5825 | |
Ampicillin | 5868 | 6728 |
ORF | Start | End | |
---|---|---|---|
ORF frame 1 | 427 | 2130 | |
ORF frame 2 | 2352 | 499 | |
ORF frame 1 | 2323 | 3081 | |
ORF frame 2 | 2954 | 3913 | |
ORF frame 3 | 5868 | 6728 |
Enzyme Name | Cut |
---|---|
PacI | 503 |
XhoI | 530 |
PmlI | 576 |
NotI | 592 |
EagI | 592 |
HindIII | 599 |
KpnI | 624 |
SalI | 626 |
PstI | 639 |
AscI | 641 |
EcoRI | 654 |
BamHI | 660 |
SacI | 672 |
SmaI | 719 |
XmaI | 717 |
SacII | 795 |
SpeI | 826 |
BglII | 941 |
MscI | 1073 |
StuI | 1126 |
NcoI | 1401 |
NdeI | 2351 |
XbaI | 2389 |
ApaI | 3392 |
HpaI | 3687 |
FspI | 6432 |
pET-44c(+)質粒 pET44c載體簡介
pET44系列載體是設計用來克隆和高水平表達含有495個氨基酸的Nus標簽的蛋白表達載體。與pET-43.1系列載體相比,pET44系列載體額外編碼一個N端的His蛋白純化標簽。 pET42a-c載體的單一的多克隆位點見上面的環狀質粒圖譜。注意:載體序列是以pBR322質粒的編碼規矩進行編碼的,所以T7蛋白表達區在質粒圖譜上面 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。對pET44a載體的單鏈測序可以使用COLIDOWN primer(Cat. No. 70845-3).< 把表達的基因插入到載體的PshAI或者SmaI限制性內切酶位點時,可以通過EK蛋白酶或者Thrombin蛋白酶將融合表達的Nus標簽完全切除。
pET-44c(+)質粒序列 pET44c載體序列
LOCUS pET-44c(+) 7309 bp DNA Circular SYN DEFINITION pET-44c(+) ACCESSION KEYWORDS SOURCE ORGANISM other sequences; artificial sequences; vectors. COMMENT This file is created by Vector NTI //gt12365.cn/ COMMENT ORIGDB|GenBank COMMENT VNTAUTHORNAME|gt12365.cn| COMMENT VNTOAUTHORAD1|Novagen Inc.| FEATURES Location/Qualifiers source 1..7309 /organism="pET-44 c(+)" /mol_type="other DNA" misc_feature 69..87 /label="T7_Terminal_primer" terminator complement(141..168) /label="rrnB_T2_terminator" terminator complement(300..343) /label="rrnB_T1_terminator" terminator complement(309..466) /label="rrnB_terminator" CDS 427..2130 /label="ORF frame 1" CDS complement(499..2352) /label="ORF frame 2" misc_feature complement(679..693) /label="EK" misc_feature complement(745..789) /label="S15" CDS 2323..3081 /label="ORF frame 1" misc_feature complement(2323..2340) /label="6xHIS" misc_feature complement(2361..2377) /label="T7_transl_en_RBS" misc_feature complement(2395..2422) /label="lacO" promoter complement(2422..2440) /label="T7_promoter" gene 2476..2739 /label="tet (300 - 563)" /gene="tet (300 - 563)" misc_feature complement(2528..2547) /label="pBRrevBam_primer" misc_feature 2822..3913 /label="lacI" CDS 2954..3913 /label="ORF frame 2" misc_feature 4485..4676 /label="ROP" misc_feature complement(4670..4692) /label="pGEX_3_primer" rep_origin complement(5091..5710) /label="pBR322_origin" promoter 5797..5825 /label="AmpR_promoter" gene 5868..6728 /label="Ampicillin" /gene="Ampicillin" CDS 5868..6728 /label="ORF frame 3" ORIGIN 1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa 61 ggggttatgc tagttattgc tcagcggata catatttgaa tgtatttaga aaaataaaca 121 aaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat 181 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt 241 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga 301 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc 361 cctactctcg catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag 421 ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc 481 agcctaggaa cgcccaactt aattaacatt agtggtggtg gtggtggtgc tcgagatcct 541 ctgggtcttc aggagcgagt tctggctggc ttgcacgtgt atacagctgt gcggccgcaa 601 gcttcgaacg cgtatcgatg gtaccgtcga cgtcctgcag gcgcgcctgt acagaattcg 661 gatccacgag ctcggactct tgtcgtcgtc atcaatcgta ccagaacccg cgctgccccg 721 ggggaccaga ccagttggcg gtggcgagtc catgtgctgg cgttcaaatt tcgcagcagc 781 ggtttcttta cccgcggagt gatggtgatg gtgatgacca gaaccactag tcgcttcgtc 841 accgaaccag caaatattac gggcagccat aatcagtgct ccggcttttt cgtcggtcaa 901 cccttcgata tcagccagat catcaatgcc ctgttcggcg agatcttcca gcgtacaaac 961 gccacgggcg gccagtttga atgccaaatc acgatctacc ccttcaaggt tcagcagatc 1021 gtcagccggt ttgttatcac cgaggctttc ttcctgggcc tgtgcaatgg tggccagtgc 1081 atttttagca cgctcgcgca gtgcttcaac ggtcggctca tcaaggcctt cgatttccaa 1141 cagctctttc atcggcacat aggccaattc ttccagcgtc gagaagcctt cttctaccag 1201 aacagtcgcg aagtcttcgt cgatgtcgag atatttggtg aaggtgtcga tcgctgcgtg 1261 cgcttccgcc tgatgcttag cttgcaggtc gtcaacggtc atcacgttga gttcccaacc 1321 gctcagttgc gaagccagac gcacgttctg accgttacgg ccaatcgcct gcgccagatt 1381 accggcttca acggcgatgt ccatggtgtg tttatcttca tccaccacga tagaagcaac 1441 gtctgccggt gccattgcgt taatcacgaa ctgcgccggg ttatcatccc acaggacgat 1501 atcgatacgc tcgccaccca gttcagtaga caccgcctga acacgcgcgc cacgcatacc 1561 tacgcaagca cctaccggat cgatacgttt atcgttggtt ttcaccgcga ttttcgcacg 1621 agaacccgga tcgcgagccg ctgctttaat ttcaatcact tcttcgccga tttctggcac 1681 ttcaatacgg aacagttcga tcagcatttc cggcttggaa cgagtgacga acagttgcgc 1741 gccacgcgct tccgggcgaa cggaatagag cacgccacga acgcggtcgc cagggcggaa 1801 gttttcacgc ggcagcatat cttcgcgcag gatcacggct tcagcgttgt tgcccagatc 1861 cagagagatg ttgtcgcggt ttactttttt caccacgccg gtgatgattt caccttcgtg 1921 ttcacggaac tgatcaacca ccatcgcacg ttcggcttca cgcactttct gcacgataac 1981 ctgttttgcc gtctgggtag tgatacggtc aaaggtaaca gactcaatct gatcttcaac 2041 gtaatcgccc aggttcaggc tttcatcttc ataacgtgcg gcttcaaggg tgatttcctt 2101 ggtcggctgg gtgacttcat caacaactaa ccagcgacgg aaagtgtcaa aatcaccgct 2161 tttgcgatcg atctgtacgc ggacgtcgat ctcttgttca tattttttct ttgttgctgt 2221 cgccagcgcg ctttccaatg cttcgaaaat cttctcgcga ggtagcgcct tttcattgga 2281 tacggcttca actacagcca aaatttcttt gttcatacta gaatggtgat ggtggtgatg 2341 gctgctgccc atatgtatat ctccttctta aagttaaaca aaattatttc tagaggggaa 2401 ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg atcgagatcg 2461 atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt 2521 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc 2581 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc 2641 gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta 2701 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat 2761 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag 2821 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta 2881 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa 2941 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc 3001 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc 3061 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc 3121 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca 3181 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga 3241 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa 3301 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt 3361 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg 3421 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga 3481 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat 3541 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat 3601 taccgagtcc gggctgcgcg ttggtgcgga catctcggta gtgggatacg acgataccga 3661 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg 3721 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca 3781 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc 3841 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga 3901 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat 3961 ctcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca 4021 tgactagcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta 4081 ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa 4141 cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg 4201 aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct 4261 ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga 4321 tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta 4381 accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta 4441 tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa 4501 aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac 4561 tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg 4621 atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 4681 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 4741 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta 4801 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt 4861 gcaccatata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg 4921 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 4981 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 5041 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 5101 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 5161 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 5221 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 5281 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 5341 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 5401 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 5461 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 5521 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca 5581 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 5641 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 5701 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 5761 ttggtcatga cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag 5821 acaataaccc tgataaatgc ttcaatcatg attgaaaaag gaagagtatg agtattcaac 5881 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc 5941 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca 6001 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc 6061 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg 6121 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac 6181 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca 6241 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg 6301 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac 6361 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg 6421 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat 6481 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg 6541 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg 6601 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc 6661 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc 6721 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt 6781 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgcca tcgtgccgcc 6841 agaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 6901 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 6961 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 7021 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 7081 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 7141 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 7201 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 7261 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgcca //
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