出品公司: | EMD Biosciences (Novagen) |
---|---|
别名: | pET44c, pET 44c |
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: | 7309 bp |
5' 测序引物: | T7 |
5' 测序引物序列: | T7: 5'-TAATACGACTCACTATAGGG-3' |
3' 测序引物: | ColiDOWN |
3' 测序引物序列: | ColiDOWN: 5'-TTCACTTCTGAGTTCGGCATG-3' |
载体标签: | C-HSV, N-His, C-His, N-Thrombin, N-Nus, N-EK |
载体抗性: | Ampicillin |
备注: |
Both Nterm and Cterm His tags; Nterm thrombin cleavage site; Nterm enterokinase cleavage site; a,b,c vary by MCS.
|
产品目录号: | 71124-3 |
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-44c(+)载体 pET44c质粒基本信息
pET-44c(+)质粒图谱 pET44c载体图谱和pET-44c(+)载体序列 pET44c质粒序列多克隆位点信息
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(+)质粒 pET44c载体简介
pET44系列载体是设计用来克隆和高水平表达含有495个氨基酸的Nus标签的蛋白表达载体。与pET-43.1系列载体相比,pET44系列载体额外编码一个N端的His蛋白纯化标签。 pET42a-c载体的单一的多克隆位点见上面的环状质粒图谱。注意:载体序列是以pBR322质粒的编码规矩进行编码的,所以T7蛋白表达区在质粒图谱上面 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。对pET44a载体的单链测序可以使用COLIDOWN primer(Cat. No. 70845-3).< 把表达的基因插入到载体的PshAI或者SmaI限制性内切酶位点时,可以通过EK蛋白酶或者Thrombin蛋白酶将融合表达的Nus标签完全切除。
pET-44c(+)质粒序列 pET44c载体序列
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 http://www.biofeng.com/ COMMENT ORIGDB|GenBank COMMENT VNTAUTHORNAME|biofeng.com| 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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