出品公司: | EMD Biosciences (Novagen) |
---|---|
别名: | pET44b, pET 44b |
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: | 7310 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: | 71123-3 |
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-44b(+)载体 pET44b质粒基本信息
pET-44b(+)质粒图谱 pET44b载体图谱和pET-44b(+)载体序列 pET44b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 | 694 | 680 | |
S15 | 790 | 746 | |
6xHIS | 2341 | 2324 | |
T7_transl_en_RBS | 2378 | 2362 | |
lacO | 2423 | 2396 | |
T7_promoter | 2441 | 2423 | |
tet (300 - 563) | 2477 | 2740 | |
pBRrevBam_primer | 2548 | 2529 | |
lacI | 2823 | 3914 | |
ROP | 4486 | 4677 | |
pGEX_3_primer | 4693 | 4671 | |
pBR322_origin | 5711 | 5092 | |
AmpR_promoter | 5798 | 5826 | |
Ampicillin | 5869 | 6729 |
ORF | Start | End | |
---|---|---|---|
ORF frame 2 | 2353 | 509 | |
ORF frame 2 | 803 | 2131 | |
ORF frame 2 | 2324 | 3082 | |
ORF frame 3 | 2955 | 3914 | |
ORF frame 1 | 5869 | 6729 |
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 | 720 |
XmaI | 718 |
SacII | 796 |
SpeI | 827 |
BglII | 942 |
MscI | 1074 |
StuI | 1127 |
NcoI | 1402 |
NdeI | 2352 |
XbaI | 2390 |
ApaI | 3393 |
HpaI | 3688 |
FspI | 6433 |
pET-44b(+)质粒 pET44b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-44b(+)质粒序列 pET44b载体序列
LOCUS pET-44b(+) 7310 bp DNA Circular SYN DEFINITION pET-44b(+) 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..7310 /organism="pET-44 b(+)" /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 complement(509..2353) /label="ORF frame 2" misc_feature complement(680..694) /label="EK" misc_feature complement(746..790) /label="S15" CDS 803..2131 /label="ORF frame 2" CDS 2324..3082 /label="ORF frame 2" misc_feature complement(2324..2341) /label="6xHIS" misc_feature complement(2362..2378) /label="T7_transl_en_RBS" misc_feature complement(2396..2423) /label="lacO" promoter complement(2423..2441) /label="T7_promoter" gene 2477..2740 /label="tet (300 - 563)" /gene="tet (300 - 563)" misc_feature complement(2529..2548) /label="pBRrevBam_primer" misc_feature 2823..3914 /label="lacI" CDS 2955..3914 /label="ORF frame 3" misc_feature 4486..4677 /label="ROP" misc_feature complement(4671..4693) /label="pGEX_3_primer" rep_origin complement(5092..5711) /label="pBR322_origin" promoter 5798..5826 /label="AmpR_promoter" gene 5869..6729 /label="Ampicillin" /gene="Ampicillin" CDS 5869..6729 /label="ORF frame 1" 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 ctccggactc ttgtcgtcgt catcaatcgt accagaaccc gcgctgcccc 721 gggggaccag accagttggc ggtggcgagt ccatgtgctg gcgttcaaat ttcgcagcag 781 cggtttcttt acccgcggag tgatggtgat ggtgatgacc agaaccacta gtcgcttcgt 841 caccgaacca gcaaatatta cgggcagcca taatcagtgc tccggctttt tcgtcggtca 901 acccttcgat atcagccaga tcatcaatgc cctgttcggc gagatcttcc agcgtacaaa 961 cgccacgggc ggccagtttg aatgccaaat cacgatctac cccttcaagg ttcagcagat 1021 cgtcagccgg tttgttatca ccgaggcttt cttcctgggc ctgtgcaatg gtggccagtg 1081 catttttagc acgctcgcgc agtgcttcaa cggtcggctc atcaaggcct tcgatttcca 1141 acagctcttt catcggcaca taggccaatt cttccagcgt cgagaagcct tcttctacca 1201 gaacagtcgc gaagtcttcg tcgatgtcga gatatttggt gaaggtgtcg atcgctgcgt 1261 gcgcttccgc ctgatgctta gcttgcaggt cgtcaacggt catcacgttg agttcccaac 1321 cgctcagttg cgaagccaga cgcacgttct gaccgttacg gccaatcgcc tgcgccagat 1381 taccggcttc aacggcgatg tccatggtgt gtttatcttc atccaccacg atagaagcaa 1441 cgtctgccgg tgccattgcg ttaatcacga actgcgccgg gttatcatcc cacaggacga 1501 tatcgatacg ctcgccaccc agttcagtag acaccgcctg aacacgcgcg ccacgcatac 1561 ctacgcaagc acctaccgga tcgatacgtt tatcgttggt tttcaccgcg attttcgcac 1621 gagaacccgg atcgcgagcc gctgctttaa tttcaatcac ttcttcgccg atttctggca 1681 cttcaatacg gaacagttcg atcagcattt ccggcttgga acgagtgacg aacagttgcg 1741 cgccacgcgc ttccgggcga acggaataga gcacgccacg aacgcggtcg ccagggcgga 1801 agttttcacg cggcagcata tcttcgcgca ggatcacggc ttcagcgttg ttgcccagat 1861 ccagagagat gttgtcgcgg tttacttttt tcaccacgcc ggtgatgatt tcaccttcgt 1921 gttcacggaa ctgatcaacc accatcgcac gttcggcttc acgcactttc tgcacgataa 1981 cctgttttgc cgtctgggta gtgatacggt caaaggtaac agactcaatc tgatcttcaa 2041 cgtaatcgcc caggttcagg ctttcatctt cataacgtgc ggcttcaagg gtgatttcct 2101 tggtcggctg ggtgacttca tcaacaacta accagcgacg gaaagtgtca aaatcaccgc 2161 ttttgcgatc gatctgtacg cggacgtcga tctcttgttc atattttttc tttgttgctg 2221 tcgccagcgc gctttccaat gcttcgaaaa tcttctcgcg aggtagcgcc ttttcattgg 2281 atacggcttc aactacagcc aaaatttctt tgttcatact agaatggtga tggtggtgat 2341 ggctgctgcc catatgtata tctccttctt aaagttaaac aaaattattt ctagagggga 2401 attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg gatcgagatc 2461 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt 2521 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct 2581 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg 2641 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact 2701 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca 2761 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca 2821 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt 2881 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa 2941 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg 3001 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt 3061 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt 3121 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc 3181 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg 3241 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca 3301 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat 3361 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc 3421 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg 3481 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca 3541 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca 3601 ttaccgagtc cgggctgcgc gttggtgcgg acatctcggt agtgggatac gacgataccg 3661 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg 3721 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc 3781 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg 3841 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg 3901 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga 3961 tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc 4021 atgactagca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt 4081 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa 4141 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg 4201 gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc 4261 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg 4321 atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt 4381 aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt 4441 atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa 4501 aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa 4561 ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct 4621 gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac 4681 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc 4741 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt 4801 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag 4861 tgcaccatat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag 4921 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 4981 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 5041 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 5101 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 5161 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 5221 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 5281 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 5341 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 5401 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 5461 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 5521 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 5581 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 5641 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 5701 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 5761 tttggtcatg acctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga 5821 gacaataacc ctgataaatg cttcaatcat gattgaaaaa ggaagagtat gagtattcaa 5881 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac 5941 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac 6001 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt 6061 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc 6121 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca 6181 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc 6241 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag 6301 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa 6361 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg 6421 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa 6481 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg 6541 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt 6601 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt 6661 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag 6721 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat 6781 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgcc atcgtgccgc 6841 cagaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct 6901 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg 6961 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact 7021 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac 7081 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga 7141 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga 7201 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca 7261 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca //
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