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产品名称：pET-44a
产品型号：
产品展商：XYbscience
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简单介绍

pET-44a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。pET-44a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。

产品描述

pET-44a

产品信息
基本信息
质粒简介
质粒图谱
质粒序列

产品信息

产品货号	产品名称	产品规格	优惠价
XY0377	pET-44a	20μl	¥请询价

使用说明

信裕质粒平台的各批次质粒菌株发货前均经过严格的多重验证，如存在质量问题，请在收到产品的三个月内通知我司。收到质粒后请短暂离心，加入20μl ddH2O溶解质粒，取2μl转化至对应感受态中，挑取单克隆重新提取质粒后使用。如需获取其他详细信息请登录我司官网查询。

基本信息

启动子:	T7/lac
复制子:	ColE1 ori，F1 ori
终止子:	T7 terminator
质粒分类:	大肠杆菌载体；PET系列表达质粒
质粒大小:	7311bp
质粒标签:	C-HSV, N-6×His, C-6×His, N-Thrombin, N-Nus, N-EK
原核抗性:	氨苄青霉素Amp
克隆菌株:	DH5α
培养条件:	37℃，有氧，LB
表达宿主:	BL21(DE3)
培养条件:	37℃，有氧，LB
诱导方式:	IPTG或乳糖及其类似物
5'测序引物:	T7:TAATACGACTCACTATAGGG
3'测序引物:	T7-ter:TGCTAGTTATTGCTCAGCGG

质粒简介

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.
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。

质粒图谱

质粒序列

LOCUS       Exported                7311 bp ds-DNA     circular SYN 11-8-2015
DEFINITION synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled
SOURCE      synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE   1 (bases 1 to 7311)
AUTHORS   .
TITLE     Direct Submission
JOURNAL   Exported 2015-8-11   from MLCC
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..7311
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          2362..2384
                     /organism="Enterobacteria phage T7"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:10760"
     terminator      26..73
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     terminator      141..168
                     /note="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      260..346
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(512..529)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             complement(536..568)
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /note="HSV tag"
                     /translation="QPELAPEDPED"
     CDS             complement(681..695)
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /note="enterokinase site"
                     /translation="DDDDK"
     CDS             complement(714..731)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /note="thrombin site"
                     /translation="LVPRGS"
     CDS             complement(747..791)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
                     /note="S-Tag"
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(801..818)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             complement(834..2318)
                     /codon_start=1
                     /gene="nusA"
                     /product="transcription elongation protein (N utilization
                     substance protein A)"
                     /note="NusA"
                     /note="highly soluble in E. coli"
                     /translation="MNKEILAVVEAVSNEKALPREKIFEALESALATATKKKYEQEIDV
                     RVQIDRKSGDFDTFRRWLVVDEVTQPTKEITLEAARYEDESLNLGDYVEDQIESVTFDR
                     ITTQTAKQVIVQKVREAERAMVVDQFREHEGEIITGVVKKVNRDNISLDLGNNAEAVIL
                     REDMLPRENFRPGDRVRGVLYSVRPEARGAQLFVTRSKPEMLIELFRIEVPEIGEEVIE
                     IKAAARDPGSRAKIAVKTNDKRIDPVGACVGMRGARVQAVSTELGGERIDIVLWDDNPA
                     QFVINAMAPADVASIVVDEDKHTMDIAVEAGNLAQAIGRNGQNVRLASQLSGWELNVMT
                     VDDLQAKHQAEAHAAIDTFTKYLDIDEDFATVLVEEGFSTLEELAYVPMKELLEIEGLD
                     EPTVEALRERAKNALATIAQAQEESLGDNKPADDLLNLEGVDRDLAFKLAARGVCTLED
                     LAEQGIDDLADIEGLTDEKAGALIMAARNICWFGDEA"
     CDS             complement(2325..2342)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     RBS             2362..2384
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2399..2423
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(2424..2442)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2755..2832
                     /gene="lacI"
                     /note="lacI promoter"
     CDS             2833..3915
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     CDS             4487..4678
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    4780..4922
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5108..5696)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     promoter        5777..5869
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             5870..6730
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(6845..7300)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
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 ctcccggact cttgtcgtcg tcatcaatcg taccagaacc cgcgctgccc
      721 cgggggacca gaccagttgg cggtggcgag tccatgtgct ggcgttcaaa tttcgcagca
      781 gcggtttctt tacccgcgga gtgatggtga tggtgatgac cagaaccact agtcgcttcg
      841 tcaccgaacc agcaaatatt acgggcagcc ataatcagtg ctccggcttt ttcgtcggtc
      901 aacccttcga tatcagccag atcatcaatg ccctgttcgg cgagatcttc cagcgtacaa
      961 acgccacggg cggccagttt gaatgccaaa tcacgatcta ccccttcaag gttcagcaga
     1021 tcgtcagccg gtttgttatc accgaggctt tcttcctggg cctgtgcaat ggtggccagt
     1081 gcatttttag cacgctcgcg cagtgcttca acggtcggct catcaaggcc ttcgatttcc
     1141 aacagctctt tcatcggcac ataggccaat tcttccagcg tcgagaagcc ttcttctacc
     1201 agaacagtcg cgaagtcttc gtcgatgtcg agatatttgg tgaaggtgtc gatcgctgcg
     1261 tgcgcttccg cctgatgctt agcttgcagg tcgtcaacgg tcatcacgtt gagttcccaa
     1321 ccgctcagtt gcgaagccag acgcacgttc tgaccgttac ggccaatcgc ctgcgccaga
     1381 ttaccggctt caacggcgat gtccatggtg tgtttatctt catccaccac gatagaagca
     1441 acgtctgccg gtgccattgc gttaatcacg aactgcgccg ggttatcatc ccacaggacg
     1501 atatcgatac gctcgccacc cagttcagta gacaccgcct gaacacgcgc gccacgcata
     1561 cctacgcaag cacctaccgg atcgatacgt ttatcgttgg ttttcaccgc gattttcgca
     1621 cgagaacccg gatcgcgagc cgctgcttta atttcaatca cttcttcgcc gatttctggc
     1681 acttcaatac ggaacagttc gatcagcatt tccggcttgg aacgagtgac gaacagttgc
     1741 gcgccacgcg cttccgggcg aacggaatag agcacgccac gaacgcggtc gccagggcgg
     1801 aagttttcac gcggcagcat atcttcgcgc aggatcacgg cttcagcgtt gttgcccaga
     1861 tccagagaga tgttgtcgcg gtttactttt ttcaccacgc cggtgatgat ttcaccttcg
     1921 tgttcacgga actgatcaac caccatcgca cgttcggctt cacgcacttt ctgcacgata
     1981 acctgttttg ccgtctgggt agtgatacgg tcaaaggtaa cagactcaat ctgatcttca
     2041 acgtaatcgc ccaggttcag gctttcatct tcataacgtg cggcttcaag ggtgatttcc
     2101 ttggtcggct gggtgacttc atcaacaact aaccagcgac ggaaagtgtc aaaatcaccg
     2161 cttttgcgat cgatctgtac gcggacgtcg atctcttgtt catatttttt ctttgttgct
     2221 gtcgccagcg cgctttccaa tgcttcgaaa atcttctcgc gaggtagcgc cttttcattg
     2281 gatacggctt caactacagc caaaatttct ttgttcatac tagaatggtg atggtggtga
     2341 tggctgctgc ccatatgtat atctccttct taaagttaaa caaaattatt tctagagggg
     2401 aattgttatc cgctcacaat tcccctatag tgagtcgtat taatttcgcg ggatcgagat
     2461 cgatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
     2521 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
     2581 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
     2641 gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac
     2701 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc
     2761 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     2821 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
     2881 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
     2941 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
     3001 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     3061 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     3121 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     3181 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     3241 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     3301 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     3361 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     3421 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     3481 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     3541 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     3601 attaccgagt ccgggctgcg cgttggtgcg gacatctcgg tagtgggata cgacgatacc
     3661 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     3721 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     3781 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
     3841 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     3901 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcaccggg
     3961 atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     4021 catgactagc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct
     4081 tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa
     4141 aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct
     4201 ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg
     4261 ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt
     4321 gatttttctc tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag
     4381 taaccgggca tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg
     4441 tatcattacc cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga
     4501 aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa
     4561 actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc
     4621 tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     4681 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     4741 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
     4801 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
     4861 gtgcaccata tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     4921 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4981 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5041 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5101 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5161 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5221 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5281 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5341 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5401 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5461 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5521 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     5581 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5641 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5701 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5761 ttttggtcat gacctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5821 agacaataac cctgataaat gcttcaatca tgattgaaaa aggaagagta tgagtattca
     5881 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5941 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     6001 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     6061 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     6121 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     6181 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6241 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     6301 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6361 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6421 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6481 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6541 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     6601 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6661 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6721 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     6781 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatgc catcgtgccg
     6841 ccagaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     6901 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     6961 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     7021 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     7081 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     7141 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     7201 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     7261 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc a
//

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pET-44a

pET-44a

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