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pProEXHTA

pProEXHTA
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簡單介紹
pProEXHTA是一種原核表達載體, N端含有一個6×His標簽和一個TEV位點。pProEXHTA該質粒含有多個常用的酶切位點,便于不同基因克隆。
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pProEXHTA

  • 產(chǎn)品信息
  • 基本信息
  • 質粒簡介
  • 質粒圖譜
  • 質粒序列

產(chǎn)品信息

產(chǎn)品貨號 產(chǎn)品名稱 產(chǎn)品規(guī)格 優(yōu)惠價
XY0053 pProEXHTA

5μg

¥請詢價

使用說明

信裕質粒平臺的各批次質粒菌株發(fā)貨前均經(jīng)過嚴格的多重驗證,如存在質量問題,請在收到產(chǎn)品的三個月內(nèi)通知我司。收到質粒后請短暫離心,加入20μl ddH2O溶解質粒,取2μl轉化至對應感受態(tài)中,挑取單克隆重新提取質粒后使用。如需獲取其他詳細信息請登錄我司官網(wǎng)查詢。

基本信息

啟動子: Trc/Lac
復制子: ColE1 ori,F(xiàn)1 ori
終止子: rrnB T1 terminator
質粒分類: 大腸系列質粒;大腸表達質粒;pET系列質粒
質粒大小: 4751bp
質粒標簽: N-6×His,N-TEV
原核抗性: 氨芐青霉素Amp(100μg/ml)
克隆菌株: DH5α等大腸桿菌
培養(yǎng)條件: 37℃,有氧,LB
表達宿主: BL21(DE3)等大腸桿菌
培養(yǎng)條件: 37℃,有氧,LB
誘導方式: IPTG或乳糖及其類似物
5'測序引物: M13/pUC everse-F( AGCGGATAACAATTTCACACAGG)
3'測序引物: pTrcHis Reverse-R(GATTTAATCTGTATCAGG)
備注: His,rTEV蛋白酶切位點

質粒簡介

        pProEXHTA質粒是一種原核表達載體, N端含有一個6×His標簽和一個TEV位點。該質粒含有多個常用的酶切位點,便于不同基因克隆。
        pProEX HT系列原核表達質粒是用來在大腸桿菌中表達外源蛋白的載體。使用該質粒表達的目的蛋白含有6個His純化標簽。目的基因可以克隆到載體的多克隆位點處,可以選擇的載體有pProEX HTa, b, c.蛋白表達后,His純化標簽就位于目的蛋白的N端,同時His純化標簽的使用可以用于蛋白純化。載體上同時含有rTEV蛋白酶識別位點,可以使用rTEV蛋白酶進行酶切,從而去除融合的His標簽。

質粒圖譜



質粒序列

LOCUS       Exported                4751 bp ds-DNA     circular SYN 06-SEP-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 5
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4751)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, September 6, 2016 from SnapGene Viewer 3.1.4
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..4751
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    230..246
                     /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)."
     CDS             279..296
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             318..338
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /note="TEV site"
                     /translation="ENLYFQG"
     terminator      642..728
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      820..847
                     /note="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        867..958
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             959..1819
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1990..2578
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     rep_origin      complement(2826..3254)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3552..3629
                     /gene="lacI (mutant)"
                     /note="lacIq promoter"
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             3630..4712
                     /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"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gtccgtataa tctgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagaccatgt cgtactacca tcaccatcac catcacgatt
      301 acgatatccc aacgaccgaa aacctgtatt ttcagggcgc catggatccg gaattcaaag
      361 gcctacgtcg acgagctcaa ctagtgcggc cgctttcgaa tctagagcct gcagtctcga
      421 ggcatgcggt accaagcttg gctgttttgg cggatgagag aagattttca gcctgataca
      481 gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc
      541 ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag
      601 tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
      661 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
      721 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
      781 caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg
      841 gcctttttgc gtttctacaa actctttttg tttatttttc taaatacatt caaatatgta
      901 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
      961 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     1021 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     1081 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     1141 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     1201 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     1261 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     1321 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1381 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1441 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1501 tacagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1561 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1621 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     1681 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     1741 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1801 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1861 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1921 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1981 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2041 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2101 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     2161 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2221 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2281 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2341 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2401 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2461 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2521 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2581 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2641 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2701 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2761 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcataatttt
     2821 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     2881 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     2941 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3001 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     3061 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     3121 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     3181 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     3241 gctacagggc gcgtcccatt cgccattcag gctgctatgg tgcactctca gtacaatctg
     3301 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     3361 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3421 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     3481 ccgtcatcac cgaaacgcgc gaggcagcag atcaattcgc gcgcgaaggc gaagcggcat
     3541 gcatttacgt tgacaccatc gaatggtgca aaacctttcg cggtatggca tgatagcgcc
     3601 cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag
     3661 agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt
     3721 ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc
     3781 gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc
     3841 tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg
     3901 gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg
     3961 tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc
     4021 aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct
     4081 ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg
     4141 tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt
     4201 ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc
     4261 agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc
     4321 aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc
     4381 tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag
     4441 tgggatacga cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac
     4501 aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc
     4561 aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa accaccctgg
     4621 cacccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     4681 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagcgc
     4741 gaattgatct g
//
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