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> pET-23b

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pET-23b

pET-23b
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  • 產(chǎn)品名稱:pET-23b
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簡單介紹
pET-23b是一種原核表達(dá)載體,N端含有一個(gè)T7標(biāo)簽、C端的6×His標(biāo)簽。該質(zhì)粒含有多個(gè)常用的酶切位點(diǎn),便于不同基因克隆。pET-23b表達(dá)由宿主細(xì)胞提供的T7 RNA聚合酶誘導(dǎo),目的基因被克隆到質(zhì)粒載體上,受噬菌體強(qiáng)轉(zhuǎn)錄及翻譯信號(hào)控制。
產(chǎn)品描述
 

pET-23b

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

產(chǎn)品信息

產(chǎn)品貨號(hào) 產(chǎn)品名稱 產(chǎn)品規(guī)格 優(yōu)惠價(jià)
XY0018 pET-23b

5μg

¥請(qǐng)?jiān)儍r(jià)

使用說明

信裕質(zhì)粒平臺(tái)的各批次質(zhì)粒菌株發(fā)貨前均經(jīng)過嚴(yán)格的多重驗(yàn)證,如存在質(zhì)量問題,請(qǐng)?jiān)谑盏疆a(chǎn)品的三個(gè)月內(nèi)通知我司。收到質(zhì)粒后請(qǐng)短暫離心,加入20μl ddH2O溶解質(zhì)粒,取2μl轉(zhuǎn)化至對(duì)應(yīng)感受態(tài)中,挑取單克隆重新提取質(zhì)粒后使用。如需獲取其他詳細(xì)信息請(qǐng)登錄我司官網(wǎng)查詢。

基本信息

啟動(dòng)子: T7
復(fù)制子: ColE1 ori,F(xiàn)1 ori
終止子: T7 terminator
質(zhì)粒分類: 大腸系列質(zhì)粒;大腸表達(dá)質(zhì)粒;pET系列質(zhì)粒
質(zhì)粒大小: 3665bp
質(zhì)粒標(biāo)簽: N-T7,C-6×His
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(dá)宿主: BL21(DE3)等大腸桿菌
培養(yǎng)條件: 37℃,有氧,LB
誘導(dǎo)方式: 無需誘導(dǎo)
5'測序引物: T7(TAATACGACTCACTATAGGG)
3'測序引物: T7-ter(TGCTAGTTATTGCTCAGCGG)
備注: 組成型表達(dá)

質(zhì)粒簡介

         pET-23b質(zhì)粒是一種原核表達(dá)載體,N端含有一個(gè)T7標(biāo)簽、C端的6×His標(biāo)簽。該質(zhì)粒含有多個(gè)常用的酶切位點(diǎn),便于不同基因克隆。表達(dá)由宿主細(xì)胞提供的T7 RNA聚合酶誘導(dǎo),目的基因被克隆到質(zhì)粒載體上,受噬菌體強(qiáng)轉(zhuǎn)錄及翻譯信號(hào)控制。
        pET系統(tǒng)是有史以來在大腸桿菌中表達(dá)重組蛋白的功能*強(qiáng)大的系統(tǒng),也是現(xiàn)今原核表達(dá)方面使用*廣泛的系統(tǒng)。該系列質(zhì)粒能很容易的通過降低誘導(dǎo)物的濃度來削弱蛋白表達(dá)。在非誘導(dǎo)條件下,可以使目的基因處于完全沉默狀態(tài)而不轉(zhuǎn)錄。

       The pET-23a-d(+) vectors carry an N-terminal T7?Tag® sequence plus an optional C-terminal His?Tag® sequence. These vectors differ from pET-21a-d(+) by the “plain” T7 promoter instead of the T7lac promoter and by the absence of the lacI gene. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circular map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand. Therefore, single-stranded sequencing should be performed using the T7 terminator primer.

質(zhì)粒圖譜



質(zhì)粒序列

LOCUS       Exported                3665 bp ds-DNA     circular SYN 14-JUN-2017
DEFINITION  synthetic circular DNA
KEYWORDS    pET-23b
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3665)
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, June 14, 2017 from miaoling plasmid
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..3665
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      12..467
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        494..598
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             599..1459
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1630..2218
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(2648..2839)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        3348..3366
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3428..3460
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /note="T7 tag (gene 10 leader)"
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             3509..3526
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     terminator      3593..3640
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
      601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1141 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     1321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     1381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     1801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2281 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2341 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2401 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg
     2461 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     2521 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     2581 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2641 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     2701 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     2761 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     2821 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     2881 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     2941 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     3001 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     3061 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     3121 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     3181 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     3241 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     3301 cccgtggcca ggacccaacg ctgcccgaga tctcgatccc gcgaaattaa tacgactcac
     3361 tatagggaga ccacaacggt ttccctctag aaataatttt gtttaacttt aagaaggaga
     3421 tatacatatg gctagcatga ctggtggaca gcaaatgggt cgggatccga attcgagctc
     3481 cgtcgacaag cttgcggccg cactcgagca ccaccaccac caccactgag atccggctgc
     3541 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     3601 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     3661 cggat
//
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