PET- 28A plasmid - 2ug

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    pET-28a

    PVT0072   2ug

     

    pET-28a Informaiton

    Use:pET Plasmid

    Alias: pET28a, pET-28a+

    Promoter: T7/lac Promoter

    Replicon: pBR322 ori

    Terminator: T7 Terminator

    Plasmid classification: large intestine plasmid; large intestine expression plasmid; pET series plasmid.

    Plasmid size: 5369bp

    Plasmid tagging: N-6 * His, N-Thrombin, N-T7, C-6 * His

    Prokaryotic resistance: kanamycin Kan (50 g/ml)

    Clone strain: Escherichia coli DH5 alpha

    Culture conditions: 37 C, aerobic, LB

    Expression vector: Escherichia coli BL21 (DE3)

    Culture conditions: 37 C, aerobic, LB

    Induction: IPTG or lactose and its analogues.

    5'sequencing primers: T7 (TAATACGACTCACTATAGGG)

    3'sequencing primers: T7-ter (TGCTAGTTATTGCTCAGCGG)

     

    pET-28a Description

    pET-28a is a prokaryotic expression vector. The C-terminal contains a 6*His tag and the N-terminal contains a 6*His tag, a thrombin digestion site and a T7 tag. The plasmid contains several commonly used restriction sites to facilitate cloning of different genes. The expression was induced by T7 RNA polymerase from host cells. The target gene was cloned into plasmid vector and controlled by strong phage transcription and translation signals. The single polyclonal site of the pET28a vector is seen above the circular plasmid map. Note: The vector sequence is encoded according to the coding rules of the pBR322 plasmid, so the T7 protein expression region is reversed on the plasmid map. The cloning and expression regions initiated by T7 RNA polymerase were also labelled in plasmid profiles. The F1 replicon of the plasmid is directed, so the viral particles containing the protein coding sequence can be produced by the T7 phage polymerase, and the protein expression can be initiated. The protein expression will be terminated by the T7 terminator sequence. The pET system is the most powerful system ever used to express recombinant protein in E. coli. It is also the most widely used system in prokaryotic expression. The plasmids can easily reduce protein expression by decreasing the concentration of inducers. Under non inducible conditions, the target gene can be completely silenced without transcribing.

     

    pET-28a Multiple cloning site


    pET-28a restriction site

     

    pET-28a Sequence

    LOCUS       Exported                5369 bp ds-DNA     circular SYN 16-JUN-2017
    DEFINITION  synthetic circular DNA.
    KEYWORDS    pET-28a
    SOURCE      synthetic DNA construct
      ORGANISM  synthetic DNA construct
    REFERENCE   1  (bases 1 to 5369)
      TITLE     Direct Submission
      JOURNAL   Exported Wednesday, July 12, 2017 
    FEATURES             Location/Qualifiers
         source          1..5369
                         /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"
         CDS             complement(560..1375)
                         /codon_start=1
                         /gene="aph(3')-Ia"
                         /product="aminoglycoside phosphotransferase"
                         /note="KanR"
                         /note="confers resistance to kanamycin in bacteria or G418
                         (Geneticin(R)) in eukaryotes"
                         /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                         KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                         TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                         SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                         ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
         rep_origin      1497..2085
                         /direction=RIGHT
                         /note="ori"
                         /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                         replication"
         misc_feature    2271..2413
                         /note="bom"
                         /note="basis of mobility region from pBR322"
         CDS             complement(2515..2706)
                         /codon_start=1
                         /gene="rop"
                         /product="Rop protein, which maintains plasmids at low copy
                         number"
                         /note="rop"
                         /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                         DELYRSCLARFGDDGENL"
         CDS             complement(3515..4597)
                         /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"
         promoter        4598..4675
                         /gene="lacI"
                         /note="lacI prom"
         promoter        4984..5002
                         /note="T7 prom"
                         /note="promoter for bacteriophage T7 RNA polymerase"
         protein_bind    5003..5027
                         /bound_moiety="lac repressor encoded by lacI"
                         /note="laco"
                         /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)."
         RBS             5042..5064
                         /note="efficient ribosome binding site from bacteriophage
                         T7 gene 10 (Olins and Rangwala, 1989)"
         CDS             5083..5100
                         /codon_start=1
                         /product="6xHis affinity tag"
                         /note="6xHis"
                         /translation="HHHHHH"
         CDS             5110..5127
                         /codon_start=1
                         /product="thrombin recognition and cleavage site"
                         /note="thrombin"
                         /translation="LVPRGS"
         CDS             5131..5163
                         /codon_start=1
                         /product="leader peptide from bacteriophage T7 gene 10"
                         /note="T7 tag"
                         /note="promotes efficient translation in E. coli"
                         /translation="MASMTGGQQMG"
         misc_feature    5167..5212
                         /note="MCS"
         CDS             5213..5230
                         /codon_start=1
                         /product="6xHis affinity tag"
                         /note="6xHis"
                         /translation="HHHHHH"
         terminator      5297..5344
                         /note="T7 term"
                         /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 aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
          601 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
          661 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
          721 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
          781 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
          841 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
          901 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
          961 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
         1021 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
         1081 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
         1141 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
         1201 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
         1261 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
         1321 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
         1381 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
         1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
         1501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
         1561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
         1621 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
         1681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
         1741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
         1801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
         1861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
         1921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
         1981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
         2041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
         2101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
         2161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
         2221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
         2281 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta
         2341 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
         2401 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
         2461 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
         2521 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
         2581 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
         2641 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
         2701 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
         2761 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
         2821 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
         2881 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
         2941 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
         3001 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
         3061 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
         3121 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
         3181 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
         3241 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
         3301 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
         3361 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
         3421 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
         3481 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
         3541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
         3601 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
         3661 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
         3721 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
         3781 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
         3841 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
         3901 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
         3961 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
         4021 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
         4081 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
         4141 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
         4201 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
         4261 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
         4321 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
         4381 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
         4441 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
         4501 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
         4561 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
         4621 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
         4681 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
         4741 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
         4801 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
         4861 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
         4921 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
         4981 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
         5041 ttttgtttaa ctttaagaag gagatatacc atgggcagca gccatcatca tcatcatcac
         5101 agcagcggcc tggtgccgcg cggcagccat atggctagca tgactggtgg acagcaaatg
         5161 ggtcgcggat ccgaattcga gctccgtcga caagcttgcg gccgcactcg agcaccacca
         5221 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
         5281 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
         5341 tttgctgaaa ggaggaacta tatccggat
    //


    Caution:
    1.  This product is FOR RESEARCH USE ONLY!
    2.  The item is lyophilized form, Please take the powder plasmid by centrifugation at 5000rpm/min for 1min. Add 20μl ddH2O in to the tube of plasmid.
     

    Search name

    pET-28a,Plasmid pET-28a,pET-28a vector