pET- 48b Plasmid


  • Model: PVT0365
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pET-48b Plasmid

PVT0365         2ug


pET-48b Plasmid Information

Promoter: T7/lac

Replicator: ColE1 ori, F1 ori

Terminator: T7 Terminator

Plasmid classification: Escherichia coli vector; PET series expression plasmid

Plasmid size: 5605bp

Plasmid Tags: N-Trx, N-6 x His, N-HRV 3C, C-S, C-Thrombin

Prokaryotic resistance: kanamycin Kan

Cloned strain: DH5 alpha

Culture conditions: 37 centigrade, aerobic, LB

Expression host: BL21 (DE3)

Culture conditions: 37 centigrade, aerobic, LB

Inducement: IPTG or lactose and its analogues

5'sequencing primers: T7:TAATACGACTCACTATAGGG

3'sequencing primers: T7-ter:TGCTAGTTATTGCTCAGCGG


pET-48b Plasmid Description

The pET-48b Plasmid carrier contains the N terminal Trx and His tags, and the HRV 3C protease cleavage site follows the label. HRV LEVLFQ: GP 3C protease can identify protein sequences with high specificity, can effectively cut off the fusion label sequences in low temperature. The pET-48b vector also contains a selectable C terminal Thrombin protease cut site, followed by the site of the S tag.

The single polyclonal site of the pET48b vector is found in the atlas of the above loop plasmid. Note: the vector sequence is encoded by the encoding rules of the pBR322 plasmid, so the expression area of the T7 protein is reversed on the plasmid profile.

The cloning and expression regions of the T7 RNA polymerase have also been labeled in the plasmid profiles. The F1 replicon of plasmid is directed. Therefore, under the action of T7 phage polymerase, virus particles containing protein coding sequence can produce and initiate protein expression, and protein expression will be terminated by the termination of T7 termination sequence. S 18 primers should be used for single strand sequencing of the carrier.


pET-48b Plasmid Sequence

LOCUS       Exported                5605 bp ds-DNA     circular SYN 11-8-2015
DEFINITION  synthetic circular DNA
KEYWORDS    Untitled 3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5605)
  TITLE     Direct Submission
  JOURNAL   Exported 2015-8-11  
FEATURES             Location/Qualifiers
     source          1..5605
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          775..797
                     /organism="Enterobacteria phage T7"
                     /mol_type="genomic DNA"
     terminator      26..73
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
     CDS             complement(168..212)
                     /product="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
     CDS             complement(213..230)
                     /product="thrombin recognition and cleavage site"
                     /note="thrombin site"
     CDS             complement(309..332)
                     /product="recognition and cleavage site for human
                     rhinovirus 3C and PreScission proteases"
                     /note="HRV 3C site"
     CDS             complement(342..359)
                     /product="6xHis affinity tag"
     CDS             complement(441..767)
                     /product="E. coli thioredoxin"
     RBS             775..797
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    812..836
                     /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(837..855)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1168..1245
                     /note="lacI promoter"
     CDS             1246..2328
                     /product="lac repressor"
                     /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             2900..3091
                     /product="Rop protein, which maintains plasmids at low copy
     misc_feature    3193..3335
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3521..4109)
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
     CDS             4231..5046
                     /product="aminoglycoside phosphotransferase"
                     /note="confers resistance to kanamycin in bacteria or G418
                     (Geneticin(R)) in eukaryotes"
     rep_origin      complement(5139..5594)
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgtttagcag cctaggttaa ttaagcctcg agagcagcag aagtagagct gtccatgtgc
      181 tggcgttcga atttagcagc agcggtttct ttactaccgc gtggcaccag agcgagctct
      241 gcggccgcaa gcttgtcgac ggacgtcggg cgcgccaagg cctgtacaga attcggatcc
      301 tggtacccgg gtccctgaaa gaggacttca agagccgcgg agtgatggtg atggtgatga
      361 ccagaaccac tagatggagt aggagtagga ggattgttat tagaaccgcc accaccacta
      421 gtatggccag aaccagaacc ggccaggtta gcgtcgagga actctttcaa ctgaccttta
      481 gacagtgcac ccactttggt tgccgccact tcaccgtttt tgaacagcag cagagtcggg
      541 ataccacgga tgccatattt cggcgcagtg ccagggtttt gatcgatgtt cagttttgca
      601 acggtcagtt tgccctgata ttcgtcagcg atttcatcca gaatcggggc gatcattttg
      661 cacggaccgc accactctgc ccagaaatcg acgaggatcg ccccgtccgc tttgagtaca
      721 tccgtgtcaa aactgtcgtc agtcaggtga ataattttat cgctcatatg tatatctcct
      781 tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac aattccccta
      841 tagtgagtcg tattaatttc gcgggatcga gatcgatctc gatcctctac gccggacgca
      901 tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca
      961 ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta
     1021 tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc
     1081 ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt
     1141 cgcataaggg agagcgtcga gatcccggac accatcgaat ggcgcaaaac ctttcgcggt
     1201 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
     1261 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
     1321 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
     1381 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     1441 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     1501 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     1561 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     1621 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     1681 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     1741 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     1801 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     1861 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     1921 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     1981 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     2041 gcggacatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     2101 ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     2161 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     2221 agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     2281 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     2341 taatgtaagt tagctcactc attaggcacc gggatctcga ccgatgccct tgagagcctt
     2401 caacccagtc agctccttcc ggtgggcgcg gggcatgact agcatgatcg tgctcctgtc
     2461 gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat
     2521 acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg
     2581 aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag cgccctgcac
     2641 cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc
     2701 tgtattaacg aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca
     2761 taccgccagt tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac
     2821 ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc
     2881 cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt
     2941 tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca
     3001 ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc
     3061 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     3121 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     3181 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt
     3241 aactatgcgg catcagagca gattgtactg agagtgcacc atatatgcgg tgtgaaatac
     3301 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
     3361 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3421 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3481 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3541 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3601 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3661 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3721 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3781 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3841 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3901 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3961 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4021 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4081 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4141 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgaacaat aaaactgtct
     4201 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcttgc
     4261 tctaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     4321 gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca
     4381 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     4441 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     4501 cctgatgatg catggttact caccactgcg atccccggca aaacagcatt ccaggtatta
     4561 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     4621 ttgcattcga ttcctgtttg taattgtcct tttaacagtg atcgcgtatt tcgtctcgct
     4681 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     4741 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     4801 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     4861 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     4921 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     4981 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     5041 ttctaagaat taattcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5101 aggggttccg cgcacatttc cccgaaaagt gccacctgaa attgtaaacg ttaatatttt
     5161 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     5221 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     5281 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     5341 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     5401 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     5461 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     5521 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5581 gctacagggc gcgtcccatt cgcca


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pET-48b,Plasmid pET-48b,pET-48b vector

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