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
ACCESSION .
VERSION .
KEYWORDS Untitled 3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5605)
AUTHORS .
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"
/db_xref="taxon:10760"
terminator 26..73
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(168..212)
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/note="S-Tag"
/translation="KETAAAKFERQHMDS"
CDS complement(213..230)
/codon_start=1
/product="thrombin recognition and cleavage site"
/note="thrombin site"
/translation="LVPRGS"
CDS complement(309..332)
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/note="HRV 3C site"
/translation="LEVLFQGP"
CDS complement(342..359)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(441..767)
/codon_start=1
/gene="trxA"
/product="E. coli thioredoxin"
/note="TrxA"
/translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
DANLA"
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
/gene="lacI"
/note="lacI promoter"
CDS 1246..2328
/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 2900..3091
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 3193..3335
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(3521..4109)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 4231..5046
/codon_start=1
/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 complement(5139..5594)
/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 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
//
Caution:
Product is for research use only!
Search name
pET-48b,Plasmid pET-48b,pET-48b vector
