pET-28b
PVT0081 2ug
pET-28b Information
Promoter: T7/laco promoter
Copier: ColE1 ori, F1ori
Terminator: T7 Terminator
Plasmid Classification: E. coli Vector; PET Series Expression Plasmid
Plasmid size: 5368 BP
Plasmid labels: N-6 *His, N-Thrombin, N-T7, C-6 *His
Prokaryotic resistance: kanamycin Kan
Cloning strain: Escherichia coli DH5alpha
Culture conditions: 37 C, aerobic, LB
Expression host: Escherichia coli BL21 (DE3)
Culture conditions: 37 C, aerobic, LB
Induction: IPTG or lactose and its analogues
5'Sequencing Primers: T7: TAATACGACTCACTATAGG
3'Sequencing primers: T7-ter: TGCTAGTTATTGCTCAGCGG
Use:pET Plasmid
pET-28b Description
pET-28b contains an N-terminal His/Thrombin/T7 protein tag and an optional C-terminal His tag. The single polyclonal site of pET28b vector is shown in the circular plasmid map above. Note: The vector sequence is encoded by the coding rules of the pBR322 plasmid, so the expression region of T7 protein is reversed on the plasmid map. The cloning and expression regions initiated by T7 RNA polymerase were also labeled in the plasmid map. The F1 replicon of the plasmid is directed, so viral particles containing protein coding sequence can be produced by T7 phage polymerase, and the protein expression will be terminated by T7 terminator sequence.
pET-28b Sequence
LOCUS Exported 5368 bp ds-DNA circular SYN 03-8-2015
DEFINITION synthetic circular DNA
KEYWORDS Untitled
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5368)
TITLE Direct Submission
JOURNAL Exported 2015-8-3
FEATURES Location/Qualifiers
source 1..5368
/organism="synthetic DNA construct"
/mol_type="other DNA"
source 305..327
/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(140..157)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(206..238)
/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 complement(242..259)
/codon_start=1
/product="thrombin recognition and cleavage site"
/note="thrombin site"
/translation="LVPRGS"
CDS complement(269..286)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
RBS 305..327
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 342..366
/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(367..385)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 694..771
/gene="lacI"
/note="lacI promoter"
CDS 772..1854
/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 2663..2854
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 2956..3098
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(3284..3872)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 3994..4809
/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(4902..5357)
/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 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
181 cgacggagct cgaattcgga tcccgaccca tttgctgtcc accagtcatg ctagccatat
241 ggctgccgcg cggcaccagg ccgctgctgt gatgatgatg atgatggctg ctgcccatgg
301 tatatctcct tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac
361 aattccccta tagtgagtcg tattaatttc gcgggatcga gatctcgatc ctctacgccg
421 gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg
481 acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg
541 tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgcatgcac
601 cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc
661 aggagtcgca taagggagag cgtcgagatc ccggacacca tcgaatggcg caaaaccttt
721 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca
781 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg
841 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg
901 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
961 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
1021 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
1081 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc
1141 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt
1201 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
1261 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
1321 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
1381 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
1441 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
1501 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
1561 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
1621 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
1681 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
1741 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc
1801 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
1861 cgcaattaat gtaagttagc tcactcatta ggcaccggga tctcgaccga tgcccttgag
1921 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
1981 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat
2041 tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt
2101 cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg
2161 cgagaagcag gccattatcg ccggcatggc ggccccacgg gtgcgcatga tcgtgctcct
2221 gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga atgaatcacc
2281 gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac
2341 atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt cagcgccctg
2401 caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac
2461 atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt cccgccgcat
2521 ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt catcatcagt
2581 aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca tgaacagaaa
2641 tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta acatggcccg
2701 ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg acgcggatga
2761 acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc gcagctgcct
2821 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
2881 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
2941 tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg
3001 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatatg cggtgtgaaa
3061 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
3121 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
3181 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
3241 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
3301 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
3361 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
3421 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
3481 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
3541 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
3601 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
3661 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
3721 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
3781 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
3841 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
3901 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaac aataaaactg
3961 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtct
4021 tgctctaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
4081 cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg
4141 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
4201 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
4261 actcctgatg atgcatggtt actcaccact gcgatccccg ggaaaacagc attccaggta
4321 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
4381 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
4441 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
4501 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
4561 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
4621 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
4681 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
4741 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
4801 tttttctaag aattaattca tgagcggata catatttgaa tgtatttaga aaaataaaca
4861 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gaaattgtaa acgttaatat
4921 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
4981 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
5041 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
5101 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
5161 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
5221 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
5281 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
5341 gccgctacag ggcgcgtccc attcgcca
//
Caution:
Product is for research use only!
Search name
pET-28b,Plasmid pET-28b,pET-28b vector
