Search name
pET28a-EGFP,Plasmid pET28a-EGFP,pET28a-EGFP vector
pET28a-EGFP Informaiton
Promoter: T7/lac
Copier: ColE1 ori, F1 ori
Terminator: T7 Terminator
Plasmid Classification: Colon Series Plasmids; Colon Fluorescent Plasmids; Colon Green Plasmids
Plasmid size: 6050bp
Plasmid tags: N-6 *His, N-Thrombin, N-EGFP, C-6 *His
Prokaryotic resistance: kanamycin Kan (50 ug/ml)
Cloning strains: Escherichia coli such as DH5alpha
Culture conditions: 37 C, aerobic, LB
Expression host: Escherichia coli such as BL21 (DE3)
Culture conditions: 37 C, aerobic, LB
Induction: IPTG or lactose and its analogues
5'Sequencing Primer: T7 (TAATACGACTCACTATAGG)
3'Sequencing primers: T7-ter (TGCTAGTTATTGCTCAGCGG)
pET28a-EGFP Description
pET28a-EGFP is a prokaryotic expression vector. The C-terminal contains a 6*His tag, and the N-terminal contains a 6*His tag, thrombin digestion site and GFP tag. The plasmid contains several commonly used restriction sites, which facilitates the cloning of different genes. The expression was induced by T7 RNA polymerase provided by host cells. The target gene was cloned into plasmid vector and controlled by strong transcription and translation signals of bacteriophages.
PET system is the most powerful system to express recombinant proteins in E. coli, and it is also the most widely used system in prokaryotic expression nowadays. This series of plasmids can easily weaken the expression of protein by reducing the concentration of inducers. Under non-induced conditions, the target gene can be completely silenced without transcription
pET28a-EGFP Multiple cloning sites
pET28a-EGFP Sequence
LOCUS Exported 6050 bp ds-DNA circular SYN 12-JUL-2017
DEFINITION synthetic circular DNA
KEYWORDS pET28a-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6050)
TITLE Direct Submission
JOURNAL Exported 2015-4-17
REFERENCE 2 (bases 1 to 6050)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Wednesday, July 12, 2017
FEATURES Location/Qualifiers
source 1..6050
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 12..467
/direction=RIGHT
/note="f1 ori"
/note="f1 ori
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="aph(3')-Ia"
/note="KanR
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="ori
high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 2271..2413
/note="bom"
/note="bom
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="lacI
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 complement(4598..4675)
/gene="lacI"
/note="lacI lacI promoter"
/note="lacI promoter"
promoter 4984..5002
/note="T7 promoter"
/note="T7 promoter
promoter for bacteriophage T7 RNA polymerase"
CDS 5083..5100
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS 5131..5847
/codon_start=1
/product="enhanced GFP"
/note="enhanced GFP"
/note="EGFP
mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
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