Search name
pET28a-DsRed2,Plasmid pET28a-DsRed2,pET28a-DsRed2 vector
pET28a-DsRed2 Information
Promoter: T7/lac
Replicator: ColE1 ori, F1 ori
Terminator: T7 Terminator
Plasmid classification: Escherichia coli plasmid, large intestine fluorescent plasmid, large intestine red plasmid.
Plasmid size: 6025bp
Plasmid tag: N-6 * His, N-Thrombin, N-T7, N-DsRed2, C-6 * His.
Prokaryotic resistance: kanamycin Kan (50 mu g/ml)
Cloned strains of Escherichia coli, DH5 A and other Escherichia coli
Culture conditions: 37 centigrade, aerobic, LB
Expression of host: BL21 (DE3) and other Escherichia coli
Culture conditions: 37 centigrade, aerobic, LB
Inducement: IPTG or lactose and its analogues
5'sequencing primers: T7 (TAATACGACTCACTATAGGG)
3'sequencing primers: T7-ter (TGCTAGTTATTGCTCAGCGG)
pET28a-DsRed2 Description
pET28a-DsRed2 plasmid is a prokaryotic expression vector. The C terminal contains a 6 x His label, and the N end contains a thrombin enzyme cut site, a 6 x His label, a T7 label and a DsRed2 label. The plasmid contains several commonly used restriction sites to facilitate cloning of different genes. Expression of T7 RNA polymerase provided by the host cell is induced, and the target gene is cloned into plasmid vector, which is controlled by the strong transcription and translation signal of phage.
PET system is the most powerful system to express recombinant protein in Escherichia coli, and it is also the most widely used system in prokaryotic expression. The plasmid can reduce protein expression easily by decreasing the concentration of inducers. Under the condition of non inducement, the target gene can be completely silenced without transcribing.
pET28a-DsRed2 Sequence
LOCUS Exported 6025 bp ds-DNA circular SYN 16-JUN-2017
DEFINITION synthetic circular DNA
KEYWORDS pET28a-DsRed2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6025)
TITLE Direct Submission
JOURNAL Exported Wednesday, July 12, 2017
FEATURES Location/Qualifiers
source 1..6025
/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
/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 complement(4598..4675)
/gene="lacI"
/note="lacI promoter"
promoter 4984..5002
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 5003..5027
/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)."
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 site"
/translation="LVPRGS"
CDS 5131..5163
/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 5173..5847
/codon_start=1
/product="improved tetrameric variant of DsRed fluorescent
protein"
/note="DsRed2"
