pET-39b
Search name
pET-39b,Plasmid pET-39b,pET-39b vector
Bacterial Resistance:Kanamycin
Growth Strain:DH5α
Expression: Bacterial
Use:pET Plasmid
pET-39b Plasmid informaiton
Promoter: T7/lac
Replicator: ColE1, ori, F1, ori
Terminator: T7 Terminator
Plasmid classification: Escherichia coli vector, PET series of expressed plasmids
Plasmid size: 6106bp
Plasmid labels: N-6 * His, N-DsbA, N-S, N-EK, N-Thrombin, C-6 * His
Prokaryotic resistance: kanamycin Kan
Clone strain: DH5 alpha
Culture conditions: 37 DEG C, aerobic, LB
Expression host: BL21 (DE3)
Culture conditions: 37 DEG C, aerobic, LB
Induction: IPTG or lactose and its analogues
Primers for 5'sequencing: T7:TAATACGACTCACTATAGGG
Primers for 3'sequencing: T7-ter:TGCTAGTTATTGCTCAGCGG
Note: DsbA fusion protein is expressed
pET-39b Plasmid Description
The pET-39b(+) vector is designed for expression of DsbA fusion proteins. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circle map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single stranded DNA that corresponds to the coding strand. Therefore, single stranded sequencing should be performed using the T7 terminator primer .
pET-39b Plasmid Sequence
LOCUS Exported 6106 bp ds-DNA circular SYN 10-8-2015
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6106)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-8-10
FEATURES Location/Qualifiers
source 1..6106
/organism="synthetic DNA construct"
/mol_type="other DNA"
source 1039..1061
/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(150..173)
/codon_start=1
/product="8xHis affinity tag"
/note="8xHis"
/translation="HHHHHHHH"
CDS complement(252..266)
/codon_start=1
/product="enterokinase recognition and cleavage site"
/note="enterokinase site"
/translation="DDDDK"
CDS complement(282..326)
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/note="S-Tag"
/translation="KETAAAKFERQHMDS"
CDS complement(342..359)
/codon_start=1
/product="thrombin recognition and cleavage site"
/note="thrombin site"
/translation="LVPRGS"
CDS complement(369..386)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(408..1031)
/codon_start=1
/gene="dsbA"
/product="bacterial periplasmic oxidoreductase"
/note="DsbA"
/translation="MKKIWLALAGLVLAFSASAAQYEDGKQYTTLEKPVAGAPQVLEFF
SFFCPHCYQFEEVLHISDNVKKKLPEGVKMTKYHVNFMGGDLGKDLTQAWAVAMALGVE
DKVTVPLFEGVQKTQTIRSASDIRDVFINAGIKGEEYDAAWNSFVVKSLVAQQEKAAAD
VQLRGVPAMFVNGKYQLNPQGMDTSNMDVFVQQYADTVKYLSEKK"
RBS 1039..1061
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 1076..1100
/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(1101..1119)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 1432..1509
/gene="lacI"
/note="lacI promoter"
CDS 1510..2592
/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 3401..3592
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
&