Basic Vector Information
- Vector Name:
- pHisJM
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4506 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Masuda J, Takayama E, Satoh A, Kojima-Aikawa K, Suzuki K, Matsumoto I.
pHisJM vector Map
pHisJM vector Sequence
LOCUS 40924_24673 4506 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pHisJM DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4506)
AUTHORS Masuda J, Takayama E, Satoh A, Kojima-Aikawa K, Suzuki K, Matsumoto
I.
TITLE A novel expression vector, designated as pHisJM, for producing
recombinant His-fusion proteins
JOURNAL Biotechnol. Lett. 26 (20), 1543-1548 (2004)
PUBMED 15604794
REFERENCE 2 (bases 1 to 4506)
AUTHORS Masuda J.
TITLE Direct Submission
JOURNAL Submitted (05-AUG-2003) Contact:Junko Masuda Ochanomizu University,
Graduate School of Humanities and Sciences; 2-1-1 Otsuka, Bunkyo-ku,
Tokyo 112-8610, Japan
REFERENCE 3 (bases 1 to 4506)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4506)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol.
Lett."; date: "2004"; volume: "26"; issue: "20"; pages: "1543-1548"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(05-AUG-2003) Contact:Junko Masuda Ochanomizu University, Graduate
School of Humanities and Sciences; 2-1-1 Otsuka, Bunkyo-ku, Tokyo
112-8610, Japan"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4506
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 183..211
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 219..235
/label=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)."
CDS 270..287
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
CDS 291..323
/codon_start=1
/label=T7 tag (gene 10 leader)
/note="leader peptide from bacteriophage T7 gene 10"
/translation="MASMTGGQQMG"
CDS 327..350
/codon_start=1
/label=Xpress(TM) tag
/note="Xpress(TM) epitope tag, including an enterokinase
recognition and cleavage site"
/translation="DLYDDDDK"
terminator 472..519
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 809..913
/label=AmpR promoter
CDS 914..1771
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1945..2533
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 2777..2854
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS 2855..3934
/codon_start=1
/label=lacI
/note="lac repressor"
/translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
protein_bind 3950..3971
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3986..4016
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4024..4040
/label=lac operator
/bound_moiety="lac repressor encoded by 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)."
CDS 4060..4233
/codon_start=1
/label=lacZ-alpha
/note="LacZ-alpha fragment of beta-galactosidase"
/translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
DRPSQQLRSLNGE"
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