pHisJM vector (V005538)

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

pHisJM4506 bp600120018002400300036004200tac promoterlac operator6xHisT7 tag (gene 10 leader)Xpress(TM) tagT7 terminatorAmpR promoterAmpRorilacIq promoterlacICAP binding sitelac promoterlac operatorlacZ-alpha

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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