LOCUS       Exported                5945 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range expression vector pMHE6, complete sequence.
ACCESSION   AY303672
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5945)
  AUTHORS   Fodor BD, Kovacs AT, Csaki R, Hunyadi-Gulyas E, Klement E, Maroti G,
            Meszaros LS, Medzihradszky KF, Rakhely G, Kovacs KL.
  TITLE     Modular broad-host-range expression vectors for single-protein and 
            protein complex purification
  JOURNAL   Appl. Environ. Microbiol. 70 (2), 712-721 (2004)
  PUBMED    14766546
REFERENCE   2  (bases 1 to 5945)
  AUTHORS   Fodor BD.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-MAY-2003) Biophysics, BRC Hungarian Academy of 
            Sciences, Temesvari, Szeged, Csongrad 6701, Hungary
REFERENCE   3  (bases 1 to 5945)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5945
                     /organism="Broad host range expression vector pMHE6"
                     /mol_type="other DNA"
                     /db_xref="taxon:244947"
     primer_bind     94..112
                     /label=T7 promoter primer
                     /note="T7 promoter primer"
     promoter        94..112
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      94..110
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     misc_feature    111..136
                     /label=lac operator
                     /note="lac operator"
     protein_bind    113..137
                     /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)."
     RBS             152..174
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    182..214
                     /label=T7 tag
                     /note="T7 tag"
     CDS             182..214
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     misc_feature    278..301
                     /label=FLAG-tag
                     /note="FLAG-tag"
     CDS             278..301
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    287..301
                     /label=enterokinase cleaveage site
                     /note="enterokinase cleaveage site"
     misc_feature    305..328
                     /label=Strep-tag II
                     /note="Strep-tag II"
     CDS             305..328
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(382..399)
                     /label=T7 terminator primer
                     /note="T7 terminator primer"
     regulatory      395..442
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      396..443
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1074..1736
                     /label=pBBR1 oriV
                     /note="pBBR1 oriV"
     CDS             complement(1074..1736)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      1737..2506
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     misc_feature    2730..3723
                     /label=RP4 mob, oriT
                     /note="RP4 mob, oriT"
     regulatory      complement(3813..3928)
                     /regulatory_class="terminator"
                     /note="T4 terminator"
     CDS             complement(4188..4979)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside-3:adenyltransferase fusion"
                     /label=aadA
                     /note="streptomycin resistance"
                     /protein_id="AAQ82545.1"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            complement(4188..4979)
                     /gene="aadA"
                     /label=aadA
     regulatory      5715..5830
                     /regulatory_class="terminator"
                     /note="T4 terminator"