LOCUS       Exported                6290 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range expression vector pMHE3, complete sequence.
ACCESSION   AY299694
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6290)
  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 6290)
  AUTHORS   Fodor BD.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-MAY-2003) Biophysics, BRC Hungarian Academy of 
            Sciences, Temesvari krt., Szeged, Csongrad 6701, Hungary
REFERENCE   3  (bases 1 to 6290)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6290
                     /organism="Broad host range expression vector pMHE3"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:244052"
     primer_bind     111..129
                     /label=T7 promoter primer
                     /note="T7 promoter primer"
     promoter        111..129
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      111..127
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     misc_feature    128..153
                     /label=lac operator
                     /note="lac operator"
     protein_bind    130..154
                     /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             169..191
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      184..190
                     /regulatory_class="ribosome_binding_site"
     misc_feature    199..231
                     /label=T7-tag
                     /note="T7-tag"
     CDS             199..231
                     /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    280..297
                     /label=His-tag
                     /note="His-tag"
     CDS             280..297
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(350..367)
                     /label=T7 terminator primer
                     /note="T7 terminator primer"
     regulatory      363..410
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      364..411
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      448..876
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      1436..2098
                     /label=pBBR1 oriV
                     /note="pBBR1 oriV"
     CDS             complement(1436..2098)
                     /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      2099..2868
                     /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    3092..4085
                     /note="RP4 mob; oriT"
     regulatory      complement(4175..4290)
                     /regulatory_class="terminator"
                     /note="T4 terminator"
     CDS             complement(4550..5341)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside-3:adenyltransferase fusion"
                     /label=aadA
                     /note="streptomycin resistance"
                     /protein_id="AAQ64665.1"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            complement(4550..5341)
                     /gene="aadA"
                     /label=aadA
     regulatory      6077..6192
                     /regulatory_class="terminator"
                     /note="T4 terminator"