LOCUS       Exported                6032 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range expression vector pMHE5Tc, complete sequence.
ACCESSION   AY299697
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6032)
  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 6032)
  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 6032)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6032
                     /organism="Broad host range expression vector pMHE5Tc"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:244055"
     primer_bind     187..205
                     /label=T7 promoter primer
                     /note="T7 promoter primer"
     promoter        187..205
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      187..203
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     misc_feature    204..229
                     /label=lac operator
                     /note="lac operator"
     protein_bind    206..230
                     /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             245..267
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      260..266
                     /regulatory_class="ribosome_binding_site"
     misc_feature    274..276
                     /label=fusion protein start codon
                     /note="fusion protein start codon"
     misc_feature    280..297
                     /label=His-tag
                     /note="His-tag"
     CDS             280..297
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    298..309
                     /label=Factor Xa cleavage site
                     /note="Factor Xa cleavage site"
     CDS             298..309
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    364..387
                     /label=FLAG-tag
                     /note="FLAG-tag"
     CDS             364..387
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    373..387
                     /label=enterokinase cleavage site
                     /note="enterokinase cleavage site"
     misc_feature    391..414
                     /label=Strep-tag II
                     /note="Strep-tag II"
     CDS             391..414
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(468..485)
                     /label=T7 terminator primer
                     /note="T7 terminator primer"
     regulatory      481..528
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      482..529
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1160..1822
                     /label=pBBR1 oriV
                     /note="pBBR1 oriV"
     CDS             complement(1160..1822)
                     /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      1823..2592
                     /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    2816..3809
                     /note="RP4 mob; oriT"
     regulatory      complement(3899..4014)
                     /regulatory_class="terminator"
                     /note="T4 terminator"
     CDS             complement(4320..5510)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline resistance protein"
                     /label=tet
                     /protein_id="AAQ64668.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            complement(4320..5510)
                     /gene="tet"
                     /label=tet
     regulatory      5895..6010
                     /regulatory_class="terminator"
                     /note="T4 terminator"