LOCUS       40924_30840        5945 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range expression vector pMHE6, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5945)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2004"; volume: "70"; issue: "2"; pages:
            "712-721"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-MAY-2003) Biophysics, BRC Hungarian Academy of Sciences, 
            Temesvari, Szeged, Csongrad 6701, Hungary"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        94..112
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    113..137
                     /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)."
     RBS             152..174
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             182..214
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
     CDS             278..301
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             305..328
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     terminator      396..443
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1077..1736)
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(1737..2506)
                     /direction=LEFT
                     /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)
                     /label=T4 terminator
                     /note="T4 terminator"
                     /regulatory_class="terminator"
     CDS             complement(4191..4979)
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     regulatory      5715..5830
                     /label=T4 terminator
                     /note="T4 terminator"
                     /regulatory_class="terminator"
ORIGIN
        1 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
       61 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt
      121 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatataca
      181 tatggctagc atgactggtg gacagcaaat gggtcgggat ccgaattcga gctccgtcga
      241 caagcttgat atcgaattcc tgcagctcga gggatccgac tacaaggacg acgacgacaa
      301 gaactggagc catccgcagt tcgagaagtg aagatccggc tgctaacaaa gcccgaaagg
      361 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
      421 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggatcg cccttcccaa
      481 cagttgcgca gcctgaatgg cgaatggaaa ttgtaagcgt taatattttg ttaaaattcg
      541 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgactgc gatgagtggc
      601 agggcggggc gtaatttttt taaggcagtt attggtgccc ttaaacgcct ggtgctacgc
      661 ctgaataagt gataataagc ggatgaatgg cagaaattcg aaagcaaatt cgacccggtc
      721 gtcggttcag ggcagggtcg ttaaatagcc gcttatgtct attgctggtt taccggttta
      781 ttgactaccg gaagcagtgt gaccgtgtgc ttctcaaatg cctgaggcca gtttgctcag
      841 gctctccccg tggaggtaat aattgacgat atgatcattt attctgcctc ccagagcctg
      901 ataaaaacgg tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc
      961 ggctccatgc accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgaggcggc
     1021 tacagccgat agtctggaac agcgcactta cgggttgctg cgcaacccaa gtgctaccgg
     1081 cgcggcagcg tgacccgtgt cggcggctcc aacggctcgc catcgtccag aaaacacggc
     1141 tcatcgggca tcggcaggcg ctgctgcccg cgccgttccc attcctccgt ttcggtcaag
     1201 gctggcaggt ctggttccat gcccggaatg ccgggctggc tgggcggctc ctcgccgggg
     1261 ccggtcggta gttgctgctc gcccggatac agggtcggga tgcggcgcag gtcgccatgc
     1321 cccaacagcg attcgtcctg gtcgtcgtga tcaaccacca cggcggcact gaacaccgac
     1381 aggcgcaact ggtcgcgggg ctggccccac gccacgcggt cattgaccac gtaggccgac
     1441 acggtgccgg ggccgttgag cttcacgacg gagatccagc gctcggccac caagtccttg
     1501 actgcgtatt ggaccgtccg caaagaacgt ccgatgagct tggaaagtgt cttctggctg
     1561 accaccacgg cgttctggtg gcccatctgc gccacgaggt gatgcagcag cattgccgcc
     1621 gtgggtttcc tcgcaataag cccggcccac gcctcatgcg ctttgcgttc cgtttgcacc
     1681 cagtgaccgg gcttgttctt ggcttgaatg ccgatttctc tggactgcgt ggccatgctt
     1741 atctccatgc ggtagggtgc cgcacggttg cggcaccatg cgcaatcagc tgcaactttt
     1801 cggcagcgcg acaacaatta tgcgttgcgt aaaagtggca gtcaattaca gattttcttt
     1861 aacctacgca atgagctatt gcggggggtg ccgcaatgag ctgttgcgta cccccctttt
     1921 ttaagttgtt gatttttaag tctttcgcat ttcgccctat atctagttct ttggtgccca
     1981 aagaagggca cccctgcggg gttcccccac gccttcggcg cggctccccc tccggcaaaa
     2041 agtggcccct ccggggcttg ttgatcgact gcgcggcctt cggccttgcc caaggtggcg
     2101 ctgccccctt ggaacccccg cactcgccgc cgtgaggctc ggggggcagg cgggcgggct
     2161 tcgccttcga ctgcccccac tcgcataggc ttgggtcgtt ccaggcgcgt caaggccaag
     2221 ccgctgcgcg gtcgctgcgc gagccttgac ccgccttcca cttggtgtcc aaccggcaag
     2281 cgaagcgcgc aggccgcagg ccggaggctt ttccccagag aaaattaaaa aaattgatgg
     2341 ggcaaggccg caggccgcgc agttggagcc ggtgggtatg tggtcgaagg ctgggtagcc
     2401 ggtgggcaat ccctgtggtc aagctcgtgg gcaggcgcag cctgtccatc agcttgtcca
     2461 gcagggttgt ccacgggccg agcgaagcga gccagccggt ggccgctcgc ggccatcgtc
     2521 cacatatcca cgggctggca agggagcgca gcgaccgcgc agggcgaagc ccggagagca
     2581 agcccgtagg gcgccgcagc cgccgtaggc ggtcacgact ttgcgaagca aagtctagtg
     2641 agtatactca agcattgagt ggcccgccgg aggcaccgcc ttgcgctgcc cccgtcgagc
     2701 cggttggaca ccaaaaggga ggggcaggca tggcggcata cgcgatcatg cgatgcaaga
     2761 agctggcgaa aatgggcaac gtggcggcca gtctcaagca cgcctaccgc gagcgcgaga
     2821 cgcccaacgc tgacgccagc aggacgccag agaacgagca ctgggcggcc agcagcaccg
     2881 atgaagcgat gggccgactg cgcgagttgc tgccagagaa gcggcgcaag gacgctgtgt
     2941 tggcggtcga gtacgtcatg acggccagcc cggaatggtg gaagtcggcc agccaagaac
     3001 agcaggcggc gttcttcgag aaggcgcaca agtggctggc ggacaagtac ggggcggatc
     3061 gcatcgtgac ggccagcatc caccgtgacg aaaccagccc gcacatgacc gcgttcgtgg
     3121 tgccgctgac gcaggacggc aggctgtcgg ccaaggagtt catcggcaac aaagcgcaga
     3181 tgacccgcga ccagaccacg tttgcggccg ctgtggccga tctagggctg caacggggca
     3241 tcgagggcag caaggcacgt cacacgcgca ttcaggcgtt ctacgaggcc ctggagcggc
     3301 caccagtggg ccacgtcacc atcagcccgc aagcggtcga gccacgcgcc tatgcaccgc
     3361 agggattggc cgaaaagctg ggaatctcaa agcgcgttga gacgccggaa gccgtggccg
     3421 accggctgac aaaagcggtt cggcaggggt atgagcctgc cctacaggcc gccgcaggag
     3481 cgcgtgagat gcgcaagaag gccgatcaag cccaagagac ggcccgagac cttcgggagc
     3541 gcctgaagcc cgttctggac gccctggggc cgttgaatcg ggatatgcag gccaaggccg
     3601 ccgcgatcat caaggccgtg ggcgaaaagc tgctgacgga acagcgggaa gtccagcgcc
     3661 agaaacaggc ccagcgccag caggaacgcg ggcgcgcaca tttccccgaa aagtgccacc
     3721 tggcggcgtt gtgacaattt accgaacaac tccgcggccg ggaagccgat ctcggcttga
     3781 acgaattgtt aggtggcggt acttgggtcg atagctttat gcttgtaaac cgttttgtga
     3841 aaaaattttt aaaataaaaa aggggacctc tagggtcccc aattaattag taatataatc
     3901 tattaaaggt cattcaaaag gtcatccacc ggatcagctt agtaaagccc tcgctagatt
     3961 ttaatgcgga tgttgcgatt acttcgccaa ctattgcgat aacaagaaaa agccagcctt
     4021 tcatgatata tctcccaatt tgtgtagggc ttattatgca cgcttaaaaa taataaaagc
     4081 agacttgacc tgatagtttg gctgtgagca attatgtgct tagtgcatct aacgcttgag
     4141 ttaagccgcg ccgcgaagcg gcgtcggctt gaacgaattg ttagacatta tttgccgact
     4201 accttggtga tctcgccttt cacgtagtgg acaaattctt ccaactgatc tgcgcgcgag
     4261 gccaagcgat cttcttcttg tccaagataa gcctgtctag cttcaagtat gacgggctga
     4321 tactgggccg gcaggcgctc cattgcccag tcggcagcga catccttcgg cgcgattttg
     4381 ccggttactg cgctgtacca aatgcgggac aacgtaagca ctacatttcg ctcatcgcca
     4441 gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc aaatagatcc
     4501 tgttcaggaa ccggatcaaa gagttcctcc gccgctggac ctaccaaggc aacgctatgt
     4561 tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg ctcgaagata
     4621 cctgcaagaa tgtcattgcg ctgccattct ccaaattgca gttcgcgctt agctggataa
     4681 cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg gagaatctcg
     4741 ctctctccag gggaagccga agtttccaaa aggtcgttga tcaaagctcg ccgcgttgtt
     4801 tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg cttcaggccg
     4861 ccatccactg cggagccgta caaatgtacg gccagcaacg tcggttcgag atggcgctcg
     4921 atgacgccaa ctacctctga tagttgagtc gatacttcgg cgatcaccgc ttccctcatg
     4981 atgtttaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct gctccataac
     5041 atcaaacatc gacccacggc gtaacgcgct tgctgcttgg atgcccgagg catagactgt
     5101 accccaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt accaccgctg
     5161 cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca ttacagctta
     5221 cgaaccgaac aggcttatgt ccactgggtt cgtgccttca tccgtttcca cggtgtgcgt
     5281 cacccggcaa ccttgggcag cagcgaagtc gaggcatttc tgtcctggct ggcgaacgag
     5341 cgcaaggttt cggtctccac gcatcgtcag gcattggcgg ccttgctgtt cttctacggc
     5401 aaggtgctgt gcacggatct gccctggctt caggagatcg gaagacctcg gccgtcgcgg
     5461 cgcttgccgg tggtgctgac cccggatgaa gtggttcgca tcctcggttt tctggaaggc
     5521 gagcatcgtt tgttcgccca gcttctgtat ggaacgggca tgcggatcag tgagggtttg
     5581 caactgcggg tcaaggatct ggatttcgat cacggcacga tcatcgtgcg ggagggcaag
     5641 ggctccaagg atcgggcctt gatgttaccc gagagcttgg cacccagcct gcgcgagcag
     5701 gggaattgat ccggtggatg accttttgaa tgacctttaa tagattatat tactaattaa
     5761 ttggggaccc tagaggtccc cttttttatt ttaaaaattt tttcacaaaa cggtttacaa
     5821 gcataaagct caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     5881 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     5941 tgtcg
//
