LOCUS       40924_30815        5878 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range expression vector pMHE2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5878)
  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 5878)
  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 5878)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5878)
  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 
            (16-MAY-2003) Biophysics, BRC Hungarian Academy of Sciences, 
            Temesvari krt., Szeged, Csongrad 6701, Hungary"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5878
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        207..225
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    226..250
                     /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             265..287
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    294..296
                     /label=fusion protein start codon
                     /note="fusion protein start codon"
     CDS             300..317
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             318..329
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     terminator      442..489
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1123..1782)
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(1783..2552)
                     /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    2776..3769
                     /note="RP4 mob; oriT"
     regulatory      complement(3859..3974)
                     /label=T4 terminator
                     /note="T4 terminator"
                     /regulatory_class="terminator"
     CDS             complement(4237..5025)
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     regulatory      5761..5876
                     /label=T4 terminator
                     /note="T4 terminator"
                     /regulatory_class="terminator"
ORIGIN
        1 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
       61 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
      121 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
      181 ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
      241 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accatggggc
      301 atcatcatca tcatcatatc gagggaaggc ctgtcgacgg tatcgataag cttgatatcg
      361 aattcctgca gcccggggga tccggctgct aacaaagccc gaaaggaagc tgagttggct
      421 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
      481 ggttttttgc tgaaaggagg aactatatcc ggatcgccct tcccaacagt tgcgcagcct
      541 gaatggcgaa tggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
      601 taaatcagct cattttttaa ccaataggcc gactgcgatg agtggcaggg cggggcgtaa
      661 tttttttaag gcagttattg gtgcccttaa acgcctggtg ctacgcctga ataagtgata
      721 ataagcggat gaatggcaga aattcgaaag caaattcgac ccggtcgtcg gttcagggca
      781 gggtcgttaa atagccgctt atgtctattg ctggtttacc ggtttattga ctaccggaag
      841 cagtgtgacc gtgtgcttct caaatgcctg aggccagttt gctcaggctc tccccgtgga
      901 ggtaataatt gacgatatga tcatttattc tgcctcccag agcctgataa aaacggtgaa
      961 tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct ccatgcaccg
     1021 cgacgcaacg cggggaggca gacaaggtat agggcggcga ggcggctaca gccgatagtc
     1081 tggaacagcg cacttacggg ttgctgcgca acccaagtgc taccggcgcg gcagcgtgac
     1141 ccgtgtcggc ggctccaacg gctcgccatc gtccagaaaa cacggctcat cgggcatcgg
     1201 caggcgctgc tgcccgcgcc gttcccattc ctccgtttcg gtcaaggctg gcaggtctgg
     1261 ttccatgccc ggaatgccgg gctggctggg cggctcctcg ccggggccgg tcggtagttg
     1321 ctgctcgccc ggatacaggg tcgggatgcg gcgcaggtcg ccatgcccca acagcgattc
     1381 gtcctggtcg tcgtgatcaa ccaccacggc ggcactgaac accgacaggc gcaactggtc
     1441 gcggggctgg ccccacgcca cgcggtcatt gaccacgtag gccgacacgg tgccggggcc
     1501 gttgagcttc acgacggaga tccagcgctc ggccaccaag tccttgactg cgtattggac
     1561 cgtccgcaaa gaacgtccga tgagcttgga aagtgtcttc tggctgacca ccacggcgtt
     1621 ctggtggccc atctgcgcca cgaggtgatg cagcagcatt gccgccgtgg gtttcctcgc
     1681 aataagcccg gcccacgcct catgcgcttt gcgttccgtt tgcacccagt gaccgggctt
     1741 gttcttggct tgaatgccga tttctctgga ctgcgtggcc atgcttatct ccatgcggta
     1801 gggtgccgca cggttgcggc accatgcgca atcagctgca acttttcggc agcgcgacaa
     1861 caattatgcg ttgcgtaaaa gtggcagtca attacagatt ttctttaacc tacgcaatga
     1921 gctattgcgg ggggtgccgc aatgagctgt tgcgtacccc ccttttttaa gttgttgatt
     1981 tttaagtctt tcgcatttcg ccctatatct agttctttgg tgcccaaaga agggcacccc
     2041 tgcggggttc ccccacgcct tcggcgcggc tccccctccg gcaaaaagtg gcccctccgg
     2101 ggcttgttga tcgactgcgc ggccttcggc cttgcccaag gtggcgctgc ccccttggaa
     2161 cccccgcact cgccgccgtg aggctcgggg ggcaggcggg cgggcttcgc cttcgactgc
     2221 ccccactcgc ataggcttgg gtcgttccag gcgcgtcaag gccaagccgc tgcgcggtcg
     2281 ctgcgcgagc cttgacccgc cttccacttg gtgtccaacc ggcaagcgaa gcgcgcaggc
     2341 cgcaggccgg aggcttttcc ccagagaaaa ttaaaaaaat tgatggggca aggccgcagg
     2401 ccgcgcagtt ggagccggtg ggtatgtggt cgaaggctgg gtagccggtg ggcaatccct
     2461 gtggtcaagc tcgtgggcag gcgcagcctg tccatcagct tgtccagcag ggttgtccac
     2521 gggccgagcg aagcgagcca gccggtggcc gctcgcggcc atcgtccaca tatccacggg
     2581 ctggcaaggg agcgcagcga ccgcgcaggg cgaagcccgg agagcaagcc cgtagggcgc
     2641 cgcagccgcc gtaggcggtc acgactttgc gaagcaaagt ctagtgagta tactcaagca
     2701 ttgagtggcc cgccggaggc accgccttgc gctgcccccg tcgagccggt tggacaccaa
     2761 aagggagggg caggcatggc ggcatacgcg atcatgcgat gcaagaagct ggcgaaaatg
     2821 ggcaacgtgg cggccagtct caagcacgcc taccgcgagc gcgagacgcc caacgctgac
     2881 gccagcagga cgccagagaa cgagcactgg gcggccagca gcaccgatga agcgatgggc
     2941 cgactgcgcg agttgctgcc agagaagcgg cgcaaggacg ctgtgttggc ggtcgagtac
     3001 gtcatgacgg ccagcccgga atggtggaag tcggccagcc aagaacagca ggcggcgttc
     3061 ttcgagaagg cgcacaagtg gctggcggac aagtacgggg cggatcgcat cgtgacggcc
     3121 agcatccacc gtgacgaaac cagcccgcac atgaccgcgt tcgtggtgcc gctgacgcag
     3181 gacggcaggc tgtcggccaa ggagttcatc ggcaacaaag cgcagatgac ccgcgaccag
     3241 accacgtttg cggccgctgt ggccgatcta gggctgcaac ggggcatcga gggcagcaag
     3301 gcacgtcaca cgcgcattca ggcgttctac gaggccctgg agcggccacc agtgggccac
     3361 gtcaccatca gcccgcaagc ggtcgagcca cgcgcctatg caccgcaggg attggccgaa
     3421 aagctgggaa tctcaaagcg cgttgagacg ccggaagccg tggccgaccg gctgacaaaa
     3481 gcggttcggc aggggtatga gcctgcccta caggccgccg caggagcgcg tgagatgcgc
     3541 aagaaggccg atcaagccca agagacggcc cgagaccttc gggagcgcct gaagcccgtt
     3601 ctggacgccc tggggccgtt gaatcgggat atgcaggcca aggccgccgc gatcatcaag
     3661 gccgtgggcg aaaagctgct gacggaacag cgggaagtcc agcgccagaa acaggcccag
     3721 cgccagcagg aacgcgggcg cgcacatttc cccgaaaagt gccacctggc ggcgttgtga
     3781 caatttaccg aacaactccg cggccgggaa gccgatctcg gcttgaacga attgttaggt
     3841 ggcggtactt gggtcgatag ctttatgctt gtaaaccgtt ttgtgaaaaa atttttaaaa
     3901 taaaaaaggg gacctctagg gtccccaatt aattagtaat ataatctatt aaaggtcatt
     3961 caaaaggtca tccaccggat cagcttagta aagccctcgc tagattttaa tgcggatgtt
     4021 gcgattactt cgccaactat tgcgataaca agaaaaagcc agcctttcat gatatatctc
     4081 ccaatttgtg tagggcttat tatgcacgct taaaaataat aaaagcagac ttgacctgat
     4141 agtttggctg tgagcaatta tgtgcttagt gcatctaacg cttgagttaa gccgcgccgc
     4201 gaagcggcgt cggcttgaac gaattgttag acattatttg ccgactacct tggtgatctc
     4261 gcctttcacg tagtggacaa attcttccaa ctgatctgcg cgcgaggcca agcgatcttc
     4321 ttcttgtcca agataagcct gtctagcttc aagtatgacg ggctgatact gggccggcag
     4381 gcgctccatt gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct
     4441 gtaccaaatg cgggacaacg taagcactac atttcgctca tcgccagccc agtcgggcgg
     4501 cgagttccat agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg
     4561 atcaaagagt tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt
     4621 cagcaagata gccagatcaa tgtcgatcgt ggctggctcg aagatacctg caagaatgtc
     4681 attgcgctgc cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat
     4741 gtcgtcgtgc acaacaatgg tgacttctac agcgcggaga atctcgctct ctccagggga
     4801 agccgaagtt tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat caagccttac
     4861 ggtcaccgta accagcaaat caatatcact gtgtggcttc aggccgccat ccactgcgga
     4921 gccgtacaaa tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac
     4981 ctctgatagt tgagtcgata cttcggcgat caccgcttcc ctcatgatgt ttaactttgt
     5041 tttagggcga ctgccctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc
     5101 cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata gactgtaccc caaaaaaaca
     5161 gtcataacaa gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt
     5221 tctggaccag ttgcgtgagc gcatacgcta cttgcattac agcttacgaa ccgaacaggc
     5281 ttatgtccac tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc cggcaacctt
     5341 gggcagcagc gaagtcgagg catttctgtc ctggctggcg aacgagcgca aggtttcggt
     5401 ctccacgcat cgtcaggcat tggcggcctt gctgttcttc tacggcaagg tgctgtgcac
     5461 ggatctgccc tggcttcagg agatcggaag acctcggccg tcgcggcgct tgccggtggt
     5521 gctgaccccg gatgaagtgg ttcgcatcct cggttttctg gaaggcgagc atcgtttgtt
     5581 cgcccagctt ctgtatggaa cgggcatgcg gatcagtgag ggtttgcaac tgcgggtcaa
     5641 ggatctggat ttcgatcacg gcacgatcat cgtgcgggag ggcaagggct ccaaggatcg
     5701 ggccttgatg ttacccgaga gcttggcacc cagcctgcgc gagcagggga attgatccgg
     5761 tggatgacct tttgaatgac ctttaataga ttatattact aattaattgg ggaccctaga
     5821 ggtccccttt tttattttaa aaattttttc acaaaacggt ttacaagcat aaagctca
//
