LOCUS       40924_30850        5920 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range expression vector pMHE7, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5920)
  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 5920)
  AUTHORS   Fodor BD.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-MAY-2003) Biophysics, BRC Hungarian Academy of 
            Sciences, Temesvari, Szeged, Csongrad 6701, Hungary
REFERENCE   3  (bases 1 to 5920)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5920)
  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 
            (21-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..5920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        195..213
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    214..238
                     /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             253..275
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    282..284
                     /label=start codon of fusion protein
                     /note="start codon of fusion protein"
     CDS             288..311
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             324..347
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     CDS             348..359
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     terminator      472..519
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1153..1812)
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(1813..2582)
                     /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    2806..3799
                     /label=RP4 mob, oriT
                     /note="RP4 mob, oriT"
     regulatory      complement(3889..4004)
                     /label=T4 terminator
                     /note="T4 terminator"
                     /regulatory_class="terminator"
     CDS             complement(4267..5055)
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     regulatory      5791..5906
                     /label=T4 terminator
                     /note="T4 terminator"
                     /regulatory_class="terminator"
ORIGIN
        1 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
       61 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
      121 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
      181 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
      241 tctagaaata attttgttta actttaagaa ggagatatac catggacgac tacaaggacg
      301 acgacgacaa gctcgagggc aactggagcc atccgcagtt cgagaagatc gagggaaggc
      361 ctgtcgacgg tatcgataag cttgatatcg aattcctgca gcccggggga tccggctgct
      421 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
      481 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
      541 ggatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggaaattgt aagcgttaat
      601 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
      661 gactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg gtgcccttaa
      721 acgcctggtg ctacgcctga ataagtgata ataagcggat gaatggcaga aattcgaaag
      781 caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt atgtctattg
      841 ctggtttacc ggtttattga ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg
      901 aggccagttt gctcaggctc tccccgtgga ggtaataatt gacgatatga tcatttattc
      961 tgcctcccag agcctgataa aaacggtgaa tccgttagcg aggtgccgcc ggcttccatt
     1021 caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat
     1081 agggcggcga ggcggctaca gccgatagtc tggaacagcg cacttacggg ttgctgcgca
     1141 acccaagtgc taccggcgcg gcagcgtgac ccgtgtcggc ggctccaacg gctcgccatc
     1201 gtccagaaaa cacggctcat cgggcatcgg caggcgctgc tgcccgcgcc gttcccattc
     1261 ctccgtttcg gtcaaggctg gcaggtctgg ttccatgccc ggaatgccgg gctggctggg
     1321 cggctcctcg ccggggccgg tcggtagttg ctgctcgccc ggatacaggg tcgggatgcg
     1381 gcgcaggtcg ccatgcccca acagcgattc gtcctggtcg tcgtgatcaa ccaccacggc
     1441 ggcactgaac accgacaggc gcaactggtc gcggggctgg ccccacgcca cgcggtcatt
     1501 gaccacgtag gccgacacgg tgccggggcc gttgagcttc acgacggaga tccagcgctc
     1561 ggccaccaag tccttgactg cgtattggac cgtccgcaaa gaacgtccga tgagcttgga
     1621 aagtgtcttc tggctgacca ccacggcgtt ctggtggccc atctgcgcca cgaggtgatg
     1681 cagcagcatt gccgccgtgg gtttcctcgc aataagcccg gcccacgcct catgcgcttt
     1741 gcgttccgtt tgcacccagt gaccgggctt gttcttggct tgaatgccga tttctctgga
     1801 ctgcgtggcc atgcttatct ccatgcggta gggtgccgca cggttgcggc accatgcgca
     1861 atcagctgca acttttcggc agcgcgacaa caattatgcg ttgcgtaaaa gtggcagtca
     1921 attacagatt ttctttaacc tacgcaatga gctattgcgg ggggtgccgc aatgagctgt
     1981 tgcgtacccc ccttttttaa gttgttgatt tttaagtctt tcgcatttcg ccctatatct
     2041 agttctttgg tgcccaaaga agggcacccc tgcggggttc ccccacgcct tcggcgcggc
     2101 tccccctccg gcaaaaagtg gcccctccgg ggcttgttga tcgactgcgc ggccttcggc
     2161 cttgcccaag gtggcgctgc ccccttggaa cccccgcact cgccgccgtg aggctcgggg
     2221 ggcaggcggg cgggcttcgc cttcgactgc ccccactcgc ataggcttgg gtcgttccag
     2281 gcgcgtcaag gccaagccgc tgcgcggtcg ctgcgcgagc cttgacccgc cttccacttg
     2341 gtgtccaacc ggcaagcgaa gcgcgcaggc cgcaggccgg aggcttttcc ccagagaaaa
     2401 ttaaaaaaat tgatggggca aggccgcagg ccgcgcagtt ggagccggtg ggtatgtggt
     2461 cgaaggctgg gtagccggtg ggcaatccct gtggtcaagc tcgtgggcag gcgcagcctg
     2521 tccatcagct tgtccagcag ggttgtccac gggccgagcg aagcgagcca gccggtggcc
     2581 gctcgcggcc atcgtccaca tatccacggg ctggcaaggg agcgcagcga ccgcgcaggg
     2641 cgaagcccgg agagcaagcc cgtagggcgc cgcagccgcc gtaggcggtc acgactttgc
     2701 gaagcaaagt ctagtgagta tactcaagca ttgagtggcc cgccggaggc accgccttgc
     2761 gctgcccccg tcgagccggt tggacaccaa aagggagggg caggcatggc ggcatacgcg
     2821 atcatgcgat gcaagaagct ggcgaaaatg ggcaacgtgg cggccagtct caagcacgcc
     2881 taccgcgagc gcgagacgcc caacgctgac gccagcagga cgccagagaa cgagcactgg
     2941 gcggccagca gcaccgatga agcgatgggc cgactgcgcg agttgctgcc agagaagcgg
     3001 cgcaaggacg ctgtgttggc ggtcgagtac gtcatgacgg ccagcccgga atggtggaag
     3061 tcggccagcc aagaacagca ggcggcgttc ttcgagaagg cgcacaagtg gctggcggac
     3121 aagtacgggg cggatcgcat cgtgacggcc agcatccacc gtgacgaaac cagcccgcac
     3181 atgaccgcgt tcgtggtgcc gctgacgcag gacggcaggc tgtcggccaa ggagttcatc
     3241 ggcaacaaag cgcagatgac ccgcgaccag accacgtttg cggccgctgt ggccgatcta
     3301 gggctgcaac ggggcatcga gggcagcaag gcacgtcaca cgcgcattca ggcgttctac
     3361 gaggccctgg agcggccacc agtgggccac gtcaccatca gcccgcaagc ggtcgagcca
     3421 cgcgcctatg caccgcaggg attggccgaa aagctgggaa tctcaaagcg cgttgagacg
     3481 ccggaagccg tggccgaccg gctgacaaaa gcggttcggc aggggtatga gcctgcccta
     3541 caggccgccg caggagcgcg tgagatgcgc aagaaggccg atcaagccca agagacggcc
     3601 cgagaccttc gggagcgcct gaagcccgtt ctggacgccc tggggccgtt gaatcgggat
     3661 atgcaggcca aggccgccgc gatcatcaag gccgtgggcg aaaagctgct gacggaacag
     3721 cgggaagtcc agcgccagaa acaggcccag cgccagcagg aacgcgggcg cgcacatttc
     3781 cccgaaaagt gccacctggc ggcgttgtga caatttaccg aacaactccg cggccgggaa
     3841 gccgatctcg gcttgaacga attgttaggt ggcggtactt gggtcgatag ctttatgctt
     3901 gtaaaccgtt ttgtgaaaaa atttttaaaa taaaaaaggg gacctctagg gtccccaatt
     3961 aattagtaat ataatctatt aaaggtcatt caaaaggtca tccaccggat cagcttagta
     4021 aagccctcgc tagattttaa tgcggatgtt gcgattactt cgccaactat tgcgataaca
     4081 agaaaaagcc agcctttcat gatatatctc ccaatttgtg tagggcttat tatgcacgct
     4141 taaaaataat aaaagcagac ttgacctgat agtttggctg tgagcaatta tgtgcttagt
     4201 gcatctaacg cttgagttaa gccgcgccgc gaagcggcgt cggcttgaac gaattgttag
     4261 acattatttg ccgactacct tggtgatctc gcctttcacg tagtggacaa attcttccaa
     4321 ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca agataagcct gtctagcttc
     4381 aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg cagcgacatc
     4441 cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg taagcactac
     4501 atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg tttcatttag
     4561 cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg ctggacctac
     4621 caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt
     4681 ggctggctcg aagatacctg caagaatgtc attgcgctgc cattctccaa attgcagttc
     4741 gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg tgacttctac
     4801 agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt cgttgatcaa
     4861 agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat caatatcact
     4921 gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg
     4981 ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata cttcggcgat
     5041 caccgcttcc ctcatgatgt ttaactttgt tttagggcga ctgccctgct gcgtaacatc
     5101 gttgctgctc cataacatca aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc
     5161 ccgaggcata gactgtaccc caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc
     5221 gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta
     5281 cttgcattac agcttacgaa ccgaacaggc ttatgtccac tgggttcgtg ccttcatccg
     5341 tttccacggt gtgcgtcacc cggcaacctt gggcagcagc gaagtcgagg catttctgtc
     5401 ctggctggcg aacgagcgca aggtttcggt ctccacgcat cgtcaggcat tggcggcctt
     5461 gctgttcttc tacggcaagg tgctgtgcac ggatctgccc tggcttcagg agatcggaag
     5521 acctcggccg tcgcggcgct tgccggtggt gctgaccccg gatgaagtgg ttcgcatcct
     5581 cggttttctg gaaggcgagc atcgtttgtt cgcccagctt ctgtatggaa cgggcatgcg
     5641 gatcagtgag ggtttgcaac tgcgggtcaa ggatctggat ttcgatcacg gcacgatcat
     5701 cgtgcgggag ggcaagggct ccaaggatcg ggccttgatg ttacccgaga gcttggcacc
     5761 cagcctgcgc gagcagggga attgatccgg tggatgacct tttgaatgac ctttaataga
     5821 ttatattact aattaattgg ggaccctaga ggtccccttt tttattttaa aaattttttc
     5881 acaaaacggt ttacaagcat aaagctcaag gagatggcgc
//
