LOCUS       40924_30820        6290 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range expression vector pMHE3, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6290)
  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..6290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        111..129
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    130..154
                     /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             169..191
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             199..231
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
     CDS             280..297
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      364..411
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      448..876
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(1439..2098)
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(2099..2868)
                     /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    3092..4085
                     /note="RP4 mob; oriT"
     regulatory      complement(4175..4290)
                     /label=T4 terminator
                     /note="T4 terminator"
                     /regulatory_class="terminator"
     CDS             complement(4553..5341)
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     regulatory      6077..6192
                     /label=T4 terminator
                     /note="T4 terminator"
                     /regulatory_class="terminator"
ORIGIN
        1 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
       61 cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat taatacgact
      121 cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
      181 taagaaggag atatacatat ggctagcatg actggtggac agcaaatggg tcgggatccg
      241 aattcgagct ccgtcgacaa gcttgcggcc gcactcgagc accaccacca ccaccactga
      301 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
      361 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
      421 ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
      481 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
      541 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
      601 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
      661 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
      721 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
      781 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
      841 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatattt tgttaaaatt
      901 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgact gcgatgagtg
      961 gcagggcggg gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc ctggtgctac
     1021 gcctgaataa gtgataataa gcggatgaat ggcagaaatt cgaaagcaaa ttcgacccgg
     1081 tcgtcggttc agggcagggt cgttaaatag ccgcttatgt ctattgctgg tttaccggtt
     1141 tattgactac cggaagcagt gtgaccgtgt gcttctcaaa tgcctgaggc cagtttgctc
     1201 aggctctccc cgtggaggta ataattgacg atatgatcat ttattctgcc tcccagagcc
     1261 tgataaaaac ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc
     1321 ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgaggcg
     1381 gctacagccg atagtctgga acagcgcact tacgggttgc tgcgcaaccc aagtgctacc
     1441 ggcgcggcag cgtgacccgt gtcggcggct ccaacggctc gccatcgtcc agaaaacacg
     1501 gctcatcggg catcggcagg cgctgctgcc cgcgccgttc ccattcctcc gtttcggtca
     1561 aggctggcag gtctggttcc atgcccggaa tgccgggctg gctgggcggc tcctcgccgg
     1621 ggccggtcgg tagttgctgc tcgcccggat acagggtcgg gatgcggcgc aggtcgccat
     1681 gccccaacag cgattcgtcc tggtcgtcgt gatcaaccac cacggcggca ctgaacaccg
     1741 acaggcgcaa ctggtcgcgg ggctggcccc acgccacgcg gtcattgacc acgtaggccg
     1801 acacggtgcc ggggccgttg agcttcacga cggagatcca gcgctcggcc accaagtcct
     1861 tgactgcgta ttggaccgtc cgcaaagaac gtccgatgag cttggaaagt gtcttctggc
     1921 tgaccaccac ggcgttctgg tggcccatct gcgccacgag gtgatgcagc agcattgccg
     1981 ccgtgggttt cctcgcaata agcccggccc acgcctcatg cgctttgcgt tccgtttgca
     2041 cccagtgacc gggcttgttc ttggcttgaa tgccgatttc tctggactgc gtggccatgc
     2101 ttatctccat gcggtagggt gccgcacggt tgcggcacca tgcgcaatca gctgcaactt
     2161 ttcggcagcg cgacaacaat tatgcgttgc gtaaaagtgg cagtcaatta cagattttct
     2221 ttaacctacg caatgagcta ttgcgggggg tgccgcaatg agctgttgcg tacccccctt
     2281 ttttaagttg ttgattttta agtctttcgc atttcgccct atatctagtt ctttggtgcc
     2341 caaagaaggg cacccctgcg gggttccccc acgccttcgg cgcggctccc cctccggcaa
     2401 aaagtggccc ctccggggct tgttgatcga ctgcgcggcc ttcggccttg cccaaggtgg
     2461 cgctgccccc ttggaacccc cgcactcgcc gccgtgaggc tcggggggca ggcgggcggg
     2521 cttcgccttc gactgccccc actcgcatag gcttgggtcg ttccaggcgc gtcaaggcca
     2581 agccgctgcg cggtcgctgc gcgagccttg acccgccttc cacttggtgt ccaaccggca
     2641 agcgaagcgc gcaggccgca ggccggaggc ttttccccag agaaaattaa aaaaattgat
     2701 ggggcaaggc cgcaggccgc gcagttggag ccggtgggta tgtggtcgaa ggctgggtag
     2761 ccggtgggca atccctgtgg tcaagctcgt gggcaggcgc agcctgtcca tcagcttgtc
     2821 cagcagggtt gtccacgggc cgagcgaagc gagccagccg gtggccgctc gcggccatcg
     2881 tccacatatc cacgggctgg caagggagcg cagcgaccgc gcagggcgaa gcccggagag
     2941 caagcccgta gggcgccgca gccgccgtag gcggtcacga ctttgcgaag caaagtctag
     3001 tgagtatact caagcattga gtggcccgcc ggaggcaccg ccttgcgctg cccccgtcga
     3061 gccggttgga caccaaaagg gaggggcagg catggcggca tacgcgatca tgcgatgcaa
     3121 gaagctggcg aaaatgggca acgtggcggc cagtctcaag cacgcctacc gcgagcgcga
     3181 gacgcccaac gctgacgcca gcaggacgcc agagaacgag cactgggcgg ccagcagcac
     3241 cgatgaagcg atgggccgac tgcgcgagtt gctgccagag aagcggcgca aggacgctgt
     3301 gttggcggtc gagtacgtca tgacggccag cccggaatgg tggaagtcgg ccagccaaga
     3361 acagcaggcg gcgttcttcg agaaggcgca caagtggctg gcggacaagt acggggcgga
     3421 tcgcatcgtg acggccagca tccaccgtga cgaaaccagc ccgcacatga ccgcgttcgt
     3481 ggtgccgctg acgcaggacg gcaggctgtc ggccaaggag ttcatcggca acaaagcgca
     3541 gatgacccgc gaccagacca cgtttgcggc cgctgtggcc gatctagggc tgcaacgggg
     3601 catcgagggc agcaaggcac gtcacacgcg cattcaggcg ttctacgagg ccctggagcg
     3661 gccaccagtg ggccacgtca ccatcagccc gcaagcggtc gagccacgcg cctatgcacc
     3721 gcagggattg gccgaaaagc tgggaatctc aaagcgcgtt gagacgccgg aagccgtggc
     3781 cgaccggctg acaaaagcgg ttcggcaggg gtatgagcct gccctacagg ccgccgcagg
     3841 agcgcgtgag atgcgcaaga aggccgatca agcccaagag acggcccgag accttcggga
     3901 gcgcctgaag cccgttctgg acgccctggg gccgttgaat cgggatatgc aggccaaggc
     3961 cgccgcgatc atcaaggccg tgggcgaaaa gctgctgacg gaacagcggg aagtccagcg
     4021 ccagaaacag gcccagcgcc agcaggaacg cgggcgcgca catttccccg aaaagtgcca
     4081 cctggcggcg ttgtgacaat ttaccgaaca actccgcggc cgggaagccg atctcggctt
     4141 gaacgaattg ttaggtggcg gtacttgggt cgatagcttt atgcttgtaa accgttttgt
     4201 gaaaaaattt ttaaaataaa aaaggggacc tctagggtcc ccaattaatt agtaatataa
     4261 tctattaaag gtcattcaaa aggtcatcca ccggatcagc ttagtaaagc cctcgctaga
     4321 ttttaatgcg gatgttgcga ttacttcgcc aactattgcg ataacaagaa aaagccagcc
     4381 tttcatgata tatctcccaa tttgtgtagg gcttattatg cacgcttaaa aataataaaa
     4441 gcagacttga cctgatagtt tggctgtgag caattatgtg cttagtgcat ctaacgcttg
     4501 agttaagccg cgccgcgaag cggcgtcggc ttgaacgaat tgttagacat tatttgccga
     4561 ctaccttggt gatctcgcct ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg
     4621 aggccaagcg atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct
     4681 gatactgggc cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt
     4741 tgccggttac tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc
     4801 cagcccagtc gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat
     4861 cctgttcagg aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat
     4921 gttctcttgc ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga
     4981 tacctgcaag aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat
     5041 aacgccacgg aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct
     5101 cgctctctcc aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg
     5161 tttcatcaag ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc
     5221 cgccatccac tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct
     5281 cgatgacgcc aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca
     5341 tgatgtttaa ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctccata
     5401 acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact
     5461 gtaccccaaa aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc
     5521 tgcgttcggt caaggttctg gaccagttgc gtgagcgcat acgctacttg cattacagct
     5581 tacgaaccga acaggcttat gtccactggg ttcgtgcctt catccgtttc cacggtgtgc
     5641 gtcacccggc aaccttgggc agcagcgaag tcgaggcatt tctgtcctgg ctggcgaacg
     5701 agcgcaaggt ttcggtctcc acgcatcgtc aggcattggc ggccttgctg ttcttctacg
     5761 gcaaggtgct gtgcacggat ctgccctggc ttcaggagat cggaagacct cggccgtcgc
     5821 ggcgcttgcc ggtggtgctg accccggatg aagtggttcg catcctcggt tttctggaag
     5881 gcgagcatcg tttgttcgcc cagcttctgt atggaacggg catgcggatc agtgagggtt
     5941 tgcaactgcg ggtcaaggat ctggatttcg atcacggcac gatcatcgtg cgggagggca
     6001 agggctccaa ggatcgggcc ttgatgttac ccgagagctt ggcacccagc ctgcgcgagc
     6061 aggggaattg atccggtgga tgaccttttg aatgaccttt aatagattat attactaatt
     6121 aattggggac cctagaggtc ccctttttta ttttaaaaat tttttcacaa aacggtttac
     6181 aagcataaag ctcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat
     6241 acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
//
