LOCUS       40924_30830        5938 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range expression vector pMHE5, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5938)
  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 5938)
  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 5938)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5938)
  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..5938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        193..211
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    212..236
                     /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             251..273
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    280..282
                     /label=fusion protein start codon
                     /note="fusion protein start codon"
     CDS             286..303
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             304..315
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     CDS             370..393
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             397..420
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     terminator      488..535
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1169..1828)
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(1829..2598)
                     /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    2822..3815
                     /note="RP4 mob; oriT"
     regulatory      complement(3905..4020)
                     /label=T4 terminator
                     /note="T4 terminator"
                     /regulatory_class="terminator"
     CDS             complement(4283..5071)
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     regulatory      5807..5922
                     /label=T4 terminator
                     /note="T4 terminator"
                     /regulatory_class="terminator"
ORIGIN
        1 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
       61 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
      121 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
      181 atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac aattcccctc
      241 tagaaataat tttgtttaac tttaagaagg agatatacca tggggcatca tcatcatcat
      301 catatcgagg gaaggcctgt cgacggtatc gataagcttg atatcgaatt cctgcagctc
      361 gagggatccg actacaagga cgacgacgac aagaactgga gccatccgca gttcgagaag
      421 tgaagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
      481 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
      541 aggaggaact atatccggat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
      601 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
      661 ttttaaccaa taggccgact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag
      721 ttattggtgc ccttaaacgc ctggtgctac gcctgaataa gtgataataa gcggatgaat
      781 ggcagaaatt cgaaagcaaa ttcgacccgg tcgtcggttc agggcagggt cgttaaatag
      841 ccgcttatgt ctattgctgg tttaccggtt tattgactac cggaagcagt gtgaccgtgt
      901 gcttctcaaa tgcctgaggc cagtttgctc aggctctccc cgtggaggta ataattgacg
      961 atatgatcat ttattctgcc tcccagagcc tgataaaaac ggtgaatccg ttagcgaggt
     1021 gccgccggct tccattcagg tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg
     1081 gaggcagaca aggtataggg cggcgaggcg gctacagccg atagtctgga acagcgcact
     1141 tacgggttgc tgcgcaaccc aagtgctacc ggcgcggcag cgtgacccgt gtcggcggct
     1201 ccaacggctc gccatcgtcc agaaaacacg gctcatcggg catcggcagg cgctgctgcc
     1261 cgcgccgttc ccattcctcc gtttcggtca aggctggcag gtctggttcc atgcccggaa
     1321 tgccgggctg gctgggcggc tcctcgccgg ggccggtcgg tagttgctgc tcgcccggat
     1381 acagggtcgg gatgcggcgc aggtcgccat gccccaacag cgattcgtcc tggtcgtcgt
     1441 gatcaaccac cacggcggca ctgaacaccg acaggcgcaa ctggtcgcgg ggctggcccc
     1501 acgccacgcg gtcattgacc acgtaggccg acacggtgcc ggggccgttg agcttcacga
     1561 cggagatcca gcgctcggcc accaagtcct tgactgcgta ttggaccgtc cgcaaagaac
     1621 gtccgatgag cttggaaagt gtcttctggc tgaccaccac ggcgttctgg tggcccatct
     1681 gcgccacgag gtgatgcagc agcattgccg ccgtgggttt cctcgcaata agcccggccc
     1741 acgcctcatg cgctttgcgt tccgtttgca cccagtgacc gggcttgttc ttggcttgaa
     1801 tgccgatttc tctggactgc gtggccatgc ttatctccat gcggtagggt gccgcacggt
     1861 tgcggcacca tgcgcaatca gctgcaactt ttcggcagcg cgacaacaat tatgcgttgc
     1921 gtaaaagtgg cagtcaatta cagattttct ttaacctacg caatgagcta ttgcgggggg
     1981 tgccgcaatg agctgttgcg tacccccctt ttttaagttg ttgattttta agtctttcgc
     2041 atttcgccct atatctagtt ctttggtgcc caaagaaggg cacccctgcg gggttccccc
     2101 acgccttcgg cgcggctccc cctccggcaa aaagtggccc ctccggggct tgttgatcga
     2161 ctgcgcggcc ttcggccttg cccaaggtgg cgctgccccc ttggaacccc cgcactcgcc
     2221 gccgtgaggc tcggggggca ggcgggcggg cttcgccttc gactgccccc actcgcatag
     2281 gcttgggtcg ttccaggcgc gtcaaggcca agccgctgcg cggtcgctgc gcgagccttg
     2341 acccgccttc cacttggtgt ccaaccggca agcgaagcgc gcaggccgca ggccggaggc
     2401 ttttccccag agaaaattaa aaaaattgat ggggcaaggc cgcaggccgc gcagttggag
     2461 ccggtgggta tgtggtcgaa ggctgggtag ccggtgggca atccctgtgg tcaagctcgt
     2521 gggcaggcgc agcctgtcca tcagcttgtc cagcagggtt gtccacgggc cgagcgaagc
     2581 gagccagccg gtggccgctc gcggccatcg tccacatatc cacgggctgg caagggagcg
     2641 cagcgaccgc gcagggcgaa gcccggagag caagcccgta gggcgccgca gccgccgtag
     2701 gcggtcacga ctttgcgaag caaagtctag tgagtatact caagcattga gtggcccgcc
     2761 ggaggcaccg ccttgcgctg cccccgtcga gccggttgga caccaaaagg gaggggcagg
     2821 catggcggca tacgcgatca tgcgatgcaa gaagctggcg aaaatgggca acgtggcggc
     2881 cagtctcaag cacgcctacc gcgagcgcga gacgcccaac gctgacgcca gcaggacgcc
     2941 agagaacgag cactgggcgg ccagcagcac cgatgaagcg atgggccgac tgcgcgagtt
     3001 gctgccagag aagcggcgca aggacgctgt gttggcggtc gagtacgtca tgacggccag
     3061 cccggaatgg tggaagtcgg ccagccaaga acagcaggcg gcgttcttcg agaaggcgca
     3121 caagtggctg gcggacaagt acggggcgga tcgcatcgtg acggccagca tccaccgtga
     3181 cgaaaccagc ccgcacatga ccgcgttcgt ggtgccgctg acgcaggacg gcaggctgtc
     3241 ggccaaggag ttcatcggca acaaagcgca gatgacccgc gaccagacca cgtttgcggc
     3301 cgctgtggcc gatctagggc tgcaacgggg catcgagggc agcaaggcac gtcacacgcg
     3361 cattcaggcg ttctacgagg ccctggagcg gccaccagtg ggccacgtca ccatcagccc
     3421 gcaagcggtc gagccacgcg cctatgcacc gcagggattg gccgaaaagc tgggaatctc
     3481 aaagcgcgtt gagacgccgg aagccgtggc cgaccggctg acaaaagcgg ttcggcaggg
     3541 gtatgagcct gccctacagg ccgccgcagg agcgcgtgag atgcgcaaga aggccgatca
     3601 agcccaagag acggcccgag accttcggga gcgcctgaag cccgttctgg acgccctggg
     3661 gccgttgaat cgggatatgc aggccaaggc cgccgcgatc atcaaggccg tgggcgaaaa
     3721 gctgctgacg gaacagcggg aagtccagcg ccagaaacag gcccagcgcc agcaggaacg
     3781 cgggcgcgca catttccccg aaaagtgcca cctggcggcg ttgtgacaat ttaccgaaca
     3841 actccgcggc cgggaagccg atctcggctt gaacgaattg ttaggtggcg gtacttgggt
     3901 cgatagcttt atgcttgtaa accgttttgt gaaaaaattt ttaaaataaa aaaggggacc
     3961 tctagggtcc ccaattaatt agtaatataa tctattaaag gtcattcaaa aggtcatcca
     4021 ccggatcagc ttagtaaagc cctcgctaga ttttaatgcg gatgttgcga ttacttcgcc
     4081 aactattgcg ataacaagaa aaagccagcc tttcatgata tatctcccaa tttgtgtagg
     4141 gcttattatg cacgcttaaa aataataaaa gcagacttga cctgatagtt tggctgtgag
     4201 caattatgtg cttagtgcat ctaacgcttg agttaagccg cgccgcgaag cggcgtcggc
     4261 ttgaacgaat tgttagacat tatttgccga ctaccttggt gatctcgcct ttcacgtagt
     4321 ggacaaattc ttccaactga tctgcgcgcg aggccaagcg atcttcttct tgtccaagat
     4381 aagcctgtct agcttcaagt atgacgggct gatactgggc cggcaggcgc tccattgccc
     4441 agtcggcagc gacatccttc ggcgcgattt tgccggttac tgcgctgtac caaatgcggg
     4501 acaacgtaag cactacattt cgctcatcgc cagcccagtc gggcggcgag ttccatagcg
     4561 ttaaggtttc atttagcgcc tcaaatagat cctgttcagg aaccggatca aagagttcct
     4621 ccgccgctgg acctaccaag gcaacgctat gttctcttgc ttttgtcagc aagatagcca
     4681 gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag aatgtcattg cgctgccatt
     4741 ctccaaattg cagttcgcgc ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa
     4801 caatggtgac ttctacagcg cggagaatct cgctctctcc aggggaagcc gaagtttcca
     4861 aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag ccttacggtc accgtaacca
     4921 gcaaatcaat atcactgtgt ggcttcaggc cgccatccac tgcggagccg tacaaatgta
     4981 cggccagcaa cgtcggttcg agatggcgct cgatgacgcc aactacctct gatagttgag
     5041 tcgatacttc ggcgatcacc gcttccctca tgatgtttaa ctttgtttta gggcgactgc
     5101 cctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
     5161 cttgctgctt ggatgcccga ggcatagact gtaccccaaa aaaacagtca taacaagcca
     5221 tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg gaccagttgc
     5281 gtgagcgcat acgctacttg cattacagct tacgaaccga acaggcttat gtccactggg
     5341 ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc aaccttgggc agcagcgaag
     5401 tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt ttcggtctcc acgcatcgtc
     5461 aggcattggc ggccttgctg ttcttctacg gcaaggtgct gtgcacggat ctgccctggc
     5521 ttcaggagat cggaagacct cggccgtcgc ggcgcttgcc ggtggtgctg accccggatg
     5581 aagtggttcg catcctcggt tttctggaag gcgagcatcg tttgttcgcc cagcttctgt
     5641 atggaacggg catgcggatc agtgagggtt tgcaactgcg ggtcaaggat ctggatttcg
     5701 atcacggcac gatcatcgtg cgggagggca agggctccaa ggatcgggcc ttgatgttac
     5761 ccgagagctt ggcacccagc ctgcgcgagc aggggaattg atccggtgga tgaccttttg
     5821 aatgaccttt aatagattat attactaatt aattggggac cctagaggtc ccctttttta
     5881 ttttaaaaat tttttcacaa aacggtttac aagcataaag ctcaaggaga tggcgccc
//
